STUDIES ON IMPROVING SALTED-FERMENTED FISH

PRODUCTS IN SUDAN

By
Ghada Ahmed El Haj

Supervised By: Professor Dr. Yousif Babiker Abu Gideiri

Dr. Mohamed EL Tahir Ali

B. Sc. (Fisheries Science) 1997

University of Juba
M.Sc. (Aquaculture) 2000
University of Putra Malaysia

A Thesis Submitted for the Fulfillment of Requirements for the

Degree of Doctor of Philosophy
Department of Zoology

Faculty of Science
University of Khartoum
May, 2006
 DEDICATION

To my parents

For their true love, constant trust and principles that guided my life

To my husband

For his devotion, understanding and support during all difficulties

To my children

For making everything worthwhile

And

To many researchers whose works have not given due recognition for the

many hours spent in the laboratory and fields to provide humanity with

solutions to better life.

 ACKNOWLEDGEMENTS

Words are not enough to express my sincere appreciation for the

people who were involved in my training towards obtaining this degree. I
am very thankful to my supervisor, Professor Dr. Yousif Babiker Abu
Gideiri for his guidance, objective discussions, criticisms, encouragement
and support throughout the course of the study. I am also indebted to my
co-supervisor Dr. Mohamed EL Tahir Ali for his constructive
suggestions, and invaluable advice.
Thanks are extended to Dr. El Zubeir Darag, Head, Department of
Microbiology, and Dr. Ali Shimat, Head, Department of Biochemistry,
and Toxin, Veterinary Research Center, and their technical staff for
providing some of their facilities for the microbiology and biochemistry
studies.
Furthermore, I would like to acknowledge the staff of the Fisheries
Research Center, Khartoum, and Mr. Ahmed Abed Rahim, Faculty of
Veterinary Science, University of Khartoum, for their valuable practical
assistance.
Finally a special word of thanks is due to my friends, Zo Alfagar,
Sara and Wesal for their assistance, companionship, support and concern.
 TABLES OF CONTENTS

Page
DEDECATION.. ii
ACKNOLWLEDGMENTS.. iii
TABLES OF CONTENTS iv
LIST OF TABLES. vi
LIST OF FIGURS. viii
ABSTRACT ix
ABSTRACT IN (ARABIC) xi

CHAPTERS

1 INTRODUCTION. 1

11 LITERATURE REVEW 4
2. 1 Spoilage of fish.. 4
2. 2 Preservation of Fish.. 6
2. 3 The Application of Technology. 8
2. 4 Salting. 8
2. 4. 1 The Salting Process 10
2. 4. 2 Salting Methods. 10
2. 4. 3 Salt Quality. 12
2. 5 Fermentation. 14
2. 6 Nutrition and Safety Considerations.. 16
2. 7 Drying.. 17
2. 8 Smoking 21
2. 9 Colour 23
2. 10 Oxidative Rancidity... 24
2. 11 Salted Fermented Fish and Fish Type 26
2. 12 Microbiology of Fermented Fish Products. 27
2. 13 Synthesis of Findings of Information Related 29
to The Subject Matter of This Thesis

III MATERIALS AND METHODS 39

3. 1 Sample Collection. 39
3. 2 Treatment.. 31
 3. 3 Chemical Analysis. 40
3. 3. 1 Preparation of the Sample.. 40
3. 3. 2 Moisture analysis. 40
3. 3. 3 Protein Content. 41
3. 3. 4 Crude Fat 42
3. 3. 5 Crude Fiber 44
3. 3. 6 Ash Content.. 45
3. 3. 7 pH measurment. 46
3. 5 Determinations of Minerals. 46
3. 5. 1 Determinations of Phosphorus 46
3. 5. 2 Determinations of Iron 47
3. 5. 3 Copper Extraction 48
3. 5. 4 Determinations of Calcium. 48
3. 5. 5 Determinations of Sodium and Potassium 50
3. 6 Colour Measurement.. 50
3. 7 Oxidative Rancidity. 51
3. 8 Organoleptic Analysis. 52
3. 9 Microbial Groups Studies 52
3. 9. 1 Total Viable Bacterial Count 52
3. 9. 2 Isolation and Identification of the Colonies
Culture. 53
3. 10 Statistical Analysis 53

IV RESULTS. 54
4. 1 Biochemical composition. 54
4. 2 General magnitude of change between fresh
and treated samples.. 55
4.3 Effect of the quality of the treated
Samples 64
4. 4 Total viable bacterial counts.. 57

V DISCUSSION. 78

VI CONCLUSIONS.. 80

VII REFERANCES 83

VIII APPENDICES 98
 LIST OF TABLES

TABLE Page
1 Chemical composition of fresh samples during all season of 56
experiments..

2 The effect of Salt concentration and time on C.P, C.F, E.E, Ash, 57
D.M and Moist. of Kawara & Debs during summer

3 The effect of Salt concentration and time on P, Ca, Fe, Na, 58

K, and Cu. of Kawara & Deds during summer....

4 The effect of Salt concentration and time in C.P, C.F, E.E,

Ash, D.M and Moist. Of Kawara & Debs SP. during
autumn..
59

5 The effect of Salt concentration and time on P, Ca,

Fe, Na, K, and Cu. of Kawara & Debs during
autumn 60

6 The effect of Salt concentration and time on C.P,

C.F, E.E, Ash, D.M and Moist. of Kawara & Debs
during winter.. 61

7 The effect of Salt concentration and time on P, Ca,

Fe, Na, K, and Cu. of Kawara & Debs during
Winter. 62

8 The magnitude of change between fresh and

treated materials during summer.. 63

9 Summery of levels of chemical parameters

obtained at different salt concentrations for the
two species in the different seasons (tables 2-
7)... 70

10 Total Viable Bacteria count of fresh samples during

all season of experiments... 73
11 Bacteria Identification of fresh samples during
different seasons. 74

12 Bacteria Identification of treated samples in summer

season. 75

13 Bacteria Identification of treated samples in autumn

season. 76

14 Bacteria Identification of treated samples in winter

season. 77
 LIST OF FIGURES

FIGURE Page
1 Crude protein of Kawara and Debs during summer
season 66

2 Crude protein of Kawara and Debs during autumn

season 66

3 Crude protein of Kawara and Debs during winter

season 67

4 Oxidative Rancidity of Kawara during seasons... 67

5 Oxidative Rancidity of Debs during seasons 68

6 Yellowness of Kawara during seasons. 68

7 Yellowness of Debs during seasons. 69

 ABSTRACT

In attempts targeting preservation of fish and their products for

consumption at subsequent times and places far away from the source,

this programme was arranged for a number of preservation methods to

address the issues. Those methods have duly included drying, salting,

smoking, fermentationetc. to give a good coverage for various types of

fish. It arises from the above, that there is a need to evaluate traditional

and improved methods of preservation to reach an acceptable product for

the consumer.

For the fulfillment of the above, this programme was set to address the

quality of fish from chemical, physical and biological viewpoint. Special

emphasis was attached to salting and fermentation and their impact.

However, the result of this study indicated that fresh samples of two

species of fish Kawara (Alestes sp.) and Debs (Labeo sp.) during three

seasons are comparable in both content and values. These values of

change in varying degrees when subjected to different salt concentrations

during short and prolonged time. General magnitude change between

fresh and treated samples was obtained for both sp. Significant variations

for all parameters of two species during the first twelve days of

treatments were detected, It has to be emphasized that the sixth months

marks the onset of further deterioration.



.... .

.
 CHAPTER I

INTRODUCTION

Fish is one of the main sources for the provision of animal protein

for a growing demand in a world of ever-growing population and

increasing consumption. In this respect, the Sudan is no exception and the

various aquatic resources (marine, freshwater, brackish, groundwater and

others) are tapped in order to fulfill the needs in this direction. To this

effect, various methods of fishing are encountered and appropriate gears

are employed to match with the requirement of the environs. However,

the ultimate goal being the maximization of the products and their

utilization for the best and economic satisfaction within the framework of

a well-defined exploitation strategy.

The current Sudanese fisheries practices are focused on the

exploitation of the fishes of the inland waters exemplified by the River

Nile and its tributaries, the Red Sea coast, the Aquaculture activities and

seasonal water spots. But it has to be realized that contribution in the final

product varies from one sector to the other. Equally, handling of the fish

and subsequent treatments vary as well. The in-common weakness seems

to be attached to the wastes during sorting out of catch, transportation and

marketing. The latter results in losses in the product and equal economic

losses should be taken into considerration. For this reason, there is the

need for paying attention to the magnitude of spoilage that occurs and

required safeguards in order to cut it to the minimum within the post-

harvest phase.

It is unfortunate that the prevalent traditional preservation methods

employed throughout the country are defective and need efforts

pertaining to their improvement and development. In this connection,

reference is made to drying, salting, fermentation, smoking, chilling and

freezing, which are practised. However, this research program is designed

to concentrate on salting and fermentation of fish and attempts to carry

out exhaustive investigation on two species of fish: Kawara sp. (Alestes

sp.) and Debs sp. (Lebeo sp.), leading hopefully to achieve a promoted

status to be placed at the disposal of practitioners who enter competition

on quality.

