G. Janukeviien, G. Zaborskien A.

Kabainskien

Evaluation of meat physical, chemical and

technological quality

Kaunas, 2012

PREFACE

This teaching book is useful training tool for Veterinary medicine and Veterinary food safety
students, learning meat processing technologies and hygiene and also for specialists, working in
meat shops. The book is well-timed and necessary, motivating students to work independantly and
preparing for practical classes in advance. In this teaching book there are described investigative
methods of meat maturing indexes, and of technological, sensual, fhysical and chemical indexes of
meat, meat products ad meat fat.

There can be found some mistakes of form and content in this book. The authors sincerelly ask
to send remarks and offerings to grazina.januskeviciene@lva.lt

Referees:
assoc. prof. dr. E. Bartkiene, Lithuanian University of Health Sciences (LUHS),
Veterinary academy, Department of Food safety and quality,
senior scientist dr. G. Garmiene, Kaunas University of Technology, Food institute.

Content
1.

Analysis of meat maturation characteristics and technological qualities

1.1

Determining pH value

1.2

Determining the amount of lactic acid

1.3

Determining the amount of bound water

11

1.4

1.3.1 Determining the amount of bound water by pressing method

11

1.3.2 Determining the amount of bound water by centrifugation method

12

Determining the solubility of muscle proteins

1.4.1 Determining the solubility of muscle proteins in a boratic solution

2.

3.

13

1.5

Determining the solubility of sarcoplasm proteins

14

1.6

Determining the solubility of myofibril proteins

14

1.7

Determining protein solubility in salt solutions

15

Sensual analysis of raw materials and meat products

15

2.1

Sensual evaluation of raw meat

19

2.2

Sensual evaluation of ready-to-cook meat

23

2.3

Sensual evaluation of meat products

24

Identifying meat of various types of animals

25

3.1

26

3.2

4.

13

Sensual analysis for detection of different animal meat

3.1.1 Determination of animal species according carcass configuration

26

3.1.2 Determining the type of animal according to anatomical structure of

bones
3.1.3 Determining the type of animal according to characteristics
of internal organs anatomical structure
3.1.4 Determining odour and flavour of meat applying boiling test

29

Laboratory tests for determination of different types of animals

33
34
35

3.2.1 Hair test

35

3.2.2 Determining the melting temperature of fat

36

3.2.3 Determining the refractive index of fat

37

3.2.4 Reaction of glycogen

37

3.2.5 Meat fluorescence

38

3.2.6 Precipitation reaction

38

Evaluation of sick animals meat

39

4.1

Visual carcass analysis

39

4.2

Determination of blood outflow degree

41

5.

4.3

Changes in limphonodes

43

4.4

Meat boiling test

44

4.5

Bacterioscopy of meat

44

4.6

Evaluation of meat pH

44

4.7

Peroxidase (benzidine) reaction

45

4.8

Formalin reaction

46

4.9

Evaluation of acidity-oxidation coefficient

46

4.10

Luminescence study

48

Evaluation of meat and poultry freshness

48

5.1

Evaluation of meat freshness

50

5.2

Evaluation of poultry freshness

57

5.2.1 Sensual analysis of poultry

57

5.2.2 Chemical analysis of poultry

58

5.2.3 Evaluation of poultry fats

61

5.3
6.

62

Detection of Trichinella in meat

64

6.1

64

6.2

7.

Freshness evaluation of rabbit and nutria meat

Diagnostics of trichinosis
6.1.1 Magnetic stirrer method for pooled sample digestion

65

6.1.2 Mechanically assisted pooled sample digestion method/sedimentation

technique using stomacher lab-blender 3 500
6.1.3 Mechanically assisted pooled sample digestion method/on filter
isolation technique
6.1.4 Detection of Trichinella in meat using digestion method
(using strong iodine and sodium thiosulphate solutions)
Trichinoscopic examination

68
68
69
70

6.2.1 Trichinoscopy additionally processing clippings

73

6.2.2 Examination of frozen meat

74

6.2.3 Examination of salted and smoked meat

74

6.2.4 Examination of lard

75

6.3

Examination of Trichinella in meat of other animals

75

6.4

Prophylaxis of trichinosis

75

Fats of animal origin and their quality evaluation

76

7.1

Animal fat stock

76

7.2

Characteristic of animal fat

77

7.3.

Evaluation of melted fat quality, physical and chemical indexes

79

8.

9.

7.3.1 Sensual fat evaluation

79

7.3.2 Evaluation of fat physical and chemical characteristic

80

7.3.2.1 Evaluation of fat melting temperature

81

7.3.2.2 Evaluation of fat stiffening temperature

81

7.3.2.3 Evaluation of fat light rays breaking index

82

7.3.2.4 Evaluation of fat moisture

83

7.3.2.5 Evaluation of fat iodine number

83

7.3.2.6 Evaluation of fat acids number

84

7.3.3 Qualitative method of antioxidant buthiloxianizolum evaluation

84

7.3.4 Evaluation of oxidative fat spoilage

85

7.3.4.1 Evaluation of peroxides number

85

7.3.4.2 Reaction with solution of neutral red

86

7.3.4.3 Reaction with 2 thiobarbituric acid

86

Composition and changes of blood

87

8.1

Evaluation of blood composition and characteristics

89

8.2

Influence of physical and chemical factors on blood haemolysis

89

Meat products

10 Evaluation of food alimentary and energetic value

89
92

10.1

Evaluation of nitrogen

92

10.2

Evaluation of humidity

94

10.3

Determination of general fat amount

94

10.4

Determination of general ash amount

96

10.5

Calculation of food products energetic value

97

11 Evaluation of salted-smoked meat products

98

11.1

Evaluation of sensual sausages characteristic

98

11.2

Laboratory analysis of sausages

99

11.2.1 Evaluation of humidity

99

11.2.2 Evaluation of salt

100

11.2.3 Evaluation of nitrites amount (reference method)

100

11.2.4 Evaluation of nitrates amount (LST ISO 3091:1997) (reference method)

101

11.2.5 Evaluation of Calcium amount (titrimetric method)

104

12 Defects of meat products

12.1

Defects of boiled sausages

105
105

12.2

Defects of dried and cold-smoked sausages

106

12.3

Defects of liverwursts

107

12.4

Defects of blood-puddings

107

13 Variety meat of cattle, pigs, sheep, horses and their processing

13.1

Chemical composition and nutritional value of variety meat

14 Canned goods

108
111
112

14.1

Evaluation of cans

114

14.2

Evaluation of canned meat quality

117

14.2.1 Sensual analysis of can content

117

14.2.2 Chemical analysis of canned meat quality

118

14.2.2.1

Evaluation of canned meat general acidity

118

14.2.2.2

Evaluation of salt in canned meat

119

14.2.3 Evaluation of meat amount in canned beef

15 References

119
121

1. Analysis of meat maturation characteristics and technological qualities

Meat quality is defined by many nutritional, biological and technological characteristics.
Meat is one of the most important sources of high biological value proteins. It contains a lot of
irreplaceable amino acids, fat and water soluble vitamins, minerals and trace elements. Meat
quality is defined by the chemical composition and physical properties (pH, colour, water holding
capacity, hardness, thermal treatment losses, usefulness of meat proteins, digestibility, etc.).
These characteristics are influenced by animal species, breed, individual characteristics, sex, age,
nurture technologies, fattening as well as other factors. Knowing them and purposeful organizing
of human activities helps improve meat characteristics of animals and meat quality. Livestock
productivity and product quality depends on the genotypic and phenotypic factors, which are
inseparable. While improving livestock genotype, we also must create appropriate conditions for
its realization. Effectiveness of improving any feature by means of selection depends on
selection intensity and perfection of evaluation methods; therefore, lately there has been a search
for ways to assess product quality by methods available to the field of production.
Form example, evaluation of meat quality according muscle electrical conduction is good and
comfortable method, which can be easy used in meat processing plants. External and internal
stressors causes stress. Sometimes small stress has positive influence, it stimulates activity, but very
strong and repeatable or prolonged stress before slaughter, can influence different changes in meat.
In rest condition, on the contrary, breathing slowly and deep, some physiological processes in
organism go on: declines heart contraction rate, breathing rate, declines amount of used oxygen,
less carbon dioxide is excreted, declines muscles electrical conductivity and tension.
Meat quality is often evaluated according maturity indexes. Meat maturity is influenced by its
type, composition, environmental factors: the more connective tissue is, the longer is meat
maturation. Because of greater amount of hydrolytic enzymes, processes of metabolism in
organisms of young animals is more intensive, therefore their meat matures more quickly if to
compare with older ones. Because of similar causes bulls meat matures slower than cows. Meat
gets soft consistence in 10-12 days and meat of young animals in 3-4days, if it is stored at 0-2C
temperature.
Meat quality can be evaluated according these maturation indexes: meat pH, amount of lactic
acid, volatile fatty acids, bounded water, solubility of proteins.
Meat pH enables to choose meat with the best water bounding characteristic for processing of
boiled sausages and to decline loss of bouillon and fat in product. Water bounding feature is the
7

main technological characteristic of meat, used for processing of sausages and hot smoked products.
Product richness, soft consistence and output depend on this meat stock characteristic. Water
binding characteristic depends not only on autolysis process, but also on many other factors, such as
age and sex of slaughtered animal, ratio of water and fat in meat, amount of proteins, solubility of
miofibrilic proteins, meat freezing, storing conditions and other. Increasing amount of fat and
connective tissue, lowers meat water binding. Also it is set, that intramuscular fat reduces meat
microstructure and it becomes able to bind more water.
Quality of meat products (especially of those, in composition of which there are added water)
depends on amount of fibrilic proteins, because these proteins have good swelling characteristic. It
is estimated, that warm meat better binds not only water but also fats. In such meat there are more
soluble proteins, which emulgate fat and during roasting process form thin elastic skin on the
surface of the product.

1.1. Determining pH value

The pH measurements are carried out using colorimetric or potentiometric method.
Colorimetric method is based on the ability of characteristics to change colour depending on the
hydrogen ion concentration. Many indicators are selective and change colour within the limits of
certain pH range. Liquid indicators or indicator paper are used. Depending on the selected
indicator and the pH meat extract colour changes. To measure pH of meat and its products,
various indicators can be used (Table 1).

Table 1. Indicators for meat extract pH measuring

Name
pH range
Methilenum red
4.2-6.2
Bromcresolum purple
5.2-6.8
Bromothimolum blue
6.0-7.6
Phenol red
6.8-8.4
Universal indicator
4.0-11.0

Colour of indicator
red-yellow
yellow-purple-violet
yellow-blue
yellow-red
red-yellow-green-blue-violet

Usually, for the measurement of the pH of meat and its products, aqueous extract of meat of
1:10 or 1:5 is prepared.
Preparing aqueous extract of meat: with accuracy of 0.01 g weigh 5 g of minced meat, put
the meat in 100 ml conical flask, and fill the flask with 50 ml distilled water. Leave the content of
the flask for 30 minutes (stirring periodically), then filter it.
Measuring the pH by potentiometric method the pH-meter readings are verified in
accordance with a standard buffer. Afterwards, electrodes are washed with distilled water, dried
8

with paper filter and soaked in an analyzed aqueous extract of meat. After the scale index is
steadied, the pH-meter readings are read. Then, electrodes are washed again and soaked in
distilled water.
1.2. Determining the amount of lactic acid
Lactic acid can be determined by analyzing meat muscle tissue or liver. The analysis is
carried out gradually, i.e. first, proteins are precipitated; then, carbohydrates are precipitated; the
solution is heated with H2SO4, colour reaction with veratrol is carried out and the intensity of
solution colour is measured.
Proteins are precipitated with the solution of 5 % metaphosphoric acid, and carbohydrates as
well as intermediate products of protein degradation - with CuSO4 and CaO x H2O. When heated
with concentrated H2SO4, lactic acid produces acetal aldehyde which while reacting with veratrol
turns the solution's colour to red. The intensity of the solution colour is measured by
photocolorimeter at 520 mm wave length (green filter).
Procedure. With accuracy of 0.01 g weigh 7-10 g of minced sample and place it in 100 ml
measurement flask, which is also filled with 40 ml distilled water. The content of the flask is
shaken for 5 minutes. Then, 10 ml of 5 percent metaphosphoric acid is poured into the flask and
the solution is shaken again. Distilled water is poured up to the mark, the content of the flask is
mixed and, after 15 minutes, it is to be filtered. Filtrate has to be clear. 10 ml of filtrate is poured
into 50 ml conical flask, to which 2-5 ml of CUSO4 solution and 3 g of CaO x H2O powder is
added. After adding CaO x H2O, the colour of the solution changes to greenish-blue. If the
solution does not turn greenish-blue (alkaline reaction is partial), more CaO x H2O powder is
added. The solution is left for 1 hour (shaking occasionally), then residue is filtered through
paper filter. The filtrate has to be clear and colourless.
The CuSO4 solution is prepared by attenuating saturated CuSO4 solution (prepared without
heating) with water at the ratio of 1:1.
Using a pipette, pour into the test-tube 0.1 ml of the filtrate and 0.4 ml of distilled water. In
parallel, carry out a control test by using 0.1 ml of water instead of the filtrate. The test-tubes are
placed into the glass with ice-cold water and 3 ml of concentrated H2SO4 is added into each testtube using a microburette. The, the test-tubes are left for 4 minutes in a boiling water bath and 2
minutes in ice-cold water. Using a micropipette, add 0.1 ml of 0.125 veratrol solution with 96 %
ethyl alcohol into each test-tube. After 20 minutes, optical density of the solutions is measured,
using water as the reference system. 1 ml cuvettes are used.
The filtrate with veratrol produces stable colour, therefore, after proteins are precipitated,
such solution can be stored in a refrigerator for a couple of days.
9

Optical density D (nm) of a sample is determined by the difference of optical densities of

the analyzed and control solution. In accordance with optical density D, using the calibration
graph, the amount of lactic acid is measured.
The calibration graph is made by using solutions made of lactic acid or zinc lactate. In the
former case, standard solution is made by dissolving 25 g of lactic acid in 100 ml of distilled
water; in the latter case, the solution is made by dissolving 0.169 mg of zinc lactate in 500 ml
of distilled water. To make the graph, working solutions are used (Table 2).

Table 2. Working solutions used to make a calibration graph of lactic acid

The amount of lactic
The amount of
acid, mg
standard solution of
lactic acid, ml
0.100
0.40
0.060
0.24
0.020
0.08
0.010
0.04

The amount of
water, ml

The amount of
sulphuric acid, ml

0.10
0.26
0.42
0.46

3.0
3.0
3.0
3.0

After optical densities of working solutions are measured, a calibration graph is drawn, on
the axis of abscissas of which values of solution concentration are marked, and on the axis of
ordinates values of optical densities are marked.
According to the optical densities D measured and using the calibration graph drawn, the
amount of lactic acid in 3.6 ml of solution is determined. The amount of lactic acid (mg/100 g)
in the analyzed product is measured using formula:

x=

c 12,5 100 100

n 10 m

c the amount of lactic acid in 3.6 ml of solution, mg;

12.5 the amount of the solution processed with CaO x H2O, ml;
100 dilution;
100 conversion to percent;
n the amount of filtrate taken for colour analysis after hydrocarbon precipitation, ml;
10 the amount of filtrate taken after protein precipitation, ml;
m the sample weight, g.

10

1.3. Determining the amount of bound water

1.3.1. Determining the amount of bound water by pressing method
This method for determining the amount of bound water in meat is based on evaluation
of liquid discharged from meat under light pressing and absorbed by filter paper in accordance
with the area of wet filter stain. The area of such stain depends on the ability of meat to bind and
retain water. Experiment showed that 1 cm2 of wet stain corresponds to 8.4 mg of water.
The method is sufficiently accurate. However, greater errors are obtained when analyzing
products with large amount of moisture (more than 90 %) or fat (more than 30 %). Three or four
parallel tests are to be carried out.
Procedure. With accuracy of 0.01 g weigh 0.3 g of minced sample placed on circular
polyethylene film and cover it with small-pore filter paper (with white stripe) or medium-pore
ashless filter (with blue strip) of size equal to that of the film. The sample between the film and
the filter is placed on a glass or ceramic plate with the filter paper side facing the plate and the
surface of the plate being covered with the same polyethylene film. The sample is covered from
the above with another plate of known weight and pressed with a weight, the overall weight of
which and the plate being 1 kg. After 10 minutes, the weight, the plate and the upper
polyethylene circle are removed. Using a soft pencil delineate the contour of pressed meat on the
filter, carefully remove the sample, dry the filter in air. When the filter becomes dry, the contour
of the entire stain shows up. Areas of pressed meat and discharged moisture are measured with
planimeter or calculated in accordance with their average diameter. Area of discharged moisture
is calculated in accordance with the difference obtained by subtracting the area of pressed meat
from the overall area of the stain.
The amount of bound water is expressed in percent from the amount of meat (xi) or
percent from the overall moisture content in meat (x2) using formulas:

x2 =

( a 8,4b )
100;
a

x1 =

(a 8,4b)
100;
m

a overall moisture content in the sample, mg;

b area of loose moisture, cm2;
m weight of meat sample, mg.
The amount of moisture in the filters has to be 8-9 %; therefore, prior to testing, they have
to be stored for three days in a desiccator.

11

1.3.2. Determining the amount of bound water by centrifugation method

When centrifugal force is applied, the analyzed meat sample placed in a fixed position
discharges the liquid phase, the amount of which depends on the strength of a link between a
liquid and the main phase of the sample. This method is not accurate; therefore, reliable results
can be obtained only after three or four repeated tests.
Procedure. With accuracy of 0.02 g weigh 4 g of well-minced sample and place it into
polyethylene test-tube with a perforated partition of known weight. The partition has to be fixed in
such a way that centrifuged liquid could easily escape. Centrifugation is carried out for 20 minutes
(centrifugation speed 100 s-1). Then, the sample is weighed together with the perforated
partition. Weight of solid particles of separated liquid is added to the weight of the centrifuged
sample.
Liquid collected after the centrifugation is poured without losses to a preheated dish of
constant weight and is dried in 105C to constant weight.
When calculating the amount of bound water, it is necessary to know the overall amount of
moisture of the sample, which is determined in accordance with one of the methods described
in subsection 2.1. According to overall amount of moisture, weight of dry materials is
determined.
The amount of bound water expressed in percent is calculated using formula:

x=

(m1 + m3 m2 ) 100
m0

m0 weight of the sample before the centrifugation, g;

m1 weight of the sample after the centrifugation, g;
m2 weight of dry materials of the sample, g;
m3 weight of particles of the centrifuged liquid, g.
Modified centrifugation method. 10 g of well-minced meat sample are weighed with

accuracy of 0.01 g. Such a sample of meat is placed on a plastic funnel which is lined with largepore filter of known weight. The funnel with the sample is placed into centrifugation test-tube.
Centrifugation is carried out for 20 minutes (centrifugation speed 66.6 s-1). Then, the sample is
weighed on the filter paper.
The amount of bound water is expressed in percent from the amount of meat (xi) or
percent from overall amount of moisture in meat (x2) using formula:
12

x = 100

(m0 + m2 m1 )
100
m0

m0 weight of the sample, g;

m1 weight of the sample and the filter after the centrifugation, g;
m2 weight of the filter before the centrifugation, g;
D overall amount of moisture in the sample, percent.
Reliable results can be obtained after carrying out three or four repeated tests.

1.4. Determining the solubility of muscle proteins

1.4.1. Determining the solubility of muscle proteins in a boratic solution
Procedure. To carry out a test, take 5 g of minced muscle tissue sample and extract it for 20

minutes with 25 ml of boratic buffer solution ( = 1.0, pH = 7.4). The suspension is centrifuged
for 15 minutes (centrifugation speed - 100 s-1). Supernatant is filtered through paper filter into
250 ml measurement flask. Further extraction, centrifugation and filtration are repeated four
times, collecting supernatant into the same measurement flask. Then, measurement flask is filled
with boratic buffer solution up to the mark. 10 ml of the solution is used for determining the
amount of proteins using the Kjeldahl method. The amount of nitrogen is calculated by formula:

x=

0,00014 (V V0 ) 100 250

2 m 100 10

V the amount of 0.01 N HC1 used for titration of the sample distillate, ml;
V0 the amount of 0.01 N HC1 used for titration of "empty" sample distillate, ml;
100 dilution volume of the digest, ml;
250 dilution volume of the sample, ml;
2 the amount of the digest taken for the distillation, ml;
m weight of the sample, g;
10 volume of the extract taken for the mineralization, ml;
0.00014 the amount of nitrogen, equivalent to 1 ml of 0.01 N HC1.
To determine soluble proteins, use the conversion factor 6.25.
Reagents: pH = 7.4 buffer solution prepared from 0.065 M H2BO3 solution, 0.0012 M

CuSO4 solution, 0.606 M KC1 solution.

13

1.5. Determining the solubility of sarcoplasm proteins

Proteins are extracted from a muscle tissue sarcoplasm by low ionic strength jo, buffer
solutions. The amount of sarcoplasm proteins is determined according to the difference between
overall amount of nitrogen and the amount of non-protein nitrogen in the sample. The conversion
factor 6.25 is used.
Procedure. 5 g of minced muscle tissue is extracted for 20 minutes with 25 ml of phosphate

buffer solution ( = 0.16, pH = 7.4). The suspension is centrifuged for 15 minutes (centrifugation
speed - 100 s-1). Supernatant is filtered through paper filter into 250 ml measurement flask. Further
extraction, centrifugation and filtration are repeated four times, collecting supernatant into the
same measurement flask. Then, measurement flask is filled with phosphate buffer solution up to
the mark.
Overall amount of nitrogen in sarcoplasm is determined using the Kjeldahl method and 10
ml of extract obtained.

x=

0,00014 (V V0 ) 100
2 m 100

V the amount of 0.01 N HC1 used for titration of the sample distillate, ml;
V0 the amount of 0.01 N HC1 used for titration of control sample distillate, ml;
100 dilution volume of the digest, ml;
2 the amount of the digest taken for the distillation, ml;
m weight of the sample, g;
0.00014 the amount of nitrogen, equivalent to 1 ml of 0.01 N HC1.
When determining non-protein nitrogen, the same amount of trichloroacetic acid is added
to 25 ml of extract. Protein precipitates are filtered through paper filter, and clear filtrate is
mineralized. The amount of nitrogenous material is determined using the Kjeldahl method.

1.6. Determining the solubility of myofibril proteins

Sarcoplasm and myofibril proteins are extracted from muscle tissue of meat by high ionic
strength buffer solutions. The amount of myofibril proteins is determined according to the
difference between overall amount of proteins in high ionic strength solutions and the amount of
sarcoplasm proteins.
Procedure. 5 g of minced muscle tissue sample is mixed in centrifugation test-tube for 20

minutes with 40 ml of buffer solution ( = 0.59, pH = 8.25). The suspension is centrifuged for 15
14

minutes (centrifugation speed 100 s-1). Supernatant is filtered through paper filter into 250 ml
measurement flask. Further extraction, centrifugation and filtration are repeated four times,
collecting supernatant into the same measurement flask. Then, measurement flask is filled
with buffer solution ( = 8.25) up to the mark. The amount of proteins in the obtained extract is
determined using the Kjeldahl method (Formula 2.5; subsection 2.3). The amount of soluble
myofibril proteins is calculated according to the difference between overall amount of proteins in
high ionic strength solution and the amount of sarcoplasm proteins in low ionic strength buffer
solution. The amount of myofibril proteins can be directly calculated in high ionic strength
solutions, if prior to that, sarcoplasm proteins were removed from the muscle tissue.
Reagents: pH=7.4 buffer solution prepared from 0.048 M sodium dihydrogen

orthophosphate (NaH2PO4- 2H2O) and 0.006 M sodium hydrophosphate (Na2HPO4 12H2O);

pH = 8.25 buffer solution prepared from 0.65 M sodium chloride and 0.03 M sodium bicarbonate;
70 percent trichloracetic acid solution.

1.7. Determining protein solubility in salt solutions

Procedure. 5 g of minced muscle tissue sample is mixed in centrifugation test-tube for 30

minutes with 30 ml of 5 % NaCl solution. The suspension is centrifuged for 15 minutes

(centrifugation speed - 100 s-1). Supernatant is filtered through large-pore paper filter into 250 ml
measurement flask. Further extraction, centrifugation and filtration are repeated four times,
collecting supernatant into the same measurement flask. Then, measurement flask is filled with 5
percent NaCl solution up to the mark. Similarly, proteins are extracted with NaCl solutions of
concentration of 10 %, 15 % and 20 %.
Amounts of proteins in the obtained extracts are determined using the Kjeldahl method. The
amount of proteins soluble in salt solutions is expressed in percent from overall amount of
proteins in the sample. Dependence between the amount of proteins extracted with salt solutions
and concentration of NaCl solution is determined.

2. Sensual analysis of raw materials and meat products

To ensure food safety, products are controlled during their raw, production and realization
phases. To determine food quality and safety, the following methods are used: sensual analysis
and measurement.

15

Sensual analysis method is based on the senses of human sensory organs: sight, hearing,
smell, taste and touch, as well as their analysis. Measurement methods are based on using special
equipment and chemical substances. Measurement methods can be physical, chemical,
biochemical, biological (physiological and microbiological), trade technological (when
characteristics of substances are analyzed by producing assays from them), etc. Sensual and
measurement methods are also called laboratory or traditional methods.
Using sensual analysis method, the quality is tested by tasting (degustation) or observing
(viewing). Degustation (Lat. degustare tasting) is determination of quality according to the
taste. When tasting a product, attention is also paid to its appearance, smell. Tasting objectivity,
accuracy depends on the following factors: degustation conditions; preparation of the product;
order of degustation; selection of persons.
The sensual method is also called sensory analysis (Lat. sensus feeling, sense), because it
is based on the senses of human sensory organs. The sensory test method is rather simple and
quick. This method is irreplaceable for testing most food products. It is true to say that
characteristics of wine, tea, butter, bread, pie, their products and other products are determined by
sensory method alone. However, quality of other products, in this case, meat and its products, is
evaluated using this method first, and only then other tests are carried out. If according to the
sensory evaluation the product is poor-quality, then further tests are not carried out the
product is declared defective. Sensory test method is used quite often in ensuring food (raw
material, substances) safety, especially during the phase of their collection.
For example, upon receiving raw material (carcass meat, spices, entrails, etc.) the best way
to evaluate physical risk factors surface contamination, dirt, and foreign objects is to
carefully observe the batch received, i.e. to evaluate sensory characteristics of the products.
Sensual characteristics of a product are evaluated in the following sequence: appearance,
smell, consistence, taste.
Appearance. Certain properties are characteristic to the appearance of individual products.

Some properties, such as clarity, are characterised by a wide scale of attributes.

Colour. When evaluating this characteristic, attention should be paid to the fact whether

it is typical to the product. Primary colours of the light spectrum are red, yellow and blue;
others are only combinations and shades of these colours. When the colour is determined,
proper illumination and background is important. Product colour shall seem lighter against dark
background, and darker against light background.
Odour. Evaluating odour of a product, standard LST ISO 6564 Sensual analysis.

Methodology. Methods of flavour and odour profiles, evaluated odour characteristics, intensity,
16

harmony of odours, external odours. There are many smells and their classification is very
complicated. It is offered to divide smells into 7 groups: camphor, muscat, flower, mint, ether,
spicy, decay. Moreover, to describe smells, the following terms are used: "aroma" and
"bouquet". To date, there is no unified concept of these terms. "Aroma" is understood as the
totality of pleasant odors or natural aroma compounds of initial raw material. "Bouquet" is
described as combination of smell and taste or as a complex of aroma compounds obtained
during the technological process.
A human can smell about 1000 scents, and experienced taster 10 times that. The
minimum concentration of various smells in the air which is smelled by a human differs. Strong
smells shut in weaker ones. Smell perception is influenced by indoor air temperature and relative
humidity.
Consistence. Consistence is a state of substance mobility, degree of density and

toughness. When evaluating consistence, the state, typicality and deviations of the material are
determined. These characteristics are determined by touch or actions using flatware or other
tools by pouring, spreading, grinding, squeezing, thrusting, cutting, brushing, etc. For
example, by squeezing elasticity of softness of meat and bread is determined. To determine
consistence, experience is also necessary. Various states of materials are determined by
squeezing. If when squeezed the material resists, it is rigid; if it fails to resist and returns to
previous state quickly, it is elastic; if it fails to resist and return to previous state it is not elastic.
Meat texture is also often determined, using standards LST ISO 11036 Sensual analysis.
Methodology. Profile of texture; LST ISO 4121 Sensual analysis. General instruction for usage of
qualitative response scales. Performing analysis of texture profile, such characteristic, as: firmness,
coherency, toughness, adhezity, tensility, elastisity are evaluated.
Flavour. Evaluating flavour standard LST ISO 6564 Sensual analysis. Methodology.

Methods of flavour and odour profiles can be used. When evaluating the flavour, its type and
typicality, extraneous taste and spices are determined. The main types of taste are: sweet, sour,
salty and bitter. When determining any sensory characteristic of a product appearance, colour,
smell, consistence typicality of a characteristic, i.e. whether it is typical to particular product,
is determined (Pict. 1-2).

17

General odour intensity

Intensity of boiled beef odour

Fatness

Intensity of acid odour

Chewingness

Succulence

Intensity of external odour

Intensity of cut colour

Fibrousness
Firmness in mouth

dairy
meat
ecological

Picture 1. Intensity of odour, appearance and texture in meat samples of different cattle types

General flavour intensity

Intensity of residual
flavour

Intensity of external flavour

General boiled beef

flavour intensity

Intensity of acid flavour

dairy
meat
ecological

Picture 2. Intensity of flavour features in meat samples of different cattle types

18

2. 1. Sensual evaluation of raw meat

Meat is a complex of muscle, fatty, connective and bone tissue, the quality of which is
determined by quantitative proportion of its components. Morphological composition of meat
depends on animal species, age, sex, fattening, technological processing. Meat quality is also
influenced by transport, pre-slaughter holding, and slaughter method of animals. Meat quality
mostly depends on its storing conditions.
External appearance. External appearance is characterized by overall appearance of a

product or carcass meat; attention is paid to presence of mould on the surface, extraneous inserts
and drawing of a cut (of products), drying of carcass meat. Bruising, clots, residues of skin and
the internal organs not only impairs commercial appearance of meat but also decreases its
durability for storing.
Meat colour. It is one of the main quality characteristics evaluated by a consumer first:

according to this characteristic, opinion about commercial appearance of meat is formed. Meat
colour depends on various factors: first, on muscle tissue pigments haemoglobin and myoglobin
as well as the amount of their compounds; also on pH, quantitative proportion of fatty and
connective tissue, mode of technological processing, storing conditions, etc.
Odour and flavour. These are also important quality characteristics which are

determined by chemical compounds characteristic to a product. Meat odour and flavour

depends on animal age, sex, the amount of fatty tissue and the nature of its distribution. Meat of
previously sick, scraggy animal will always taste worse. Flavour of fresh meat is specific,
somewhat sweetish. Meat of get does not have strong taste or smell, and meat of cattle has
stronger specific smell and less pleasant taste.
Consistence. It is one of the most important quality characteristics of meat and meat

products which is evaluated quite strictly by a consumer. The concept of consistence is rather
wide and difficult to define. When evaluating meat consistence, its softness, tenderness,
juiciness, etc. is evaluated. Meat consistence highly depends on the condition myofibril
proteins, ratio of fatty and connective tissue, pH of muscle tissue, level of hydration of muscle
proteins and other meat characteristics. Meat consistence changes while storing it refrigerated,
corning, after thermal processing. When storing meat, its quality characteristics change;
intensity and character of change depends on storing conditions and mode, also on composition
and characteristics of meat.
Putrefaction of meat. It is the main and most frequent deterioration of meat. Putrefaction of

meat is caused by proteolytic microorganisms, growth of which requires proteins or products of

19

partial protein hydrolysis. This process can take place under both aerobic and anaerobic
conditions, and its progress depends on the following factors:
1. Type of meat;
2. Initial microbial contamination and micro flora composition;
3. Meat pH;
4. Water content in the upper layers of meat or water activity;
5. Destructive properties of proteins;
6. External environmental factors (storing conditions).
Usually, putrefaction processes begin on the surface of meat when aerobic microorganisms
from the environment start acting.
The first sign of aerobic putrefaction is formation of mucus on the surface of meat. At the
initial stage of putrefaction, previously red colour of meat gradually turns gray. At the deep stage
of putrefaction, the colour of meat turns greenish. In the event of aerobic decomposition, the meat
pH is between 7.0 and 8.0, the meat has unpleasant smell, however less strong than that of
aerobic putrefaction.
Anaerobic putrefaction of meat begins in thick muscle layers, near bones, joints. During
this process, organic compounds are discharged which accumulate among muscle plates, the
structure of meat becomes porous, unpleasant smell is present, meat colour changes to blue-red
or gray-green. Alkaline pH values (8.0 < pH > 9.0) are typical for such meat.
When carrying out sensory evaluation of raw material (meat), it must be ensured that:
1. Raw material was obtained from animals corresponding to health requirements
established in the EU legislative acts;
2. Raw material was obtained from enterprises following HACCP or the principles of
good hygiene practices;
3. Raw material was transported using vehicles corresponding to requirements
applied to such raw material;
4. Raw material received veterinary certificate with health mark or other identification mark
in accordance with requirements provided for in legislative acts;
5. Raw material is fresh.
When compliance of documents received with raw material is assessed, the most important
thing is to determine whether raw material (meat) is fresh.

20

Evaluating freshness of raw material (meat)

When determining the freshness of meat by means of sensory method, the following
characteristics are determined: external appearance, colour, consistence, smell of meat,
condition of fat and tendons, broth transparency and aroma as well as condition of broth fats.
External appearance is evaluated by observing its surface and deeper layers. Dry surface of
meat indicates long term of meat storing. Appearance and colour of muscles is viewed at the fresh
cut of muscles. Moreover, when appearance of meat is evaluated, viscosity of meat by touching
it and dampness of the cut by applying filter paper (when filter paper is placed on the fresh cut of
muscles, no wet stain appears on the paper) are also evaluated.

Determining colour of meat

The colour is determined by observing the whole carcass meat upon taking a sample. When
the colour of meat is evaluated, attention is paid to the fact whether the coulour is typical for that
type of meat. The colour of meat depends on the changes of pigments haemoglobin and
myoglobin. The colour of meat surface (up to 3 cm deep) is determined by the pigment
oxymyoglobin. The colour of deeper layers of meat depends on purple-red myoglobin. As
microorganisms develop, the colour of meat gets darker (sometimes it turns green) because
pigment reactions with various compounds formed in meat take place. Therefore, various
pigment derivatives are formed: metmyoglobin, methaemoglobin, sulphohaemoglobin.
Under effect of hydrogen peroxide (which is formed due to microorganism activities),
myoglobin may decompose to yellow or green pigments. The colour of meat can also change due
to blue-green, pink pigments which are discharged by various types of microorganisms, for
example, some mould may colour meat black, white or blue-green. The colour of muscles of fresh
meat has to correspond to the type of meat, i. e., pork from pale pink to red, beef from pale to
dark red, etc.

Determining consistence

Consistence is determined at the fresh cut of meat by gently pressing with a finger and
observing the pit equilibrating: the cut of fresh meat is tight, elastic, the pit equilibrates quickly
(up to 1 minute). When meat deteriorates, its consistence becomes soft (as opposed to tight).

Determining odour

First, the smell of the surface of carcass meat or a sample taken is determined. Then, a cut is
made with a clean knife and the smell of deeper layers is determined immediately. Especial
21

attention is paid to the smell of connective tissue which is around the bones. The smell has to be
specific, typical to particular type of meat. Changes of the smell of meat appear in great part
due to aromatic and decomposition of sulfur-containing amino acids, when aromatic
compounds of unpleasant smell are formed (anaerobic putrefaction).

Determining condition of fat

It is determined by taking samples, determining the colour, smell and consistence of fat,
in case of poultry the number of peroxides and the number of acidity as well. Fat of fresh meat
must be typical to that particular type of fat, it cannot be limp or rancid.

Determining condition of tendons

It is determined by touching them when samples are taken. Solidity and condition of their
surface is evaluated. Tendons must be elastic, solid. Moreover, attention should be paid to the
condition of the surface of joints it must be smooth, glossy.

Determining transparency and smell of broth

The broth has to be transparent, fragrant. Broth turbidity is caused by primary products of
protein degradation dissolving in the hot water. Cold-processed meat and poultry are tested
when defrosted. When freshness of meat is evaluated, it should not be based on any single
characteristic alone. A complex test defines the condition of meat quite well. The given
characteristics of the freshness of meat are given an evaluation in scores and in accordance with a
total value of scores the meat is attributed to one of the three categories:
1. Fresh;
2. Questionable freshness;
3. Not fresh.
If any doubts as to the freshness of meat arise, chemical tests and tests of microbiological
contamination are carried out. Meat has to be clean, without any visible additives, without
extraneous smells, bruising, blood residues.
Evaluation of the results. Meat freshness is evaluated in points according to the

requirements of "Regulation on the Evaluation of Meat and Poultry Freshness", which provides
that sensory characteristics (appearance, colour, consistence, broth quality) of meat are
evaluated up to 13 points. When meat freshness is evaluated, the amount of points is reduced in
accordance with changes determined (Table 3).

22

Table 3. Evaluation of sensual characteristics of meat

Characteristics
Number of points deductet
Mild sliminess of the surface without any other deviation of
2
sensory characteristics.
The colour of meat surface and fat is changed, single spots of white
7
mould, slightly sour smell, and darker surface.

Dry or extremely wet surface of meat, grey or greenish colour

of the surface, small amount of mucus; slightly slimy, darkened
surface of a fresh cut; significant moistening of filter paper applied
to the surface; turbid meat juice; slow (more than 1 minute) or
partial equilibrating of a pit after pressing; slight smell of
deterioration on the surface, not felt in deeper layers; grayish
opaque shade of fat and smell of decomposing fat; staining tallow;
turbid broth with unpleasant smell, small drops of fat on the surface.

13

Grey or greenish colour of the surface of meat, the surface is slimy

or mouldy; the surface of cut is highly sticky, greenish or grey;
meat is softened at the cut; pits do not level after pressing; strong,
sour smell of putrefaction in deeper layers of muscles as well; fat is
greenish with dirty shade, staining, with strong smell of
decomposing fat; turbid broth with flakes and smell of putrefaction,
there are almost no drops of fat on the surface.

According
to
such
sensory
characteristics,
meat is qualified as
defective without giving it
any points.

2.2. Sensual evaluation of ready-to-cook meat

The main objective of the sensual evaluation is to make sure whether a product
corresponds to requirements of normative documents. Moreover, sensory characteristics of
newly made (prepared) samples could be determined.
The sensual evaluation is carried out by experts-tasters who have experience in evaluating
quality of meat preparations and products. Tasters evaluate individually or tasting commission is
constituted.
Products are evaluated by applying a scoring system, if it is provided for in technical
documents, or by describing how product characteristics correspond to standard requirements or
specifications.
The sensual evaluation of preparations is carried out in particular order. First, solid (uncut)
samples are evaluated, then, cut samples and their taste after thermal processing are evaluated.

Determining sensual characteristics of solid samples:

1. Appearance, colour and surface condition are determined by observing the sample.
2. The odour is determined by smelling the surface of the sample. If it is necessary to
determine the odour inside of the sample, the product is prickled with special wooden or
metal needle which is swiftly taken out and the product is smelled. Moreover, in case of
23

products with bones, the odour of muscles around the bone is determined.
3. Consistence is determined by pressing the sample with fingers or spatula.

Determining sensual characteristics of cut product:

1. Appearance (structure, arrangement of components, degree of breakup, uniformity of

mixing) and colour of the cut are determined by observing meat samples and ready-to-serve
products cut breadthwise and lengthwise;
2. The odour (aroma) is determined by smelling; attention should be paid to any extraneous
smells.
After thermal processing of ready-to-cook meat, the following are determined: external
appearance, flavour and odour of ready-to-serve products. These characteristics are evaluated for
warm products (the temperature of a product should be not lower than 65C).

2.3. Sensual evaluation of meat products

When carrying out the sensual evaluation of prepared meat products, it is determined
whether sensual characteristics correspond to requirements set for particular product.
The sensual evaluation is carried out with both: solid and cut products.

Sensual characteristics of solid products are determined in the following order:

1. External appearance, colour and surface appearance are determined by observing

externally;
2. Odour (aroma) of product surface is determined. The smell on the inside of the product
(if necessary) is determined by means of needle. Moreover, in case of products with bones, the
odour of muscles around the bone is determined.
3. Consistence is determined by gently pressing the surface of the product with a finger or
spatula.
Sensual characteristics of cut products are determined in the following order:

1. Appearance (structure and arrangement of components, degree of breakup, uniformity

of mixing) and colour of the cut are determined by observing sausages, meat breads, brawns,
meat jelly, minced meat preparations and ready-to-serve products cut breadthwise and
lengthwise; and pork, beef, mutton, poultry, etc. products cut breadthwise.
2. Odour (aroma), taste and juiciness are determined by smelling immediately after the
cut was made and tasting the cut piece. Attention is paid to any extraneous odours, tastes. Aroma
of spices and smoke (of smoked products) is also evaluated.
24

The odour, flavour and juiciness of all types of sausages are determined after the product is
heated to 65-70C (inside the product).
Juiciness of sausages in natural casing is determined by piercing the casing - in the spot of
piercing, a drop of liquid must appear.
3. Consistence is determined by pressing, cutting, chewing, spreading (in case of paste) the
product. When evaluating consistence, solidity, softness, tightness, pourability, uniformity
of the mass are also evaluated.

Sensual characteristics of canned meat are tested only after appropriate results of

microbiological analysis are received.

When odour, flavour and consistence of products are evaluated, maximum of three
products can be presented. Observation (of colour, appearance of a cut) can be carried out with
maximum of six products at a time.
According to the results of the sensual evaluation, the conclusion that a product is allowed
to be realized is drawn up. Defective products, products with signs of deterioration, etc. are not
allowed to be realized. For example, sausages are not allowed to be realized if casing is cracked,
with traces of stuffing, they are stained, there are bright spots on the casing which appeared
when sausages were in contact with each other during thermal processing, grey spots, large
cavities, etc. are present at the cut.

3. Identifying meat of various types of animals

Replacing meat of one type of animal with that of another (less valuable or not used in
human food) is falsification. Usually, the following types of falsification are to be determined:
distinguishing between the meat of bovine animals or moose meat and horsemeat, between
mutton as well as goat meat and dog meat, and between rabbit and cat meat.
The type of animal is determined by analyzing the appearance of carcass meat and muscles,
anatomical structure of bones and the internal organs, microstructure of medullary and cortical
layer of hair, chemical and physical properties of fat, the amount of glycogen in the meat,
precipitation reaction.
To identify meat of various types of animals, sensory and laboratory tests may be carried
out (Table 4).

25

Table 4. Methods of identification of meat of various types of animals

Sensual tests
Carcass arrangement, anatomical structure
of bones and the internal organs (visually).
Determining colour and consistence of
muscles and fat (visually).

Laboratory tests
Hair testing

Testing physico-chemical characteristics of fat,

i.e., the melting temperature, refractive index,
density, iodine value (physico-chemical methods).
Determining glycogen content in meat.
Determining smell and taste of muscles fat Precipitation reaction
(sensual analysis).
Testing proteins typical to type of animals or
their group (raw meat extract serologically).
Characteristic structure of enzymes, i.e., esterase,
lactate dehydrogenase testing (raw meat extract
by means of electrophoresis).
Testing characteristic structure of myoglobin
fractions (raw meat extract by means of
electrophoresis).
Chromosome analysis to determine type and sex
of animals (cytogenetic analysis).
Identification of animal types according to DNA
tests.
3.1. Sensual analysis for detection of different animal meat
3.1.1. Determination of animal species according carcass configuration

Neck of bovine animals is short, thick and wide. Horse's neck is long, narrow, there is a layer
of fat at the upper part. A sheep's neck is long, thin. Loins of bovine animals are pointy (not
prominent). Horse's loins are prominent. The rear part of sheep carcass is wide, massive; breast is
round, ridge is small, neck is round. The rear part of goat carcass is narrow, breast is not as
round, ridge is protruding, and neck is ovally pressed (Pict. 3-6).
Pork. Meat of young pigs is pale pink, and that of old pigs is red, soft, finely grained, with

visible fat inserts, muscle fibers are long, fascicles are thin, connective tissue is spongy with fat
inserts, boar meat is very tight. Fat are white (Pict.7).
Cow meat. Meat is red, grain (cross-sectional view) is pronounced, marble characteristics

depend on animal's age, breed and fattening, muscle fibers are long, muscle fascicles are thin,
intramuscular connective tissue layers are slightly spongy. Fat are yellowish.
Bull meat. Meat is dark red, hard, coarse-grained, muscle fibers are short, fascicles are thick,

connective tissue is developed, dense. Fat are white (Pict. 8).

26

Pict. 3. Carcass of sheep

Pict. 4. Carcass of goat

Pict. 5. Carcass of horse

Pict. 6. Carcass of bovine animal

27

Meat of young cattle. Meat is pale red, muscle fibers are thin, connective tissue is spongy.

Fat are white.

Veal. Meat is white or grey-pink, soft, finely grained, muscle fibers are very thin,

connective tissue is spongy. Fat are white.

Rabbit. Meat is white or pale pink, soft, tender, finely grained, muscle fibers are thin,

connective tissue is not developed, spongy. Fat are white.

Nutria meat. Meat is pale pink, spongy, overgrown with fat. Fat between 5-8 appendages

of thoracic vertebrae is a specific characteristic. Fat are white.

Mutton. Meat is pale or dark red, finely grained, muscle fibers are short, connective tissue is

dense. Fat are white.

Goat meat. Meat is pale or dark red, coarse-grained, connective tissue is dense, developed.

Fat are grey-white.

Horsemeat. Meat is dark red (later, bluish or black shade appears), tight, coarse-grained.

Fat are yellow.

Meat of wild animals is dark red, sometimes with a shade of blue, poorly bled, connective

tissue is developed, and the meat is lean.

Boar. Meat is from pale to dark red, of coarse structure, somewhat hard. Meat of grown males

is hard and has specific smell. Muscle fibers of young wild boars are thin, meat is soft, juicy. Fat
aggregates under the skin, around kidney, seldom among muscles. Fat are white.
Moose meat. Meat is red, fibers are thick, muscles are covered with well-developed fasciae

of connective tissue, there are no streaks of fat. Small areas of fat can be present around
breastbone, girdle and pelvis.
Venison. Meat is from pale to intense red, muscle fibers are thin. Streaks of fat are seldom

found. Fat are white, hard.

Hare meat. Meat is dark red with a shade of blue, relatively hard and dry. Meat of young

hare reminds that of rabbit. Fat are white and is found around kidney.
Meat of wild sheep and goats is dark red, fibers are thin. Often, the meat is somewhat hard

and dry (depending on the age). There are very thin layers of fat aggregates between muscles.
Yak meat is dark red and coarse; has a lot of connective tissue, without streaks of fat. When

cooked, the meat is dry and hard.

Bear meat is dark red with a shade of blue-violet; of solid consistence; muscle fibers are

coarse, dry; there is a lot of connective issue; the surface of carcass meat is covered with thick
layer of fat, in the autumn, there can be as much as 30-35 kg of fat.

28

Badger meat is pale pink and has specific smell. Muscle fibers are thin. There are many

layers of fat between muscles (marble).

Dog meat. Meat is red, dark red. Meat of young dogs is soft and that of old dogs is firm.

Muscle fibers are thin, short; connective tissue is soft with streaks of fat. Muscle structure at
the cross-section is finely grained. The meat is characterized by specific smell of dog. Fat are
white, greyish.

Pict. 7. Pork (ham)

Pict. 8. Meat of bovine animal (bull ham)

When boiling a meat, its specific smell and appearance show up. Boiled veal, pork and rabbit
are white or pale grey. Boiled horsemeat and meat of sheep and bovine animals is dark grey.
When boiling mutton, goat meat and meat of wild animals, stronger smell is felt. When boiling
boar, goat or dog meat, strong specific smell is felt. Horsemeat and boar meat foam strongly.
Sensory characteristics of meat of various animals are quite similar. Moreover, they depend
not only on the type of animal, but also on its age, sex, fattening and other factors. Therefore,
they are not reliable characteristics for determining the type of animal.

3.1.2. Determining the type of animal according to anatomical structure of bones

If the analyzed meat contains bones, then after removing muscles (it can be done by boiling),
the structure of bones is analyzed. According to comparative anatomical characteristics, it can be
quickly and accurately determined to which type of animal the analyzed meat belongs. Main
differences of bones of some animals are presented in Tables 5-7.

29

Table 5. Differences of anatomical structure of cattle and horse bones

Bone
Atlantis (atlas)

Cattle
Short, massive, there are no
transversal hole (for. transversarium)
on the wings
Axial
or
the Short; form of dental process is of
second
neck cloven along cylinder; spine process is
vertebra (axis)
tall, straight
Chest
vertebrae Instead of caudal bight there is lateral
(vertebrae
hole; spine processes are very tall,
thoracales)
wide, with sharp edges, dont touch
one another. There are 13 (14)
vertebrae

Ribs (costae)
Breastbone
(sternum)

Sacrum
sacrum)
Scapula

Horse
Wings big, slightly bended down.
Caudally of the center transversal
hole transfix the wings
Long; dental process is flat; spinal
process is divided into 2 parts
caudally
There is caudal bight; spinal
processes almost touch one another,
they are bended into caudal side.
There are 18 (17-19) vertebrae;
spinal processes of vertebrae are
shorter and thicker than of cattle
Wide with sharp edges, widens Tight and with more rounded edges
distally (13)
than of cattle (18)
Flat, flattened dorsoventrally. Consists There is clear comb in the ventral
of 7-8 rhombic segments
surface crista ventralis; it is
flattened from the sides. Consists of
7 segments
(os Prominent, spinal processes join
Flat; spinal processes are not joined

Triangular; spine of scapula becomes

taller in distal end, forming vertical
angle acromion
Humerus
Not long, massive, has 2 condyles
the great and the little; deltic
roughness is low; the great condyle is
tall (bulging 2.5-5 cm above bone
level)
Carpus bones (ossa Consist of 6 bones; there are 4 bones
carpales)
in proximal part and 2 in distal.
Foreleg
bones Ulna is developed well and reaches
(ossa antebrachii) distal end of radius.
Femur (os femoris)

Shinbones
cruris)

Cross-section
tubular bone

Corpus of bone is cylindric; the big

tuber is not divided

(ossa There is only shinbone (tibia); splintbone (fibula) is reduced, it forms a

hook on the head of shinbone.
of It is filled with medullas, after
removing of which cavity appears

Spina of scapula lowers in distal end

and disappears not reaching the neck
of scapula
Less massive, but longer; has 3
condyles: the great, the medium and
the little; deltic roughness is
developed well.
Consist of 7-8 bones; there are 4
bones in proximal part and 3-4 in
distal.
Ulna becomes more thin at the distal
end and disappears at the middle of
radius
The big tuber is divided into 2 parts
because of bight; distally from the
big tuber, the third tuber is
developed.
The head of splint-bone is flat, and
corpus becomes more thin at the
distal end and finishes at the upper
third of shinbone
Inside of bone is filled with bony
septa net.

30

Table 6. Differences of anatomical structure of sheep, dog and pig bones

Bone
Atlantis
(atlas)

Sheep
Wings are short, thick,
there
are
no
for.
transversarium, but there
is for. alarae (priekyje)
Axial or the It is similar to cattle,
second neck spinal process is tall,
vertebra (axis) straight, becoming a little
taller caudally; dental
process is of splitted
cylinder form
Chest
Corpus of vertebra is
vertebrae
long, there are 13
(vertebrae
vertebrae
thoracales)

Loin vertebres Spinal processes are

(vertebrae
wide, square to corpus of
lumbales)
vertebra;
ends
of
transversal processes are
widened and slightly
bented forward; consist of
6 vertebrae
Sacrum
Long; consists of 4
(os sacrum)
joined segments
Breastbone
(sternum)

Wide, flat; consists of 7

segments

Dog
Pig
Wings are wide, directed Wings
are
narrow,
caudolaterally; there is straight,
for.
for. transversarium
transversarium transfixes
along the wings
Spinal process is long; the Spinal process is tall,
primary end hangs at the cross-sectioned,
dental process; dental becoming taller to the
process is rounded and caudal
end;
dental
long
process is long, blunt, of
cone form
Corpus of vertebra is Spinal processes are tall;
short; spinal processes are there are 14-15 vertebrae;
low; there are13 vertebrae during proc. transversus
from above to under
there
is
for.
transversarium
Spinal
processes
are The height of spinal
narrowing
above; processes is bigger than
transversal processes are width;
thansversal
narrow,
bented processes have sharp
cranioventrally; consist of edges and bented down,
7 vertebrae
widened ends, consist of
6 vertebrae
Short, consists of 3 Without spinal processes,
segments; spinal processes consists of 4 segments
are short, separated
Very narrow; consists of 8 Flat; consists of 6
segments
segments

Scapula

Triangular; spina ends with Long, narrow and rounded if

spiky angle - acromion at to compare; spina is
the distal end
developed well, breaks at the
articular surface, forming
hook like process proc.
hamatus
Foreleg bones Ulna becomes thinner to Ulna and radius are almost
(ossa
distal end
of the same thickness along
antebrachii)
all length; the bones are
joined with articular join
Femur
(os Almost straight, cylindric; Long,
thin,
hooked;
femoris)
suddenly widens at the gradually widens at the distal
distal end
end

Wide, spina lowers to both

ends, and the middle of it is
tall, turned to caudal side

Radius is short, ulna is

massive if to compare; they
are joined with connective
tissue
Short, massive; corpus is
almost quadrilateral in
distal part

Bones of goat are more slender and longer, outgrows are longer and have sharper edges than
of sheep. Goats scapula is longer than of sheep. Thickness of spine edge is equal all along and
outgrow of sheep scapula is thinned from the back side.

31

Table 7. Differences of anatomical structure of rabbit, nutria and cat bones

Bone
Atlantis (atlas)

Rabbit
There are no holes at the
front and back wings, there
are only bights (incisura
alaris)

Nutria
The corpus is short, thin,
wings are narrow, long
enough, there are no back
bight, but the front one is
clear
Axial or the Spinal process is prominent The corpus is short, dental
second
neck forward
process is cylindric; spinal
vertebra (axis)
process is directed caudally
Breastbone
Consists of 6-7 segments;
(sternum)
handle
of
breastbone
(manubrium sterni) is blunt,
flattened from the sides
Scapula
Long (twice longer than Form
of
inequilateral
width)
acromion
is triangular; cranial edge is
elongated, it has caudally oval; from the middle third
directed long process spine
forms
acromion
proc. suprahamatus
process, which from more
than half of scapula height is
separated from it and
finishes below articular
surface of scapula
Humerus
The head is clearly Short, turned along its axis at
separated from the corpus the distal end; there is for.
and is of the same height as supracondylicum, the great
the great condyle; there are condyle is developed, but
lateral and medial ones are
no for. supracondylicum
not clear

Foreleg bones Radius and ulna are of the

(ossa
same length, touched one to
antebrachii)
another; the bones are of
hook form

Sacrum
(os sacrum)

Long and has 4 long spinal

processes

Femur
(os femoris)

Under trochanter major

there is trochanter minor
and t. tertius

Shinbones

Splint-bone is free at the

proximal end, then it joints
with shinbone at its lower
third

Radius and ulna are of hook

form, not joined, articular
join joins them at the
proximal end, and fibre
cartilage at the distal end;
there is wide spacing
between these bones
Consists of 4 well developed
and joined vertebrae; there
are 4 spinal processes
Head of the neck is clearly
separated from the corpus;
trochanter
major
is
developed well, t. minor is of
clear trochanter shape, t.
tertius is not developed
Splint-bone and shinbone are
of the same length, their
ands are joined with articular
joints

Cat
The same as of rabbit

Spinal process is prominent

forward
Consists of 9 segments,
handle of breastbone is
spiky
Short (the lenght exceeds
the width times); proc.
suprahamatus is not clear

The head is separated from

the corpus not clearly; the
proximal end is little
hooked; the great condyle
is higher than the head;
there
is
for.
supracondylicum
above
condylus lateralis
Both bones are of the same
length, there is spacing
between them, they are not
joined; articular join joins
them at the proximal end,
and fibre cartilage at the
distal end
Short, has 3 low spinal
processes
There is only one great
trochanter

Splint-bone is developed
and it is of the same length
as the shinbone; their ands
are joined with articular
joints

32

3.1.3. Determining the type of animal according to characteristics

of internal organs anatomical structure

Anatomical structure of different animals internal organs (lungs, liver, kidneys, spleen, heart,
brain, tongue and other) is not the same.
Liver. The right and the left lobes of cattle, sheep and goat liver are not divided. There is no

impression of esophagus on cattle liver, and caudal outgrow of caudal lobe is overhanged the liver
edge. Caudal lobe of sheep and goats is not big. The left lobe of horses liver is divided into medial
and lateral lobe. At the obtuse edge, there is impression of esophagus. There is no gall-bladder in
horses. Impression of esophagus is clear in pig liver and the left and the right lobes are divided into
medial and lateral lobes. At the surface of pigs liver lobating is seen clearly (Pict. 9-10).
Lungs. The right cattle lung is divided into 5 lobes: apical (which is divided into 2 parts),

hearts, diaphragms and additional; the left lung is divided into 3 lobes: apical, hearts and
diaphragms. Lung lobules are very clear, therefore their surface looks like marble. Horses lungs are
little lobed: the right side is divided into 3 lobes: apical, diaphragms and additional, the left is
divided into 2 lobes: apical and diaphragms. Lungs lobes are not seen through the pleura. Pig lungs
are lobed, like of cattle, only the right upper lobe is not divided into 2 parts (Pict. 11-12).
Kidneys. At the surface of cattle kidneys between lobes there are deep striae; kidneys are

oval. Kidneys of horses are not the same form: the left kidney is like bean form and the right like
heart form. Lobes of horses kidneys are grown between each other, the surface is smooth. Kidneys
of sheep, goats and pigs are like bean form and their surface is smooth too (Pict. 13-14).
Referring to anatomical structure of internal organs is possible only if they are not separated
from the carcass.

Picture 17. Liver of bovine animal

Picture 18. Liver of pig

33

Picture 11. Lungs of bovine animal

Picture 12. Lungs of pig

Picture 13. Kidney of bovine animal

Picture 14. Kidney of pig

3.1.4. Determining odour and flavour of meat applying boiling test

The boiling test takes up an intermediate position between the sensory and laboratory
analyses. Meat of various animals can be identified according to its colour, smell and appearance
after boiling. Boiled pork, rabbit and veal are white or pale grey; boiled meat of bovine animals,
sheep and horses is dark grey. When mutton, goat meat or meat of wild animals is boiled,
delicate specific odour can be felt. When meat of boar, male goat or dog is boiled, quite strong
specific and unpleasant odour can be felt. Foaming appears when horsemeat and boar meat is
boiled.
However, the odour and flavour of meat can be not typical to particular type of animal
due to various reasons: metabolic disorders, poisoning, drug injections, improper diet, etc. Meat
of boars, cryptorchids, hermaphrodites often has strong odour and flavour of urine.
Odour and flavour of meat is determined by sensory method. This analysis can be more
accurate if it is carried out by few people. The main requirement is that people carrying out the
34

analysis are healthy, i.e. can evaluate the smell and taste. Prior to the analysis, the carcass meat
has to be stored for at least 24 hours in a cold, well-ventilated room. The analysis is carried out
by 2 methods:
1. a sample of minced meat is placed in cold water and the water is heated until it starts
boiling. Smell and taste components are transferred to the broth;
2. a sample of minced meat is placed in boiling water and is boiled for some time. In
this case, due to denaturated proteins on the surface of meat samples, odour and flavour
components remain inside the samples and after cutting such sample, they show up very well.
If the analysis is carried out applying both methods, more accurate results are obtained. If
mild sex odor is present, in case of pork, fat sample can be melted or a sample of salivary gland
can be boiled. In such case, the odour will be stronger.
3.2. Laboratory tests for determination of different types of animals
3.2.1. Hair test

Microstructure of medullary and cortical layer of hair of various animals is different. On the
surface of carcass meat, some hair can almost always be found. Especially often, the hair is found
on carcass meat of goats and wild animals. Hair is collected and analyzed with microscope. When
identifying the type of animal according to the hair, descriptions and drawings of microstructure
of the hair provided in special literature and hair sets are used (Fig. 15).