It is anticipated that the fulfillment of this goal requires a certain

procedure that includes:

Determination of physical, biochemical composition and nutritive

values of the fresh fish in order to adopt as control for subsequent

treatments.
 Determination of the terminology and specific products, which

will be incorporated e.g. Fasseikh, Terkeen, and Muloha etc.

Full investigation of salt quality, methods of application, the

impact and concentration on flesh.

Follow up of the changes and the correlation with other factors,

which will end-up with the acceptable formulae.

The adoption of the standards and specifications, which are

approved locally, regionally or internationally, that render the products

the quality for marketing.

In the light of the above, the research program will be designed to

lead to the final result on those two species and register them in order to

reflect the improvement on the traditional methods used now.

The specific objectives comprise:

1- The determination of the magnitude of losses in quality resulting

from spoilage.

2- The targeting of the progressive promotion of the traditional

treatment in the direction of adoption of scientific approaches.

3- The adoption of standards and specifications currently applied

domestically, regionally, and internationally, wherever appropriate.

4- The determination of the onset of spoilage time.

 5- Advice on a recommended range of salt concentration which gives

the best preservation.

CHAPTER II

LITERATURE REVIEW

This chapter is handled through a number of subheadings as given in the

following:

2.1 Spoilage of fish

Freshly caught fish have a shining, iridescent surface with bright,

characteristic colouration and markings. The surface is covered with a
thin, whitish but transparent, smooth, homogeneous layer of mucus or
slime. The eyes are clear, bright and full, with a prominent jet-black pupil
and transparent cornea, the gill is bright pinkish red, the skin and flesh are
firm, moist, and elastic. Hess (1950) stated that most fish flesh, however,
is considered more perishable than meat because of more rapid autolysis
by the fish enzymes and because of less acid reaction of fish flesh that
favors microbial growth. Fish, in general, usually spoil more rapidly than
other muscle foods, particularly when mishandled, and such spoilage is
primarily bacterial in nature.

A delicate fresh fish odor is typical although sometimes it is

described as a feed odour, once associated with the feed of the fish. In the
marketplace fresh fish odours are masked by the bacterial growth on
decomposing refuse and slime. Although the fish begins to deteriorate as
soon it is removed from water, the spoiled condition develops gradually
(John, 1994).
 Fish becomes spoiled within 12 hours at tropical regions when a
complicated series of chemical and bacterial changes triggered by high
temperature, take place within the fish. (ILO, 1989).

Rigor mortis is especially important in the preservation of fish, for

it retards post mortem autolysis and bacterial decomposition. Therefore,
any procedure that lengthens rigor mortis lengthens keeping time. It is
longer if the fish have had less muscular activity before death and have
not been handled roughly and bruised during catching and later
processing, and is longer in some kinds of fish than in others (Akamatsu,
1959). Reducing the holding temperature will lengthen the period. Hess
(1950) reported that the bacterial spoilage of fish does not begin until
after rigor mortis, then juices are released from the flesh of fibers.

Fresh whole fish are decomposed by several types of enzymatic

and microbial activities. While the fish is still in a state of rigor, autolytic
enzymes begin breaking down nitrogenous compounds in areas adjacent
to the visceral cavity. Meanwhile, bacterial decomposition of the slime
layer, the gills, and the intestinal tract get underway and, after rigor is
complete, proceeds rapidly (Evelyn and McDermott, 1961). In the stages
of spoilage, the highly unsaturated fat which is especially abundant in
fatty fish becomes oxidatively rancid.

Spoilage depends on the temperature and water activity of the fish

and the presence or absence of the various spoilage microorganisms
(FAO, 1981). The rate of spoilage is related directly to time and
temperature. During postmortem autolytic and microbial activity,
numerous chemical changes occur in fish flesh, resulting in liberation of
end products of digestion or of metabolism. A number of these end
products have been suggested as indicators of spoilage.
 Spoilage of fish may take place before, during or after processing
(ILO, 1989). There are many ways to minimize the spoilage occuring
during these times such as:

1- Improvement of landing facilities and distribution: these will

reduce the period of time that may elapse before the fish can be
processed.

2- Maintaining the fish at low temperature will minimize the spoilage

when fish is kept in cool condition immediately after catching until
processing begins.

In many areas far from towns especially in tropical area ice may not
be available in sufficient quantities, in order to keep fish at low
temperature, fish must be kept in the shade, placing damp sacking over
the fish to reduce the evaporates and/or mixing the fish with wet grass or
water weeds (FAO, 1992), so that the fish should be kept continuously
wet.

3- Maintaining a hygienic environment. This is achieved by washing

the fish with clean water,keeping it cool and avoiding breaking the
skin by bad handling.

4- Keep all tools, fish boxes, boat holds, cutting tables clean by
washing with clean water.

5- Working area should be cleaned regularly

6- Use salt during drying to avoid spoilage.

7- After processing appropriate packaging must be used and cool

storage area to avoid dust, insectsetc
2.2 Preservation of Fish
Artisanal fish processing remains the predominant and most
important method of fish preservation in Africa. The principal methods
are salting, fermentation, drying and smoking. These processes may
either be used alone or combined in order to achieve the desired product.
Preservation by salting, smoking, drying is called curing of fish. This
term is defined as fish preserved without the need for refrigeration or
freezing, but excluding sterilized products in air-tight containers. Drying,
smoking, salting and combination of these treatments are the basic means
whereby such product is prepared. Cured fish are consumed mainly in
tropical countries (FAO, 1981). In India, for example, only about 5
percent of marine fish landings are frozen but 21 percent are preserved by
traditional curing (Chakraborty, 1978).
Fish is one of the most perishable foods and its preservation is
usually accomplished by combination of different techniques.
Contamination with spoilage microorganisms is almost unavoidable
because fish is a very good culture media. The primary purpose of food
preservation methods is the creation of conditions unfavorable to the
growth or survival of spoilage organisms (Gracey, 1986).
Frazier (1958) mentioned that the principle of food preservation
was the prevention of microbial decomposition, self decomposition and
prevention of damage due to insects. The author stated that the chief
methods of food preservation are keeping out microorganisms. These
methods include the anaerobic conditions as in sealed or evacuated
container using high temperature, low temperature or drying which
include the drying up of water by solutes or hydrophilic colloids. Other
methods include chemical preservation, irradiation, and mechanical
destruction of microorganisms as by high pressure or using combination
of two or more of the preservation methods.
Gracey (1986) stated that the principles of all food preserving
methods are drying, curing, cold or heat application and chemicals. While
Clucas and Ward (1996) reported that the pereservation methods include,
salting, drying, chilling, smoking and feezing. Post-harvest losses of such
fish catch on processing include material, as well as value and nutritional
losses. Preservative methods must be applied even on the fishing boat.
Evisceration should be done promptly to stop active digestive enzymes in
the gut. Advantages gained by gutting may be offset by a possible delay
in rapid cooling of the fish (Akamatsu, 1959). The need for efficient
processing of landed fishes for maximum yields with best quality, should
be emphasized (Ali et al., 1996). The processing of food has its objective
provision of means for widespread distribution from point of harvest of
products that are safe, palatable, and nutrition.
2.3 The Application of Technology
Salting and drying fish in Sudan still follows traditional methods,
and remains basically an art or craft. The fisherman becomes a food craft-
man and by his experience can make a good product when his basic
materials are normal. But any abnormality in materials or conditions
usually leaves him helpless, because he has no knowledge of the
fundamentals, which might help him to overcome his difficulties. The
evolution of food technology will provide the essential link between
fundamental knowledge and empirical practice. One of the criteria that
distinguish the technologist from the craftsman is that the latter is able to
predict the effect of his scientific knowledge, whereas the craftsman can
only do this by trial and error.
Losses in production and quality can also be a result of social and
economic factors (James and Krone, 1977). It is futile to introduce an
improved technique if it cannot be incorporated into the existing lifestyle.
One of the most difficult problems in the application of technology to
rural areas is to bring the scientist and the craftsman together. Both have
different background and different type of education. The scientist is
interested in scientific problems and the main interest of the craftsman is
pride in his craft. A better understanding must therefore be reached
between socio-economic factors and deeply entrenched conservation.
Work in small-scale (processing of fish) sectors technology should
have as its main objective: the development and use of simple, cheap
equipment for the processing of food products, to obtain salted-dried fish,
salted-fermentative, or any other food commodities on a small scale.
2.4 Salting
An alternative to lowering the water activity of fish flesh by merely
extracting the water, as in simple dehydration, is to increase the
concentration of solutes in the flesh. Common salt, is more effective than
other safe, common and cheap food solutes (Reid, 1973).
Salt is the most important of the curing ingredients. It makes up the
bulk of the curing mixture because it is not only a good preservative but it
provides the most desirable flavour and its diffusion in fish meat by the
process of osmosis (Rosenthal, 1999). Where the objective of dehydration
is to remove water from the depth of the flesh quickly enough to reduce
water activity below the minimum for microbial growth, before
significant spoilage takes place. The objective of salting is to ensure that
the salt penetration is rapid enough to similarly lower the water activity in
the deepest parts of the flesh (Doe et al., 1983). The idea of all traditional
processing methods, is reducing the water activity, which is a measure of
the amount of water in the foodstuff available to support the growth of
microorganisms, until spoilage processes are prevented or at least greatly
slowed.
Common salt (sodium chloride) has been used as a preservative for
a long time. It acts as a bacteriostatic and a bacteriocidal agent when
present in sufficient concentration (Beatty and Fougere, 1957). This
property of salt has been frequently used in food processing and is the
basis for the preservation of salted-dried or salted-fermented fish. Food
products containing the required amount of salt will keep at room
temperature for longer time than those without. Salt may be added to
safety of product. Pearson and Gillett, (1996) reported that the curing and
meat processing can be manipulated by using salts and other additives,
the curing salts presented also to improve texture and palatability. The
salt concentrations differ according to species of fish. Saisithi (1987)
stated that salt concentration differs according to different types and
batches of fermentation. It is therefore of interest to identify the optimal
salt concentration, which does not inhibit the growth of the fermenting
microorganisms and in addition contributes positively to the flavour and
texture of the product.
2.4.1 The Salting Process
According to the fish composition and size, salting may be dry,
where the fish are stacked in salt and the brine formed is allowed to run
away, or wet, where they are immersed in a strong brine, or pickle. The
most commonly employed method is a hybrid of the dry and wet
methods, the fish are placed in dry salt and eventually become immersed
in the liquid pickle formed by solution of the salt in the liquid extracted
from the fish. This is sometimes called blood pickle (Doe et al., 1983).
Size of fish determines whether fish are salted whole and
uneviscerated, eviscerated and split open, or in smaller pieces ranging
from fillets to mince. The barrier presented by fish skins to salt
penetration mean that only small specimens, can be salted whole without
gutting. Larger fish salted in this manner would deteriorate the center
before salt could exert its effects there. Sodium chloride diffuses through
the fish flesh by a dialysis mechanism and water will diffuse to the
outside due to the osmotic pressure between the brine and fish muscle
solution.
2.4.2 Salting Methods
 Four types of salting methods (kench curing, pickling, brining and
gaspe curing) will be considered:

1. Solid or kench salting: in this method dry salt is rubbed all over the
fish to leave the flesh fairly dry. Fish are split, opened out flat and
placed in layers interspersed with layers of salt, and the liquor
which exudates is allowed to drain away, along with the shelf-life
of the dry product is produced. This method is used for non-fatty
fish.

2. Pickling, refers to fish that are treated with salt brine (Josephson et
al., 1986), is similar to kench salting but here the brine form is not
allowed to drain off. Salt is spread over the first layer of fish laid
then put the second layer of fish and then another layer of salt. The
fish must be kept below the surface of the brine; this is done by a
good covering of container used. Pickle curing is recommended in
preference to kench salting as it produces a more even salt
penetration and provides a better protection of the fish against
insects and animals since they are covered with brine. Doe et al.,
(1983) reported that the immersion in concentrated brines for long
periods is generally used for longer-term preservation, mainly of
fatty fish. The restriction of oxygen access by immersion retards
rancidity reactions, although some rancidity is desirable in the
development of characteristic flavour.

3. Brine salting: this is a uniform and controllable salting technique.

In this method prepared concentrated salt solution and fish are
soaked into it. This method is preferred when a fixed concentration
of salt in the final product is needed. Brine salting method is
commonly used in developing countries when a smoked product is
to be made and the salt concentration required in the final product
 must be lower than 3% (ILO, 1986). Also in this method the
ultimate concentration of salt in the fish is required to be sufficient
only for flavouring purposes: is affected by other techniques such
as smoking, where fish are treated for several minutes only in less
than saturated brine.

4. Gaspe curing: Doe et al., (1977) stated that if, instead of allowing
the exudates liquor to run away, the dry salting is carried out in
tubs, where the brine is formed. Weights are used to keep the fish
immersed for 2-3 days, after which they are taken out and dried in
the sun.

The use of salt in fish processing may either be applied by dry salting
(kenching) or wet salting. In dry salting the granular salt is applied
directly to the fish either in the gills, on the surface or, in the case of split
fish, in the belly. The exudate from the fish may be allowed to drain off
or be retained in the latter case; the fish becomes immersed in the exudate
and this is often referred to as pickling. In wet salting, the fish immersed
in brine for up to two days or dipped for few hours. It was observed that
some processors who cure fish with brine sometimes reuse the salt
solution a number of times. This may be a potential source of bacterial
contamination to fresh fish.

Common salt (sodium chloride) has three major effects on fish and
meat products. It enhances taste, solubilizes protein to create desired
texture and controls microbial growth to enhance shelf life and inhibit
pathogens (Terrel, 1983). Rock and Ebertz (1924) stated that the
preserving effect of sodium chloride implies more than its dehydration
action on the meat.

Salt is one of the most important food additives in food

preservation, as its concentration determines what types of
microorganism if any can grow by dehydrating or by lowering the
amount of water available for growth. Salting requires a little equipment,
but the method used is important.

2.4.3 Salt Quality

There are four types of salt: solar, brine-evaporated, rock and

manufactured salt, may be used in the processing of fish:

Solar salts are prepared by the evaporation of sea or salt-lake water

by sun and wind. Lagoons constructed on the shoreline are flooded,
sealed-off and allowed to evaporate to dryness. The salt obtained is
very impure due to the presence of salts other than sodium
chloride, and when it is dug out, may be further contaminated by
sand from the bed of the lagoon.

Brine-evaporated salts are prepared by the application of heat to

evaporate strong brines pumped from deep mines. The purity of
such salts depends on the nature of the underground deposit but
they are less likely to be contaminated with sand than solar salts.

Rock salt is mined from underground deposits of varying purity

from 80 to 90% sodium chloride.

Purified manufactured salt may contain 99.9% sodium chloride and

be derived from any one of the three types of salt mentioned above,
which may contain up to one fifth of their weight of impurities.

The salt used in the salting and fermenting method of fish in the
Sudan is solar salt (FAO, 1989). It is obtained after the evaporation of sea
water. The main constituent of rock salt is sodium chloride, although it
may also contain certain amount of calcium, magnesium and potassium
salt, carbonates, sulfates, bromides, iodides and even traces of heavy
metals and oxides may also be found.

According to Beatty and Fougere (1957) and Sadjeli et al, (1974)

good salt should contain very little or only a few of these constituents in
particular, the compounds of magnesium, iron, copper and heavy metals.
A high concentration of gypsum in the salt may cause a rough whitish
crust to be deposited on the surface of the fish. Iron and copper or traces
of metal are particularly bad as they stained the fish a dirty brown or
yellow color. A high level of impurities will also reduce the rate of
penetration of sodium chloride and impart a better taste and tough texture
to the fish. For these reasons good quality salt should be used. Low-grade
salt is the cause of the inferior quality fish produced in many areas and
also the quantity of salt used affect the quality of the fish.

Salt causes the proteins in fish muscle to swell. A high

concentration of salt lead the protein become denatured (Hamm, 1994).
This leads to a product, which is tough and dry. Too much salt will also
mask the taste of the fish and the flesh take on an unpleasant white
colour. The occurrence of impurities interferes with salt penetration,
acceleration of fat oxidation and consequently lead to low quality product
(FAO, 1981).

El-tom (1989) stated that solar salt is used on large scale for
production of salting fermenting fish in Sudan. According to Beatty and
Fougere (1957), bacteria which contribute to spoilage in fresh fish cannot
survive at salt concentration above 12% (w/w). Abu Gideiri (2001) found
that the number of microorganisms increased rapidly during the first
fermentation days and then began to decrease.

Losses of salted fish due to insect damage may be appreciably

lower than those of unsalted fish (FAO, 1984). Cole (1963) found that
Dermestes sp. Larvae were unable to develop in salted fish while freely
developing in unsalted fish. Poulter (1980) confirmed that unsalted
smoked fish are highly susceptible to infestation by Dermestes maculatus
and Dermestes frischii.

The quality and quantity of salt used are important factors that
affect the organoleptic qualities of salted fish. The present random use of
salt should be checked when a satisfactory ratio of salt to fish has been
determined. Also the size and shape of the fish should be taken into
account.

A good fishery salt contains 98% of common salt known

chemically as sodium chloride. Sometimes salt contains moulds and
bacteria (ILO, 1982). These bacteria cause the pink colour seen in some
salted fish and make the fish slimy and produce an unpleasant odour.

2.5 Fermentation

Fermentation is derived from the Latin verb fervere to boil (Peter

and Allan, 1984). The boiling appearance is due to the production of
carbon dioxide bubbles caused by the anaerobic catabolism of the sugars
present in the extract. Fermentation has different meanings to biochemists
and to industrial microbiologists. Its biochemical meaning relates to the
generation of energy by the catabolism of organic compounds whereas it
means for industrial microbiologists any process for the production of
product by the mass culture of microorganisms.