Goat

Rabbit

Elk

Horse

Hind

Bovine animal

Dog

Mouse

Sheep

Pict. 15. Examples of different animals hair

35

3.2.2. Determining the melting temperature of fat

According to the melting temperature of animal fat, it is possible to determine to which type
of animals the analyzed fat belongs (Table 8). The melting temperature of fat (consistence of fat)
mainly depends on the content of saturated and non-saturated fatty acids as well as ratio of low
and high molecular weight fatty acids. If the fat contains more stearin, palmitin or other saturated
fatty acids, such fat is harder and its melting temperature is higher. If the fat contains more olein,
linolenic acid and other non-saturated fatty acids, such fat is softer and its melting temperature is
lower.
Procedure. 2 clean and dry 1.4-1.5 mm diameter glass capillaries are immersed in melted

and filtered fat (grease). The fat has to fill around 1/3 of capillary height. Then, the capillaries
are left for 1-2 hours on ice, in a refrigerator or in cold running water so that the fat freezes.
After chilling, the capillaries with the fat are fixed by means of thin rubber ring to a
thermometer so that frozen fat is located at thermometer mercury or spirit reservoir. The
thermometer with attached capillaries is placed into test-tube and is fixed by means of a cap in
such a way that neither the thermometer nor the capillaries are touching the walls of the testtube. The test-tube with the thermometer is placed into glass flask and fixed in a stand. The flask
is filled with boiled water slightly coloured with methylene blue. The capillaries are located below
upper level of water. The water in the glass is heated (slightly mixing) and the temperature has
to rise maximum 2C per minute; and in the end, around 1C per minute. The thermometer
readings and condition of fat is carefully observed. The melting temperature of analyzed fat is the
thermometer reading when the fat in the capillaries becomes transparent. A difference between the
results of individual analyses should not be greater than 0.5C. The average of the results of two
analyses is considered to be the final result.
Table 8. Fat melting temperature of different animals and birds
Animal

Bovine
Sheep
Goat
Deer
Pig
Horse

Melting
t-re (oC)
45-50
44-55
48-50
48-52
36-46
28-32

Animal

Dog
Cat
Rabbit
Hare
Bear
Badger

Melting
t-re (oC)
30-39
39
42.3
45.5
39.15
31.33

Animal

Elk
Whale
Hen
Goose
Turkey
Partridge

Melting
t-re(oC)
46-48
50
34
32.2
35.5
31.9

The melting temperature can be roughly determined by holding a piece of fat for 1-2
minutes in a hand: horse or dog fat softens quickly and starts melting, whereas the fat of
bovine animals and sheep starts crumbling.
36

3.2.3. Determining the refractive index of fat

Refractive index is a relative value indicating the refraction of light rays moving through
thin layer of fat located between two prisms of a refractometer. Refractive index of fat depends
on the content of saturated and non-saturated fatty acids. The amount of such acids in a fat of
various animals is different, therefore, their refractions also differs (Table 9). The refractive
index of a fat increases as molecular weight of fatty acids and the amount of non-saturated acids
increase.
Table 9. Fat refraction degree of different animals species (40C)
Type of animal
Bovine
Sheep
Pig
Badger
Bear
Dog
Cat
Horse

The refractive coefficient

1.4510-1.4583
1.4566-1.4583
1.4636
1.4562-1.4564
1.4611
1.4512
1.4570
1.4563-1.4590

The refractive index

45-50
44-55
36-46
57
54.1
30-39
49.1-49.2
15-39

To analyze fat, universal refractometers which show refractive coefficient of fat are used, or
special refractometers for analyzing fat which show refractive index of fat are used. Normally, fat
is analyzed when it is of such temperature which it has when it is melted, i.e. 40C. If a
thermometer of special fat refractometer does not show 40C precisely, correction of 0.55 is
added to the reading of the thermometer, and 0.55 is subtracted from each degree below 40C.
Procedure. Both prisms of a refractometer are carefully cleaned. Water of 40C is poured

through the prisms of the refractometer. 2 drops of molten filtered fat are dripped on the lower
prism. The prisms are immediately pressed together and refractive coefficient or index of
analyzed fat is determined.

3.2.4. Reaction of glycogen

Carbohydrate glycogen is assigned to polysacharides group (C6H10O5)n. The highest amount

of it is found in liver (up to 20 %) and in skeletal muscles too. Glycogen is important reserved
organism material. It easy turns into glucose, which is source of energy. Glycogen is made in liver
and muscles from monosacharides (glucose). In organism, reaction of glycogen formation from
glucose reversible: in liver the excess of glucose turns into glycogen and when there is lack of
glucose liver glycogen turns into glucose.

37

Amount of glycogen in meat of different animals is not the same. Though it depends on
animal species, but also on meat maturation degree, preslaugter animal condition, age, health and
other reasons.
Evaluating amount of glycogen in meat with iodine solution, reaction is positive, if there is
about 1 % of glycogen. Reaction is based on the fact, that polysacharide glycogen, on action of
iodine, turns red.
Procedure. Lugol solution is used: 2 g of iodine, 4 g of potassium iodide are melted in 100 ml

of distilled water. For extraction of glycogen, 15 g of analyzed meat (without fats and connective
tissue of tendons) are weighed, well grinded, put into flask, poured 60 ml of water and boiled for 30
min., after boiling moment. The bouillon is filtrated through the paper filter and is cooled. 3-5 ml of
filtrate are poured into the test-tube and 5-10 drops of Lugol solution are added. Reaction is positive
if bouillon becomes cherry red, suspicious orange, negative yellow.
In many cases positive react meat of horse, dog, camel, bear and cat. Negative reacts meat of
sheep, goat, cattle, pig and rabbit. Besides, meat of young animals often reacts positive and meat of
old or ill negative. Meat, taken from head or neck area also often reacts positive. This reaction
doesnt give absolutely right results.

3.2.5. Meat fluorescence

Different animals meat can be differentiated by fluorescence of meat, fats and medulla,
lighting with ultraviolet light. (Table 10).
Table 10. Fluorescence of different animals meat, fats and medulla
Kind of meat
Horsemeat
Beef
Elk meat
Pork
Mutton

Meat
Red-brown (like red brick)
with grey stripes
Red velvet with violet stripes
Dark brown with clear
orange shade.
Light brown (sand colour)
with light blue strips
Grey-brown with white strips

Colour of tissue lighting

Fats
Medulla
Greyish yellow
Clear grey
White
Clear yellow

Grey with violet shade

Yellow-green

Light blue

Light brown
colour)
Grey-yellow

Clear white

(sand

3.2.6. Precipitation reaction

Precipitation reaction is specific precipitation of antigen (protein) in dregs in clear solutions,

acting immune precipitating serum. Precipitation reaction is very sensitive and specific, therefore is
used evaluating of what animal meat is, what meat was used for sausages or other meat products
processing. Precipitation reaction is used in forensic medicine.
38

Precipitating serum, got after hyper-immunisation of rabbits with proteins of different

animals meat, and analyzed meat extract are used. Meat extract is poured into the test-tube and
precipitating serum is slowly released from Paster pipette, soaked to the bottom.
If there is protein (precipitinogen) in analyzed extract, which correspond to antibodies
(precipitins) of precipitating serum, at the line of fluids connection light greyish precipitate layer
forms, which is called precipitation circle. The circle is better seen at the black background.

4. Evaluation of sick animals meat

Evaluation of sick animals meat has a significant sanitary mean. In such meat food-borne
infection or other diseases agents can be found. Besides, value of sick animals meat is lower if to
compare with healthy ones, it is not stable for storage and begins to spoil quickly.
The evaluation is based on complex of pathologoanatomic, bacteriologic and biochemical
analysis. The attention is pointed on cutting/slaughter place, hypostases, bleeding degree and
different pathologoanatomic changes, visible in carcass and variety meat.

4.1. Visual carcass analysis

Carcass is examined from outside and inside: condition of subcutaneous tissue, muscles, fat
and connective tissues, bones, joints, pleural and peritoneal conditions, carcass tissue color, odour is
fixed and degree of carcass bleeding is assessed. In one case significant carcass changes may be set
while in the other case, in order to draw reasonable conclusions further analysis is necessary.
Analyzing sick animal carcasses, carcass thinning because of disease (which is not difficult to
differentiate from thinning of alimentary origin, or thinning because of high productivity) can be
determined.
Yellow colouring of tissues is typical in case of pathological icterus (which should be
differentiated from alimentary icterus, or senile tissue colouring yellow).
When there is insufficient bleeding, diffusely red subcutaneous tissue and dark red spots in
muscles is typical for cattle carcasses. Well visible blood vessels, especially at ham and shoulder
area are typical for swine carcasses. If pigs were slaughtered after death or not well bleedened after
stunning, clearly demarcated colouring of bottom body third, coloured dark red is visible.
Numerous petechial bruising in the skin of pigs, especially visible after scalding, is a
significant factor diagnosing pig fever. It is very important to differentiate this type of bruising
setting septicemical diseases and acute poisoning.

39

Carcass muscle liquidity, more or less expressed by tissue oedema is often observed in
metabolic disorders, or renal pathology, but such changes may be as a result of animal treatment
with various medicinal substances, or thyroid pathology. In addition, thyroid dysfunction in cattle
may be suspected when gelatinous subcutaneous connective tissue and pale pink carcass muscles
are discovered.
Diffuse, in spots or sections coloured red serous body cavities tapetum is often associated with
fibrinous inflammation; during pleura or peritoneum inflammation brown-red purulent deposits are
found on their surface. In cases of bone fractures deformation of individual carcass parts
(asymmetry) is visible, cutting these places blood clots or abscess are found.
If animal was overheated during transportation, experienced stress before slaughter had a high
temperature, "bent the front limb phenomenon" is seen limbs are strong bent over the elbow joint,
lower leg part stiffen and over a joints arent inflexible because premature legs muscle stiffness.
Localized changes in the carcasses are less obvious and often depend on animal species, age
and sex. Limited or spread changes of pig muscle color at internal parts of waist and ham muscles
may be seen in PSE and DFD meat case, and for pigs the long back muscles may be associated with
this muscle inflammation "banana" disease. In cow's pelvic area found inflammations, bruisings,
oedema, knittings, scarrings are often associated with violations during the calving period.
Phlegmons, necrosis, abscesses at hip, knee or tarsus joint region, are often found in old, lean,
partially paralyzed or with damaged hoofs cows.
Swollen calves hock joints and pigs wrist and tarsus joints may show local or systemic
infection. Knee joint inflammation of young animals is commonly observed during salmonella
infection, so each case must be examined bacteriologically. Bovine tarsus oedema, gelatinous or
purulent mass on the outside of the tarsus is often the result of chronic inflammatory injury
(especially if animals lying places are bad). Incorrect posture and joint pain often poses problems in
evaluating the sanitary quality of the meat. Young animals (particularly bulls) often have perverted
bones and arthrosis of the joint cartilages defect and cariosity.
Yellowish-green, pea-sized, soft or liquid consistency focus cross and lumbar vertebrae and
abscesses in the spinal canal often can be found when infection spreads through damaged tail of
pigs, as well as of bulls, kept on slatted floor when tail tips are injured. Pigs abscesses often occur at
the neck and strangles area. Under ears and in neck area of pigs, on both sides of cattle and calves
neck, shoulders and hip region, injection sites can be found, the tissue color is changed,
haemorrhagia, oedema, and pus can be found. Injection sites may also be solid, with fibrinous
papules in muscles if oil-based medicines were injected long time before; action of antibiotics
injection site is characterized by yellowish stains and typical odor; to find a deep intramuscular
40

injection is particularly difficult and experienced eye, which could see asymmetric carcass location,
is needed.
For pigs blood spills in pubic area caused by the rupture of muscle on uneven floors are not
rare, as well as blood spills in the area of pectoral vertebrae region, pelvic or spinal fractures due to
improper electrical stunning.
At well visible muscle parts, in the diaphragm, parasites can be found, particularly cattle and
pigs trichinas (rice grain-sized vesicles). In cattle carcasses, along the shoulders and chest muscle
fibers or in diaphragm, in the form of light stretches, Sarcocystis damaged muscles can be seen.
In addition to the specific changes occurring in infectious and parasitic diseases, trauma,
inflammation and other, degenerative muscle changes associated with metabolic disturbances are
often found, affecting post-mortem meat maturation and causing problems during the post-mortem
examination, when suitability of meat for human consumption is solving.
After visual observation of carcass (meat) and internal organs and if there is suspicion, that
meat is of sick or slaughtered at agony animal, it must be analyzed bacteriologically, meat ph must
be evaluated, peroxidase and formalin (for cattle) reactions and meat boiling test must be done.
Additional reactions can be done too (determination of acidity-oxidation coefficient, luminescentic
analysis and other).
Inspection of cut place. Edges of cut place of slaughtered healthy animals are rough and

infiltrated with blood more intensively than in other places. Edges of cut place of animals,
slaughtered in agony can be rough, but blood is infiltrated into surrounding tissues marginally.
Edges of cut place of dead animals (imitation of slaughter) are smooth, blood is not infiltrated into
wound muscles and can be easy cleaned.
Haemostasis. If slaughter was imitated (dead animal was slaughtered), or animal in agony or

sick animal was slaughtered, bluish red with not clear lines and not elevated spots can be seen in
hypoderma of carcass, in serosal tissue and internal organs. They appear the mostly in that side, on
which sick animal layed during slaughter (bruisings and thrashings are clearly red colour, have clear
lines; if there are haematomas after cutting we find tattered tissues and blood clots). Mucous
membranes of hardly sick or slaughtered in agony animals get lightly blue colour because of
hypoxia.
4.2. Determination of blood outflow degree

There are 4 blood outflow degrees: good, satisfactory, bad and very bad. It is considered about
this from colour of fats and muscles and from blood in small and large vessels.
Blood outflow degree could be checked quite credibly after cutting primary leg from the
carcass (from that side, on which animal was laying during slaughter) and cutting axilary vessels
41

and muscles of the lower neck third. If blood outflow degree is bad, the blood spils out the vessels
and neck muscles are full with blood. Blood badly outflows from carcasses of forced slaughtered
animals. In heart cavities of slaughtered in agony animals blood clots are found: red clots short
agony; motley, yellow or white long agony (clots in heart cavities also form if heart was injured:
miocarditis, toxicoses, white-muscles disease and other).
Not always reason of bad bleeding is animal disease or agony. The reason can be incorrect
stunning or slaughter. If animal was healthy before slaughter and blood outflowed badly because of
incorrect stunning or slaughter, then the carcass is left hanged and the next day blood plash is seen
under the carcass and the carcass is lightened. If blood outflowed badly because of animal
pathological condition before slaughter, after storage hanged carcass there arent any blood plash
and signs of bad blood outflow are more significant.
Blood outflow degree can be checked in such way: after 24 hours from animal slaughter into
fresh cut of thick muscles (m. pectoralis profundus; m. triceps brachii; m. glutaneus and other.) in 5
cm deep, strip of filter paper is put, leaving the free end at the meat surface (width of strip is 1.5 cm,
length is 10 cm). After 2 minutes the stripe is taken away and inspected. If blood outflow is good
tissue fluids are infiltrated into paper filter strip, at the contact place with muscles, marginally. If the
strip is logged with tissue fluids, but not higher than contact with muscles line blood outflow
degree is satisfactory. If paper filter strip is infiltrated with fluids and blood at the connection with
meat place, and also the part of stripe outstanding 2-3 mm above the cut is dewy too blood
outflow degree is bad. Very bad blood outflow degree is when the stripe is hardly logged with blood
at the connection place with meat and 5 cm above (this evaluation doesnt fit for defrosted meat),
table 11.
Compressoric method. Several pieces of analyzed meat are compressed between compressor

glasses and are microscopied. If blood outflow is good and satisfactory, there are no trace of blood
and muscle juices are liquid. If blood outflow is bad or very bad, cappilaries are fulfilled with
blood, muscle juices are red and blood spots are seen.
Shonberg analysis (haemoglobin-peroxidase). Small pieces of analyzed meat are put into

pits of porcelain plate and tincture of 5 % guaiacum salt is poured. Everything is mixed with glass
stick and 2 drops (0.1 ml) of 2 % hydrogen peroxide are dropped. On action of enzymes, after some
seconds, oxygen bubbles appear. If blood is outflowed well, after 1 min. thin light blue circle forms
around meat piece or no reaction appear even in 5 min. After 3 min. piece of meat is stirred several
times and is taken out. Then colour of solution is evaluated (but not later than 5 min after dropping
of hydrogen peroxide).

42

If blood outflow degree is good, the colour of solution is yellowish brown, if it is satisfactory
colour is light green or green, if it is unsatisfactory colour is dark blue or bluish violet, if it is
bad colour is dark blue, muddy.
Table 11. Evaluation the degree of animals blood outflow from carcasses
Indexes
Meat colour
Fat colour
Vessels

Muscle cut

Blood outflow degree

good
satisfactory
bad
Natural pink Intensively red
Dark red
or red
White
or White or yellow
Pink
yellow
Empty, dont There are some Blood is seen,
translucent
blood, they are they translucent
through
badly seen through through
pleura
pleura
and pleura
and and peritoneum
peritoneum
peritoneum

Blood
seen

isnt Wet

Limph node Lightly grey Lightly grey

or
lightly lightly yellow
cut
yellow
Healthy
Antemortem Healthy
animal
condition

very bad
Dark red with bluish
violet shade
Pink

Fulfilled with blood,

they are translucent
through pleura and
peritoneum,
pleura
and peritoneum are
pinky violet
After
pressing, Blood drops appear
blood and lymph
drops appear
or Lightly pink
Bluish pink

Sick

Agony or dead

Roder analysis. Roder reagent consists of 0.1 ml of Lefler methylenum blue, 40 ml of

distilled water and 0.05 ml of saturated spirituous solution of fuxin, dulluted with water for 10
times.
Procedure. 3 g of well crushed meat are put into test-tube and 5 ml of Roder reagent are

poured. The content of test-tube is shacked and after 5 min reaction is inspected.
If blood is outflowed well or satisfactory, the reagent stays blue, if bad it becomes brownish
green, if very bad the reagent becomes dark brown (Table 10).

4.3. Changes in limphonodes

Cut surface of healthy and in time processed animals limphonodes is lightly grey or lightly
yellow. Cut of limphonodes of animals slaughtered in agony is lightly violet or blue, because blood,
collected in small vessels starts to filter into sinuses of limphonodus and colours tissues of
limphonodus with pink colour. If oxidation processes of these tissues are at a loss, carbon dioxide
concentrate and blue or violet colour (of limphonodus) appears. Besides, limphonodi of sick
43

animals can be enlarged, juicy or atrophied, with haemorrhagia, necroses or other pathological
changes.
4.4. Meat boiling test

Meat sample is minced. 20 g of minced meat are put into 150-200 ml capacity flask, 60 ml of
distilled water are poured, the content is well mixed and, after closing with clock glass, it is stored
in boiling water for 10 minutes. When bouillon heats to 80-85oC, its odour is evaluated. Into
cylinder of 25 ml content and 20 mm diameter 20 ml of meat bouillon are poured and lucidity (in
white background) is evaluated visually. Meat bouillon of healthy animal is lucid and aromatic.
Bouillon of sick animals meat and of meat from animals, slaughtered in agony, is muddy, with
flakes, can have outsider odour, unrepresentative for meat.

4.5. Bacterioscopy of meat

Bacterioscopy of meat can be done in case to clear provocatives of infective diseases (anthrax,
emphysemic carbunculus, salmonellosis, swine erysipellas, pasteurellosis, Diplococcus or
Streptococcus infections and other).
Procedure. From different depth of muscular tissue little pieces are cut sterille and they are

pressed with cut side on objective glass (3 impressions are made on every objective glass).
Preparations are dried, fixed with fire, coloured by Gram and microscopied with immersion. In
preparations of healthy animals only separate microorganisms are seen or they are absent at all. In
preparations of sick, slaughtered in agony or died animals meat great amount of microorganisms is
seen. If it is suspicion, special search of toxicoinfections provocatives is made.

4.6. Evaluation of meat pH

For evaluation of meat pH, standard method must be applied: LST ISO 2917:2002 (LST ISO
2917:2002) Meat and meat products. pH evaluation. Reference method (identical to ISO
2917:1999).
Procedure. For biochemical reactions meat extract 1:4 is prepared. For testing partly

maturated meat is used, that means the time after animal slaughter is not less than 20-24 hours. 25 g
of meat (without fats and connective tissue) are weighed and well chopped with scissors. The
minced meat is put into the flask and 100 ml of distilled water (pH 7.0) are poured. The content of
flask is extracted for 15 min., during that time shaking not less than 3 times. Then the extract is
filtrated through paper filter.
Soon after animal slaughter meat pH is neutral or slightly alkaline (pH 7.0-7.2). After some
hours after animal slaughter, because of glycolytic processes in meat, lactic acid starts to
44

concentrate and meat becomes of acid reaction (pH 6.2-6.4), after 24-48 hours pH drops to 5.8-6.0.
Such pH stays for longer time. But if animal was sick, thinned or in agony before slaughter, there
are little glycogen in the muscles and activity of muscles enzymes is lowered. During maturation of
such animal meat, amount of concentrated lactic acid is little and meat becomes acid not significant.
pH measurings are made using colorimetric or potentiometric methods. Colorimetric method
is based on indicators feature to change colour depending on concentration of hydrogen iones. The
most indicators are selective and change colour only in margins of particular pH interval. Liquid
indicators or indicatory papers are used. Depending on used indicator and pH, colour of meat
extract changes. Different indicators can be used for measuring pH of meat and meat products
(Table 1).
For pH measuring of meat and meat products water extract 1:10 or 1:5 is prepared.
Preparation of meat-water extract: 5 g of minced meat are weighed at accuracy of 0. 01 g and put
into conic flask of 100 ml capacity, where 50 ml of distilled water are poured. Periodically stirring,
content of flask is stored for 30 min. Then it is filtrated.
Measuring of meat pH by potentiometric method and preparation of ph-meter for work is done
according instruction.

4.7. Peroxidase (benzidine) reaction

Activity of peroxidase enzyme depends on pH and oxidizing substances content in the extract
of meat. If pH of meat aqueous extract (1:4) is less than 6.0, peroxidase reaction is mostly positive,
if pH is 6.1-6.2 reaction is suspicious, and if pH is more than 6.2 it is negative. The reaction is
based on the fact that, on action of peroxidase, oxygen separates from hydrogen peroxide, which
oxidizes indicator benzidine and formes parachinon diimid, which, in responding to unoxidized
benzinine, forms a coloured compound coloured extract blue-greenish, later changing to brown
colour.
Procedure. 2 ml of tested meat aqueous (1:4) extract are added to a test tube, later 5 drops of

0.2% benzidine spirituous (96%) solution are added and everything is mixed. Then 2 drops of 1%
fresh hydrogen peroxide solution are dripped and the reaction is being observed. The filtrate of
healthy animal meat within 1-2 minutes becomes blue greenish, then gets to brown (if peroxidase
reaction is positive); meat filtrate of sick, weary during transport or slaughtered in agony animals
turns blue green, soon changing to brown or very quickly becomes brown (negative reaction).
Peroxidase reaction can be carried out in an accelerated (express) method. On fresh meat slice
2 drops of 1% hydrogen peroxide solution and 5 drops of 0.2% benzidine are dropped. The reaction
is considered positive if, in the place where the reagent has been applied, there appears blue-green
45

colour spot, which later gets brown; if the reaction is negative, blue-green colour spot does not
form.
4.8. Formalin reaction

In slaughtered after a long agony or seriously sick animals meat protein breakdown products polypeptides and free amino acids can appear. During their reaction with formaldehyde a clot
forms. This reaction is used to investigate the beef.
Procedure. Formalin before the usage is being neutralized with the 0.1n sodium hydroxide

solution, using mixture of 0.2% neutral red and methylenum blue aqueous solutions equal parts as
an indicator. The mixture is being titrating until the red color changes to green. 10 g of tested meat
(with no fat and connective tissue) are well chopped with scissors in a mortar, then 10 ml of saline
solution and 10 drops of 0.1n sodium hydroxide solution are added and everything are grinded with
pestle. Retrieved mash muscle, with a help of glass rod, is being placed in the flask, and in order to
precipitate proteins, everything is heated to boiling. After the content of the flask is being cooled
with water, neutralized with 5 drops of 5 % oxalic (sorrel) acid solution and filtered through a paper
filter. If filtrate is cloudy, you must filter it again. Add 2 ml of clear filtrate and 1 ml of neutral
formalin into a test tube. Reaction is seen in a minute. Meat extract of found dead, slaughtered at
the stage of agony or hardly affliction animals turns into a blood clot, or flakes occurs (positive
reaction). Meat extract of animal, which had difficult and long-term suffer, produces flocculent
sediments (suspicious reaction). A healthy animal's meat extract remains liquid and transparent,
sometimes noticeable turbidity (negative reaction).

4.9. Evaluation of acidity-oxidation coefficient

Titratable acidity of healthy animals meat and its extract significantly increases because of
accumulation of milk, and ortho-phosphoric and other acids. Titratable acidity of sick animals meat
increases slightly. Meats oxidation depends on amount of microorganisms and organic matter
degradation products (reducing substances). Reducing matters or reducers are called substances,
which reduce oxidator (potassium permanganate). In turn, potassium permanganate oxidizes
reducers to colorless oxidation products, while reducing agents reduce potassium permanganate to
colorless materials: manganese (II) sulfate, potassium sulfate and water.
In meat of sick animals, titratable acidity increases slightly during the same period (24 h),
while reducing material content is significantly higher than healthy animals. Thus, there are less
reducers in fresh meat of healthy animals. Characteristic of the meat titratable acidity and reducing
material size vary in opposite directions. Acidity-oxidation coefficient of healthy animal's chilled

46

meat is generally 2-3 times greater than of the sick animals. When the meat starts to spoil, this ratio
decreases.
Evaluation of titratable acidity. 10 ml of a meat extract (1:4) are poured to the flask No.1 and

it is diluted with 40 ml of distilled water, later 3 drops of phenolphthalein spirituous 1% solution are
added and are titrated with 0.1n of sodium hydroxide until slightly pink color appears and amount
of 0.1 n NaOH (ml) used for titrating is wrote.
Evaluation of reducing substances amount (oxydation). In acidic environment potassium

permanganate oxidizes organic substances which are in the tested meat extract to colorless
oxidation products. Thus, amount of potassium permanganate (oxydants) describes the amount of
organic reducing substances in meat extract. To the flask No. 2, 50 ml of distilled water, 5 ml of
0.4n sulphuric acid solution and 1-2 drops of 0.1n potassium permanganate solution are added until
slightly pink colour appears. Contents of the flask is heated to 40-50oC (after heating if color
disappears add an additional 1-2 drops of potassium permanganate solution). 2 ml of meat extract
(1:4) are poured into a warm flask and immediately filtered with 0.1n potassium permanganate
solution till slightly pink color appears, not disappearing for 0.5 min. 0.1n KMnO4 solution (ml)
consumed for flask No. 2 content titration is converted to 10 ml of extract, while 2 ml of meat
extract were taken for analysis. Therefore, amount of this solution, used for titrating of flask No.2,
is multiplied by 5.
Acidity-oxidation coefficient (AOC) is calculated using formula:
AOC = A/B
A amount of 0.1n NaOH solution (ml) consumed for flask No.1 content neutralization
(titration);
B amount of 0.1n KMnO4 solution (ml) consumed for titration of 10 ml of flask No.2
content.
Evaluation: AOC of fresh not chilled meat is 0.15-0.2. AOC of healthy animal mature meat is

0.4-0.6. ROK of sick animals meat and doubtful freshness meat is 0.2-0.4. AOC of spoiled meat is
from 0.05 to 0.2.
Example of calculation. For titration of 10 ml extract in the flask No.1, 3 ml of 0.1 n Na OH

solution was used, and for titration of 5 ml extract in the flask No.2 1.4 ml of 0.1 n KMnO4 solution
were used. Amount of 0.1 n KMnO4 solution (ml) used for flask No 2 content titration is converted
to 10 ml of extract. For this case amount of this solution, used for titration of flask No.2 is
multiplied by 5:
1.4 ml of 0.1 n KMnO4 x5 = 0.1 7ml N KMnO4 solution;

AOC = 3/7 = 0.43

47

4.10. Luminescence study

It was found that the meat shine is specific depending on its quality indicators which change
when animal is ill. Meat extract (1:4) is heated to coagulate the proteins. Then it is being filtered
through the paper filter. Ultraviolet radiation-emitting device is plugged 10 min. before starting
work. The analysis is conducted in the dark room. 2 ml of meat extract are added to the test tube.
The test tube with the meat extract is being placed against ultraviolet ray stream at angle of 450 on
the dark background. Colour of the tube content and fluorescence intensity are observed. Meat
extract of healthy animals shine in pink or pale purple color; extract of sick animals shine of
various intensity greenish-blue colour.

5. Evaluation of meat and poultry freshness

Changes of meat quality during storage. During storage of mature meat, so-called deep

autolysis process begins, when proteins and fats start to break down. On action of cathepsins and
peptidases, peptide junctions break and on action of lipase fat hydrolysis proceed. Changing
proteins, muscle tissue morphological structure changes too: meat juice secretes, brown surface
color and unpleasant sour taste appears. Such meat is unfit for human consumption.
Meat is perfect medium for the development of microorganisms, thus changes happening
during meat storage depends not only on the tissue enzyme, but also on the activities of
microorganisms. Microbiological deterioration of meat is directly related to the animal's anteslaughter condition: sick, hungry and tired animals meat deteriorates quickly. After slaughter
microorganisms may be found in surface and inner layers of the meat. Microorganisms get to the
inner layers when animal is alive or at slaughter, and surface contamination increases, when animal
preparing to slaughter sanitary (washing, condition of ante-slaughter environment), slaughter
(quality of bleeding, skin peeling, removal of internal organs) and meat storage (disinfection of
cameras, optimal temperature) conditions are bad.
When meat spoils because of microbiological effect, it texture, color, odour and taste change,
besides toxic substances can accumulate.
The main and most common meat failure is the putrescence. Putrescence is caused by
proteolytic microorganisms, for development of which proteins or their partial hydrolysis products
are required. This process can be both aerobic and anaerobic and depends on pecularities of
destroying proteins, external environmental factors and the type of microorganisms.

48

Anaerobic decay of meat begins in the thick muscle layers, near the bones and joints. During
this process gases are produced and accumulate between muscle fibers. Meat color changes to bluepurple or gray-green, the structure becomes porous with unpleasant smell. For such meat, specific
alkaline pH values are typical (8.0 pH 9.0). Anaerobic putrescence may occur during forced
slaughter case, as well as if there was long time period from animal is stunning till the removal of
guts during which microorganisms moved from intestines to the carcass.
The first sign of aerobic putrescence is formation of mucus on the surface of the meat. In the
initial stage of decay brown meat color gradually changes to gray during deterioration process. In
deep decay phase the meat color becomes green. Under aerobic degradation meats pH is between
7.0 and 8.0, the meat has unpleasant, but not so sharp as of anaerobic rotting odour.
During meat maturating, accumulate acids, which make adversity for proteolytic bacteria
development. However, mold fungi are active in the acidic environment too. On action of molds
enzymes, ammonia and organic hydroxides excrete, which replace the ambient pH to the alkaline
side, creating the right conditions for bacteria to develop. Proceeding aerobic and anaerobic
microorganisms (together or in succession) meat proteins hydrolysis starts, during which various
molecular weight peptides and free amino acids form. Later, in parallel with the further
decomposition of proteins and polypeptides, amino acids decompose: deamynate, decarboxylate,
oxidize or reduce. Circumstances of ones or other process are determined by microorganisms type
and conditions of meat storage in cameras. Predominant amino acid degradation processes are
deamination and decarboxylation. Deamination and decarboxylation processes formed by oxy-and
keto acids changes, formation gas, alcohols and aldehydes. During deamynation of amino acids,
fatty acids and ammonia form. More than 90% of the volatile fatty acids consist of acetic, butyric,
formic and propionic acids, giving the meat an unpleasant odor.
A number of amino acid degradation products agmatine (from arginin), cadaverin (from
lysine), phenilethylamine (from phenylalanine), tiramine (from tyrosine) are toxic, but their
degradation products are less toxic. This may explain the higher poisoning in primary meat spoilage
stages and lower if putrification is prolonged.
Because amino acids can deaminise and decarboxylise at one time, from tyrosine and
tryptophan forme cresol, phenol, indole and skatole. In decay process, the degradation of amino
acids having sulfur (cysteine, cystine, methionine) hydrogen sulphide and ammonia excretes and
mercaptan formes. Indole, skatole, cresol and mercaptan are toxic substances like hydrogen
sulphide, characterized by a specific unpleasant odour. Greater H2S content accumulates during
stages of meat spoiling deeper stages. Reacting sulfur hydrogen with myoglobin, meat gets greenish
shade (sulfomioglobin forms).
49

During early meat putrification stages, ammonia, hydrogen sulphide, carbon dioxide, volatile
fatty acids form and during later stages indole, skatole, phenol, cresol start to accumulate.
During meat storage fat break down to hydrolysis and oxidation proceed. Fat hydrolysis is
mostly stimulated by meat enzymes, but it can be caused by microorganisms (genus Pseudomonas)
too. They also have an impact on the oxidative breakdown of fat. As a result of this, free of fatty
acids, organic peroxides, and in later stages aldehydes, ketones, low molecular mass fatty and
oxyacids accumulate in meat.
Storing meat at low temperatures and increased humidity ambience, mold and pigment
microorganisms form different colour dirts or stains on the meat surface. Mature meat with pH from
5.6 to 6.0 quickly gets molded, molds mostly form on the meat surfaces where air couldnt get (eg,
in the inner side of the ribs, flank folds, etc.). Mould fungi can cause decay of meat, besides some of
the mycotoxins secreted by the molds are toxic.
On the surface of meat and phosphorescent bacteria can develop too, they contaminate stock
usually at the storage in cameras. Phosphorescence at first appears in more wet meat places (around
the joints, bones depressions in the vertebral cartilage). When firs putrification signs appear, shine
disappears, because the proteolytic bacteria inactivate phosphorescent microflora.
In aerobic conditions molds, yeast and bacteria degrades the meats carbohydrates to CO2 and
H2O, but intermediate products organic acids can form too. They little influence meats taste and
odour, but change pH to the acidic side. In cases if carcasses are not well and quickly cooled,
accumulate glycolysis products pyruvic, lactic acid, ethanol, acetic and butyric acid, CO2, caused
by tissue enzymes and some anaerobic microorganisms. Sour odor and flavor, softer texture
appears, color at the cut becomes gray, meat gets rancid. Rancid meat putrifies and becomes
mouldy more quickly.
During action of microorganisms, lipids break from lipoproteides and they broke down
further. In this way from choline, constituent lecithin part, during meat putrification trimethylamine,
dimethylamine, methylamine form. In oxidation of trimethylamine, trimethylamine oxide, with a
specific smell of fish form. From choline in such way toxic material neurine can form. During
microbiological spoilage nitrogenous extractives of meat break down, NH3, CO2, and some toxic
compounds form: metil-guanidin from creatine, histamine-from carnosine. In anaerobic conditions,
phosphine (PH3) with unpleasant odour form.

5.1. Evaluation of meat freshness

Thus, the freshness of meat and suitability for further processing depends on the initial
preparation and storage process and the quality can be estimated by the sensory characteristics and
50

the quantity and composition of nutrition substances changes primary and secondary products
generated during storage.
Meat freshness assessment is based on the "Meat and poultry freshness evaluation technical
regulation" (2002). At first, meat sensual analysis is done. If there are any doubt chemical and
microbial pollution analysis are done. Chilled meat and poultry are investigated defrosted.
Meat freshness is evaluated according sensual analysis, chemical characteristics, microbial
contamination and meat pH to 25 points.
Sensual indexes (appearance, colour, texture, odour and quality of the broth) are estimated to
13 points. Chemical characteristics are measured to 10 points, including: content of volatile fatty
acids 4 points, reaction with copper sulphate 4 points, content of ammonium nitrogen 2 points,
microbial contamination 2 points. Meat freshness indexes, in points: fresh meat from 21 to 25
points, meat of doubtful freshness from 10 to 20 points, not fresh meat from 0 to 9 points (Table
12).
Meat appearance and color, texture, odour, muscle, fat and tendon condition are evaluated.
Also meat boiling test is done.
Meat surface, colour and fat colour are evaluated. Meat is incised (in thicker muscle area) to
the bone and colour, viscosity and appearance of fresh cut are observed. Meat surface moisture is
determined touching with fingers or touching white filter paper to the surface of fresh meat incision.
Chilled and defrosted meat is wet and red juice is seen. Consistency is measured by pressing meat
with finger and observing the time required to equilibrate the pressed pit. Odour is determined by
analysing meat samples. At first surface of the meat is smelled, later deep incision is made and is
smelled immediately opening cut. Odour of meat near bones is observed especially. Pronounced
flavor is of meat bouillon.
Fat condition is evaluated by assessing fat color, odour and texture. Colour of chilled fat

change little, its texture is firm. Defrosted fat are soft, watery, with reddish shade. Chilled for a
second time fat become brownish tinge.
Bone marrow are assessed by monitoring the tubular bones cut. It is watched whether they

fill all entire hollow bone. Later marrow are removed from the bone, their color, luster and elasticity
of the fracture site are evaluated. Fresh meat bone marrow is firm, smooth, not bounded from bone,
yellow, shiny in fracture site.
Condition of tendons is evaluated observing their elasticity and strength. Bones joints and

their fluid color are monitored too. Tendons of fresh meat have to be strong, hard. Articular bone
heads and they holes have flat, smooth surface, and little amount of fluid is colorless and
transparent. Tendons of defrosted meat are dark red, bloated and soft.
51

Sensual characteristics of meat freshnes is shown in Table 13.

Table 12. Evaluation of meat sensual and chemical indexes
Indexes
Reduced points
Sensual indexes
Slight surface mucousity without other sensual changes
2
Changed colour of meat surface and fat, single white molds, slightly
5
sour odour, dark surface
Slow (not longer than 1 min.) equilibration of pit after pressing, slightly
sticky fat and slightly cloudy broth.
7
13
Meat surface is dried or very wet, gray or greenish color, small amount
of mucus, slightly slimy fresh-cut surface, darkened, significant moistening of
filter paper, attached to the surface; cloudy meat juices; slow (longer than 1
min.) or partial equilibration of pit after pressing; slight putrific odour at the
surface, not appreciable in deeper layers; fat with gray matte colour and
fissionable fat odour; beef tallow is spready; bouillon is cloudy with
unpleasant odour, small fat globules on the surface.
Meat surface is gray or greenish, slimy or moldy; cut surface is very
By such
sticky, greenish or gray; meat is softened in cut; pits after pressing dont organoleptic rates
equillibrate; strong acidic putrescent odour in deeper layers of muscles; fat are meat is discarted
greenish with dirty colour, spready, with the sharp smell of fat breakdown; unvalued points
bouillon is cloudy with flakes and putrescent odour, fat droplets on the surface
almost non-exist.
Chemical indexes
Volatile fatty acids, ml
to 0.35;
0
from 0.36 to 0.50;
1
from 0.51 to 0.65;
2
from 0.66 to 1.0;
3
more than 1.0.
4
Reaction with copper sulphate in bouillon:
- bouillon is clear or slightly cloudy;
0
- bouillon with flakes;
3
- bouillon with a bluish or greenish flakes and sediments
4
Amount of amino-ammonia nitrogen, mg/100 g:
to 80;
0
from 80 to 130;
1
more than 130.
2
Microbial contamination
There are no microorganisms in the field of view or isolated cocci and rods are
seen, there are no reminders of spoiling meat.
There are 20-30 cocci and isolated sticks in the field of view, clear signs of
muscle tissue spoiling.
There are many microorganisms and decomposing muscle tissue in the field of
view.

0
1
2

52

Table 13. Sensual characteristics of meat freshness

Indexes

Meat
Fresh
Doubtful freshness
Not fresh
Meat is encrusted with Surface is wet, sticky, The surface is very dry,
Surface
bright red dry film
darkened
encrusted with grayappearance
brown mucus or molds
and colour
Slightly wet, but wet Wet (wet filter paper), Wet (wet filter paper),
Muscle
trace is not left on slightly sticky, dark red sticky, red-brown color
incision
filtered paper, color is color
typical
for
animal
species
Firm at the incision, with Not so firm at the incision Tumbled at the incision;
Consistency
finger
pressed
pit as fresh meat; with finger with finger pressed pit
equilibrates soon
pressed pit equilibrates doesnt equilibrates
slowly (within 1 min.)
Specific, typical for Slightly acidic or tainted
Sour, tainted or weak
Odour
fresh meat
decay
Cattle - white, yellowish Dull
greyish,
slightly Gray
mat,
spotting
Fat
or yellow, hard, crumble sticky, may have rancidity consistency.
under pressure;
odour
Swine
sometimes
Swine - white or slightly
encrusted with mold,
pink, soft, smooth;
have rancidity odour.
Sheep - white, solid.
Condition of Tendons are elastic, Tendons are little soft, Tendons are soft, grayish
Articular opaque, white. Articular color. Articular surface is
tendons and resilient.
surface is smooth, glossy surface is slightly slimy
mucous.
joints
Meat boiling test. The study follows the methodology described in the chapter Evaluation of

sick animals meat. Bouillon of fresh meats is clear and fragrant; bouillon of doubtful freshness
meat is clear or cloudy, not typical for fresh meat odour can be felt; bouillon of not fresh meat is
cloudy, has unpleasant odour.
Lucidity and odour of bouillon. Weigh 20 g of crushed sample into a 100 ml conical flask

with 0.2 g nicety and add 60 ml of distilled water. Solution is mixed and heated for 10 minutes a
water bath at 80-85C temperature. At the moment when a steam appear, odour of bouillon is
evaluated. Lucidity is assessed visually in 20 mm diameter glass cylinder.
Meat bacterioscopy. The study follows the methodology described in the chapter "Evaluation

of sick animal meat. Meat freshness is evaluated by scores by the number of bacteria in the
preparation and the degree of degradation of muscle tissue. When in the visual field
microorganisms are not seen or seen isolated cocci and rods, there are no decompose meat tissue
residues 0 points; when in the field of view there are seen 20-30 individual cocci and rods and
there are clear signs of breakdown of muscle tissue 1 point; when in the field of view there are
many microorganisms and disintegrated muscle tissue 2 points.
53

Volatile fatty acid (VFA) content. During deaminysing of amino acids, fatty acids are

formed, most of which are volatile (formic, acetic, propionic, butyric, valerian, nylon, etc.). They
influence formation of meat flavor. Volatile fatty acids content is determined by steam distillation
of the acidified aqueous extract of the sample under investigation and then titrating distillate.
Investigative apparatus is shown in picture 21.

Pict. 21. Volatile fatty acid distillation apparatus: 1 - a round-bottomed flask for steam processing, 2
distilation flask, 3- Libich cooler, 4 flask for distillate collection, 5 electric heater.

Procedure. 25 g of crushed sample are weighed at accuracy 0.01 g, placed into 0.75-1 liter

round-bottomed flask and 150 ml of 2% H2S04 solution are poured. Content is well mixed, the flask
is tightly covered with plug with consolidated glass tubes. One of them is connected to the
distillation flask with evaporator (flat-bottomed flask), the other - to steam gathering dial, connected
to fridge. In evaporator water is simered and it is distilled until in 250 ml capacity flask 200 ml of
distillate are gathered. The distillate is titrated with 0.1 M NaOH or KOH solution, using
phenolphthalein as indicator. Parallel control test is done.
VFA content expressed in milligrams of KOH or NaOH used in the sample contained 25g
neutralize VFA is calculated using the formula:

x=

5,61 (V1 V2 ) K 100

m

54

V1 amount of 0.1 M NaOH (KOH) used for titrating 200 ml of distillate, ml;
V2 amount of 0.1 M NaOH (KOH) used for titrating control sample of the distillate, ml,
5.61 amount of NaOH (KOH), being in 1ml of 0.1 M solution, mg;
K the exact conversion to the 0.1 M NaOH (KOH) coefficient ,
m sample weight, g.
Meat freshness is evaluated, comparing data with the standard VFA content, expressed as mg
of NaOH (KOH) values.

Evaluation of primary protein degradation products in bouillon (copper sulfate

reaction). The method is based on copper ion reaction with protein primary interaction products,

forming of insoluble compounds of copper sulphate.

Procedure. 20 g of crushed sample are weighed at accuracy of 0.2 g into a 100 ml conical

flask. 60 ml of distilled water are added. The solution is mixed, heated for 10 minutes in a water
bath to 80-85C temperature, filtered through 0.5 cm thick layer of cotton wool into a test tube,
soaked in glass of cold water. If after filtration, protein fibers remain, bouillon is filtered again
through the paper filter. 2 ml of filtrate are poured into the test-tube and 3 drops of 5 % CuSO4
solution are added. Tube is shacked for 2 or 3 times and placed in the rack. After 5 minutes,
analysis results are set, comparing them with the results shown in Table 12.

Evaluation of Ammonia and ammonium salts. Ammonia and ammonium salts reacting

with the Nessler reagent (double salt of mercury iodide and potassium iodide, melted in potassium
hydroxide) form mercuramony iodide a substance of yellow-brown colour.
Procedure. 5 g well crushed sample are weighed at accuracy of 0.001 g and placed in a

conical flask with the 20 ml of distilled water. It is left for 15 minutes, during that time three times
shaking the flask content. Received aqueous extract is filtered through paper filter. In a test tube 1
ml of aqueous extract and 10 drops of Nessler reagent are added. Substance is mixed and it colour
change and lucidity are observed.

Peroxidase analysis method (poultry is not analyzed according this method). In healthy

animals muscle tissue peroxidase is found. In meat of sick, tired or in agony slaughtered animals
amount of this enzyme is low or completely absent. Activity of peroxidase depends on the pHindicator. When pH is lower than 6.5 (spoiled meat), the peroxidase reaction is negative.
On action of tissue enzyme peroxidase, hydrogen peroxide oxidizes benzidine to
parachinondiamid. The latter, reacting with unoxidized benzidine forms unstable meri-chinoydic
55

compound, coloured bluish-green changing to dark brown colour. When amount of peroxidase in
meat is high, all benzidine amount is oxidized to parachinondiamid and meat extract doesnt get
coloured specific bluish-green color or immediately assumes dark brown colour.
Procedure. 2 ml of an aqueous meat extract and 5 drops of 0.2% benzidine spirituous solution

are added into the test-tube. Content is mixed and 1% of H2O solution is poured. Colour changes of
meat extract are observed.

Evaluation of acid number. The method is based on titration of free fatty acids (FFA),

dissolved in fat, with aqueous KOH solution. Ether is used as fat solvent and ethanol homogenising
system, which form fat with aqueous alkali solution during titration.
For determination of meat fat condition, analyzed sample is mixed with the water absorbing
material and fat are extracted using organic solvent. For water absorption different polarity solvent
mixes are used (i.e., chloroform and methanol), and lipids usually are extracted with chloroform.
Analyzed sample is crushed, mixed in 250-350 ml flask with water absorbing NaSO3 (ratio of
product and Na2SO3 is 1:2) and the flask is plugged. Later, the sample is extracted with chloroform
(ratio 1:3). Solution in the flask is shacked intensively for 15 minutes and is filtered through paper
filter.

Evaluation of fat amount in chloroform extract. 10 ml of chloroform solution are poured

into a pre-dried to constant weight and weighed porcelain dish, which is placed for 10 minutes into
a water bath to evaporate the chloroform. Porcelain dish with fat is drying in oven at 100-105C
temperature to constant weight.

Evaluation of acid number. 10 ml of the chloroformic extract are poured into a conical flask

and dried in the drying oven at 100-105C temperature until smell of chloroform disappears. After
that 20 ml of neutral ethanol and ether mix and two or three drops of indicator (1% phenolphthalein
solution) are poured and quickly titrated with 0.01 M NaOH or KOH solution until the solution gets
coloured into pink color, not fading during 1 min.
Acid number, expressed in milligrams of KOH or NaOH used for neutralization of 1 gram of
fat in FFA, is calculated using the formula:

x=

5,61 V K
m

56

V amount of 0.01 M KOH (NaOH) (ml) used for titration;

5.61 - KOH (NaOH) content in 1 ml of 0.01 M solution (mg);
K recalculating to 0.01 M KOH (NaOH) coefficient;
m fat amount in extract g.
Meat luminescence analysis. Each material glow in ultraviolet light specifically according its

own wave spectrum. During storage fluorescence spectrum of meat changes. According adequate
spectrum changes, meat freshness can be evaluated.
Procedure. 2.5 % aqueous meat extract is prepared. For settlement of proteins, extract is

heated to 60C and filtered. Test-tube with extract is placed in 45 angle against ultraviolet light.
Colour of meat extract is observed in dark background.

Meat pH. pH values of fresh meat range from 5.6 to 6.2 (1 hour post-mortem pH is from 6.8

to 7.0; 24 hours post-mortem pH is from 5.6 to 6.0). Poultry freshness is evaluated according
sensual, chemical, microbiological indexes and according to the freshness of poultry fat (colour,
flavour, odour, acid number, number of peroxides).

5.2. Evaluation of poultry freshness

Freshness of poultry is evaluated according sensual, chemical and microbiological indexes

and according freshness of poultry fat (colour, taste, smell, number of acids, number of peroxides) .

5.2.1. Sensual analysis of poultry

Appearance and colour are evaluated at first inspecting surface of poultry carcass, deeper
layers are inspected after fresh incision. Surface moisture of incision is evaluated touching filter
paper to it. Consistence is evaluated pressing with spatula or finger, according time of pit
equilibrating. Odour is evaluated at surface and incision, nobbling to muscular tissue near bones.
Consistence of fats is evaluated pressing with spatula or finger. Quality of bouillon is evaluated
according odour, lucidity, flavour and condition of fats at the surface. For evaluation of lucidity, 20
ml of bouillon are poured into cylinder (diameter 20 mm) and observed visually. Feculence of
bouillon is caused by primary breaking products of proteins, solved in hot water. On this effect,
poultry fats are emulgated too. Indexes of poultry sensual analysis are shown in table 14.

57

Table 14. Sensual indexes of poultry

Index
Typical signs
Fresh
Doubtful freshness
Appearance
and Dry, white to yellow colour Wet and sticky in
places, sticky under
colour of poultry with pink tone.
wings and at skin
surface
wrinkles, white to
yellow colour with
grey tone.

Appearance
and
colour of muscular
incision

Consistence
muscles

of

Odour

Characteristic
bouillon

of

Bad (not fresh)

Coated with mucus,
especially
under
wings and at skin
wrinkles, white to
yellow colour with
grey tone, sometimes
light yellow or dark
spots are seen.
Lightly wet, not leaving Wet, leaving wet Wet, leaving wet
wet spots on filtrating spots on filtrating spots on filtrating
paper, white pink colour paper, lightly sticky, paper, sticky, dark.
(hens and turkeys) or red darker than of fresh
(geese and ducks) colour.
poultry.
Muscles are resilient, pit Muscles
are
less Muscles
are
not
after pressing equilibrates resilient to compare resilient, pit after
soon
with fresh poultry, pit pressing
doesnt
after
pressing equilibrate.
equilibrates
slowly
(not longer than after
1 min.).
Specific, typical for fresh Fusty
Putrid both at the
poultry
surface and at muscle
incision
Lucid, fragrant.
Lucid or light turbid, Turbid with much
with light unpleasant flakes
and
clear
odour.
unpleasant odour.
5.2.2. Chemical analysis of poultry

Evaluation of volatile fatty acids. VFA appear in meat or poultry during reactions of

deamination of amino acids, when meat or poultry are spoiling. Their amount is evaluated inducing
extract of meat or poultry with sulphur acid and distilling with steam.
Procedure. Weigh 25 g of minced poultry, put in rounded bottom flask (capacity of 0.75-1.0

L), pour 150 ml 2% solution of sulphur acid, mix, heat-seal and distille with water steam till getting
200 ml of distiller. In parallel, at the same circumstance prepare control sample. It is necessary to
evaluate VFA, which can be in sulphur acid. Then titrate the distiller with solution of 0,1 mol/l
potassium hydroxide (indicator phenolphtalein). Amount of VFA (mg) in poultry is calculated using
formula:
(0,230,05) 15,61
X=

25

= 4 mg KOH

58

5.61 titre of 0.1 mol/l potassium hydroxide solution, mg/ml,

V1 amount of 0.1 mol/l potassium hydroxide solution, used for neutralization of VFA in 200
ml of poultry extract, ml,
V2 amount of 0.1 mol/l potassium hydroxide solution, used for neutralization of VFA in 200
ml of control extract, ml,
K correction coefficient of potassium hydroxide molar concentration (1,0 for 0,1 mol/l
solution of potassium hydroxide).
g sample mass, g.
In fresh poultry, amount of VFA is not higher than 4.5 mg; poultry is of doubtful freshness if
amount of VFA is 4,5-9,0 mg and not fresh if VFA = 9,0 mg.

Reaction with copper sulphate. This reaction shows protein breaking down products in

bouillon of meat or poultry. In bouillon of fresh meat, 5 % copper sulphate solution doesnt makes
any changes or light turbidity appears. In bouillon of not fresh meat flakes or jelly sediments (light
blue or light green colour) appear. Flakes appear because of cooper sulphate reaction with primary
protein breaking down products; coloured sediments appear because of cooper sulphate reaction
with later breaking down products.
Procedure. 20 g of minced meat pour with 60 ml of distilled water. Mix content of flask and

keep in bath of boiling water for 10 min. Filtrate hot bouillon through filter of cotton-wool. If
filtrate is not lucid, repeat filtrating through paper filter. Pour into test-tube 2 ml of bouillon and 3
drops of 5 % cooper sulphate solution. Shake test-tube and after 5 min. evaluate results of the
reaction.
Evaluation of amino-amonia nitrogen. The method is based on reaction of ammonia and

amino groups with formalin. Total amount in meat extract is equivalent to amount of 0,1 mol/l
sodium hydroxide and is evaluated titrating.
Procedure. Grind 25 g of minced meat with 30-40 ml of distilled water, pour everything into

flask and dilute with distilled water to 100 ml. Shake content of flask 3 min., let to settle, shake
again for 2 min. and filtrate. To 100 ml flask pour 40 ml of extract and settle proteins with 10 %
solution of aluminium alum [AlK(SO4)212H2O] and solution of saturated barium hydroxide. Total
capacity of settled solution should be not less than capacity of extract. Amount of barium
hydroxide, which is necessary for neutralization of 10 % aluminium alum, is evaluated titrating 10
% solution of aluminium alum (adding 5 drops of 1 % spirituous solution of phenolphtalein) with
saturated solution of barium hydroxide. From for titrating used amount of barium hydroxide,
calculate amounts of aluminium alum and barium hidroxide necessary for precipitating of extract
59

proteins. After precipitation, dilute capacity of flask to 100 ml, mix and after 10 min. filtrate. In
parallel prepare control solution. Neutralise20 ml of prepared filtrate and control solution (adding
0.3 ml of indicator: equally mixed 0.1 % neutral red and methilenum blue spirituous solutions)
titrating with 0.1 mol/l solution of sodium hydroxide till light green colour. After that, into mix of
reaction pour 10 ml of formalin (titrated till neutral reaction) and add 0.5 ml of another indicator (1
part of 0.1 proc. thimolium blue mixed with 3 parts of 1 % phenolphtalein, dissolved at 50 % of
spirit). After reaction with formalin, titrate again with 0.1 mol/l solution of sodium hydroxide till
violet colour.
Amount of amino-ammonia nitrogen (mg/ 100g) is calculated using formula:

X=

1,4x100x100(V-V1)x100
=70x(V-V1)
25x40x20

X = 70 (V- V1)
V amount of 0.1 mol/l sodium hydroxide solution (ml), used for titrating the main
(investigative) sample after reaction with formalin.
V1 amount of 0.1 mol/l sodium hydroxide solution (ml), used for titrating the control
solution.

Reaction with Nesler reagent. Preparation of poultry extract.

Procedure. Cut peaces of muscle tissue (without fats and connective tissue) from different

depth of investigative sample and crush. Pour 5 g of sample in flask with 20 ml of distilled water
and leave for 15 min., shaking every 5 min. Filtrate the amount of flask through paper filter.
Increase of ammonia in meat is as a result of deamination, happening in putrescent meat. Drip 1 to
10 drops of Nesler reagent into 1 ml of extract. Shake test-tube after every drop and watch changes
of extract colour and lucidity.
If poultry is not fresh, after dripping 6 drops of Nesler reagent into poultry extract, it becomes
yellow to orange and flake sediments appear.
Peroxidase reaction using benzidine. Peroxidase in poultry tissues reacts with hydrogen

peroxide and makes complex. Hydrogen peroxide in action of peroxidase reacts with benzidine
formatting coloured compounds.
Procedure. Pour 2 ml of extract and 5 drops of 0.2 % spirituous solution of benzidine into

test-tube, shake and add 2 drops of 1 % hydrogen peroxide solution. If after 1 to 2 min. blue-green
colour, changing to dark brown appears, reaction is positive. If colour doesnt appear or appears
60

later than after 3 min., reaction is negative. Reaction of fresh poultry should be positive. If it is
negative, poultry is of doubtful freshness or not fresh (this reaction doesnt fit for meat of water
birds and chickens).
Evaluation of poultry pH is performed according LST ISO 2917 Meat and meat products.

Evaluation of pH. Reference method.

Evaluation of microbial contamination. Little pieces from different depth of muscle tissue

are cut aseptically and pressed with cutting side to objective glass (make 3 impresses on every
glass). Then they are dried, fixed with fire, painted according Gram and microscopied with
immerse. Freshness of poultry is evaluated according count of bacteria in preparate and degree of
muscle tissue degradation.
When poultry is fresh, there are no bacteria in sight or single cocci or rods are seen, there are
no signs of muscle tissue degradation. When poultry is of doubtful freshness 20 to 30 cocci and
several rods are seen in sight, there are first signs of muscle tissue degradation. When poultry is not
fresh more than 30 cocci or rods are seen and there are clear signs of muscle tissue degradation.

5.2.3. Evaluation of poultry fats

Subcutaneous and internal fats are evaluated separately. Sample of subcutaneous fats is made
of fats from back side, neck and under wings. Internal fats are taking from fatty tissue, separated
from meat and connective tissue. Fats are crushed, dissolved in water bath, filtrated through 4 layers
of gauze and cooled to 20oC temperature.
Colour is evaluated pouring 5.2 ml of fats (dissolved and cooled till room temperature) into

dry test-tube. Colour should be evaluated in day light.

Odour and flavour are evaluated by sensual analysis, mixing dissolved fats in glass with

glass stick at the room temperature. Colour of fresh fats is light yellow or yellow. Colour of
doubtful and not fresh fats is palish yellow or yellow with grey tone. Odour and flavour of fresh fats
are typical without outside odour and taste. When fats are of doubtful freshness light specific
flavour and odour of breaking down fats are felt. When fats are not fresh clear specific flavour
and odour of breaking down fats are felt.

Evaluation of fat acidity number. Acidity number is evaluated according LST EN ISO 660

Animal and vegetable fats and oils - determination of acid value and acidity.
Acidity number is amount (mg) of potassium hydroxide, necessary for neutralization of free
fatty acids in 1 g of analyzing fats. Acid value in fresh fats is from 1.2 to 2.2 mg KOH/g.

61

Evaluation of peroxides number. Peroxides number is evaluated according LST EN ISO

3960 Animal and vegetable fats and oils - determination of peroxide value.
Peroxide value is amount of active oxygen (mmol/kg arba mE/kg.) corresponding to amount
of peroxides in 1 kg of sample. Peroxide value in fresh fats is not higher 10 mE of active oxygen for
1 kg of fats. Evaluation is shown in Table 15.

Table15. Evaluation of poultry freshness according the results of fat investigation

Quality of fats

Fresh (chilled and frozen poultry)

Doubtful freshness: chilled poultry of:
hens
geese
ducks, turkeys
Frozen poultry
Not fresh poultry

Number of acids Number

of
(KOH, mg)
peroxides (J, g)
to 1.0
to 0.01

1.0 2.5
0.01 0.04
1.0 2.0
0.01 0.1
1.0 3.0
0.01 0.1
1.0 1.6
0.01 0.03
Higher than of doubtful freshness poultry

5.3. Freshness evaluation of rabbit and nutria meat

For sensual, chemical analysis and microscopy 3 carcasses of rabbits (nutrias) should be
selected from the parcel. If after sensual analysis there comes suspicion about freshness of rabbit
(nutria) meat, chemical analysis or microscopy should be done. In cases, if data of sensual analysis
are different from chemical investigation or microscopy, 5 carcasses should be evaluated according
chemical reactions, repeatedly.

Sensual analysis of rabbit and nutria carcasses. Analysing, appearance and colour of

carcasses surface and inside, also condition of fatty tissue and incision of muscles, meat odour and
consistence should be evaluated. Evaluation of rabbit meat freshness according sensual indexes is
shown in Table 16.

Laboratory analysis of rabbit and nutria meat. For evaluation of rabbit (nutria) meat

freshness, microbiological (bacterioscopy) analysis, evaluation of ammonia and ammonium salines,

amount of VFA and cooper sulphate reactions should be done.

Bacterioscopy. Methodology of investigation and evaluation of the results are the same as

analysing meat of other species.