Fermentation is considered an easy and low-energy preservation

method for meat, that results in distinctive products that have an
important part in the diet of people making them (Margy, 1992). Such
fermented meats contribute both nutritional value and pleasure to meals.
 The employment of fish fermentation differs from place to the
other. In Asia, fermented foods are popular and well liked by the general
population and so widely used, that the daily diet of the people would not
be complete without them. In south East Asia fish are highly salted and
fermented until the fish flesh is transformed into simpler components
(Huss and Valimarson, 1990). Also Bhumiratana, (1980), Del Rosario,
(1980), and Sundhaghul et al., (1975) stated that in Southeast Asia
fermented fish products are very popular. However, in West Africa there
are few records of fermented fish products. Fermented fish sauce is a
processed product that is common in South East Asia. This product is
prepared by mixing small, uneviscerated fish with salt, at a ratio of three
to one, and then fermented (Beddows et al., 1976).

In Philippines, where many fermented food products are known,

their popularity is due not only to their characteristic flavour but also to
the fact that other processing methods, such as freezing and canning are
generally expensive (Olympia et al., 1992).

Fish fermentation in the south East Asia normally lasts several

months (three to nine months) and the fish flesh may liquefy or turn into a
paste (Huss and Valimarson, 1990).

Salting and drying fish in Africa are accompanied by fermentation,

but the period is short (a few days) and the product is not transformed
into paste or sauce. Olympia et al., (1992) stated that fermented fishery
products in the Philippines can be divided into two groups, the first one
includes those containing high concentration of salt about 15-20 percent
in the final product for example Bagoong (fish paste), Patis (fish sauce).
The second group includes Burong (fermented rice fish mixture) and
Burong Hipon (fermented shirmp rice). These products become acidic
with cheese-like aroma when fermented. The characteristic smell of
fermented fish is the result of enzymatic and microbiological activity in
the fish muscle. Watanabe (1982) found that the fermented fish products
of Senegal are highly salted and semi-dried fish products with an
obnoxious odour and cheesy but rich fish flavour recovery of kusaya
from Japan. Also Toury et al., (1970) reported that the Guedi is reported
as fermented dried fish product popular in Senegal, unsalted fresh fish is
piled together for about 24h in the open air. During this period the fish
undergoes putrefactive fermentation by its own enzymes and endogenous
bacteria. Then it is eviscerated, sometimes the big species are filleted to
shorten the period of drying and soaked in salty sea water in wooden
buckets. This water is changed once a week when it has become too dirty.
Finally the fishes are spread out on straw mats to dry in the sun for 2 to 4
days.

2.6 Nutrition and Safety Considerations

The preparation of fermented foods predates the recorded history

of Man. Early humans used observation of the apparent effect of
microbial alteration of food characteristics to develop processes for food
fermentation. Fermented products normally have a different texture and
flavour compared to the unfermented starting materials, thus making
them more palatable and digestible and prolonging their shelf life
(Amano, 1962).

With the rapid advancement in understanding of the basic sciences

of microbiology and biochemistry, improving the safety and efficiency of
the bioprocess is used to manufacture traditional fermented food.

Microbial fermentation has played an important role in food

processing for thousands of years. Fermentation of fish preserves fish
products, to enhance nutritive value, to destroy undesirable factors, to
make a safer product, to improve the appearance and taste of fish and to
reduce the energy required for cooking (Beddows, 1985).

During fermentation the microorganism secretes hydrolytic

enzymes into the substrate and assimilates the fatty acids, amino acids
and simple sugars are accordingly liberated. These are converted into
microbial structural components and secondary metabolites. Lactic acid
fermentation used for meat is an ancient process whereby a varied group
of bacteria ferment carbohydrates producing lactic acid as the major end
product (Hall, 1989). The organic acid produced during the fermentation
of fish in Mali was mainly acetic acid, where as it would appear that in
Asia mainly lactic acid is produced (Zakhia and Cua, 1991).

Fermentation increases the quantity of soluble protein in foods, it

may improve the amino acid profile, and reduce the levels of certain
antinutritional factors that interfere with digestion.Many fermented foods
are produced using fungi; the risk of mycotoxin contamination is high.
During natural fermentation, food poisoning flora and coliforms may also
grow with the lactic acid (Wang and Hesseltine, 1981). The
microorganism needs to be eliminated to make fermented foods safe for
consumption. There are several factors that contribute to the safety of
fermented foods: 1- Soaking and cooking: Soaking and cooking treatment
reduce the in situ microbial contamination. 2- Salting: Various fermented
foods are made with the addition of salt, which acts as preservative. 3-
Acid fermentation: Much fermentation is carried out by acid-producing
microorganisms where these organic acids act as preservative or as
bacteriostatic agents. 4-Low moisture content: The lower water activity
may be important preservative factor.

Safe products are obtained when using hygienic conditions during,

handling, and storage (Samson et al., 1987).
 Kofi (1992) reported that there are various types of salts used for
salting and fermentation of fish. Solar salt, which is most widely used in
fish preservation, has been found to contain the largest amount of
microorganisms. The general bacterial flora of solar salt mostly
comprises bacillus type (95 percent with remainder being micrococcus
and carcina type). The most important spoilage organism always presents
in solar salt are the red halophilic bacteria.

2.7 Drying

Drying preserves food by removing the water that is needed for

microbial growth and enzyme activity. According to (Doe et al., 1983)
there are three types of processes that can be employed in the drying of
fish:

Air or contact drying, where heat is transferred to the fish from

heated air or heated surface, utilizes the air movement above the
fish to carry away the moisture.

Vacuum drying, where advantage is taken of the greater

evaporation rate of water from the fish at reduced pressure, utilizes
conduction by contact with heated surfaces or radiation to
evaporate the water, which is removed by the vacuum pump.

Freeze-drying, relies upon the attainment of very low pressures by

highly efficient vacuum pump in sealed chamber containing the
fish.

Fish can be dried with or without salt. The process of drying in

tropical countries depends on spreading the fish to sun dry allowing
moisture to evaporate from the flesh surface. In temperate countries fish
can be hung up to dry in the wind, the process depends on the wind
speed. These processes take longer time than sun drier but fish take
longer time before spoilage. This is due to lower ambient temperature
(FAO, 1986).

Fish drying rate depends on the speed with which water leaves the fish
surface and on the rate at which water diffuses from the center of the fish
to the surface. The rate of diffusion of small fish is higher compared to
large one. These are due to decreased distance from the center to the
surface of the fish (Waterman, 1976).

Dried fish is particularly suited for low-income groups who cannot

afford expensive fish products. Simple drying of fish is not practical in
many of the worlds river systems because of the high atmospheric
humidity, but in desert or Sahelian Savanna rivers the practice is
common, especially for smaller species (FAO, 1986) In Senegal basin
simple drying is the usual form of treating the fish after they have been
eviscerated, scaled and in the case of larger species, cut into strips. In the
Niger, only the smaller species such as Alestes are sun dried, and in the
Chad basin one of the traditional fish product salanga consists of
Alestes dentex and Alestes baremose which are split open ventrally and
laid on mats to dry in the sun. In the Mekong, and other Asian rivers, sun
drying is also common, although this is sometimes combined with
salting. Sun drying is the method used by all fish processors in Sudan.
However, it is ineffective for the moisture level of the fish, is not reduced
to the level that can prevent spoilage of product by microorganisms
(Moy, 1977). Moreover, the humidity in tropical countries is very high.
The average humidity reaches 85-90% in some countries and thus the
salted-dried fish will tend to absorb moisture from the atmosphere when
left on the shelf or in storage. This can increase the moisture content of
fish and will import a wet appearance to the surface of the fish.
 Simple improvement, such as the use of drying racks raised above
ground level can increase drying rates and reduce contamination, thus
helping to make products of good quality (FAO, 1981). Peter (1997)
stated that the dried food can have poorer nutritional and eating quality
than the corresponding fresh food, so the correct design and operation of
dryers are therefore needed to minimize quality changes to the food.

Tunnel drier have been recommended by FAO for the drying of

fish in tropical countries (Waterman, 1976). According to this report,
tunnel drier has been used in Barazil, Combodia and many African
countries. The air speed and humidity in the drier can be controlled to suit
the drying rates required. The temperature can be adjusted by the use of
heaters. In addition to the use of tunnel drier, polythene tent drier can also
be used without much capital outlay. They have been used in Bangladesh,
Hawaii, Australia, New Guinea and some African countries (Doe et al.,
1977, Moy, 1977). Tent drier can prevent contamination and reduce
infestation by flies. The internal temperature is about 45-50c. At this
temperature, growth of spoilage microorganisms is retarded and the rate
of drying increased. Drying times vary considerably depending on the
process and weather, but 3-10 days is generally required to prepare a
typical sun dried product (Waterman, 1976).