62

Table 16. Sensual indexes of rabbit meat freshness

Index

Typical signs
Fresh
Doubtful freshness
Bad (not fresh)
Surface is dry, covered Surface is wet in Surface is covered with
Appearance
places, lightly sticky grey to brown mucus.
and colour of with lightly pink skin
and darkened
carcass
surface
Yellow to white colour
Yellow
to
white Grey to white colour; with
Fats
colour; with pink tone brown tone of defrosted
in defrosted carcasses
carcasses
Without
brightness, Without brightness, covered
Inside
of Wet, bright
sticky, sometimes with with mucus and moulds
carcass
little
mucus
and
moulds
Incision
of Little wet, but not Wet, leaving wet spot Wet, leaving wet spot on
leaving spot on filtrating on filtrating paper, filtrating paper, sticky, red
muscles
paper; lightly pink colour dark red colour
to brown colour
with brownish tone
Muscles are stiff, elastic, Muscles are not so stiff Muscles are tremulous, pit,
Consistence
pit, made by pressing as of fresh meat, pit, made by pressing with
with
finger,
soon made by pressing with finger, doesnt equilibrate,
equilibrates, fats are stiff. finger,
equilibrates fats of defrosted carcasses
slowly (during 1 min.), are soft and rancid
fats are soft and lightly
mellow of defrosted
carcasses
Specific for fresh rabbit Musty odour, better Purulent odour, better felt
Odour
meat
felt inside carcass
inside carcass
Lucid
or
lightly Muddy, much flakes, strong
Lucidity and Lucid, aromatic
muddy,
lightly unpleasant odour.
odour
of
unpleasant odour
bouillon
Evaluation of ammonia and ammonium salines. For analysis, prepare concentrated (1:4)

meat extract. Put 5 g of analyzing meat into the flask and pour 20 ml of twice boiled distilled water.
After 15 min. filtrate solution through paper filter. Pour 1 ml of filtrate into test-tube and drip 10
drops of Nesler reagent, then shake and evaluate colour and lucidity of extract.
Extract of fresh meat gets greenish yellow colour, stays lucid or lightly muddy. Extract of
doubtful freshness colours intensive yellow and becomes muddy. In extract of frozen meat
sediments appear. Extract of bad meat becomes yellow to orange, soon large flakes appear, which
settle on the bottom of test-tube.
Evaluation of VFA. Analysis is made according methodology, described in chapter

Evaluation of meat and poultry freshness. In case to get more straight results, analysis should be

63

repeated for 3 times and arithmetic average is calculated. Difference between parallel analysis
should not exceed 9 % of average. Calculation should be done in accuracy of 0,1 mg KOH.
In fresh defrosted rabbit (nutria) meat, amount of VFA doesnt exceed 2.25, in chilled up to
4.5 mg KOH, in chilled of doubtful freshness 2.25-9.0, in frozen 4.50-13.50, in not fresh chilled
more than 9.0, in not fresh frozen more than 13.50 mg KOH.
Cooper sulphate reaction. Methodology and evaluation of reaction are described in chapter

Evaluation of meat and poultry freshness.

6. Detection of Trichinella in meat

Trichinosis is caused by Nematoda class Trichocephalata suborder Trichinellidea family
parasites. All vertebrate except fish can be sick. Trichinas are not specific for different animal
species. Especially susceptible for trichinosis are humans, pigs, rats, foxes, wolfs, mice, badgers,
bears, dogs and cats. Horses, beavers, martens, hamsters, behemoths also can be sick with
Trichinosis. Nowadays there are 8 Trichinella species known in the World: T. spiralis, T.
pseudospiralis, T. papuae, T. nativa, T. britovi, T. nelsoni, T. zimbabviensis, T. murrelli. All
trichinella species are dangerous to human. T. spiralis is spread all over the World.

6.1. Diagnostics of trichinosis

Commission regulation (EB) Nr. 2075/2005 assigns specific rules on official controls for
Trichinella in meat. The main method for diagnostics of trichinosis is Magnetic stirrer method for
pooled sample digestion. Equivalent methods are Mechanically assisted pooled sample digestion
method/sedimentation technique and Mechanically assisted pooled sample digestion method/on
filter isolation technique. In exceptional cases (using meat for own consumption) trichinoscopy can
be used. Other methods, such as serological, can be used for monitoring or species evaluation.
Serological analysis doesnt fit wishing to find Trichinella in animals, used for human food.
Samples from carcasses of domestic swines are constantly taken at slaugterhouses and they
are a part of postmortem examination. Samples of not resistant for Trichinella horses, boars, other
farm and wild animals carcasses are taken systematically at slaughterhouses or processing
enterprises of wild animals ant it is a part of postmortem examination. Pending the results of the
Trichinella examination and provided full traceability is guaranteed by the food business operator.
Such carcasses may be cut up into a maximum of six parts in a slaughterhouse or in a cutting plant
on the same premises as the slaughterhouse (the premises). Such carcasses may be cut up at a

64

cutting plant attached to or separate from the slaughterhouse provided that: the procedure is under
supervision by the competent authority; a carcass or the parts thereof will not have more than one
cutting plant as its destination; the cutting plant is situated within the territory of the Member State;
and in case of a positive result all the parts will be declared unfit for human consumption.
Carcasses and their parts may not leave the premises, before the result of the Trichinella
examination is found to be negative. Other parts of an animal intended for human or animal
consumption which contain striated muscle tissue may not leave the premises before the result of
the Trichinella examination is found to be negative. Animal waste and animal by-products not
intended for human consumption and not containing striated muscle may leave the premises before
the results of the Trichinella examination are available. However, SFVS may require a Trichinella
examination or prior treatment of animal by-products to be carried out before permitting them to
leave the premises. Where a procedure is in place in the slaughterhouse to ensure that no part of
carcasses examined leaves the premises until the result of the Trichinella examination is found to be
negative and the procedure is formally approved by the SFVS, the health mark may be applied
before the results of the Trichinella examination are available.

6.1.1. Magnetic stirrer method for pooled sample digestion

Collecting of specimens. In the case of whole carcasses of domestic swine, a specimen

weighing at least 1 g is to be taken from a pillar of the diaphragm at the transition to the sinewy
part. Special trichinae forceps can be used provided an accuracy of between 1.00 and 1.15 g can be
guaranteed. In the case of breeding sows and boars, a larger sample weighing at least 2 g is to be
taken. In the absence of diaphragm pillars, a specimen of twice the size 2 g (or 4 g in the case of
breeding sows and boars) is to be taken from the rib part or the breastbone part of the diaphragm, or
from the jaw muscle, tongue or abdominal muscles. For cuts of meat, a sample weighing at least 5 g
of striated muscle, containing little fat is to be taken, where possible from close to bones or tendons.
For frozen samples, a sample weighing at least 5 g of striated muscle tissue is to be taken for
analysis. Special attention must be paid when collecting muscle samples from the tongue in order to
avoid contamination with the superficial layer of the tongue, which is indigestible and can prevent
reading of the sediment.
Procedure. 16 0.5 ml of hydrochloric acid is added to a 3 liter beaker containing 2,0 liter of

tap water, preheated to 46 to 48oC; a stirring rod is placed in the beaker, the beaker is placed on the
preheated plate and the stirring is started. 10 0.2 g of pepsin is added. 100 g of samples collected
in accordance with point 2 is chopped in the blender. The chopped meat is transferred to the 3 liter
beaker containing the water, pepsin and hydrochloric acid. The mincing insert of the blender is
65

immersed repeatedly in the digestion fluid in the beaker and the blender bowl is rinsed with a small
quantity of digestion fluid to remove any meat still adhering. The beaker is covered with aluminium
foil. The magnetic stirrer must be adjusted so that it maintains a constant temperature of 44 to 46oC
throughout the operation. During stirring, the digestion fluid must rotate at a sufficiently high speed
to create a deep whirl without splashing. The digestion fluid is stirred until the meat particles
disappear (approximately 30 min.). The stirrer is then switched off and the digestion fluid is poured
through the sieve into the sedimentation funnel (Pict. 22). Longer digestion times may be necessary
(not exceeding 60 min.) in the processing of certain types of meat (tongue, game meat, etc.).

Picture 22. Sedimentation funnel and net

The digestion process is considered satisfactory if not more than 5 % of the starting sample
weight remains on the sieve. The digestion fluid is allowed to stand in the funnel for 30 minutes.
After 30 minutes, a 40 ml sample of digestion fluid is quickly run off into the measuring cylinder or
centrifuge tube. The 40 ml sample is allowed to stand for 10 minutes. 30 ml of supernatant is then
carefully withdrawn by suction to remove the upper layers and leave a volume of not more than 10
ml. The remaining 10 ml sample of sediment is poured into a larval counting basin or Petri dish.
The cylinder or centrifuge tube is rinsed with not more than 10 ml of tap water, which has to be
added to the sample in the larval counting basin or Petri dish. Subsequently, the sample is examined
by trichinoscope or stereo-microscope at a 15 to 20 time magnification (Pict. 23-24). Visualization
using other techniques is allowed, provided examination of positive control samples has been
shown to give an equal or better result than traditional visualization methods. In all cases of suspect
areas or parasite like shapes, higher magnifications of 60 to 100 times must be used. Where the
digests are not examined within 30 minutes of preparation, they must be clarified.

66

Evaluation of pools of less than 100 g. More than 15 g must be examined as a complete pool.

For pools of up to 50 g, the digestion fluid and the ingredients may be reduced to 1 liter of water, 8
ml of hydrochloric acid and 5 g of pepsin.
Where examination of a collective sample produces a positive or uncertain result, a further 20
g sample is taken from each pig. The 20 g samples from five pigs are pooled and examined using
the method described above. In this way samples from 20 groups of five pigs will be examined.
When Trichinella is detected in a pooled sample from five pigs, further 20 g samples are
collected from them individual pigs in the group and each is examined separately using the method
described above.
When contaminated carcass or parts of it are detected, an official veterinarian must eliminate
all contaminated carcass, parts and organs, control, that they must be arranged as secondary animal
products of I category, to report territory State Food and Veterinary Service (SFVS) about keeping
place, keeper, hunting area, hunting club of animal, contaminated with Trichinella. Not later than in
3 work days an official veterinarian must send a specimen of 50-100 g to National Food and
Veterinary Risk Assessment Institute (NFVRAI) for determining Trichinella species. After taking
specimens, positive fluids (digestion fluid, oversedimental fluid, outwash and other) are neutralized
heating at least 60oC temperature. NFVRAI informs SFVS about detected Trichinella species.
If carcasses were split to pieces and mixed with parts of other carcasses having no answer
about the testing results, an official veterinarian must take a decision about all the meat, which
could be mixed with Trichinella contaminated meat, recognizing it as not fit for human
consumption and to refer to manage it as secondary products of I category.
It is forbidden to take carcasses, their parts and internal organs from slaughterhouses before
negative answer about contamination with Trichinella would be clear.

Picture 23. Trichinella after sedimentation

Picture 24. Trichinella after sedimentation

67

6.1.2. Mechanically assisted pooled sample digestion method/sedimentation technique

using stomacher lab-blender 3 500
Procedure. The stomacher lab-blender 3 500 is fitted with a double plastic bag and the

temperature control set at 40 to 41oC. One and half a liter of water preheated to 40 to 41oC is poured
into the inner plastic bag. 25 ml of 17.5 % hydrochloric acid is added to the water in the stomacher.
100 samples weighing approximately 1 g each (at 25 to 30oC) taken from each individual sample in
accordance with 2 are added. Lastly, 6.0 g pepsin is added. This order must be followed strictly to
avoid decomposition of the pepsin. The stomacher is then allowed to pound the content of the bag
for 25 minutes. Then 300-400 g of ice are added in case that capacity of fluid increased to 2 L.
digestion fluid is mixed until the ice has melted.
The chilled digestion fluid is transferred to a 2 liter separation funnel, equipped with a vibrator
in an extra clamp. Sedimentation is allowed to proseed for 30 minutes, during which time the
sedimentation funnel is vibrated intermittently, i.e. one minute vibration followed by a one-minute
pause. After 30 minutes, a 60 ml sample of the sediment is quickly run off into a 100 ml measuring
cylinder (the funnel is rinsed with detergent solution after use). The 60 ml sample is allowed to
stand for at least 10 minutes, after which time the supernatant is withdrawn by suction to leave a
volume of 15 ml, to be examined for presence of larvae. The remaining 15 ml is poured into a larval
counting basin or two Petri dishes and examined using a trichinoscope or stereo-microscope. The
measuring cylinder is washed with 5 to 10 ml of tap water and the washings are added to the
sample.

6.1.3. Mechanically assisted pooled sample digestion method/on filter isolation technique
Aparatus and reagents. For this analysis additionally are needed: 1 liter Gelman funnel,

complete with filter holder (diameter 45 mm); filter discs, consisting of a circular stainless steel
mesh with an aperture of 35 microns (disc diameter: 45 mm), two rubber rings 1 mm thick (external
diameter: 45 mm; internal diameter: 38 mm), the circular mesh being placed between the two
rubber rings and bonded to them, an Erlenmeyer flask, capacity 3 liters, fitted with a side tube for
suction, a filter pump, Trichomatic 35 blender with filter, plastic bags, capacity at least 80 ml and
equipment for sealing them, rennilase, strength 1: 150 000 Soxhlet units per gram.
Procedure. For analysis 30 ml 8.5 0.5 % hydrochloric acid and 7 g of pepsin strength 1: 10

000 NF (US National Formulary) corresponding to 1:12500 BP (British Pharmacopoeia) and to

2000 FIP (Fdration internationale de pharmacie) are used. Mechanically assisted pooled sample
digestion method is performed analogous to sedimentation method.

68

Ice (300 to 400 g of ice flakes, scaly ice or crushed ice) is added to the digestion fluid to bring
its volume up to about 2 liters. In the case of smaller pools, the amount of ice must be reduced
correspondingly. The digestion fluid is stirred until the ice has melted. The chilled digestion fluid is
then left for at least three minutes to let the larvae coil. The Gelman funnel, fitted with a filter
holder and filter disc, is mounted on an Erlenmeyer flask connected to a filter pump. The digestion
fluid is poured into the Gelman funnel and filtered. Towards the end of filtration, the digestion fluid
can be helped to pass through the filter by applying suction with the filter pump. Suction must cease
before the filter becomes dry, i.e. when 2 to 5 ml of fluid is left in the funnel. Once all the digestion
fluid has been filtered, the filter disc is removed and placed in an 80 ml capacity plastic bag,
together with 15 to 20 ml of rennilase solution. The plastic bag is sealed twice and placed between
the inner and outer bags in the stomacher. The stomacher is allowed to pound for three minutes, e.g.
while it is working on a complete or incomplete pool. After three minutes, the plastic bag, complete
with filter disc and rennilase solution, is removed from the stomacher and opened with scissors. The
liquid contents are poured into a larval counting basin or Petri dish. The bag is washed out with 5 to
10 ml of water, which is then added to the larval counting basin for examination by trichinoscope or
to the Petri dish for examination by stereo-microscope.

6.1.4. Detection of Trichinella in meat using digestion method

(using strong iodine and sodium thiosulphate solutions)
Procedure. 10 g of analyzed muscles (diaphragm, masticatory or other) are crushed with

scissors or homogenized, put to laboratory glass and 500 ml hydrochloric acid-pepsin solution (10 g
of pepsin (3000 U/mg) + 10 ml conc. HCl + H2O up to 1 L (pH=2)) are added. Sample is digested at
38oC temperature for 30 min. During digestion, suspension is mixed and shacked regularly (if
special digesting apparatus is used this happens automatically). After digestion, content of glass is
filtered through the mesh (diameter 400-500 mm), after that through another mesh (diameter 20-30
mm). Then the mesh is put for 3 min., if meat was frozen for 4 min. (in Petri dish) to a plate with
iodine solution (strong iodine solution: 20 g J2 + 40 g KJ + H2O up to 1 L). After that, the mash is
put into sodium thiosulphate solution for 4 seconds (Sodium thiosulphate solution (30 %): 300 g Na
thiosulphate, H2O up to 1 L), for discolouring remainders of tissues and learve (except Trichinella
learve, which are black). Examination is made by stereo-microscope.

69

6.2. Trichinoscopic examination

Trichinoscopic examination is called investigation of muscle pieces, pressed between two

glass plates of compressorium using simple or projective trichinoscope looking for Trichinella
learve.
Collecting of specimens. In the case of whole carcasses, several hazelnut-size samples are

taken from each animal: in domestic swine, such samples are taken from both diaphragm pillars at
the transition of the sinewy; in wild boar samples are taken from both diaphragm pillars at the
transition of the sinewy part and in addition from the jaw, the muscles of the lower leg, the
intercostal muscles and the tongue muscles, giving a total of six samples from each individual
animal; if certain muscles are not available for sampling, a total of four samples are taken from the
muscles that are available in pieces of meat, four hazelnut-size samples of striated muscle tissue
containing if possible no fat, taken from different points, are taken from each piece, where possible
close to bones or tendons
Procedure. In general a compressorium is filled with 1.00.1 g of meat, normally

corresponding with 28 oat-kernel-size pieces. If necessary, two compressoria need to be filled to

examine 56 oat-kernel-size pieces. If both diaphragm pillars are present in a domestic swine, the
Trichinella inspector cuts 28 oat-kernel-size pieces from each of the above specimens taken from a
whole carcass, making 56 pieces in all. If only one diaphragm pillar is present, 56 pieces are cut in
different places, if possible from the transition to the sinewy part. 2 compressoria are examined at
all.
The samples collected from the other four muscles of wild boar are each cut into seven oatkernel-size pieces, giving a total of 28 additional pieces (1 compressorium). The Trichinella
inspector then compresses the 56 (or 84) pieces between the glass plates so that normal print can be
clearly read through the slide preparation.
Examining, the lower (numbered) glass plate of compressorium is put on the table near the
edge. Sample of investigative muscles is kept in the left hand, bended on the finger (forefinger).
From each muscle sample 28 pieces of 3 mm thickness (average oat grain size) are cut. Cut is made
with scissors from different places of sample along the muscle string. Pieces are cut with concave
side of scissors. This enables to put cut meat piece on the glass plate easier. After putting 28 muscle
pieces, the upper glass plate is put on them. Both glass pieces are pressed using screws at the ends
of compressorium. Pressing must be done in such way, that looking through the compressorium, we
could read small printed text. If the flesh of the specimens to be examined is dry and old, the
preparations must be softened for 10 to 20 minutes before pressing with a mixture of one part of
potassium hydroxide solution to about two parts of water.
70

The microscopic examination must be carried out by scanning each preparation slowly and
carefully at a magnification of 30 to 40 times. Taking compressorium along muscle fibers, all
preparation must be examined carefully, not leaving any muscle fiber. Right and left eyes are used
alternatively, free eye is left opened. The trichinoscopic examination must be carried out for at least
six minutes. It is easier to work with projective trichinoscope, because all pressed muscle piece is
shown on the screen. 40 to 50 compressoria are examined during 1 hour. Examination is carried out
in the dark place.
Evaluation and differential diagnosis. Incapsulated Trichinella in muscles is scrolled to

spirale and surrounded with capsule, inside of which is filled with lucid fluid. Capsule of
Trichinella can be as a lemon (in muscles of swine) or oval (in muscles of wolfs, bears, rats). Swine
Trichinella capsule is about 0.68mm of length and about 0.37 mm of width. There can be 1, 2, 3 or
even more Trichinellas in one capsule. Capsules and learvae inside them can be calcificated (Pict.
25- 30).

Picture 25. Trichinella, looking through

microscope (compressor method)

Picture 26. Calcificated Trichinella

capsules, looking through microscope
(compressor method)

Picture 27. Sarcocystis in native preparation

Picture 28. Sarcocystis in hematoxillineozin painted preparation

71

Picture 29. Cysticercus in painted preparation

Picture 30. Alaria alata painted muscle

preparation

As often as not in muscles of swines and wild boars Sarcocystis (Sarcocystis miescheriana, S.
suihominis) can be found. They localize in muscle fibers or between them and can calcificate.
Bigger cysts can lay free in intramuscular connective tissue. They can be rounded, oval or bean
form. Sarcocystis are of various size 0.44-4 mm of length and 0.3-3 mm width and has
characteristic cigar form. Despite size of Sarcocystis depends on their age and owner, the main
structures are the same. So called capsule can be striated (ridged), it partly consists of sarcolema,
cavity of Sarcocystis can be divided or not in septa, content is viscose or gelatinous, dark, opaque,
coloured like milk white or yellowish. There are sporoblasts in cists and different amount of sickle
form sporozoits, which have nuclea and are called Raineys bodies. Sarcocystis calcificate more
often than Trichinella: whiting at first concentrate at the center, they can be of S form and can be
found in heart muscle where Trichinella can be found very seldom (Pict. 27-28).
Young, undeveloped or degenerated Cysticercus, in the contrary to Trichinella, are found
between fibers of connective tissue and are greater bigger (about 2-3 mm). Examining Cysticercus
through microscope, head of parasite or it remains (chitinic hooks) are seen (Pict. 29).
In muscles of swines and wild boars mesocercaria of thrematodus Alaria alata can be found.
Body of mesocercaria is of pear form, flat at side of belly and convex at the back, size 0.31-0.50 x
0.14-0.28 mm (Pict. 30). In front there is oval so called therminal organ. Cavity of mouth is
surrounded by 6 to 7 rows of needles. Starting from the mouth, further goes muscular pharynx,
oesophagus and wide channels of intestines, not reaching the back side. At the belly side, there is a
sucker, surrounded by 3 rows of small needles. Also there are two pairs of big penetrating glands,
throws of which open at the edges of mouth cavity. Terminal cells are found between excretion
bladder and intestinal channels. Mesocercaria parasitize in animal muscles or other organs, and as
72

usual, thick-walled capsule from connective tissue forms around them, which looks like capsule of
Trichinella. This is Alaria cyst. In not painted preparations, free Alaria mesocercarias differentiate
from Trichinella easy. Pressed between glass plates of compressorium, Alaria often moves. Also
moves mesocercaria in the cyst. If there are many Alaria mesocercarias in meat of swines and wild
boars, such meat doesnt fit for human consumption because of great amount of metabolic helmint
products and toxins. Alaria is widely spread in Lithuania. Alaria mesocercarias are found in
muscles of wild boars and mousy rodents and adult helmints are found in intestines of dogs, foxes,
wolves and raccons. There are no data about human infection with Alaria. It is supposed that
humans can be reserved owners.
During trichinoscopy, Phisocephalus larvae can be found too. Swine phisocephallosis is
caused by widely spread nematode Physocephalus sexalatus. In their vital cycle several owners
participate: terminal owners despite swine and wild boars, can be camels, asses, hares and rabbits.
Physocephalus parasitize in gastric mucous of these animals. Larvae develop in intermediate
owners muckworms. Insectivorous and rodents can be reserved owners. Larvae parasitize in
muscles, where rounded or oval capsule forms around helmint. Capsule looks much like Trichinella
capsule. During this stage, if Physocephalus get to another owner, they can reincapsulate. Using
usual compressoric diagnostic methods, it is a little bit hard to notice differences between
Trichinella and Physocephallus capsules. To differentiate these helmints, histological examination
should be done.

6.2.1. Trichinoscopy additionally processing clippings

Trichinoscopy painting clippings with 0.2 % methylenum blue aquatic solution.

Clippings are put into Petri plate on metal or plastic percolator or on thin fabric. 2 % methylenum
blue aquatic solution is poured on clippings and left for 15 min. painting. After that, clippings with
percolator are taken from the paints and put on the filter paper. After paint absorbtion to filter paper,
clippings are put (together with percolator) into another Petri plate and after pouring 1.5 % vinegar
acid, are left for 3 min. Then clippings are taken from the vinegar acid solution, they are pressed
between glass plates of compressoria and examined with trichinoscope or microscope. Trichinella
and capsules become blue.
Trichinoscopy painting clippings with 7-8 % methylenum blue solution. Clippings are put

into clock glass or into porcelain plate. 2-3 drops of 7-8 % methylenum blue solution in 80 %
vinegrad acid is poured on clippings. After 1-2 min. painted clippings are washed with hot water
until flowing water becomes clear. Then clippings are put on glass plates of compressoria, they are
straightened and one drop of glycerin (diluted with water, ratio 1:1) is dripped on each of them.
73

After pressing Trichinoscopy or microscopy is performed. Trichinella and their capsules become
blue and muscle fibers become lightly blue.
Examination of meat if calcificated formations were found (Trichinella, Sarcocystis and
other). Capsule of Trichinella doesnt translucent because of whitings and it is hard to differentiate

it from other calcificated formations.

If calcificated formations (trichinas) were seen, pressed clippings are taken away from the
glass plates of compressoria and soaked for several minutes into glicerine, diluted with water (1:1)
or into the 5 % lactic acid. Later they are pressed again and examined with trichinoscope.
If in such clippings trichinas are seen badly, clippings must be taken away from the glass
plates and soaked into 10 % hypochloric and vinegar acid solution for 2-3 hours, then put on the
compressoria again and one drop of glycerin must be dripped in each clipping. After pressing,
trichinoscopy can be done. In action of acids, whitenings melt and Trichinella or their fragments
can be seen more clear.
6.2.2. Examination of frozen meat

In frozen meat, capsules become permeable. Defrosting such meat, a part of fluid flows from
capsule and it deflates. After getting to capsule, muscle juice colours it red in that case it is hard to
notice Trichinella through trichinoscopy. For Trichinella to be clearer, clippings should be coloured
with methylenum blue or put into hipochloric acid.
Procedure. Thin muscle clippings (1.5-2 mm) are cut and pressed between glass plates of

compressoria, that juice could flow. Later 1-2 drops of 0.5 % hypochloric acid or solution of
methylenum blue (5 ml saturated spirituous methylenum blue, diluted with 195 ml of distilled
water) is dript on them and after 1-2 min trichinoscopy is made. After action with hypochloric acid,
muscle fibers become greyish, but lucid. Capsules of Trichinella bloat and become visible more
clear and fluid inside them after protein coagulation, becomes more lucid. Fibers after action with
methylenum blue, becomes lightly blue, capsules become pink violet, pink or blue, fluid in capsules
becomes lightly blue and learve of trichinas dont paint, but are clearly visible.

6.2.3. Examination of salted and smoked meat

Procedure. Muscle clippings of salted or smoked meat, pressed between compressoria glass

plates, are badly translucent and looking through trichinoscope, trichinas are seen badly. Such meat
must be prepared specially. To make muscle clippings softer, they could be left in 45oC temperature
5 % potassium hydroxide solution for 10 min. Later they are put on the glass plates of comperssoria
and after dripping 1 drop of glycerin, are examined using trichinoscope.
74

Clippings of salted meat are definitely painted with methylenum blue. Clippings should by
twice thinner than of not canned meat.

6.2.4. Examination of lard

Muscle layers are cut from lard and the same method is used as examining salted pork. If there
are no muscle tissue in lard, A. Merkushev method is used.
Procedure. Lard are cut across and from the surface of atrophied muscles, using scalpel, thin

(2-3 cm length) lard piece is cut. Another piece is cut from the bottom surface. Oval 0.3-0.5 mm
lard pieces are cut from each lard sample and are pressed between compressoria glass plates. Then
the upper glass plate is taken away and 1-2 drops of 1 % methylenum blue solution are dripped on
each clipping. After that, lard clippings are pressed again and are heated above spirit lamp till
becomes lucid (10-15 s) and are examined using trichinoscope. Connective tissue of lard becomes
light blue and muscle fibers become green to blue.

6.3.Examination of Trichinella in meat of other animals

Horse meat, wild game meat and other meat that could contain Trichinella parasites must be
examined in accordance with one of the digestion methods with the following changes: specimens
weighing at least 10 g are taken from the lingual or jaw muscle of horses and from the foreleg,
tongue or diaphragm of wild boar. In the case of horse, where those muscles are lacking, a largersized specimen is to be taken from a pillar of the diaphragm at the transition to the sinewy part. The
muscle must be clean of connective tissue and fat. At least 5 g of sample is digested following the
reference method of detection or an equivalent method. For each digest, the total weight of muscle
examined must not exceed 100 g. Where the result is positive, a further 50 g specimen is taken for a
subsequent independent examination. Without prejudice to the rules on conservation of animal
species, all meat of game animals other than wild boar, such as bears, carnivorous mammals
(including marine mammals) and reptiles, are to be tested by sampling 10 g of muscle at the
predilection sites or larger amounts if those sites are not available. Predilection sites are: in bear:
diaphragm, masseter muscle and tongue, in walrus: tongue, in crocodiles: masseter, pterygoid and
intercostal muscles; in birds: muscles of the head (i. e. masseter and neck muscles). The digestion
time must suffice to ensure adequate digestion of the tissue of these animals but must not exceed 60
minutes.
6.4. Prophylaxis of trichinosis

Pork, horsemeat, nutria meat, meat of wild carnivores and omnivores, which can be infected
with Trichinella, and their carcasses parts fitting for human consumption, must be examined for
75

trichinosis according attitudes of Regulation (EB) Nr.2075/2005. It is very important to inform

animal breeders and hunters about risk of trichinosis, ways of spreading and prophylaxis measures.
It is strongly forbidden to feed swine with raw waste from hunting and slaughterhouses products,
which can be infected with Trichinella. Carcasses of hunted or died wild raptors, rodents, vagabond
dogs and cats must be buried or dug deep, that swines or other wild animals couldnt eat them. As
rodents are the main spreaders of trichinosis in nature they must be liquidated in farms. It is
forbidden to sell meat and meat products without label of wellness.