Jan et al., (2001) has found that, it is moulds rather than bacteria
that cause spoilage during the preparation of dehydrated fish. Open-air
drying method is time consuming drying times are considerably different
depending on weather condition. Slow drying leads to fish spoilage by
microbial contamination.

Attempts to improve open-air drying practices in developing

countries have not changed much from traditional open air drying,
substantial losses can occur both in quality and quantity with subsequent
financial losses (FAO, 1981). Sun drying as method of food preservation
still have many limitations, even when racks of salt are used (ILO, 1986).
Long periods of sunshine without rain are required, drying rates are low,
products are often of low quality as a result of slow drying, insect
damage, contamination of air born bacteria, dustete. It is often too
difficult to obtain uniform product.

A solar dryer was developed to maintain better result than open air-
drying. Solar dryer proved successful in killing flies and improvement in
drying times were obtain (FAO, 1981). Infestation occurred during sun
drying can be considerably reduced by subsequent solar drying (Kenddy
and Wood, 1983). The temperature control with solar dryer was found to
be effective method of reducing losses due to larvae infestation (Doe et
al., 1977).

Precaution should also be taken to maintain the level of moisture

during transportation and storage of dried fish. During transport and
storage, the fish should be wrapped in moisture-impermeable packaging
so that it does not absorb moisture from the atmosphere. The packaging is
also effective against infestation by flies.

2.8 Smoking

Smoke was used long before the reasons for its effectiveness were
understood. In preserving fish and meat with smoke, the preservation
action generally comes from a combination of factors. Smoke contains
preservative chemicals such as small amounts of formaldehyde and other
materials from the burning of wood (Desrosier and Desrosier, 1977).
Smoke is generally associated with heat, which help to kill
microorganisms. Hansen, et al., (1998) stated that smoking process brings
about changes in quality parameters such as flavour, colour and texture.
 In many countries fish may be salted or partially sun-dried before
smoking and further drying may take place after smoking, depending on
the final moisture content required (FAO, 1981). Sometimes smoking is
used more for its flavour than for its preservative action especially when
combined with other preservatives. Eddy (1958) stated that salt, precedes
smoking when serves not only to flavour the fish but also to improve its
keeping quality by reducing the moisture content. The uses of the
smoking methods alone will not prevent the post-harvest putrefaction in
fish (ILO, 1986). For better results of smoking the water level of the
product must be reduced either by salting or drying or a combination of
the two. According to Howgate (1979), the loss of weight due to
dehydration in the smoking process is around 10-25 % depending upon
the origin of the raw material, the final product characteristics and the
process parameters such as time and temperature during smoking. The
commonly observed loss in weight of fillets during the process is both
due to dehydration of muscle and lipids leaching from the muscle (Sojofn
et al., 2000). Ofstad, et al., (1995) showed that liquid loss in minced
salmon fillet during heating decreased with increasing salt concentration
from 1 to 2%. The use of smoking as a curing technique is determined by
the availability of fuel and local traditions. About 35% of the total
landings in tropical Africa being processed by smoking (FAO, 1970) but
this technique is less important in Asia and Latin America. The type of
fish smoke varies according to availability and consumer preferences
(ILO, 1986).

Generally there are three types of smoking:

1. Cold smoking, in this method the fish are not cooked and the final
product is similar in keeping qualities to fresh fish.
 2. Hot smoking, fish are cooked and this method prevents spoilage for
one day or two if the product is not dried.

3. Smoke drying method, this produces cooked dried products.

Smoke drying is the most widespread way of preserving fish in
Africa, and is practised in nearly all river systems.

In West Africa hot smoking is the main method of fish processing.

However, salting, fermentation and sun drying are also important fish
processing methods (Kofi, 1992). In this area nearly 70 percent of the
total fish supply is marketed in smoked or dried form. Atlantic salmon is
often cold smoked or marinated and cut in thin slices and consumed
without any further heat treatment (Sojofn et al., 2000). Also the author
stated that the cold smoking process includes three stages, each of which
is important to the products potential shelf-life: salting, drying and
smoking all at temperature below 30C. In Sudan 70 percent of the total
fish landing is consumed fresh, the rest is cured either by salting,
fermentation or sun drying. A little amount of the local fish supply is
smoked except in Southern Sudan, where smoked and very dry fermented
fishery products are very popular among the local community (Kofi,
1992). The use of smoking at the present time is primarily to impart a
pleasant taste, and this method commonly is used in conjunction with
some other to provide preservation.

2.9 Colour

The colour of fish is an important quality attribute, which affects

fish buying consumer. The colour of fish flesh is governed by blood,
melanin derivatives (produced from skin colouration) or arises directly
from its diet. Pink flesh colour arises from the carotenoids, which enter
food chain originally from photosynthesis (John, 1994). The chemical
groups involved in the colour of fish flesh are heam pigments, carotenoid,
and melanin. As in all foods the specific colours produced are influenced
by physical structure. Kropf, (1980) reported that the colour is most
important factor for consumers when purchasing meat.

The method of catching and handling can be responsible for flesh

bruising discolouration. Botta et al., (1987b) found that cod caught by
gillnet are significantly more bruised than those caught by trap, hand-line
or long-line. Fish which have struggled extensively before being brought
for gutting gives lower colour grades (Botta et al., 1987a). In freshly
caught fish, blood contained in the vessels is still red. Holding the fish in
ice before filleting results in a darker red or brown colour as the
haemoglobin oxidizes.

Normally within the fish itself the flesh is of two types, dark and
light on frozen storage; pigments in the darker meat are especially subject
to oxidation, becoming deep yellow or brown. The colour of pink or red
fish is sensitive to frozen storage abuse; the pigments oxidize, and the
colour can disappear (Hillman, 1983). Impurities present in common salt
can have an accelerating effect on the oxidative deterioration of frozen
fish. Anon (1970) suggested that only high quality salt should be used for
the brine dipping of species particularly susceptible to oxidation. Mixed
tocopherols with ascorbic acid have been used to prevent oxidation of red
fish (Wasson et al., 1991).

Salt from the saline deserts to the north contains nitrates and borax
as impurities, and these would be converted to the nitric oxide necessary
for the formation of red complexed haem pigments. Washing minced
flesh of fish has a beneficial effect on the color, but significant quantities
of water are needed (Martin, 1976). The presence of blood, kidney tissue,
or the black lining of the belly cavity can also cause darkening of the
flesh (King, 1973, and Dyer, 1974).
Oxidation of the blood pigments may be the cause of the yellow and
brown colour, which develops on the storage of fish (Jauegui and Baker,
1980).

Fish oil colour depends on the species, but the pigments in all types
can become yellow or brown on oxidation. While the whole fish or the
fillet is deep frozen, oil can be forced out of the tissue to collect on the
skin, these fish are rusted (Wasson et al., 1991).

2.10 Oxidative Rancidity:

The term rancidity is used to describe the task or smell of rank stale
fat. Rancidity associated with a characteristic, unpalatable odour and
flavour of the oil. Virtually all fish contain highly unsaturated fatty acids
as major components of their lipids. However, the total amount of fat may
vary greatly (Lands, 1986). Obviously fatty fish such as mackerel, and
salmon will be more susceptible to appreciable oxidation. Hess, (1950)
reported that fat and oils of many kinds of fish, especially the fatter ones,
such as herring and salmon, are composed to a great extent of unsaturated
fatty acids, and are subject to oxidative changes producing oxidative
rancidity and sometimes undesirable alteration in colour.

Oxidative spoilage of salted fish can occur during processing as

well as during storage (Van Arsdel et al., 1973). The rancidity is caused
by the changes that occur from reaction with atmospheric oxygen, is the
so-called oxidative rancidity. The off flavours are produced by hydrolytic
reactions, which are catalyzed by enzymes is the so-called hydrolytic
rancidity (Huss and Valimarson, 1990). Rancid flavour are chemically
very complex, since they are derived from any or all of the unsaturated
fatty acids originally present in the oil, each of these can oxidize through
several different mechanisms (Clucas and Ward, 1996).
 Rancid fat contains a wide varity of chemical substances, whose
structures are not all known (Hultin, et al., 1982). Human taste buds are
highly sensitive to some compounds such as lactones and free fatty acids,
so only minute amounts of these compounds need to be produced to spoil
the taste of food (Harris and Tall, 1994). Fresh and saltwater fish contain
significant levels of polyunsaturated fatty acids. Consequently, such fish
should readily go rancid on storage (Smith and Hanson, 1990). At
ambient temperatures storage, the development of rancidity is not a major
problem with fresh fish, because the normal level of microbial flora
associated with fresh fish is such that bacterial spoilage will render it
inedible before rancidity has proceeded to any great extent. However,
rancidity development is very important in the storage stability of dried or
frozen fish.

Fey and Regeustein (1982) stated that the total level of fat in fish
varies depending on the species of fish and season. Fish can be divided
into low and high fat species. Low fat species usually have less than 4%
total fat and high fat species more than 5%. In a typical fish, about 1% of
its total body weight will consist of various phosphlipids, which are
distributed through the body and perform essential function such as
regulating the properties of membranes. Any lipid in addition to this level
will be made up of triacylglycerides, which are primarily deposited in the
liver and under the skin and act as food reserve. Seasonal variation in fat
level is due to the availability of food and in mature fish, the state of the
breeding cycle, during spawning, most of the fat is converted into
reproductive tissue. This fat can contain highly unsaturated fatty acids, all
of which can make fish very susceptible to rancidity.

Lobuza (1971) found that fish tissue susceptibility to rancidity does

not only depend on the amount of lipid present, but the lipid composition
and its location in fish tissue matrix. Susceptible to oxidative rancidity,
other lipid types materials, such as tocopherol and carotenoid pigments
are also associated with autoxidative reaction in fish tissue. These may be
involved in controlling oxidative rancidity, or may be oxidized
themselves or co-oxidized with lipid (Lobuza, 1971).

Fish lipids are even more susceptible to oxidation and are probably
responsible for the rapid spoilage of oily fish such as mackerel and
herring (Sanders, 1994). The 20-22 polyunsaturated fatty acids are highly
susceptible to peroxidation. The high concentration of 20-22
polyunsaturated fatty acids are found in fish and fish oils, many fish oils
contain about 20% of their total fatty acids as these higher
polyunsaturated fatty acids. This explains the susceptibility of fish to
oxidative rancidity (FAO, 1962). Many of the Fish oils seem to be more
susceptible to oxidative deterioration than most animal fats (Hess, 1950).

2.11 Salted Fermented Fish and Fish Type:

In Sudan salted fermented product without drying is called

fessiekh. The product is also known by the same name in the Middle
East, (Campbell-platt, 1987) and in Egypt (Hassan et al., 1972, Hamed et
al., 1973). Fessiekh is a popular fermented product in Sudan when
compared with other fish products.

The technique of fassiekh probably entered to the Sudan during the

Turk-Egyptian rule (1821-1885) but its production on a large scale was
only well established during the Anglo-Egyptian condominium rule
(1898-1956).

The methodology in the preparation of the product was

traditionally carried through from one generation to another.
Commonly there are two types of fish used in fessiekh making in Sudan,
Hydrocynus species (tiger fish) known locally as Kass and Alestes
species (pebbly fish) known as kawara, both belonging to the family
Characidae (Idriss, 1981). These two types of fish are considered the first
class of fessiekh, (Agab and Bashir, 1987). The second class can contain
other fish.

The totality of fissiekh in the Sudan is made from Nile fish. Only
one kind of fish from the Red Sea can be used to make the product. This
is the mullet (Mugil cephalus), called alarabi. About 80% of fessiekh is
made from Alestes baremose and A. dentex, while Hydrocynus sp. makes
the balance, this is because kawara is always more abundant than Kass in
any catch. Fessikh processing is a seasonal process generally starting in
November and ending in June, with a peak in February and sometimes in
March.

The ratio of salt used depends on the experience of commercial

sector and it varies widely. The salted fishes allow fermenting and the
fermentation time varies according to the season. Dirar (1989) mentioned
that the first stage of fermentation on palm leaf takes about 3-4 days in
the summer and 4-7 days in the winter.

2.12 Microbiology of Fermented Fish Products:

The most important factors in consumer acceptability are flavour

and odour, although the colour can be significant. As aseptically a
product of fermented fish did not give the typical aroma associated with
them, it was anticipated that microorganisms were involved in aroma
development (Beddows et al., 1979).

A considerable number of investigators have reported on the

microbiology induced changes in spoiled and fresh fish, but the work on
changes in a high salt environment, as found in salt-fermented production
is very limited. Eddy (1958) stated that contamination comes from the
fish, which ordinarily introduces chiefly species of Pseudomonas,
Alcligenes, and Flavobacterium, from ice, which introduces these genera
plus Corynebacterium and cocci, and from mechanically introduced
sources, e.g., dust, which add cocci (Boez and Guillerm, 1930) isolated
an anaerobic spore-bearing bacterium from an early stage in the
fermentation of fish sauce (nuoc-mam), which was accredited as being a
species of Clostridium. Aseptic preparation failed to give the typical
nuoc-mam aroma, so it was concluded that bacteria must be involved in
this, (Beddows, 1985).

Krempt (1929) tried to produce a sauce more quickly with a true

aroma, on a commercial scale with some degree of success and he
concluded that microorganisms were required for aroma production.

Beddows (1985) suggested that all the factors, which can influence the
relative activities of proteolysis enzymes present in the material used, the
pH of the pickle, the temperature and the salt concentration could
influence taste quite markedly.

Saisithi et al. (1966) isolated 10 species of bacillus, 1 coryneform,

2 streptococci and 1 each of micrococcus and staphylococcus from fish
sauce (nam-pla). They stated that these bacteria could have been derived
from the solar salt used, since they found that it contained, an average,
2700 bacteria /g. The total viable count decreased to 2103 cell/cm3 after
9 months. Crisan and Sands (1975) examined the microflora of four
fermented fish. From nam-pla they isolated Bacillus cereus and a strain of
B. licheniformis after 7 months of fermentation, but at the end of the
fermentation period they isolated another strain of B. lincheniformis, B.
megaterium and B. subtilis. All the isolated species were grown on a
medium containing 10% salt to which they seem to be halotolerant and
not halophilic. No growth was achieved on a medium containing 20%
salt.

Crisan and Sands (1975) stated that investigation of patis (fish

sauces produced in Philippines) after one month showed no organism to
grow on 20% salt, but single strains of Bacillus pumilus, Micrococcus
copoyenes, M. Narians and Candida clausenii were isolated from 10%
salt medium.

2.13 Synthesis of Findings of Information Related to The Subject

Matter of This Thesis

As regards the situation of salting and fermentation, attempts

have been made by a number of researchers like Cruess (1951),
Mahmoud (1977), Johnson (1994), Abu Gideiri (2001) and Abu
Gideiri et al., (2004), who have worked on a number of aspects like
(post-harvest treatments, meat quality of Nile fish, freezing of fish,
salting and smoking of fish, drying, salting and fermentation of fish)
and the main findings so far reached are enlisted below:

The proximate composition of whole fresh fish was in the range of

63.3-75.2%, 15.0-22%, 0.4-2.5% and 0.5-1.94% for moisture, protein,
fat and ash content respectively (Mahmoud, 1977). Johnson (1994)
stated that protein content of fresh fish varied widely from species to
species and in different seasons. The crude protein of fresh fish ranges
between 14-20% and higher levels are obtained during winter season
(Clucas and Ward, 1996). Chemical composition of Debs species was
76.7,19.3, 2.1 and 1.6 for moisture, protein, fat and ash respectively,
(Ali et al., 1996). Chemical composition of fresh fish varies greatly
from one species and from one individual to another depending on
age, sex, environmental conditions and seasons (FAO, 1986). The
penetration of salt into the fish muscle is dependent on many factors;
including the thickness of the fish, osmotic pressure, temperature,
purity of the salt, freshness of the fish, and the fat content of the fish (
Ingram and Kitchell, 1967).

The rate of salt penetration varied with the thickness of flesh,

temperature and surface/ volume ratio of the flesh (Cruess, 1951).
The temperature exerted a definite influence on rate of salt penetration
and moisture loss (Narayanaswarny et al., 1980). Some fermented
salted fish products need low salt concentration to be in a good
condition like plaa-som Thailand fermented fish product (Christine
et al., 2001) in which salt concentration should not be higher than 6%
or 7% (w/w) for the fermentation to occur within 4 to 7 days and other
fermented products need higher salt concentration.

Levanidov (1958) reported that the mechanical losses taking place

during salting process, e.g. dropping of scales, tearing of internal
organs and the losses, which like differences between water and
organic substances.

Abu Gideiri et al., (2004) found a significant change in some

chemical constituents of salting fish (O.niloticus) among different salt
concentrations. Also there was a significant variation in chemical
composition of fessikh products from kass and kawara species
prepared at different salt concentrations (Abu Gideiri, 2001).

El-Sebahy and Metwali (1988) found a decrease in the level of

crude protein, fat and increase in ash content of small and large salted
fermented Bouri fish muscle (Mugil Cephlus) with increase in salt
concentration. The protein content of fassiekh was 61.2% and 58% found
by Dirar, (1993), and Kofi, (1992) respectively. The losses of protein
during processing of fish averaged 35% in the case of Makassar fish
Ngapi and pedah siam and little or no loss in Shiokara (Amano,
1962). Lawrie (1990) stated that crude protein decreased with storage of
cured meat and this was attributed to some changes during storage that
caused by maillard reaction, where in carbonyl groups of reducing
sugars react with amino groups of protein and amino acids non-
enzymtically, and might also be due to an attack of myoglobin by bacteria
during storage and changes in pH. The crude protein of dried salted Debs
species was 66.6% (FAO, 1990). Medynski et al., (2000) reported lower
pH strong effect on protein of salted fish. Salting processes was usually
accompanied by protein loss (Tarr, 1965). The ash content of Botswana
dried salted fish was 24.4% (Mmdlewa, 1990). Agab and Beshir (1987)
found that the proximate composition of traditionally salted fermented
fish of the Sudan were 18.12-28.5%, 20.7-45.5%, 10.6-22.5%, and 3.6-
5.2% for protein, moisture, fat and ash respectively. Kofi (1992) found
that the products with high moisture content tend to deteriorate faster than
dried products especially if the salt level is low. Also he found that the
treated fessiekh was low in fat than traditional samples. Lower fat content
in Nigerian salted fish and lower ash in dried salted debs species was
found by (FAO, 1990). Ruiter, (1995) stated that salted dried seafood is
affected both by loss of water and by the amount of added salt.