7. Fats of animal origin and their quality evaluation

7.1. Animal fat stock

After animal (cattle, swine, sheep) slaughter, from processed fats stock (fatty and osseous
tissues bones), we get alimentary fats and after birds (hens, ducks, geese) slaughter, we get
poultry fats. To fats of animal origin also fats from sea mammalians (whales, cachalots, seals,
dolphins) and fish fats are assigned. The main fat stock is fatty tissue, so called soft stock. This is
spongy connective tissue, consisted mainly of fatty cells. In those cells, fat inserts make one big
drop, putting cytoplasm and nucleus to periphery. Besides, there is rich connective tissue net,
consisted of elastic and reticular fibers. From chemical view, fatty tissue consists of fats (74-97 %),
proteins (0.4-7.2 %), inorganic materials (0.08-1.0 %) and water (2-36.8 %).
For processing of fats, osseous tissue (solid stock) can be used too. This connective tissue is
solid and strong, consisted of osseous cells (osteocytes) and intracellular substance, where collagen
fibers are dominated. From chemical view, osseous tissue consists of water (17.4-35.6 %), fats (630 %), proteins (in composition of ossein). Processing bones stock, we get fats, gelatine, glue,
bones flour. For that purpose, tubular (haunch, shin, shoulder, forearm, hand and other.) and tabular
(pelvic bones, scapula, ribs, head bones, vertebraes and other) bones are used. Mostly fats are found
in sternum (up to 30 %), tubular bones (22-28 %.), pelvic bones (up to 22.5 %), ribs, scapulas (up to
14 %), the least fats are found in bones of head (6-10 %). Fat stock from bines is from white to
yellow colour. Fats from bones spoil very soon, therefore if to get good quality fats, stock must be
fresh and clean. Fats from unclean bone stock fits only for technical purposes. Stock of bones can
be kept at 3-4 C temperature, not longer than for 24 hours.
Stock of fats is classified according animal species (cattle, swine, sheep). Stock of every
animal species according to it concentrating place is classified into 2 groups:

76

the first group consists of omentum, fats from mesenteric, kidneys, cattle sternum, fats

from Kurdish sheep rump and also cuttings from cattle and swine carcasses processing, lard cuttings
and strangles.

the second group consists of gastric and intestinal fats and fats cuttings from skin

processing.
The main part of fat stock belongs to omentum and renal fats (45-65 %) and mesenteric fats
(10-30 %). Ranking fat stock according concentration place, the best are internal fats: omentum,
renal fats, lard (from back, flank). Fat stock from stomach and intestines is of worse quality because
of unpleasant specific odour.
Regarding to quality, fats of animal origin are classified to the highest and first class; in case
of bids there are two classes: first and second. From fat stock, which doesnt fit requirements of the
highest and first classes, technical fats are prepared.
Fat stock spoils quickly if requirements for processing arent kept, because in fatty tissue there
are ferment lipase and plenty of microorganisms. Despite, protein substances and water in fat stock
are good environment for microorganisms multiplication. This environment determines spoilage of
fats. To avoid spoilage, fats stock should be processed as soon as possible. Such fat stock cant be
polluted with blood and remainders from other tissues (muscular tissue, limphatic nodes, cartilages
and other) because they worsen quality of production. If fat stock is polluted with blood and there
are muscle residues, haemoglobin and muscle myoglobin spoil during stock melting process and
pigmental substances excret, which influence brown to grey colour of fats. Besides, iron from
haemoglobin and myoglobin quickens fat oxidation and residues from of intestines give unpleasant
specific odour.
7.2. Characteristic of animal fat

Colour of different animal species can vary from white to yellow. Intensity of colour depends
on animal age, sex, forages. Spoiling fat become dark grey, sometimes lighten, at late spoiling
stages they can become brown or greenish. Odour and colour of good quality melted fat are typical
for every species and havent scalding taste. Sometimes scalding taste can appear because of bad
forrage. Besides, fat can absorb outside odours, so keeping them in bad environment, they can get
nonspecific for them odour.
Fat melting and gelation temperature depends on composition of fatty acids. If there are more
saturated fatty acids of big molecular mass, their melting temperature would be higher.
There is no strict fat melting temperature, because fat are heterogenic glycerids, characteristic
of which depends on composition of fatty acids. Fat refraction depends on amount and kind of fatty

77

acids too. Number of refraction increases when molecular mass of fatty acids is higher and when
there are more unsaturated fatty acids.
Differently from oils, fat have less poli-unsaturated, but more saturated fatty acids: stearic,
palmic, myristic, arachidonic. Depending on nature of fat, amount of fatty acids is different,
therefore physical and chemical fat indexes are different too.
According fat physical and chemical indexes it is possible to detect of what animal fat is and
parlty evaluate their quality.
Processing or keeping fat stock or melted fat, various changes of their chemical composition,
sensual indexes and nutritional value are going. Acting lipase, hydrolysis of fat stock starts, which
can be quickened by temperature higher than environment (10-15C), higher amount of water (more
than 40 % all fat stock mass), actine surface substances, metals and microorganisms.
There are levels of fat hydrolysis when di- and monoglycerides form. Melting of fat stock
stops process of hydrolysis, because at 60C temperature lipase is inactivated. But because of
primary changes of fat stock, formed amount of free fatty acids stays in new-melted fat too. Formed
fatty acids with high molecular mass, little influence sensual peculiarities of products, but fatty
acids with little molecular mass (acetic, propionic, antacid) worsen flavour and aroma of fat.
Besides, increased amount of free fatty acids makes optimal conditions for oxidative spoilage of
product.
Hydrolysis is influenced by other factors too (Table 17). Especially big influence is because of
microorganisms. Participating ferments from tissues and microorganisms spoil fat more quickly.
Table 17. Influence of different factors on fat spoilage
Factors

Light
Warm
Oxygen
Water
Tissue lipase
Fe, Cu, Sn iones
Microbial
ferments

Kind of spoilage
Oxidation

Hydrolysis
Increasing
of
Tallowing
acidity
+
+
+
+
+
+
+
+
+
+

Ketonic souring

+
+
+
+

Aldehidic
souring
+
+
+

+
+

Free fatty acids can form during oxidative spoilage too as secondary products of spoiling
peroxide compounds. If amount greater than described in normative documents is detected in
melted alimentary fat, they are sorted to not alimentary. Because of lower temperature of smoke
78

formation, melted fat with higher amount of free fatty acids have worse culinary characteristics:
they start to puff at 160-180C temperature, while fresh, qualified fats only at 220 C.
Interaction of air oxygen and fat causes oxidative spoilage, characteristic of which depends on
fat composition, features and keeping circumstances. During fat oxidation process, free radicals
form. They interact with molecules from environment substances and sever atoms making stabile
compounds. Unsaturated fatty acids oxidize easier, therefore fats, which have more unsaturated
fatty acids (of swine, hens) oxidize quicker.
Primary products of fat oxidation are hydroperoxides. They also change forming more stabile
oxidation compounds. During various reactions, secondary oxidation products form: epoxicompounds, aldehides, ketones, oxi- and keto- acids, low molecular mass organic acids. In
concentration of these compounds, fat colour and taste worsen.
There are two main kinds of oxidative spoilage, they are souring and tallowing (stearinsation).
During souring products of lower molecular mass (aldehides, ketones, fatty acids) which are toxic,
concentrate. Especially worsens odour of such fat. If temperature is low, tallowing of fat can occur
where dominate oxy-acids susceptible to condensate and form many molecular compounds. During
this process, fat odour and colour change, colour can fade, their consistence become more solid.
Rate of fat oxidation depends on composition of fatty acids, degree of hydrolysis, temperature
and other factors (Table 17). Technologically right processed fat, prepared for melting, can be kept
long at positive (2-4C) or negative (-18C) temperature. While swine fat, kept at minus 8C
temperature, already after 1-2 months lose their consuming peculiarities.
For slowing of oxidation process, various antioxidants are used. In Lithuania, processing
melted fats, which are going to be kept longer than one year, natural and synthetic tocopherols,
galates and butilhydroxianizole (BHA) can be used. They act destroying oxidation chain, interacting
with free radicals or stopping spoilage of hydroperoxides. Amount of these antioxidants is strictly
regulated and cant exceed amounts described in hygienic normatives (for tocopherols it is 0.2, for
other antioxidants it is 0.1g/kg).

7.3. Evaluation of melted fat quality, physical and chemical indexes

7.3.1. Sensual fat evaluation

Analysis is processed at 15-20o C temperature.

Evaluation of colour. Melted fats are poured into dry colourless glass test-tube. The test-tube

is put in a glass with cold water for 1-2 hours. Fat colour and shades are evaluated at day light.
Evaluation of lucidity. Melted at water bath fat are poured into dry colourless test-tube and

their lucidity is evaluated at day light.

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Evaluation of odour and flavour. Thin layer of melted fat is spread on objective glass and

their odour is evaluated. For evaluation of flavour, little piece of fat is tasted.
Table 18. Sensual characteristics of fat
Index
0

(18-20) C

Colour

Sort of melted fat

The highest
I
cattle from light yellow to yellow;
cattle from light yellow to yellow;
swine white;
swine white, light yellow or greyish
sheep from white to light yellow;
shade is allowed;
hens from white to yellow,
sheep, hens from white to yellow;
gees white or light yellow, greyish ducks white or light yellow, greyish
shade is allowed
shade is allowed

Odour
Typical for fat, melted from fresh fat
stock, without outside taste and odour

Typical for fat, melted from fresh fat

stock, light frying odour and taste is
allowed

In birds typical for species and melting way, without outside taste and odour, soft
frying odour and taste is allowed
Lucidity
Consistence

lucid
Cattle fat stiff or solid, swine fat soft or sticky, hens, gees and ducks fat
liquid, sticky
7.3.2. Evaluation of fat physical and chemical characteristic

Fat consist of fatty acids mix with different molecular mass, physical and chemical
characteristic. Therefore they melt and stiffen at some temperature interval, while separate
components of fatty acids mix melt or stiffen.
7.3.2.1. Evaluation of fat melting temperature

Fat melting temperature is such temperature, in which fat from solid condition become liquid
and lucid. Before analysis, fat are melted and filtrated through dry paper filter, for eliminating of
moisture which increases melting temperature.
Procedure. 1 cm of fat is sucked into two 1-1.5 mm diameter glass capillaries. It is important

that air couldnt pass. Capillaries with fat are left for 24 hours in 10oC temperature, that easy
melting triglycerides could cristalize at all. Then capillaries with fat are attached to thermometer
hydrargyrum end. Both are soaked into a glass, that 1 cm of water would be above the capillary.
After that the glass is heated permanently at the rate of 2 cm/min. Melting point is temperature
when fat become lucid and rise above the capillary. Temperature difference must not exceed 0.3oC.
Melting temperature can be measured in capillary, one end of which is closed (Pict. 32.).
Poking with open end of capillary, chopped analyzed material is taken and it is shaken to the bottom
of capillary, letting capillary to fall through the thin 50-80 cm tube, put square on laboratory table.
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Height of material column in capilarry must be 3-5 mm. The simplest apparatus for determination
of fat melting temperature consists of long-necked flask with rounded bottom and the test tube, put
into the flask. Thermometer is attached at the test-tube, at the end of which capillary with analyzed
material is fixed using rubber ring. Sulphuric acid or silicone oil is poured in to the round-bottomed
flask These fluids can be heated up to 275oC temperature. Heating is performed on spirit lamp, that
temperature would increase 5-10 C during a minute, and near the melting point 1-2 C per
minute. Usually material melts in special temperature interval: at first it moistens and softens at the
bottom of capillary (the start of melting), we must fix the temperature, later some fluid appears and
finally all the material turns into the fluid (end of melting).

Rubber ring

Thermometer

Sample

Picture 31. Curves of fat stiffening temperatures

Picture 32. Filed capilary with

thermometer for measuring fat melting
temperature

7.3.2.2. Evaluation of fat stiffening temperature

According to composition of fatty acids and glicerides, different kinetic curves (which depend
on crystallization of different components) of fat stiffening can be seen. Cooling melted fats, at first
significant fall of temperature is seen, then, when crystallization begins, temperature decreases
more quickly or sometimes even increases because of secret crystallization warm. After
crystallization, fat temperature decreases again.
81

Stiffening temperature is such temperature (Pict. 31 a), which is got cooling fat, decreasing of
which is stopped by crystallization secret warm, or that maximal temperature, to which increases
temperature of stiffening fats because of crystallization secret warm (Pict.31 b,c,d, point A).
Procedure. Put in porcelain plate fats are melted in water bath. When temperature of melted

fat becomes about 15oC higher than expected fat stiffening temperature, they are poured ti dish,
leaving 2 cm above. Thermometer is fixed in such way that end of hydrargyrum column would be at
the center of fat column. At first liquid fat are mixed with thermometer. Starting thickening process,
fat stiffening temperature is checked. Showings of thermometer are fixed every minute. According
analysed data, fat stiffening curve is drawing, at x axis marking time of analysis (min), at y axis
showings of thermometer (in degrees).

7.3.2.3. Evaluation of fat light rays breaking index

Breaking index depends on composition of fatty acids and other factors. According this it is
possible to decide about fat clearness, nature and oxidation degree. Fat breaking point increases,
when molecular mass of fats and amount of unsaturated acids increases too. The more unsaturated
acids in fat are, the higher their breaking point is. Breaking index is described as a relation of light
rays falling angles sin with outcome from analyzed substance layer, rays falling angles sin :

n = sin /sin
Procedure. 2-3 points of analyzed melted fats with glass stick are dripped on rephractometer

prism. Prism is lightened with lamp or day light. Changing prism position, rephractometer mark is
set that light flow would go through the center of ocular (athwart to crossing lines). Deductions of
scale are made at accuracy of 0.0002. Rephractometer scale is gradated for measuring deductions at
20oC analyzing temperature, therefore analyzing in other temperature, breaking index is calculated
using formula:
n 20 = n T + (T-20) 0.00035
n 20 breaking point, analyzed at 20oC temperature;
n T breaking point, analyzed in real temperature;
0.00035 average error of fat breaking index analysis, when temperature increases in one
degree.

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7.3.2.4. Evaluation of fat moisture

Procedure. At accuracy of 0.002, 2-3 g of fat are weighed into heated to fixed mass empty

dish and are drying in drying fire (at 102-105oC temperature) to fixed mass. First time, sample is
weighed after 1 val., later weightings are repeated every 30 min. Before weightings, dishes are
cooled in exicator. Amount of moisture is calculated using formula:

w = ((m1-m2)/(m1-m0))x100
m0 mass (g) of dish with sand and stick;
m1 mass (g) of dish with sand and stick, before drying;
m2 mass (g) of dish with sand and stick, after drying.
The results are of accuracy of 0.1 %. Results of parallel analysis should differ no more than
0.05%. During drying, fat oxidation may occur, which increases sample mass, there fore calculating
is made according smaller dried sample mass.

7.3.2.5. Evaluation of fat iodine number

Iodine number describes amount of unsaturated fatty acids in fat. Unsaturated fatty acids,
affected with hallogenes, easy attach them at the places of double joints. From amount of attached
hallogenes it is decided about composition of fatty acids and nature of fat. According iodine number
and other indexes there is possibility to decide about fat clearness. Iodine number is it amount in
grams which 100 g of fat attach at the place of double joint.
Quicker method of iodine amount evaluation is based on spirituous iodine reaction with joints
of unsaturated fatty acids, participating sodium thiosulphate.
Procedure. 0.3-0.4 g of fat are weighted into cone flask at accuracy 0.001g and 30-40 ml of

96o ethylic alcohol is poured. Flask is covered with plug which has a glass tube (30-50 cm of
length) and left at 40-50oC temperature water bath, till fat melt. Mix is cooled to environment
temperature, then 25 ml of 0.2 N spirituous iodine solution is dripped with pipette and 50 ml of
distilled water is poured. Flask is plugged again and mix is shacked well. After 5 min. 0.5 ml 1 %
starch solution is poured and it is titrated with 0.1 N Na 2 S 2 O 3 , till blue colour disappears. Parallel
control analysis is made, using the same amount of water instead of fat.
Iodine number (%) is calculated using formula:

X=0.01269 (a b) 100/ m
83

0.01269 amount of iodine, corresponded to 1 ml of 0,1 N Na 2 S 2 O 3 solution, g/ml;

a Amount of 0.1 N Na 2 S 2 O 3 solution (ml), used to titrate control sample;
b Amount of 0.1 N Na 2 S 2 O 3 solution (ml), used to titrate analyzed sample;
m fat amount, g

7.3.2.6. Evaluation of fat acids number

Acids number is one of the most important indexes of fat quality. During fat processing, this
index describes stickness of hydrolytic fat spoilage, and analyzing quality of keeping fat shows
oxidative spoilage (together with other more important indexes). Method is based on titrating of
fatty acids, which are in fat-spirituous-ether solution with aquatic hydroxide solution. Ether is used
as fat solvent and ethanolum homogenizes fat and aquatic hydroxide system, formed during
titrating. If spirit isnt used, reaction takes place only at the surface of phases connection, therefore
system must be homogenized mixing ethanolum with water and organic solvents.
Procedure. 3-5 g of fat are weighed into 250 ml konus flask at accuracy 0.01g. Fat are melted

in water bath. 50 ml of neutral mix (1:2) of ethyl and ether, which is neutralized in advance with 0.1
M of hydroxide solution to pink colour using phenolphtalein, are poured into the flask. Solution is
mixed in flask, 2-3 drops of 1 % phenolphtalein are dripped and immediately titrated with 0.1
NaOH (KOH) till pink colour. If solution in flask becomes muddy, 5-10 ml of ethylic alcohol and
ether mix are added. If feculence doesnt disappear, flask is lightly warmed in water bath and is
titrated after cooling. Number of fatty acids, described as KOH or NOH amount (mg), used for
neutralization of free fatty acids in 1 g of fat, is calculated using formula:

X= 0.561 V K/m

0.561 KOH (NaOH ) amount (mg) which is in 1 ml of solution;

V titrated amount (ml) of 0.01N KOH (NaOH) solution;
K re-counting coefficient to straight 0.1M KOH (NaOH ) solution;
m fat amount, g.

7.3.3. Qualitative method of antioxidant buthiloxianizolum evaluation

Procedure. 1 g of fat is weighed to glass test-tube and 2ml of 72 proc. Ethylic alcohol are

poured. Fat are melted in water bath. Also 5 ml of mix (1 part of NaNO 3 and 99 parts of sulphanilic

84

acid) are poured into test-tube, everything is mixed and 1.2 ml of 1 N NaOH is added. If there is
buthiloxianizolum in fat, solution becomes purple red.

7.3.4. Evaluation of oxidative fat spoilage

One kind of fat spoilage is oxidation with air oxygen. Hydroperoxides are primary products of
oxidation. Formed peroxides later take part in various chemical reactions, during which secondary
fat spoilage products form: aldehides, ketones, little molecular mass acids, alcohols. Part of these
products have unpleasant odour and taste and are toxic. Degree of oxidative spoilage is established
using Table 18.
Table 18. Indexes, characterising oxidative fat spoilage
Reaction with solution of neutral red
Colour
Swine or sheep From yellow with green shade to yellow
From dark yellow to brown
fat
From brown to pink
From pink to red
From yellow to brown
Cattle fat
From brown to browny pink
From browny pink to pink
From pink to red

Evaluation of fat condition

fresh
fresh but not for storage
doubtful freshness
spoiled
fresh
fresh but not for storage
doubtful freshness
spoiled

Number of peroxides, % of iodine

to 0.03
from 0.03 to 0.06
from 0.06 to 0.10
more than 0.10

fresh
fresh but not for storage
doubtful freshness
spoiled

7.3.4.1. Evaluation of peroxides number

During fat processing or keeping not melted fat stock for long, at influence of air oxygen,
light, high temperature and other peroxides can form. Number of peroxides is related with sensual
fat indexes: worse fat taste and odour is typical for fats with high peroxides number (Table 18).
Procedure. 1 g of fats are weighted with accuracy of 0.002 g into konus flask with polish plug

and are melted in water bath. From measuring cylinder along flask wall, 10 ml of chlorophorm, 10
ml of icy acetic acid and 0.5 ml of fresh prepared saturated solution of KJ are poured, that attached
fat parts would be washed. Flask is plugged, solution is mixed accurately and is left for 5 min. in
cool dark place. Then 100 ml of distilled water and 1 ml of 1 % starch are poured, mixed and
separated iodine is titrated with 0.01 M Na 2 S 2 O 4 solution till blue colour disappear. Control
analysis is made in parallel.
85

Reagents are right for analysis if for titration of control sample no more than 0.07 ml of 0.01
MNa 2 S 2 O 4 solution are used.
Number of peroxides, expressed as amount of iodine (g), which peroxides present in 100 g of
fat separate from potassium iodine in acidy environment, is calculated using formula:

X = 0.00127 K( V 1 V 2 )100/ m

V 1 amount of 0.1 M Na 2 S 2 O 4 solution (ml), used for titration of analyzed sample;

V 2 amount of 0.01 M Na 2 S 2 O 4 solution (ml), used for titration of control sample;
K re-counting coefficient to straight 0.01M Na 2 S 2 O 4 solution;
m amount of fat in extract, g.

7.3.4.2. Reaction with solution of neutral red

Procedure. 0.5-1.0 g. of melted fat are put into porcelain plate and solution of neutral red is

poured (fresh prepared aquatic 0.01 % solution of neutral red, 7,0 pH 7,2, is used. Solution can
be used for some hours). Using porcelain pistil, fat and solution of neutral red are rubbed for1 min.
Then solution is poured out and fat colour is watched. If left solution drops trouble for watching,
they are washed with water. Degree of oxidation spoilage is assessed using table 19.

7.3.4.3. Reaction with 2 thiobarbituric acid

Method is based on measuring of colour intensity of compound, formed during reaction of fat
oxidation products with thiobarbituric acid (TBA). Point of this reaction is not known till the end,
but experimentally it is improved that spectrical characteristic of TBA and oxidated fat reactions
product are the same as of product, got during reaction of TBA and malonic dialdehid.
Procedure. 3 g of melted fats are weighed to test-tube with polish plug and 10 ml of benzolum

(free from thiophen) or carbon tetrachloride are poured. Using pipette 10 ml of TBA reagent are
poured. Test-tube is plugging and is shaking intensively for 4 min. Got emulsion is poured to
dividing funnel and aquatic phase is separated to a test-tube (25 mm x 200). Test-tube is immersed
to boiling water bath, later it is cooled with flowing cold water. Part of this solution is poured into
photocolorimetric cuvette. According control sample (3 ml of distilled water instead of fat), at wave
length 530 m, colour intensiveness of this solution is measured and optical density is calculated.
Note: TBA reagent is prepared from 0.67 g of chemically clean TBA, which is melted in
water in 100 ml measuring flash, gently heating in water bath. Solution is cooled, distilled water is
86

poured to the flask till the mark and solution is mixed. Got solution is mixed well with icy acetic
acid, ratio 1:1. TBA reagent is prepared at the analysis day.

8. Composition and changes of blood

Blood in organism of cattle composes 7.5 % of body mass, in pigs 4.5 %, in sheep 7 %, in
hens 8 %. During bleeding process 40-50 % of blood flow from the carcass, the rest amount stays
in capillaries of tissues, organs and skin. pH of fresh blood, as buffer system, is stabile. Different
pH values are specific for each animal species. However, during storage and on influence of
ferments, lactic acid and compounds of inorganic phosphorus form, which lower blood pH from
7.3-7.4 to 6.8-7.0.
Blood consists of cells: formal elements and intracellular fluid (plasma). Ratio of formal
elements (erythrocytes, leucocytes, thrombocytes) and plasma depends on animal type and age
(Table 20).
Table 20. Ratio of formal elements and plasma in animal blood
Blood

Cattle
Calves
Pigs
Sheep

Amount, %
plasma

Formal elements

63
72
56.4
72.0

37
28
43.6
28.0

Blood plasma consists of 90 % of water and 10 % of dry substances (proteins, nitrogen and
not nitrogen substances, lipids, mineral substances). There are about 40 sorts of plasma proteins
assessed, the greatest amount of which consist serum proteins: albumin, globulin and fibrinogen.
Fibrinogen takes part in blood coagulation process. On action of ferments, fibrinogen changes to
unsoluble protein fibrin and polymerizes, making a net, where blood cells are cached.
Thromboplastin, prothrombin and calcium take part in blood coagulation process too. Rate of
various animals blood coagulation process is different. Cattle blood coagulates during 6.5-10 min
(pH 7.36-7.50), pigs blood during 3.5-8.5 min (pH 7.85-7.95), sheep during 4-8 min (pH 7.40795), birds < 1 min.
Seeking that blood wouldnt coagulate, it is stabilized. Sodium pyrophosphate, sodium
chloride, citrates, oxalates, heparin and others are used as stabilizers. They bind one of components,
which takes part in coagulation process and stop formation of blood clots in such way. Blood

87

stabilization relieves its further processing and fibrinogen stays in blood. Sometimes alimentary
blood is defibrinated fibrin is separated. Plasma without fibrinogen is called blood serum.
The main erythrocytes protein is haemoglobin (83 %), amount of which depends on animal
type, keeping circumstances and forages. Haemoglobin consists of protein globin and four prostetic
groups, which have carbon atoms. Depending on animal type, amount of carbon in blood can vary
from 0.035 % to 0.05 %.
Blood density is 1.055 kg/m 3 , plasma density is 1.03 kg/m 3 , density of erythrocytes is 1.09
kg/m 3 . Erythrocytes are heavier than plasma therefore they can precipitate in it. This feature is used
separating formal elements from plasma applying methods of precipitation, centrifugation and
separation. Haemoglobin is separated together with formal blood elements, however, a part of it can
solve in plasma, painting it with red colour.
Passing of haemoglobin to plasma is called haemolysis. Haemolysis could be indicated by
organic solvents (ethylic alcohol), active surface substances (soaps, bile acids), mechanical action
and freezing. Haemolysis can also occur when blood is diluted with water. Passing of haemoglobin
to plasma stops physiological solution. Depending on field of blood products usage, haemolysis can
be both: negative and desirable process. Light albumin, intended for confectionery, should be
processed from light blood plasma, whether for processing of boiled sausages plasma with partial
haemolysis is used. Processing liquid smooth haemathogen, blood haemolysis is indicated by
ethanol.
Blood breakdown depends on action of microorganisms and blood enzymes. During this
process, amounts of nonalbuminous nitrogen (amino acids, formations of purin) lactic acid,
inorganic phosphorus increase and compounds with unpleasant odour (indole, scatole, mercaptanes
and other.) separate form products of haemoglobin breakdown (bilirubin, bilverdin). During
concentration of organic substances breakage products, blood haemolysis occur. Separated
haemoglobin oxidizes to methaemoglobin, which, together with it breakage products, colours blood
dark with brown, green or other shades.
Conservation slows process of microbiological breakdown. Stabilized blood, plasma and
formal elements usually are conserved using NaCl, sodium citrate, ammonia, phosphates and other.
Mastitic blood and plasma are recommended to conserve with cold. At 0-2oC temperature cooled
plasma can be stored for 4-5 days, whereas frozen at minus 10oC temperature by 6 months.
Blood is used for processing of alimentary and nor alimentary albumin, haematogen, dry
lightened blood, blood powder. Sensual, physical, chemical and microbiological indexes of blood
products must correspond to values, determined in standards.

88

8.1. Evaluation of blood composition and characteristics

Colour of plasma is evaluated visually, analyzing sample (20-50 ml) poured into measuring
cylinder. Colour of fresh plasma is from yellow to orange without putrific odour. Colour and odour
of frozen plasma are analyzed after defrosting. There can be flakes like precipitates in defrosted
plasma.
Evaluation of blood pH. Blood pH is analyzed according potentiometric method, using not

diluted blood. Showings of pH-meter are checked according standard buffer. Then, electrodes are
washed with water, dried using paper filter and poured to analyzed blood. After setting of scale
cursor, ph-meter showings are checked. Then, electrodes are washed again and poured at distilled
water. Blood sort is determined according blood pH and blood freshness is determined according
sensual indexes.
Evaluation of dry blood substances amount. Amount of dry blood substances is one of the

technological key indexes, which enables to predict possible outputs of blood and its products. It
can be rated according density.
Procedure. Clean and dry hydrometer is soaked carefully into wide and large (250- 300 ml

capacity) cylinder with blood watching that it wouldnt touch walls and bottom of cylinder. When
hydrometer sets, showings of its scale are checked according blood level. According density, from
Table 21 amount of dry blood substances is found.

Table 21. Amount of dry blood substances depending on blood density

Density, measured with
hydrometer, at 20 0 C t.

1.059
1.055
1.054
1.052
1.051
1.050
1.048
1.046
1.045
1.044
1.043
1.042

Amount of dry substances,

%

20.85
19.40
19.48
18.38
18.27
17.81
17.68
16.41
16.25
16.12
15.92
15.24
8.2. Influence of physical and chemical factors on blood haemolysis

Various physical and chemical factors differently influence haemolysis process.

89

Influence of mechanic action. 30-40 ml of fresh stabilized blood are poured into centrifugal

test-tube. Haemolysis process is observed increasing blood centrifugation rate from 60 s 1 to 90 s 1

every 10 s 1 . Rate, at which haemolysis occurs, is marked.
Influence of chemical factors. 10 ml of fresh stabilized blood are poured into 50 ml capacity

measuring cylinder, and 60 % ethanolum is dropped from burette. Amount of ethanolum, causing
haemolysis is determined. Analogous analysis is made with water and 0.95 % NaCl solution.

9. Meat products
Meat products must be processed using technological instruction, according requirements of
Rules of meat and other animal origin products processing and delivering for marked places and
HN 15.
Pollution of stocks with chemicals and pesticides used for processing of meat products must
not exceed allowable levels, described in HN 54, microbial contamination must not exceed
allowable levels, described in HN 26. Water, used for processing of meat products must correspond
to requirements, described in HN 24. Food additives and fragrant substances for processing of meat
products must be used according rules, described in HN 53, HN 53-1, HN 53-2.
Meat products according processing are classified in to these groups:
- heat-treated meat products,
- not heat-treated meat products.
Heat-treated meat products are classified into these subgroups:
- boiled sausages (boiled sausages and bangers, roasted sausages),
- other boiled sausages (boiled variety meat sausages and bangers, boiled liverwursts, boiled
liver pies, boiled blood-puddings, boiled jelly sausages and bangers),
- hot smoked sausages (hot smoked sausages and bangers),
- boiled smoked sausages (boiled smoked sausages and bangers, boiled smoked variety meat
sausages and bangers),
- smoked liwer-wursts, boiled smoked liver pies, boiled smoked blood-puddings,
- roasted sausages (roasted sausages and bangers, roasted liver-wursts, blood-puddings and
variety meat sausages),
- boiled meat products (boiled rolls, boiled products in jelly, boiled meat breads, boiled formal
hams, boiled formal meat, boiled brawns, boiled nubbly meat and variety meat products),

90

- roasted meat products (roasted meat breads, roasted liver-pies, roasted nubbly meat and
variety meat products, roasted rolls, roasted formal hams, roasted formal meat),
- hot smoked meat products (hot smoked nubbly meat and variety meat products, hot smoked
rolls, hot smoked formal hams).
Not heat-treated meat products are classified into these subgroups:
- cold smoked sausages (cold smoked sausages and bangers, smoked spread sausages and
bangers),
- withered sausages (withered sausages and bangers),
- spread sausages (spread sausages and bangers),
- cold smoked meat products (cold smoked formal meat or variety meat products, cold
smoked rolls, cold smoked lard),
- lightly smoked meat products (lightly smoked formal meat products),
- withered meat products (withered formal meat products),
- salted meat products (salted lard, salted formal meat products).

For heat-treated meat products there are some regulated parameters: amount of meat proteins
without collagen (%), humidity (%), fat amount (%), amount of albuminous meat substitutes (%),
starch amount (%). For not heat-treated meat products there are some regulated quality parameters:
amount of meat proteins without collagen (%), humidity (%), fat amount (%), amount of
albuminous meat substitutes (%).
In processing of the highest sort meat products it is unallowed to use food fillers,
mechanically separated meat, soya proteins flour. For processing of the highest sort saussages and
bangers no more than 3 % of albuminous milk and egg products are allowed to use.
In processing of the first sort meat products it is unallowed to use soya protein flour. In
processing of the first sort meat products it is allowed to use no more than 3% of fillers.
In processing of not heat-treated meat products (except smoked-spread and spread sausages
and bangers) it is not allowed to use fillers.
Meat products, which are labeled using traditional qualified meat productsnames: Panerio
dera, pienika dera, daktarika dera, pienikos derels, servelatas, mediotoj derels,
lietuvika dera, grdtoji dera, Alytaus dera, Panevio fil, Vilniaus fil, mgj kepsnys,
ikylautoj ukanda, aromatinis kumpelis, Kauno kumpelis must correspond only for the highest
meat products sort indicated quality indexes. These names of traditional meat products are not
allowed to be changed. Either similar to these or other names, which can be originated from
traditional name roots or new words are not allowed to use

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Meat products with exclusive, high quality describing words extra, prima, premium, lux,
The highest class, delicacy, special, perfect, golden, for gourmands or words, originating from
roots of those words must correspond only for the highest sort indicated quality indexes.
Microbial meat products contamination must not exceed levels, described in HN 26.
Packing of meat products. Material and the main package, used for packing must correspond

to HN 16 requirements. Meat products must be packed according requirements of Rules for meat
and other animal origin products processing and delivering to market. Meat products can be
packed on polymeric pads, which are covered with wrap, not permeable to air and moisture, to
polymeric bags using vacuum packing or in packing gas atmosphere on polymeric pads, which are
covered with wrap, not permeable to air and moisture or in any other primary package. Packed in
primary package meat products can be packed in secondary package (cardboard, polymeric, metal
boxes or other secondary package).

10. Evaluation of food alimentary and energetic value

10.1. Evaluation of nitrogen

Meat sample, no smaller than 200 g, is taken. It cant spoil during storage and it composition
cant change. Used preservatives cant consist of nitrogen. The sample is homogenized twice
grinding it in grinder and is mixed. It should be stored hermetically, that it composition couldnt
change. Homogenization analysis must be done as soon as possible but necessarily in 24 hours.
Procedure. Boiling regulating substance, 15 g of not aqueous potassium sulphate and 0.5 g of

copper (II) sulphate are poured into Kjeldal flask. 2 g of sample (or 1.5 g if fat products are
analysed) are weighed at accuracy of 0.001 g, on a piece of paper, not permeable to fat. Paper with
sample is put into Kjeldal flask. 25 ml of sulphate acid are poured in to Kjeldal flask too.
Everything is mixed slowly twirling. If it is necessary, pear formed plug is inserted into flask neck
down peaked tip. Flask in oblique position (about 40o from vertical position) is put on heating
device. At first heating is light, till content of flask stops foaming and liquefies. Later it is boiled,
sometimes twirling the flask, till the fluid liquefies and becomes lightly blue-green colour. Content
of flask is boiled for another 90 min. General boiling time must be not less than 2 h. It is necessary
to avoid, that fluid condensate shouldnt flow outside the flask. When heating is very intensive, it is
necessary to avoid, that sulphate wouldnt evaporate too much, because this can determine
decreasing of nitrogen amount. Solution is cooled to about 40o C temperature, 50 ml of water are
poured carefully, mixed and allowed to cool. 50 ml of boratic acid using measuring cylinder, are

92

poured into Erlenmejer flask, capacity of 500 ml. 4 drops of indicator are dropped, mixed and the
flask is put under distillers condenser. Tip of condensers adapter must be soaked to fluid in the
flask.
Content of Kjeldal flask is analysed in direct distilling way. Content of Kjeldal flask is
carefully diluted with 500 ml of water and is mixed twirling. If it is necessary, 1 L flask can be
used. After 15 min, 100 ml of sodium hydroxide solution are poured using measuring cylinder. This
solution must be poured carefully through the neck of oblique flask in such way, that two liquid
layers could not mix. Flask with distillated prepared mixture is attached to distiller immediately.
105 ml of distillate must be gathered, despite the mixture sometimes jumps. Distillation is
prolonged till mixture stops jumping and 205 ml of distillate are gathered. During distillation
procedure, the distiller must be cooled well and heating of boratic acid must be avoided. Finishing
distillation, Erlenmejer flask must be lowered, in case that tip of adapter was over the fluid. Internal
and external sides of adapter tip are washed with little amount of water, keeping it above the flask
fluid. The end of ammonia distillation is confirmed using red litmus paper, immersed with distilled
water. It colour must not change from the drops from condenser. Heating is finished. If it is
supposed that distillation is not over, analysis must be repeated more carefully according the
description.
The solution in Erlenmejer flask is titrated with acid of hydrogen chloride. Amount of acid,
used for titrating is marked at accuracy of 0.02 ml.
In parallel, two independent samples of the same sample are analyszd.
Empty analysis. 2 empty analyses are performed, when new parcel of reagents or fresh

prepared reagents are used. Empty analyses are performed if used reagents were prepared long ago
too. Empty analysis is performed according reference method, only using not permeable for fat
piece of paper (meat is not used).
Calculation of the results. Amount of nitrogen in mass % is calculated using formula:

0.0014x(V1-V2)x100/m
V2 amount of 0.1 mol/l concentration hydrogen chloride acid (ml), used for titrating of
empty analysis solution;
V1 amount of 0.1 mol/l concentration hydrogen chloride acid (ml), used for titrating of
sample solution;
m mass of sample (g).
Note: if concentration of ethalonic hydrogen chloride acid is not the same as described,
calculating the results corrective coefficient must be used.
93

10.2. Evaluation of humidity

Procedure. Not less than 200 g of representative sample is taken. It is stored in such way, that

wouldnt spoil and it content wouldnt change. The sample is homogenized using right equipment.
It is important, that temperature of sample wouldnt up-rise more than 25oC. The sample is
homogenized twice grinding it in grinder. It should be stored hermetically, that composition
couldnt change. After homogenization analysis must be done as soon as possible but necessarily in
24 hours.
Preparation of cruet and sand. Sand is put into a cruet. Mass of sand must be 3 to 4 times
higher than mass of sample. Cruet with sand ant glass stick is heated for 30 min. in shelf at 103o C
temperature. Later cruet with sand and glass stick is cooled to room temperature in exicator and is
weighed at accuracy of 0.001 g.
Content of cruet is mixed with glass stick (if it is hard to mix sample with sand, ethanol can be
poured, but ethanol must be evaporated before drying of sample in the shelf). Cruet with content
and glass stick is heated for 2 hours in shelf at 103oC temperature. After that it is taken, put into
exicator, cooled to room temperature and weighed at accuracy of 0.001 g. Heating, cooling and
weighing are repeated according analysis till after 1 hour heating difference between two weighing
results (m2), made one after another would be not higher than 0.1 % of sample mass.
Calculation of the results. Amount of humidity (w), expressed in mass percentage, is

calculated using formula:

w = ((m1-m2)/(m1-m0))x100
m0 mass (g) of cruet with sand and glass stick;
m1 mass (g) of cruet with sand and glass stick before drying;
m2 mass (g) of cruet with sand and glass stick after drying.
Results are got at accuracy of 0.1 %.

10.3. Determination of general fat amount

Representative sample of weight not less than 200 g is taken. It is stored in such way, that
wouldnt spoil and it content wouldnt change. The sample is homogenized twice grinding it in
grinder and mixed. It should be stored hermetically, that wouldnt spoil and composition wouldnt
change. After homogenization analysis must be done as soon as possible but necessarily in 24
hours.

94

Analyzing sample: according the predicted fat amount 3 to 5 g of minced sample are weighed
at accuracy of 0.001 g and put into 250 ml Erlenmejer flask.
Exicator flask with boiling controlling material in it, is dried for 1 hour in drying shelf at
1032oC temperature. Then flask is cooled in exicator to room temperature and weighed at
accuracy of 0.001 g. 50 ml of saline acid are poured into the sample and Erlenmejer flask is covered
with clock glass. Flask is heated on gassy fuse with asbestos net, till content of flask begin to boil.
Further it is heated on light fire for another 1 hour sometimes shaking and 150 ml of hot water are
poured. Corrugate paper, put into glass funnel is moistened and hot flask content is poured through
the filter. Flask and clock-glass are washed with hot water for three times and are dried in drying
shelf. Filter paper is washed with hot water until blue litmus paper stops changing colour. Filter
paper is put on clock glass or Petri dish and is dried for 1 hour in drying shelf at 1032o C
temperature. After that it is cooled.
Filter paper is rolled and put into extraction shell. Cotton-wool is moistened with extraction
solvent and is used for cleaning any fat residues from clock glass or Petri dish. Cotton-wool is put
into extraction shell too. Shell is put into extractor. Filter paper is taken either with cleaned pincers
or putting paper fingerstalls. Extraction solution is poured into dried extractors flask. Inside of
Erlenmejer flask is washed with saline acid, clock glass is washed with extraction solution and
everything is poured to extraction flask.
General amount of solvent should be from 1,5 to 2 times higher than volume of extractors
extraction tube. Flask is put into extractor. Extraction flask is heated for 4 hours at sand or water
bath or in similar equipment. After extraction, flask with fluid is taken away from extractor and
solvent is evaporated using for ex., sand or water bath. The last solvents drops are evaporated in
water bath and, if it is necessary, air ventilator is used. Extraction flask is dried for 1 hour in drying
shelf at 1032o C temperature, then it is cooled in exicator to room temperature and weighed at
accuracy of 0.001 g. This operation is repeated until difference between results of two last
weightings would be not higher than 0.1 % of general sample mass. End of extraction is checked
with another extraction flask and with new solvent portion, extracting 1 hour longer. Increase of
mass must not exceed 0.1 % of sample mass. The second analysis is performed with the same
prepared sample.
Calculation of the results. General fat amount, expressed in mass percentage is calculated

using formula:
(m2-m1)x(100/m0)

95

m0 sample mass (g);

m1 mass (g) of extraction flask with boiling regulating material;
m2 mass (g) of extraction flask with boiling regulating material and fat, after drying.

10.4. Determination of general ash amount

Representative sample, not less than 200 g, is taken. It is stored in such way, that wouldnt
spoil and it content wouldnt change. The sample is homogenized twice grinding it in grinder and
mixed. It should be stored in refrigerator, hermetically, fulfilled. Sample analysis must be done as
soon as possible but necessarily in 24 hours after homogenisation.
Melting-pot is heated for 20 min in muffel heater at 550o C temperature. The melting-pot is
cooled in exicator to room temperature and is weighed with analytic balance at accuracy of 0.1 mg.
From 1.5 to 2 g of prepared sample are put into the melting-pot and are overspread gradually. The
melting-pot with the sample is weighed immediately at accuracy of 0.1mg.
Analysis, using muffel heater without time and temperature regulator. Melting-pot with

sample is stored for 1 h in drying shelf at 103o C temperature. Melting-pot is taken out from the
shelf and put on electric stove or gassy fuse. Gradually temperature is elevated, till the carboning
material separates smoke. Carefully it is carboning till smoke dont appear. The sample cant heat to
glowing or burning. The melting-pot is put into cold muffel heater and temperature is elevated to
55025o C. After 4 h the melting-pot with the sample is taken out from the muffel heater and is
cooled in exicator to room temperature. Moving the melting-pot with ash from muffel heater to
exicator and from exicator to analyzing balance, it is necessary to avoid loss of ash. Ah are
observed carefully. If they are still black, they are treated with some drops of hydrogenum peroxide
or water and analysis is repeated according this description. If ash are greyish white, the melting-pot
with the sample is weighed with analytic balance at accuracy of 0.1 mg.
Calculating of the results. Sample ash amount is calculating using formula:

wa=((m2 m0)/(m1 m0))x100%

wa sample ash amount in mass percentage;
m0 mass of empty melting-pot (g);
m1 mass of melting-pot with the sample (g);
m2 mass of the melting-pot with ash (g).
The results are written at accuracy of 0.01%.

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Analysis, using muffel heater with time and temperature regulator. Melting-pot with the

sample is put into a cold muffle heater and temperature of the heater is gradually elevated from 6 h
to 7 h to 55025C temperature. Later it is heated at 55025C temperature, till ash become greyish
white.
The melting-pot is taken out of the muffle heater and is cooled in exicator to room
temperature. Ah are observed carefully. If they are still black, they are treated with some drops of
hydrogenum peroxide or water and analysis is repeated according this description. If ash are greyish
white, the melting-pot with the sample is weighed with analytic balance at accuracy of 0.1 mg.
Calculating of the results. Ash amount of the sample is calculating using formula:

wa =

( m2 m0 ) 100%;
( m1 m0 )

wa sample ash amount in mass percentage;

m0 mass of empty melting-pot (g);
m1 mass of melting-pot with the sample (g);
m2 mass of the melting-pot with ash (g).
The results are written at accuracy of 0.01%.

10.5. Calculation of food products energetic value

After laboratory evaluation of dry matter, mineral matter, humidity, fat and after calculating
proteins according coefficients, we can calculate energetic value. It is calculated using formula:

E = 4 x P + 9 x F + 4 x C (1)
E energetic value, kcal;
P, F, C amounts of proteins, fat and carbohydrates, g;
4, 9, 4 coefficients of energetic value, kcal/g.
Energetic value can be expressed in kcal or kJ, showing their amount in 100 g or 100 ml of
product, in one meal or portion, in day ratio or separate eating.
For calculating of carbohydrates the next formula is used:

C = D. m. (P + F + M.m)

(2)

C carbohydrates, g;
D. m., P, F, M. m. respectively: dry matter, fat, proteins, mineral matter, g.
Energetic value according laboratory analysis is calculated using formula:
97

E = 4 x P + 9 x F + 4 [D. m. (P + F + D. m.)] (3)

E energetic value, kcal must be indicated for concrete amount;
4, 9, 4 energetic value coefficients, kcal/g;
D. m., P, F, M. m. respectively: dry matter, fat, proteins, mineral matter, g.
Determination of errors

Errors are determined comparing amounts of proteins, fat, carbohydrates and energetic value,
which were got using laboratory analysis with their calculated amounts. Errors are calculated using
formulas:
XP =

XF =
XC =

XE =

(P1

P 2 ) 100
P2

(4)

(F1 F2 ) 100 (5)

F2

(C 1

C 2 ) 100
C2

(6)

(E 1

E 2 ) 100
E2

(7)

XP, XF, XC, XE errors for proteins, fat, carbohydrates and energetic value, %;
B1, R1, A1, E1 amounts of proteins, fat, carbohydrates (g) and energetic value (kcal, kJ)
determined using laboratory methods;
B2, R2, A2, E2 theoretically calculated amounts of proteins, fat, carbohydrates (g) and
energetic value (kcal, kJ).

11. Evaluation of salted-smoked meat products

11.1. Evaluation of sausages sensual characteristics

Analyzing sausages, they are observed visually, taking attention, how they are corded, what is
their form, how does they look like and so on. The sausage is cut along in twain. Wrap is taken out
from one side and odour outside and under the wrap is observed. Meat condition, size of lard pieces
and their spreading at the sausage surface and inside are noted. Sensual evaluation is assessed
according next indexes.
External appearance. Surface of sausages must by dry, clean, not tattered, without molds,

not covered with fat. At the surface of boiled sausages there should not be any dark spots. Wrap
must be elastic, not mucous, not separated from meat. If surface of sausage is covered with molds,
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mucous, is sticky, wrap is separated from meat, the sausage is of doubtful quality. Sausages with
such appearance cant be realized for human consumption, and semi-smoked and smoked sausages
must be processed. Sausages, covered with ash, smoky, with tattered wraps and so on, cant be
realized. Smoked products (hams, ribs) must be smoked nice, dry, clean, not covered with salt.
Moldy, especially if molds are penetrated into muscle tissue, also with grubs, especially near bones
or in meat cords, mucous products cant be realized.
Consistence and colour. Sausages must be tight, not softened. Their consistence in cut must

be juicy, meat pressed well, not crumbling, without air caves. Cut of good quality sausage is pink,
without grey spots. Lard is all white. In periphery of not fresh sausages and inside in places, there is
greenish grey colour, their consistence is flabby (especially of boiled sausages, blood-puddings and
liver wursts), lard or fat are greenish. Fly grubs can be found in such sausages. Smoked meat
products must be tight, elastic, muscles in cut must be pink and fat white. There cant be greenish
or rusty colour near bones.
Flavour and odour of good sausages is pleasant, specific for each sort, without sour or musty

signs. Out-of-date sausages have sour, scalding or putrescent (especially quickly such become
boiled sausages, blood puddings and liver wursts, stored badly) flavour and aroma. Fat are soured
and yellowed. Flavour and aroma of additives is lowered.
Spoiled smoked products have putrific odour near bones, lard become yellow and soured.

11.2. Laboratory analysis of sausages

Samples of sausages and smoked products are homogenized and put into bottles or flasks,
with lapping plugs.
11.2.1. Evaluation of humidity
Procedure. Glass cruet with 7-8 g of clean sand with glass stick are dried to constant weight

and cooled in exicator. About 3 g of prepared meat are weighted with analytic balance in to cruet
and is mixed well with the sand. Drying is assessed in drying shelf or thermostat at 150oC
temperature for 1 h from that moment, when temperature inside the shelf after putting of analyzing
samples reaches 150oC. Later cruet is cooled in exicator and weighed. Humidity (%) is calculated
using formula:
X = (g1 g2) 100/g

g1 mass (g) of cruet with products before drying,

g2 mass (g) of cruet with products after drying
g mass (g) of analyzed product.
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11.2.2. Evaluation of salt

Procedure. 3 g of prepared analyzed minced meat are weighed into 200 ml content glass and

100 ml of distilled water are poured. Analyzing boiled sausages, meat is mixed well (10 min.) using
glass stick with rubber tag, that bigger pieces of meat didnt left and salt was melted in water. Later
mix is left to stand for 5 min.
Analyzing semi-smoked and smoked sausages and other smoked meat products, after pouring
needed amount of water (100 ml), everything is heated, that mix would be 30oC, and it is stirred
well with glass stick with rubber tag for 10 min., that bigger meat pieces would split and salt would
better melt in water. After that mix is left for 5 min to precipitate.
15 ml of precipitated fluid are taken with pipette and they are titrated with 0.05 N silver nitrate
solution using indicator solution of potassium chromate.
Amount of salt (%) in analyzed sample is calculated using formula:

x = 0.0029 v 100 100/ b g

v amount of 0.05 N silver nitrate (ml), used for titration,

b amount of solution (ml), taken for titration,
g amount of minced meat (g), taken for analysis,
0.0029 titre of 0.05 N silver nitrate solution.

11.2.3. Evaluation of nitrites amount (reference method)

Meat extract is being got using hot water, proteins are precipitated and it is filtered.
Sulphanilamide and N -1-naftilendiamine dihidrochloride are added into filtrate, and red colour
appears. Photometric measures are performed, when length of wave is 538 nm.
Representative sample, not less than 200 g, is taken. It is prepared at once, or if it is
impossible, it is stored from 0oC to 5oC temperature, not longer than 4 days.
The sample is homogenized, grinding it with grinder at least two times and mixing. It is stored
in refrigerator in fulfilled and hermetic tare. Prepared sample must be analyzed as soon as possible,
but necessary in 24 h. When products are raw, after grinding they are analyzed at once.
Procedure. 10 g of sample are weighed at accuracy 0.001 g. For elimination of proteins, the

sample is put into conic flask, and straight are added 5 ml of saturated borax solution and 100 ml of
water, not less than 70oC temperature. The flask with the content is heated in boiling water bath for
15 min. and is shacked constantly. The flask with the content are left to cool to room temperature
and straight are added 2 ml of the 1-st reagent and 2 ml of the 2-nd reagent. After pouring each of
100

them, everything is mixed carefully. The content is poured into 200 ml measuring flask, is diluted
with water and is mixed. The flask is left for 30 min. in room temperature. The surface fluid is
poured out carefully and it is filtered through wrinkly paper filter in case to get lucid solution.
Needed amount of filtrate V (ml), but not more than 25 ml, is dripped with pipette into 100 ml
measuring flask and such amount of water is added, that about 60 ml of solution would be got. 10
ml of the 1-st solution, later of the 3-rd solution are poured, everything is mixed and the solution is
stored dark in room temperature for 5 min. 2 ml of the 2-nd solution are added, everything is mixed
and solution is stored from 3 min. to 10 min. dark in room temperature. Later it is diluted with
water to the mark. Optical density of the solution is measured in ciuvete of 1 cm thickness with
photoelectrical colorimeter or spectrophotometer, when wave length is about 538 nm.
Note. If optical density of coloured solution, got from the sample, exceeds optical solution of
the highest concentration standard solution, described operations of colorimethric measuring are
repeated with less amount of filtrate. Two independent analyses are made with separate samples
from the same sample.
Preparing of calibration curve. 10 ml of water are poured with pipette into one of four 100

ml measuring flasks. Into other three flasks 10 ml of other standard nitrite solutions, where there are
2.5 g, 5.0 g, 10.0 g of nitrite in 1 ml.
Such amount of water is poured into each flask, that we could get about 60 ml of solution and
it is done as described in last paragraphs of colorimetric measurement method.
Marking measured optimal densities in diagramme, corresponding to concentrations of
standard solutions (g/ml), calibration curve is drawing.
Calculating of the results. Amount of sample nitrites Q(NaNO2), expressed as 1mg of sodium

nitrite for 1 kg of product, is calculated using formula:

Q (NaNO2) = c 2000 / m V
m mass of sample, g;
V capacity of filtrate, used for photometric measurements, ml;
c sodium nitrite concentration (g) for 1 ml, deducted from calibration curve, corresponding
to optical density of solution, prepared from sample.

11.2.4. Evaluation of nitrates amount (LST ISO 3091:1997) (reference method)

Using hot water, meat extract is got, proteins are precipitated and it is filtered. Nitrates from
extract are reduced to nitrites using metallic cadmium. Sulphanilamide and N-1-naftilendiamine
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dihydrochloride are added into filtrate and red colour appears. Photometric measures are performed,
when length of wave is 538 nm.
Representative sample, not less than 200 g, is taken. It is prepared at once, or if it is
impossible, it is stored from 0oC to 5oC temperature, not longer than 4 days.
The sample is homogenized, grinding it with grinder at least two times and mixing. It is stored
in refrigerator in fulfilled and hermetic tare. Prepared sample must be analyzed as soon as possible,
but necessary in 24 h.
Preparing of cadmium column. From 3 to 5 zinc sticks are put into cadmium sulphate

solution, in laboratory glass.

Note. 1 liter of cadmium sulphate solution is enough to prepare one cadmium column.
Every 1 h or 2 h spongy metallic cadmium precipitates are moved from zinc sticks. For that
purpose, sticks are twirled in solution or they are rubbed one to each other. After 6-8 h solution is
poured out and precipitates are washed twice with 1 liter of water, checking, that cadmium would
constantly be sank in fluid.
Precipitates of cadmium and 400 ml of hydrochloric solution are poured into laboratory mixer
and are mixed for 10 s. After that content of mixer is returned into laboratory glass.
Cadmium precipitates sometimes are stirred with glass stick. Leaving them for night in
hydrochloric solution, they are stirred once more, that all gassy bubles would be moved from
cadmium.
Solution is poured out and pasta of cadmium is washed twice, each time using 1 liter of water.
Glass-wool plug is put on bottom of glass column, where cadmium would be put. Cadmium is
washed with water into glass column, till height of cadmium becomes about 17 cm. Filling the
column, sometimes it can stream, but cadmium layer must be sank. Gassy inserts are moved out,
for. ex., using needle. The fluid must flow not faster than 3 ml/min.
Procedure. 10 ml of sample are weighed at accuracy of 0.001 g. The sample is put into conic

flask and straight 5 ml of saturated borax solution and 100 ml of not less than 70oC temperature
water are poured. The flask with the content is heated at boiling water bath, constantly shaking for
15 min. Then flask with the content are left to cool to room temperature, and straight 2 ml of the 1st reagent and 2 ml of the 2-nd reagent are poured. After pouring each of them, everything is mixed
carefully. The content is poured into 200 ml measuring flask, diluted with water to the mark and
mixed. Flask is left for 30 min. in room temperature. Then carefully the surface fluid is poured out,
it is filtered through paper filter, to get lucid solution.
Note. If it is necessary to evaluate amount of nitrates and nitrites, analyzing the same sample,
the same filter for each analysis can be used.
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The final preparation of cadmium column. Cadmium column is straight washed with 25 ml

of hydrochloric solution, 50 ml of water and 25 ml of diluted buffer ammonia solution. Fluid level
in funnel cant fall lower than top of cadmium column capiliary incoming tube.
Evaluation of cadmium column reductive ability. 20 ml of standard potassium nitrate

solution and 5 ml of buffer ammonia solution are poured with pipette into reservoir which is at the
top of cadmium column. Outflow is gathered into 100 ml measuring flask.
When reservoir is almost empty, it walls are washed with about 15 ml of water and washing is
repeated once more. When the second portion flows to column, the reservoir is fulfilled with water.
When near 100 ml of outflow is gathered, the flask is taken out from column and the content is
refilled with water to the mark.
10 ml of column outflow is poured with pipette into 100 ml measuring flask and everything is
performed as described in last stages.
If concentration of nitrites in column outflows, deducted from calibration curve is less than
0,9 g of 1 ml sodium nitrite, analysis results, got with this cadmium column are eliminated.
Reduction of nitrates into nitrites. 20 ml of filtrate, together with 5 ml of buffer ammonia

solution are poured into reservoir at the top of column. Column outflow are gathered into 100 ml
measuring flask. Later washing is made as it was described above.
Colorimetric measuring. Needed amount of filtrate V (ml), but not less than 25 ml, with

pipete are poured into 100 ml measuring flask and such amount of water is used, that we could get
about 60 ml of solution. 10 ml of the 1-st and 3-rdsolutions are poured, mixed and left dark in the
room temperature for 5 min. 2 ml of the 2-nd solution are poured, mixed and solution is left from 3
min. to 10 min. dark, in room temperature. It is diluted with water to the mark. Optical density of
solution is measured in 1 cm thickness column with photoelectric colorimeter or spectrophotometer,
when wave length is about 538 nm.
Note. If optical density of coloured solution, got from the sample exceeds optical density of
the highest concentration standard solution, described colorimetric measuring operations are
repeated with less amount of filtrate. Two independent analysis using different samples from the
same sample are made.
Calibration curve. Into one of four flasks of 100 ml capacity 10 ml of water are poured with

pipette and into each other 10 ml of other three standard nitrite solutions in 1 ml of which there
are 2.5 g, 5.0 g and 10.0 g of nitrite, are poured.
Such amount of water is poured into each flask, that we could get about 60 ml of solution and
it is done as described at the last paragraphs of colorimetric measuring.

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Calculating of the results. Amount of sample nitrates Q, expressed in mg of potassium nitrate

for 1 kg of product is calculated using formula:

Q (KNO3) = 1,465 [ c 10000 / m V Q (NaNO2)]

m mass of sample, g;
V capacity of column outflow, taken for colorimetric measuring, ml;
c concentration of sodium nitrite g/ ml, deducted in calibration curve, corresponding
optical density of solution, prepared from the sample.
Q (NaNO2) amount of nitrites in sample, expressed as sodium nitrite mg/kg, evaluated
according ISO 2918.
The result is written at accuracy of 1 mg/kg.

11.2.5. Evaluation of Calcium amount (titrimetric method)

Analysis sample is mineralized using dry method. The ash is melted in hydrochloric acid.
Calcium, which is in solution, is precipitated as calcium oxalate, which is melted in sulphate acid.
Solution of calcium oxalate is titrated with reference potassium permanganate solution. Amount of
calcium is calculated according formed amount of oxalic acid.
Procedure. About 5 g (or more, if needed) of analysis sample are weighed in melting-pot at

accuracy of 1mg. The melting-pot with the analysis sample are put into muffel heater and is burned
about 4 h at 55020C temperature, till all organic matter burn.
If after 4 h burning there are left not burned material particles in the melting-pot (black,
charred particles), some drops of concentrated nitrogen acid are dropped into the melting-pot and it
is dried on hot electric stove. Then once more, the melting-pot is put into muffel heater and burned
for 30 min at 55020C temperature. This procedure is repeated several times, till all organic matter
burn. Ash from melting-pot analysis sample is poured into 250 ml capacity glass.
40 ml of hydrochloric acid solution, 60 ml of water and some drops of concentrated nitrate
acid are poured into the glass with ash. Content of glass is heated to boiling and is boiled 30 min.
Later it is cooled and poured into 250 cm3 measuring flask. The glass is washed with water which is
poured into the same flask. Then, content of measuring flask is diluted with water till the marking
place, it is mixed well and filtrated. The got filtrate is solution of analysis sample.
Aliquotic part of analysis sample solution, where must be 10-40 mg of calcium, is poured with
pipette into 250 ml glass. 1 ml of citric acid monohydrate solution and 5 ml of ammonium chloride
solution are poured into the glass, diluted to 100 ml and put to boil. After that, 10 drops of
104

bromcresolum green solution and 30 ml of warm ammonia oxalate solution are poured. If
precipitate appears, they are melted, dropping some drops of hydrochloric acid solution. Glass
content, constantly mixing, is neutralised with ammonia hydroxide solution to pH 4.4-4.6 (changes
colour of indicator). Then the glass is put into boiling water bath and left for 30 min. in case to form
precipitates of calcium oxalate. The glass, taken from the boiling water is stored for 1 h in room
temperature. The solution is filtrated through the glass filter.
The glass and the filter with precipitates are washed with water, till ammonium oxalate
disappears at all. This is proved by negative chloride test in outflow water.
The filter is put into 250 ml glass or open-mouthed flask. 80 ml of sulphate acid are poured
and heated to 70-80C temperature till precipitates melt.
Hot solution is titrated with reference potassium permanganate solution, till colour changes
into light pink not changing for 1 min.
Calculating of the results. Amount of calcium in grams for 1 kg of sample is calculated using

formula:
20, 04 V v 250

;
m
V1

V amount of potassium permanganate solution, used for titration, ml;

V 1 aliquotic part of analysed solution, ml;
c the right concentration of reference potassium permanganate solution, mol/L;
m mass of analysis sample, g;

12. Defects of meat products

12.1. Defects of boiled sausages

Forming of jelly appears when old meat is consumed or too much water is added into
sausages.
Dry consistence too much beef or too less pork is added. There is too less fat in mincemeat.
Tasteless, pale, dry consistence insufficient amount of salt, additives and water in sausage
minced meat.
Separated fat mincemeat heated too much during cuteration. Fat layer forms between
intestine and mincemeat during boiling.
Crumbling of large inserts bad mixing of large inserts with sausage meat.
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Lack of coherence too less salt is added into sausage meat or it is absent at all. Too much
water or lard was used.
Spongy consistence too much water were added into sausage-meat or used watery meat.
Dark cut too much beef was used.
Pale colour too much poultry in the sausage meat, lack of pork, blood plazma, milk proteins
and colour stabilizers.
Grey core sausages were smoked too short or smoking temperature was too low.
Acidity sausages were smoked too long.
Not clear taste meat stock was salted for too long.
Taste of old sausage overtimed stock and spice.