The physical characteristics of fresh fish (Labeo coubie) from lake

Nubia showed that, the lightness, redness and yellowness were 47.19,
6.23, and 8.4 respectively (Adam, 1996). The absence of oxygen in
dehydrated salted meat caused millard reaction where the carbonyl
groups of reducing sugars react with the amino groups of protein and
amino acids non-enzymatically giving a dark-brown colour (Sharp,
1957). The storage of cured meat at high temperature converts myoglobin
to bile pigments, thus causing the meat to become pale and yellow
(Lawrie, 1990). Also he stated that the loss of moisture by curing meat
have a significant darkening effect on the surface of fresh meat and the
production of metmyoglobin from myoglobin or oxymyoglobin is
accelerated by different conditions that include low pH, heat, and salt.

Ishag (1998) stated that an increase in yellowness of salted meat

compared to unsalted one. Jane et al., (1987) studied the characteristics of
longissimus dorsi muscle, heated to seven internal temperature between
55-85C. He reported that the lightness value become higher with
increasing temperature, redness value reduced with increasing
temperature from 75 to 80C and yellowness values were not affected. The
reaction of oxidation is faster at lower pH values and is markedly slowed
by lowering the temperature ( Jane et al., 2001). Reiss and Tappel (1973)
reported that the oxidative rancidity of meat tend to increase over storage
period towards reaching a maximum value before starting to decline.
Igene et al., (1979a) reported that there are differences among the meats
from different species in their susceptibility to lipid oxidation as well as
among different tissues in the same species. Interaction of oxidation with
muscle food components may in turn bring about changes in their colour,
texture and nutritional value (Spanier et al., 1992). Younathan and Watts
(1959) stated lower oxidative rancidity for cured as compared to uncured
meats. Oxidative rancidity affected by quantity of oxygen present,
temperature, light and peroxidants such as salt (Kramlich, 1975).

Abdalla (1989) reported that the pH of salted fish product ranged

from 6.4 to 6.9. The pH should be below 5-4.5 in order to inhibit the
pathogenic bacteria and allow the lactic acid bacteria to ferment the
available carbohydrates and thereby cause a decrease in pH (Owens and
Mendoza, 1985). Also they stated that the lower the pH of the fish flesh,
the slower in general bacterial decomposition would be. Lowering the pH
of the fish flesh results from the conversion of muscle glycogen to lactic
acids. Rosenthal (1999) reported that most bacteria that cause food
spoilage have a relatively low tolerance to salt (NaCl), and salt acts as a
dehydrating agent, consequently lowering the water content of bacterial
organisms and thus limiting their ability to thrive and reproduce.
However, there are numerous spoilage and pathogenic bacteria (2 to 4
percent). These organisms are collectively referred to as halophilic (salt-
loving) organisms. Ingram and Kitchell (1967) reported that salts were
preservative means that inhibit dangerous microorganisms. Also Owens
and Mendoza, (1985) stated that pH should be below 5-4.5 to inhibit
pathogenic and spoilage bacteria. Crutchfield (1959) stated that, salting
improved the quality of fish product through inhibiting microbial and
mould growth. Also (Anonymous, 1982, Saisithi, 1987) reported that the
salt concentration might range from 1 to 10% (w/w) in different types and
batches of fermentation fish. This is likely to have a pronounced
influence on the microbial growth and the rate of fermentation and
thereby on the sensory quality and safety of the product. Halleck et al.,
(1958) reported that salt serves to prevent microbial growth during and
after curing.

Where the moisture content of the food was lowered, the amount of
water available for supporting microbial growth decreased (Scott, 1957).
The total viable bacterial count increased during early stages of salting
and then decreased with time (EL Tom, 1989). Also Beatty and Forgere
(1957) stated that bacteria, which contribute to spoilage in fresh fish,
could not survive at salt concentration above 12%.

Cold temperature is generally beneficial for the cured meats

product in that they retard bacterial degradation and also slow the action
of the indigenous muscle enzyme (Locker and Hagyard, 1963).
 Staphylococcus sp. were isolated from thai-fermented fish with salt
concentration above 5% (Tanasupawat et al., 1991, Tanasupawat et al.,
1992) and from Korean fermented (hydrolyzed) fish with salt
concentrations ranging from 8% to 26% NaCl (Um and Lee, 1996).
Akanda and Oladosu (1988) stated low incidence of Staphylococcus
aurous in salted fish at 30 C storage for eight weeks. At low water
activity in the presence of high salt concentration microbiological
problem should be non-existent (Zapata et al., 1990). Also Scott (1957)
reported that the growth of microorganisms was much dependent on
water activity and temperature.

The composition of salt has a great importance in determining the

physical qualities of the final product (Tressler, 1950). Texture in flesh
food is usually referred to as tenderness, which is the most important
quality factor in meat and fish products (Chrystall et al., 1994). Also
Boleman et al., (1997) stated that consumers are able to identify meats
that are tender and are willing to pay a premium for improved tenderness.
Processing of fish such as curing (Shahidi, 1992) and smoking (Maga,
1987) bring about a characteristic flavour in the products. Allen and
Angelo, (1991) stated that the degradation of texture, flavour and odour
of stored seafood were attributed to the oxidation of unsaturated fatty
lipids. Processing operations as salting, cooling and mincing promote
oxidation while smoking, dehydration and freezing retarded oxidation.

The characteristic flavour and odour of oxidized fat is caused by

the presence of low molecular weight aldehydes, acids and ketones that
form during the oxidation and decomposition of the fatty acids molecules
Judge et al., (1990). Processing can have a dramatic impact on the aroma
of fish (Josephson et a., 1986). Microorganism and fat oxidation have
been considered as possible aroma. Also they stated that bacteria play an
important role in development of the aroma of fermented fish (Beddows
et al., 1979).

With all the above information in the background, it is appropriate

here to dwell on the contribution of the findings of this research to
knowledge. This contribution is attached to the fulfillments of the
objectives, which were enlisted in the introduction with special reference
to the determination of the magnitude of losses resulting from spoilage,
targeting of progressive promotion of traditional treatment by applying
scientific approaches, adoption of standards and specification approaches,
the onset of spoilage and recommend range of salt concentration that
secures the best preservation.

The set program and findings obtained from studies made on both
kawara and debs have addressed the above- mentioned issues and largely
the objectives are met with specifically in the areas of the determination
of the basic biochemical composition of the two species in both treated
and non-treated stages, the changes in the nutritive values as coverned by
season, salt concentration and microbiological attack. Both the keeping
time and preservation through salting range have been determined. These
areas can be forwarded as contribution to knowledge and in particular as
marketing of the two species are of lower economic importunes in the
country.

Apart from the above there are areas of full agreement with those
obtained by Hess (1950), Cruess (1951), Shahine (1956), Scott (1957),
Beatty and Fongero (1957), Eddy, (1958), Halleck et al., (1958),
Younathan and Watts (1959), Locker and Hagyard, (1963), Tarr (1965),
Ingram and Kitchell (1967), Green (1969), Reiss and Tappel (1973),
Beddows et al., (1976), Mahmoud (1977), Narayanaswarny et al., (1980),
Akande and Oladosu (1988), EL- Sebahy and Metwalli, (1988), EL Tom
(1989), Lawrie (1990), Zapata et al., (1990), Tanasupawat et al.,(1991),
Spanier et al., (1992), Tanasupawat et al., (1992), Johnson (1994), Ruiter
(1995), Clucas and Ward (1996), Ishag (1998), Rosenthal (1999),
Christine et al, (2001), Jan et al., (2001), Abu Gideiri et al., (2004), ,
while disagreement is registered in case of Amano (1962), Anonymous
(1982), Agab and Beshir (1987), Jane et al., (1987), Saisithi (1987), FAO
(1990), Mmdlewa (1990), Kofi, (1992), Dirar (1993), Adam (1996), Ali
et al., (1996).

Perhaps the differences from those of this piece of reasarch could

be attributed to a number of reasons such as time factor, varying
techniques, inconsistency in procedure and bias or pre-determined
thoughts.
 CHAPTER III

MATERIALS AND METHODS

These studies were conducted at the Fisheries Research Center,

Ministry of Science and Technology, University of Khartoum and

Veterinary Research Center, Soba.

3.1 Sample Collection

Sample of fresh fish, namely, Kawara (Alestes spp.) and Debs

(Labeo spp.) were obtained from El Mawrada market. The samples were

transported immediately (early in the morning) to the laboratory at

Elshagra Fisheries Research Center. The microbiological, physical and

chemical analysis were immediately carried out for fresh sample.