12.2. Defects of dried and cold-smoked sausages

Not clear cut overweighted grinder content, meat wasnt cooled enough before grinding.
Wrinkly sausages wrap too intensive smoking, too wet sausage-meat was crammed.
Sausage-meat was crammed not tight.
Weeping processed sausage-meat was heated too much, too little salt was used.
Soft consistence sausage-meat was prepared from wet meat, meat wasnt cooled enough,
mincemeat was heated too much during grinding.
Not adhered sausages wrap sausage-meat was crammed into wet intestines or it was too
intensive air movement in the room, therefore sausage-meat shrank too much and became not
adhered.
Cavities improper sausages maturing.
Moulding cramming of sausages into too fresh natural intestines or if after smoking
sausages were stored in badly ventilated room.
Colour of sausages is not clear enough.
Too fat cut.
Crumbling cut meat was wet during processing and during maturing moisture evapoured
intensively.
Too tight consistence there is too much beef in sausage-meat.
Browning too long or hot smoking or too short maturing.
Greying badly dried intestines, too high humidity or too low temperature in sausages dryer.
Greyish green shade.
Whiteness, enlarging towards the cord.
Quickly whitening cut.
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Urine smack meat of not castrated boar was added into sausage-meat.
Fusty odour bad ventilation during maturing or storage of sausages.
Bitterness high environmental temperature and light affected stored sausages, old lard and
fresh intestines were used. Improper filing was used for sausages smoking.

12.3. Defects of liverwursts

Dry consistence too much fat-free stock wase used and cattle internal organs were added
into sausage-meat.
Soft consistence there is maximal amount of fat and too little amount of salt in sausagemeat.
Insufficient tightness too much stock were used in sausage-meat.
Insufficient spreading there are too little fat in sausage meat and too much cooked liver and
fat-free meat.
Tasteless sausages cooked stock was cooled for too long at cold water or was placed into
cold water for cooking.
Sharp taste and odour natural intestines were stored without defatening for too long.
Scalding flavour not all bile ducts were removed from liver.
Souring too long sausages processing, not enough or too late reached proper cooking
temperature. Microorganisms are appearant in sausage meat.

12.4. Defects of blood-puddings

Bloody lard lard were not cooked enough before processing.

Souring microorganisms ferment carbohydrates (which are in grits), not enough cooked
pancreas was added (action of ferments).
These defects can be controlled, introducing new technologies, which can improve
consistence or even flavour. They also can improve all technological process and it can be
controlled. One of the newest methods for improvement of sausages quality is functional food and it
ingredients. The most important ingredients are:
Nutritional fibers polydextrins, fructoolygosacharids, cyclodextrins;
Peptides and proteins soya globalin, caseinphosphopeptides;
Probiotic cultures Lactobasillus alimentarius and other;
Poli-unsaturated fatty acids linolum, linolenum, arachidone.
Antioxidants vitamins A, E, C, biophlavanoids.

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One of the methods to process functional food is using of probiotic cultures alive good
bacteria. Lactobacillus spp (L. acidophillus, L. casei Shirota strain, L. plantarum and other) are
used more frequently. During sausages smoking process they act like catalyst mature sausages.
There are no preservatives in such sausages. Besides, probiotic cultures positive act on human
organism: they fortify immune system, lower gastric acidity, fight with infections. Furthermore,
they help to restore normal intestinal activity, which can work bad using antibiotics, being ill with
infective gastrointestinal diseases, being stressed or even because of poor nutrition.
Probiotic cultures in sausage products perform function of starter cultures, also improve
nutritional value, stops proliferation of pathogenic microorganisms. Prebiotics are used too, as a
novelty, improving technological process. Various alimentary fibers are used as prebiotics. They are
added into emulgated sausage products, which dont influence amount of proteins and connective
tissue in product. Prebiotics absorb water (expand output), change matrix pH (6.4-5.8), change
products colour into darker brown shade. Big concentration of fiber suspension (29 %) worsens
consistence indexes. However in fermented sausage products they dont influence amount of
proteins and connective tissue in the product, lower (35 %) energetic value, improve nutritional
value, change pH (6.4-5.8). Fiber concentration of 3% worsens consistence indexes.

13. Variety meat of cattle, pigs, sheep, horses and their processing
Variety meat is called animals parenchymic and other internal organs and some carcass parts,
got during technological process. Parenchyma is specific tissue of organ, which performs the main
function. Variety meat of cattle composes 24 %, of sheep 20 %, of pigs 17 % of total carcass
mass.
Comparing morphological structure of variety meat, we can notice, that ones much differ
from carcass, because they consist of parenchyma organs (liver, brain, lungs, kidneys, udder and
other). Connective tissue divides them into separate parts.
Other group consists of organs and tissues, which permanently move (function intensively) if
animal is alive. These are: heart, tongue, stomach, diaphragm. Besides connective tissue there is
muscular (striated and smooth muscles) tissue in them.
One more group consists of variety meat, which is close to carcass, but differ quantitative
ratio of different tissues (muscular, fatty and connective). These are heads, ears, lips, legs and tails.
According using, variety meat is classified into two groups: alimentary and technical. Variety
meat must correspond requirements of Standard LST 1245: 2008 Alimentary Variety meat of pigs,

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cattle, sheep and horses. The general requirements. There are two groups of alimentary variety
meat:
pigs tongues, livers, kidneys, heads, ears, heads meat, masseters, hearts; tails, legs, lungs,

stomachs, meat from oesophagi, larynx, tracheas, head and spinal brain, spleens, diaphragms, meat
cuts, skins and vessels;
cattle tongues, livers, kidneys, ears, heads meat, hearts; tails, legs, tarsi, lungs, pregasters,

meat from oesophagi, larynx, tracheas, head and spinal brain, spleens, diaphragms, meat cuts, skins
and vessels, udders, lips and testicles.
Variety meat is used directly or as components, processing high quality sausages, pastes, canned
meat, ready-to-cook products, delicacies and other.
Specific dangerous material of animal origin, described in Regulation (Regulation (EC) Nr.
999/2001) and assessing order prevention, control and elimination of contagious spongiphormic
encephalopaties are not assigned to variety meat.
Heads (after removing of brain, muscles and tongue), ears, larynx and udders are classified as
technical variety meat.
According morphological structure and way of technological processing, variety meat is
divided into soft, bony, mucous and hairy (bristly).
Soft: tongues, livers, brains, hearts, kidneys, udders, meat cuts, muscles from oesophagi,
lungs, spleens.
Bony: heads, tarsi, tails.
Mucous: rumens, omasi, abomasi, stomachs.
Hairy (bristly): heads, tails, ears, tarsi, lips, legs.
According animal type, there are cattle, pigs, sheep and poultry variety meat.
According freezing way, variety meat can be:

cooled (internal temperature is not higher than 3 C and not lower than -1 C);

freeze (internal temperature is not higher than -12 C and not lower than -18 C);

deeply freeze (internal temperature is -18 C or lower).

Variety meat cant be processed in any other way but freezing, they can be packed in
vacuum or modified gas atmosphere.
Quality and sensual characteristics of processed variety meat must correspond to those,
described in Table 22.

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Table 22. Characteristics of alimentary variety meat

Name
Characteristics
Tongues of pigs and Clean, of natural form, not chopped, without sublingual meat or with it, without
cattle
main limphonodes, sublingual bone, pharynx and larynx offal, without blood
and mucus.
Warrantable:
three not deep (to 1cm.) incisions;
tongues without epithelium.
Liver of pigs and cattle
Clean without surface vasa, bile ducts, main limphonodes and other tissue offal.
From light to dark brown with shades. Serosa of liver must be not broken.
Warrantable:
to leave conglutinated bile ducts offal: not longer than 5 cm for pigs liver and
not longer than 10 cm for cattle liver.
three not deep (to 1cm) incisions.
Kidneys of pigs and Of natural form, clean, without fat and fibrosa, without surface vasa, ureters and
cattle
limphonodes. From light to dark brown colour.
Warrantable:
only three not deep (to 1cm) incisions.
Head and spinal brain of Of natural form, clean, not broken capsule, without blood clots and bone chips.
pigs and cattle
From light to dark pink colour.
Ears of pigs and cattle
Cut at the base, clean, internal meatuses must be removed. Skin lacerations
must not exceed 15% of the surfice.
Warrantable: to realize only ear pinne.
Heads of pigs
Not cut in half or cut in half, without brain, without tonques and ears, clean,
without burns, bristles, blood clots, yellow brownish colour.
Warrantable: to realise with ears or without masseters.
Head meat of pigs and Of various mass and form, pieces of muscles with connective tissue, without
cattle
blood clots, the main limphonodes, salivary glands and other additives. Colour
of muscles is from light red to red with brown shade. Colour of fat is from white
to light yellow.
Masseters of pigs
With skin or without it, without bristle, haemorrhagies, the main limphonodes,
clean, not yellowed.
Hearts of pigs and cattle
Without pericardium and surfice vasa, with auricles and fat layer or without fat
layer at the heart surfice. Ventricles are cut along, without blood, clean.
Warrantable not longer than 1.5cm remainders of aorta, accreted to muscles.
Tails of pigs and cattle
Without hair or bristles, clean. Lacerations of pig tails must not exceed 15 % of
surfice. Cattle tails must be without skin remainders.
Udders of cows
Incised or cut inpieces, without skin and hair, blood remainders, limphonodes,
clean.
Legs of pigs and cattle
Without hair, bristles, hoofs, clean, without burnings, light yellow or brownish
colour. Lacerations of pig legs skin must not exceed 15 % of surface.
Tarsi of cattle
Without skin, clean, without burnings.
Lips of cattle
Of natural form, clean, without hair and skin remainders, grey, light yellow or
pink.
Rennets of cattle
Cut, clean, without mucous membrane, without fat, light yellow or grey colour,
without dark spots.
Lungs of pigs and cattle
Clean, without blood and mucous, without remainders of trachea and bronchi,
light or dark brownish colour with grey shade.
Stomachs of pigs and Cut, clean, without fat and dark spots, light pink, yellow or grey colour.
cattle
Esophagus meat of pigs Clean, without blood clots, pink or red. Internal skin can be left.
and cattle

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Name
Larynx of pigs and cattle
Tracheas of pigs and
cattle
Spleens of pigs and
cattle

Characteristics
Clean without blood clots, mucous, glands and remainders of limphonodes.
Clean, without mucous, blood clots, glands and remainders of limphonodes.

Of natural form, clean, without fat and blood clots, from pink to dark red colour
with light blue, grey or violet shade.
Warrantable: a narrow fat strip.
Vasa and tendons of pigs Clean. Warrantable remainders of meat and fat.
and cattle
Skins of pigs
Clean, without bristles and burnings.
Meat cuts of pigs and Clean, without the main limphonodes, remainders of hair and bristles, blood
cattle
clots.
Sublingual meat is ascribed too.
Diaphragms od pigs and Clean, without remainders of internal organs, blood clots, skin.
cattle
Cattle testicles
Of natural form, without incisions, serous skin is removed.

Microbial contamination of alimentary variety meat must not exceed described in HN 26 ones.
Contamination with chemical pollutants and pesticides must not exceed described in HN 54.
Drinking water, used for variety meat processing must correspond to requirements, described
in HN 24.

13.1. Chemical composition and nutritional value of variety meat

Chemical composition of variety meat depends on ratio of their different tissues, therefore
their nutritional value is not the same. Chemical composition of variety meat depends on animal
species, sex, age and condition.
Variety meat consists of:

proteins (9.7-25.2 %);

fat (2.3-39.4 %);

carbohydrates (0.5-5.8 %.);

inorganic material (0.4-1.5 %);

water (43.2- 82.7 %).

Besides, there are vitamins and ferments in variety meat.

According nutritional value, variety meat of I-st category is equal to meat and some of them
according amount of inorganic material and vitamins (liver, kidneys, brain) are valuable even more.
The variety meat is used both: directly or as components, processing high quality sausages, pastes,
canned meat, ready-to-cook products, delicacies and other.
Amount of proteins of mostly variety meat (liver, kidneys, tongue, heart, brain) correspond to
amount of proteins in meat.
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In most variety meat of II-nd category (mucous and hairy) dominates connective tissue, where
is not valuable protein, collagen. Although variety meat with much collagen, are of less nutritional
value than meat, they have positive characteristic too. When there are collagen breaking products in
food, they stimulate peristalsis of digestive tract and positively act on intestinal microflora.
Therefore, according present nutrition view, food material, which have connective tissue, are
assigned to necessary food components. So it is desirable, that in recipes of meat products there
would be processed variety meat, from II-nd category too. Besides, in this case variety meat is used
more rationally.
In some variety meat (tongue, brain) there are more fat than in meat. The mostly part of
variety meat fat composition consist of neutral fatty acids with high amount of lipids (phosphatides,
cholesterol, cerebrosides).

14. Canned goods

There are about 60-70 different canned goods products processed in Lithuania. They are
processed in three enterprises: the mostly part in Panevys (about 50 different kinds) and also in
Klaipda and Vilkavikis. The canned meat is appreciated because these reasons:
- it is stored easy, because are processed hermetically and thermally;
- everything is suited for eating, all not eaten and not suitable parts are removed;
- it has good flavour, because different spices and sometimes fatty additives are added;
- it is comfortable to use, because doesnt take long time for preparing,
- it makes meat products assortment more different.
There are some peculiarities in processing of canned meat, but the mane processing scheme is
similar for many canned products:
1. Preparing of stocks: it is done according technological requirements for canned meat
processing and consists of meat cutting, grinding and, if necessary, boiling, grilling or blanching.
This operation needs much work, hand work too.
2. Preparing of tare: washing, sterilizing etc.
3. Exhautiration of tare (air removing from tare): pouring of stock into tare and hermetical
closing. This operation is performed by equipment, which fill to 1-2 thousands of cans during 1
hour.
4. Thermal processing of canned meat: sterilization or pasteurization. It is made in special
autoclaves. The time depends on can mass and stock preparing method.

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5. Thermal storing: canned meat is stored at about 30oC temperature in thermostats for 1 to 3
days. During storing some microorganisms are destroyed and defects become clear.
6. Quality evaluation.
7. Packing into transportation (external) tare: wooden, polymeric, cartoon, metal boxes.
Depending on used stock and processing peculiarities, the canned goods are divided into such
groups:
1. Canned meat depending on the processing of the main stock meat is divided into such

groups:
a) Braise. Cut in pieces meat is put into cans and salt, spices and fats are added. Cans are
closed hermetically and sterilized for 1.5-2 hours.
b) Cooked meat. Cooked meat is cut in pieces, put into cans and concentrated bouillon, spices
and fats (if necessary) are added. Hermetical cans are sterilized for 0.5-1 hour.
c) Roasted meat. Cut in pieces meat is roasted in bones fats with onions. Roasted meat pieces
are put into cans and sterilized for 0.5-1 hour. Canned roasted meat can have another name, for ex.:
goulash, small meat pieces, meat balls, minced meat balls.
d) Salted-pressed meat. They can be like braise type. In Lithuania Tourist breakfast canned
meat is popular which is prepared from pork or beef. Salted-pressed canned meat has up to 3 % of
salt, they can be stored long and has strategic mean.
2. Canned variety meat. Depending on used stock and preparing method, they can be of such

kind:
a) Tongue in jelly. Salted, blanched, cut in pieces tongues of cattle, pigs and sheep are put into
cans and poured with thick sterilized bouillon.
b) Roasted liver. Beef liver, cut in pieces, roasted in pork fats with onions.
c) Roasted brain. Beef brain, roasted in pork fats.
d) Kidneys in tomato sauce. Different animals kidneys: cut, roasted in lard and poured with
tomato sauce.
3. Canned poultry. Depending on used stock and processing way, there are such canned

poultry:
a) Cooked poultry. Poultry in jelly, turkey in jelly, duck in jelly, poultry ragout in jelly.
b) Cooked poultry fillet. Poultry fillet. Anserine fillet. Bones are removed from poultry.
c) Cooked poultry fillet with grits. Rise with poultry are made the mostly.
4. Pastes. They are made from cuterated minced meat. Variety meat, meat, butter, egg yolk,

milk and spices are used. According composition, they are divided into 2 groups:
a) natural (liver paste with butter, meat paste),
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b) special (there are not only meat and variety meat in their composition, but also other
products).
5. Meat-vegetable preserves. The mostly of them are prepared like braise or cooked meat,

according technological process of these products. According composition of stocks, they are
divided into such kinds: meat with vegetables, meat with macaroni, meat with grits. Braise canned
meat with vegetables are sterilized for 2 hours. Cooked canned meat with vegetables is sterilized for
1 hour.

Picture 33. Analysis of canned meat

Picture 34. Analysis of canned meat content

Picture 35. Analysis of can

Picture 36. Labeling of canned meat

14.1 Evaluation of cans

Inspecting tin can various defects can be observed: absence of hermetic (not hermetic closing,
perforating, corrosion and other), bunching, folding and other defects of can corps, cover, bottom or
juncture (Pict. 35).
Changes of canned meat are influenced be changes of cans and content, sometimes both of
them.
Marbling is quite often changing of can internal wall. Spots, strips or rays from brown to

black colour are seen at internal can wall. Surface of painted places is smooth and shining. Marbling
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of white tin cans internal surface appears because of formation of zinc oxide, in both: covered with
glaze and not covered cans. Sterilizing albuminous products (meat, fish, peas) proteins break and
sulphur compounds form, which go through the glaze and reacting with tin zinc form zinc sulphide.
Marbling can be seen soon after processing of preserves. The degree of marbling depends on
organic compounds, having sulphur, found in content, on content pH, on peculiarities of tin surface
and on permeability of glaze, covering inside of can. Poliphosphates which are in meat products,
have especially great mean. Pork preserves are more susceptible to marble inside of can than beef
preserves. Marbling is also held by garlic, onions and mustard granules. Marbling doesnt influence
content and it is called as appearance defect.
Not adhered glaze. For covering of cans inside different glaze are used, which consist of

alcidic resin and poliamidfenolum resin. Despite glaze stability and resistance for sterilization, in
some circumstances glaze can become not adhered. At first blebs form, then glaze slivers. Passed
into the content, glaze can change flavour and reduce value of preserves.
Corrosion is called changes of cans, influenced of chemical and electrochemical reactions,

when tare surface is destroyed, forming oxides and hydroxides. It is impossible to cover tin with
glaze in such way, that no pores appeared at all. Covering metal surface with glaze, primary
(structures determined of microscopic amount) and secondary (mechanical nature) pores can form.
Hard-molecullar compounds, which enter composition of preserves (neutral fats, carbohydrates)
slowen corrosion effect, whereas oxygen, acids, free fatty acids stimulate corrosion. Corrosion is
also stimulated by pH and product electrical conduction.
Compound preserves of meat and sausages, which pH is near neutral, but have relative much
salt and sulphur, are average aggressive for corrosion. Partially influencing corrosion factors are:
high heating temperature, storing of preserves at high temperature and high humidity.
The most visible kind of corrosion is forming of rust. Even holes can form and cans become
not hermetic.
Because of corrosion of internal can side, sensory indexes of content change. Preserves get
sour, metal savour (especially because of iron and zinc compounds). If aluminium salts appear in
content alkaline or light metal savor can be felt. Melted iron salts determine blackening of surface.
Besides, iron ions catalyze spoiling processes.
Because of corrosion, content of preserves become polluted with iron, zinc, aluminium and
plumbum.
Kind and size of corrosion and content condition must be considered. When external corrosion
and only spotty damage is seen, preserves can be stored not longer than one month in dry place.

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When there is great, deep external corrosion, but not damaged hermetic, preserves are realized as
less value.
When there is internal corrosion without content changes, preserves must be realized as soon
as possible. When there are changes of content, they are evaluated and amount of metals in content
is detected. Depending on these indexes, preserves are used as chaffy or are not suited for human
consumption at all.
Bloating of cans (bombage). Not suitable choosing of stock and not enough accurate

preparing of it (processing, not keeping of sanitary rules, bad covering, not keeping heating mode)
can influence changes of preserves and increasing of internal cans pressure, there fore cans or their
covers bloat that means bombage occurs.
Bloating can be of bacterial, chemical or physical nature. Bacterial (biological, real) bloating
occurs, when, because of microorganisms action, gas (CO2, NH3, H2S, N2) start to concentrate in
can, therefore cans cover and bottom (or one of them) bloat and dont return to previous position
after pressing. Knocking the can, tympanic sound is heard, because there is gas inside. This bloating
often occurs when not enough heating is used or if secondary contamination occurs after
sterilization, when cans become not hermetic. In such way not only Clostridia (C. botulinum, C.
sporogenes, C. bifermentans, C. Perfringens) but also spores not forming bacteria (Proteus,
Escherichia, Pseudomonas, Streptococcus, Lactobacilus) and bacili (B. cerius, B. subtilis, B.
pamilus) can be found.
Content of such preserves is spoiled and contaminated with microorganisms. Especially
danger appears because of bloating, which occurs because of anaerobic bacteria Clostridium
botulinum, which produce toxin, influencing deadly human intoxication.
Physical (not real) bloating occur when can is filled too much or it is filled with very cold
product, also, because of freezing of can content. Bloating of frozen preserves disappear after
unfreezing their content.
Physical bloating can occur because of content swelling. It can be seen in all kind preserves of
sausages or in preserves, where are used rise, beans and other legumes, when tare is fulfilled.
Frozen preserves can bloat, especially if there are much fluid in their content. Because of
changed structure, the content becomes dry, straw, looking tasteless. After defrosting such
preserves, bloating disappears.
Preserves can bloat because of covering air too when much air having products are heated
(for ex., crushed raw meat sausage for roasting, sausage mince and other). Air expands and goes out
of the product. After cooling air doesnt come back.

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Therefore, cans bloat if preserves are stored in regions, where atmospheric pressure is lower
than in their processing place.
Cans bloat because of deformation too when one or both sides of fulfilled can are pressed.
Chemical bloating most often occurs when preserves (especially mixed) are stores for long
time, when hydrogen gass accumulate. They appear because of content acids interaction with cans
metals. Content of such preserves can look normal, but often gets metal savour.
If the real bloating was assessed, the preserves are utilized.
Rusted cans, after cleaning of rusts (if can is hermetic) must be realized as soon as possible.
Cans leakage test. One of the simplest cans leakage tests is their soaking into hot water.

Label is taken out of can and it surface is washed. Then can is soaked into hot water. Water
temperature must fall not lower than 85oC. Water layer above the preserves must be not thinner than
25-30 mm. Cans are left in hot water for 5-7 minutes. Cans are put in one layer: at first on the
bottom, later they are turned on the cover. If can is not hermetic, gas bubbles go out from the
perforation. Not hermetic cans are utilized.

14.2. Evaluation of canned meat quality

Quality of canned meat depends on meat quality: meat, which is used for canning cant have
external flavour and odour, it must be gradually salted and easy for cutting. The concept Meat
doesnt mean variety meat, fat (including meat fat) and bones.

14.2.1 Sensual analysis of can content

After opening the can, content odour and appearance are evaluated at first (Pict. 33). Then the
content is put into the can and appearance, colour, flavour (if there are no signs of Cl. botulinum or
spoiling), odour, consistence, quality of bouillon and meat, number of pieces and features are
evaluated (pict. 34).
Canned meat is evaluated being warm or heated, depending on their consuming peculiarities.
If testing cold canned meat, it is of suspicious availability, it is heated and analyzed once more. If
different kind canned meat is analyzed, at firs are tested of milder flavour and later tang (more
salted, soured) canned meat.
Sensual characteristics of different canned meat are specific. It must be described in Standards
of each sort. At first, mass of can, it content and separate content parts are evaluated. They must
correspond to the requirements, described in Standard. Then internal can surface is observed. Inside
of good canned meat can is bright and clean. It is analyzed if there are no black spots, which could
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appear after melting of tin, which covers inside of the can, if there are no rusts or other changes.
Sometimes at internal surface of can dark bluish brown spots or strips can be seen. When
albuminous products (meat, fish, peas) are sterilized, sulphur compounds form, with react with tin
and cover inside of the can. Such skin is not noxious, because it doesnt melt in water and is hardly
sticked to the can.

14.2.2 Chemical analysis of canned meat quality

Fore each kind of canned meat this ratio is rationed, i. e.: amount of meat and fats must be not
less than 72 % in cooked canned meat, fats not more than 10 %, concentrated bouillon not more
than 27 %, salt not more than 1.4%; another example in canned poultry bones must compose not
more than 14 %.

14.2.2.1 Evaluation of canned meat general acidity

Acidity is evaluated of such preserves, filler of which is sour, for ex., meat-vegetable
preserves and other. Too high content acidity increases corrosion of cans.
Procedure. 20 g of product are weighed into porcelain plate and without loss, washing with

distilled water, are put into 250 ml measuring flask. After filling the flask to of it content with
80oC temperature distilled water, it is shacked well and left for 30 min. sometimes shaking. After
that, the flask is cooled with water flow to room temperature, adding distilled water to the mark, it
is shacked again and the content is filtrated through the paper filter.
50 ml of filtrate are measured with pipette, poured into the flask, dropped 3-5 drops of 1 %
phenolphtalein solution and titrated with 0.1 N alkali solution till the pink colour. The general
acidity, expressed in % of lactic acid, is calculated using the formula:

X=

0.009 n 250 100 k g

50 a

N amount of 0.1 N alkali solution, used for titration (ml)

a mass of preserves (g)
k correction of 0.1 N alkali solution
0.009 amount of lactic acid (g), equivalent to 1 ml of 0.1 N alkali
Difference between two parallel analysis must be not higher than 0.02 %. Acidity of
preserves, calculating for lactic acid must be not higher than 0.4 %.

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14.2.2.2 Evaluation of salt in canned meat

Analysis is similar like investigating sausages and salted meat products. Extract is prepared
analogous as in analysis of the general preserves acidity. The same extract, left after acidity
evaluation, can be used.
Process analysis. 25-50 ml of filtrated extract are measured with pipette and after pouring of

phenolphtalein, are neutralized with potassium hydroxide. Later 1 ml of 10 % potassium chromate

solution are poured and is titrated with 0.05 N argent nitrate solution. The further analysis is the
same as analyzing salted meat products. In different canned meat there are 1.2-2.2 % of salt.

14.2.3 Evaluation of meat amount in canned beef

Canned salted beef is packed into hermetic tare and thermal processed after packing meat from
different cattle kinds and also cattle hearts. Hermetic tare is such tare, which is absolutely closed and
not permeable, made from material, which fits for storing canned meat in it. The tare must
correspond the requirements of Lithuanian hygiene norm HN16 Materials and products, contacting
with food products. The general requirements.
The main indexes of composition and control

The product is prepared using large cut and heat-treated beef or mixture of heat-treated beef
and no more than 5 % of raw beef. Tare is closed hermetically and sterilized in such temperature and
for such time, that product wouldnt spoil, it composition wouldnt change during storage and there
wouldnt be any danger for health of consumers.
Composition of the product: not salted beef, alimentary salt and sodium or potassium nitrite,
spices, other components, which can be coordinated with correct institutions of Ministry of Health
of Republic of Lithuania. Used additives are: saccharose, inverted sugar, glucose, lactose, maltose,
glucose syrup, maize syrup.
Chemical composition: general amount of proteins in canned beef must be not less than 21 %
(mass %). Meat amount in salted canned beef can be from 80 to 90 % (mass %) and more.
Amount of meat in canned beef is calculated using formula:
(Nt Nx) 100
M=

3.55

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Nt general nitrogen amount (%), estimated during analysis;

Nx nitrogen of not meat origin (%);
General nitrogen amount is calculated according methodology, described in chapter 10.1.
Nitrogen of not meat origin Nx is calculated when each product component, in composition of
which there is nitrogen and amount of nitrogen in each of them is known. In table 23 average
amount of not meat origin nitrogen in components, used for canned beef processing is shown.
Table 23. Amount of nitrogen in components, used for canned beef processing
Components
Pulled bread
Casein
Sodium caseinate
Soya proteins isolate
Textured soya proteins
Soya flour
Monosodium glutamate
Cattle kidneys
Cattle tongues

Amount of nitrogen, %
2.0
15.8
14.8
14.5
8.0
8.0
8.3
2.7
3.0

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References
1. Council Regulation (EB) Nr. 2075/2005, laying down specific rules on official controls for
Trichinella in meat.
2. Council Regulation (EB) Nr. 700/2007 on the marketing of the meat of bovine animals aged 12
months or less.
3. Feiner G., Quality Ingredients, Australia. Meat products handbook: Practical science and technology.
September 2006. 672 p.
4. Gracey J. F., Collins D. S., Huey R. J. Meat hygiene. 10th edition. Elsevier Health Sciences. 1999. P.
21 758.
5. Herenda D. C., Don Franco A. Poultry diseases and meat hygiene a color atlas. Wiley-Blackwell.
1996. 337 p.
6. Lerche M., Rievel H., Goerttler V. Lehrbuch der Tierarztlichen Lebensmitteluberwachung. Jena,
1957. 1078 p.
7. Liutkeviius A., Speiien V., Mieelien A., Alenikien G., Zaborskien G. prastomis slygomis
uaugint msini, pienini bei ekologikai uaugint pienini galvij msos kokyb. ISSN 13920227. MAISTO CHEMIJA IR TECHNOLOGIJA. 2009. T. 43. Nr. 2. P. 47-56.
8. LST 1245:2008. Valgomieji kiauli, galvij, avi ir arkli subproduktai. Bendrieji reikalavimai.
9. LST 1919. Msos gaminiai.
10. LST ISO 1442. Drgms kiekio nustatymas.
11. LST ISO 1443. Bendrojo riebal kiekio nustatymas.
12. LST ISO 2917. Msa ir msos produktai. pH nustatymas. Pamatinis metodas.
13. LST ISO 2918. Nitrit kiekio nustatymas.
14. LST ISO 3091. Nitrat kiekio nustatymas.
15. LST ISO 6490-1. Kalcio kiekio nustatymas (titrimetrijos metodas).
16. LST ISO 936. Bendrojo pelen kiekio nustatymas.
17. LST ISO 937. Azoto kiekio nustatymas.
18. Mead G. C. Poultry meat processing and quality Cambridge: Woodhead Pub. Boca Raton: CRC
Press, cop. 2004. Series: Woodhead publishing in food science and technology.
19. Rules of labeling and price ordering of goods, sold in Republic of Lithuania.
20. Rules of meat and other animal origin products processing and providing to market places.
21. Sasnauskait L. Grdtos varks gamybos technologijos tobulinimas. Magistrantros studij
baigiamasis darbas. Akademija, 2010. 61 p.
22. imkeviien Z., Kaemkaityt D. Msos ir jos produkt kokybs bei technologijos laboratoriniai
darbai. 81 p.
23. Vaitkus J. Veterinarins sanitarins ekspertizs darbai I dalis. Vilnius. 1989. 84 p.

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