Traditionally treated samples (Fasseikh), were obtained from Bahari

market after giving the traditional processor fresh fishes to treat as they

treat their fishes. The salt used in the processing of the experimental

fessiekh was obtained from Khartoum market.

During processing of fessiekh in the laboratory, samples were

withdrawn at random from different salt concentrations from different

containers of different species for microbiological, physical and chemical

analyses. Sampling was carried out every four days for the first 2 weeks

and then was taken monthly until 6 month.

3.2 Treatment

Fresh fishes were weighed, washed, eviscerated, washed again and

transferred to baskets to dry up. Then weighed again to obtain the actual

weight, which will be treated with salt. Fishes were then divided into 3

groups each one divided into 4 containers (small plastic barrels with lids)

each has an equal weight of 3 kg.

Each group was thoroughly treated with commercial salt according

to the following percentage:

Group 1- treated with 20% salt (w/w) (3kg of fish +0.6kg of salt)

Group 2- treated with 25% salt (w/w) (3kg of fish +0.75kg of salt)

Group 3- treated with 30% salt (w/w) (3kg of fish +0.9kg of salt)

In each group, salt was applied by brushing and rubbing of the fish

surface, the gill chamber and the inner lining of eviscerated abdominal

cavity. All of these treatments were repeated during the three seasons of

the year (summer, autumn and winter).

3.3 Chemical Analysis

3.3.1 Preparation of the Sample

The samples of fresh and treated fish were minced through a meat

mincer, and then mixed several times to be homogenized before analysis.

3.3.2 Moisture Analysis

The methods of A.O.A.C. (1990) were used to determine the

moisture content of the sample and dry matter. Sample was weighted into
the clean dish and heated in the oven for 3 hours. Transferred to a

desiccator with lid on and allowed to cool at room temperature,

reweighing. Calculation was done according to the following equation:

% Moisture = (A-B)*100
A

% Dry matter=B*100
A
Where: A is the original sample weight,

B is the sample weight after heating.

3.3.2 Protein Content

Protein content of the sample was determined according to Kjeldahl

method. The samples were weighed (0.25 g for each sample) by using 9

cm 540 filter paper folded then the sample was transferred to a tube and

then added 2 A.A.O.A.C. kjeltabes (potassium sulfate +mercuric oxide)

were added to the sample plus 20 cm conc.sulphate acid (swirl to mix).

Tubes were transferred to digestion unit (model Buchi 426) vacuum pump

was turned gently on.

When the samples were digested and tubes appeared clean, the

vacuum pump turned off and left for an hour then tubes were transferred

and exhaust cap to side trays. The digested samples were distilled on an

automatic distillation unit model (Buch B-323). One thiosulphate tablet is

added to the digested sample (swirl to mix), and then the tube is placed
on the distillation unit. 70 ml of 40% NaOH are automatically dispensed

into the tube by the machine along with steam. The distillate was

collected in a flask containing 50 ml 2% Boric acid and 4 drops of

screened methyl red indicator and then titrated with 0.1 N H2SO4.

The percentage of total nitrogen of the sample was obtain by:

% Total nitrogen = (titre-blank) normality of acid 0.00140

________________________________
sample weight (g)0.1

0.00140= g of N equivalent to 1.0 ml of 0.1 N sulphuric acid.

The percentage of curde protein were obtained by:

% Crude protein = %N by 6.25 (i.e. there is an average of 16%

nitrogen in proteins, therefore, 100/16 = 6.25 = factor to convert nitrogen

to crude or generalized protein.

3.3.3 Crude Fat

Crude fat was determined according to the method of the fertilizer

and feeding stuff regulation (1976). Five grams of sample were weighed

(weighed to the nearest 0.001 g) into an extraction thimble using a beaker

as support. The thimble marked with a ball-point pen for identification. A

small plus of cotton wool was pressed into the top to prevent loss of

sample and the thimble was inserted into an extractor. The sample

weighed to nearst 0.001g (M1). 80 ml of petroleum spirit was poured into

the extractor, and attached to the condenser and refluxed on a water bath

for 4 hours. The apparatus was removed from the water bath, retaining

the solvent in the flask. The thimble then removed and placed fume

cupboard to evaporate the solvent. Lightly grind the defatted material in a

mortar and pestle and return it to the thimble as follows:

Clamp upright the extractor and flask and wash the defatted

material (using a small amount of petroleum spirit from a wash bottle)

into the thimble via the 100 mm funnel supported so that its stem is 1/2

below the neck of the thimble, continue extracting for one hour using the

original solvent in the flask. Disconnect the apparatus, remove the

thimble and allow the solvent to evaporate. Allow the solvent in the flask

to evaporate (or collect in a side-arm soxhlet extractor for re-distillation).

The flask with the defatted material dried at 103 C for 2 hours,

cooled at room temperature in a desiccator and weighed. Reheated for 30

minutes, cooled and weighed. Reheated this until a difference of 0.002g or

less was achieved. The defatted material was transfered to a glass sample

bottle for determination of crude fiber. Finally calculate the % of crude fat

using the following formula:

% crude fat = weight of oil

_______________100
sample weight (g)
3.3.4 Crude Fiber

About 3.0g of the dried and defatted sample (the material originates

from the estimation of oil/fat on the same sample). After deffating, content

of the soxhlet thimble were removed and briefly air dried to allow most of

the solvent to evaporate. The sample is then transferred to a small sample

jar and dried in an oven at 100 C for 2 hours and transferred to a 600 cm

tall-form beaker with a few antifoaming granules. 200 ml of 0.25 N

sulphuric acid was added in portions, using the first portion to wet the

sample and the rest to rinse off sample sticking on the sides of the beaker.

Turn condenser water on. Turn the heating on and when it starts to

boil, set the stopwatch and boil for exactly 30 minutes. Meanwhile pour

200 cm of 0.313 N Na OH into a quick fit washed-bottle and heat it

gently. The buchner funnel was prepared, a piece of cotton wool anf filter

paper smoothly inside the funnel ( under slight vacuum ). After paper disk

was soaked on both sides with distilled water to remove loose fiber and lay

it onto the cotton support, excluding all air bubbles.

When the 30 minutes boiling period lapse, the beaker was removed

and allowed it to stand for one minute before pouring the contents onto the

filter paper in the buchner funnel (unde low vacuum). The fiber remaining

in the beaker was rinsed out with distilled water. Lift the filter paper out

and lay flat inside the 6 glass funnel and wash the fiber back into the

beaker with the new boiling Na OH, Sodium hydroxide remaining inside
the wash-bottle must be poured into the beaker. The beaker was heated for

exactly 30 minutes and then filter through a new filter paper through the

buchner. The fiber was washed with 1% hydrochloric acid to neutralise the

Na OH and removed from the buchner, layed inside the glass funnel and

the fiber scraped,washed into the scintered crucible with hot water. The

fiber rinsed with 3. 5 ml portions of acetone (under slight vacuum). The

scintered crucible was put into a nickel dish and dried at 103 C for one

hour. Cooled at room temperature in a desiccator and weighed. Incinerated

at 500 C over night, cooled and re-weighted. The difference in weight is

reported as crude fiber. The fertilizer and feeding stuff regulation (1976).

Crude fiber % = loss in weight (g) (100-% moisture- % oil)

sample weight (g)

3.3.5 Ash Content

A.O.A.C. (1990) method was used to determine the ash content of

the samples. Two grams of the samples were weighed into a clean pre-

dried and weighed porcelain dish. The dish containing the sample was

placed in a muffle furnace at 550 C and left burning for 5 hours at this

temperature. Then, the dish with its content was weighed again after
cooling in a desiccator at room temperature. Ash contents was calculated

as follows:

Ash % = weight gained by the dish

100
sample weight (g)

3.4 pH Measurement

The pH was read using digital pH-meter (model Jenway

3015). The pH-meter was calibrated using standerd buffer solutions of pH

= 4 and pH = 7. Two grams of sample was minced with 9 ml distilled

water and was transferred to test tub. The pH was taken as a mean of 3

readings.

3.5 Determinations of Minerals

All minerals studies of fresh or salted fish were determined

according to the fertilizer and feeding stuffs regulations (1976).

3.5.1 Determinations of Phosphorus

Prepared an ash extraction of the sample, diluted the ash

extract solution and take a suitable aliquot to give a phosphorous

concentration of between 4.0 and 24.0 ug cm3.pipetted the final

aliquot into a 25 ml volumetric flask. Prepared standard solutions

by transfer approx 50ml of the phosphorus stock solution to a clean

beaker and pipette 2.0,4.0, 6.0,8.0,10.0 and 12.0 ml aliquots of the

stock solution into a series of 25.0ml flask, then commenced

 addition of A. V. M. added 10ml of A. V. M. reagent to all sample

flasks and to the standard flask. Diluted all flasks to the mark with

water, stopper and mix well, then prepared a blank solution by

diluting 10.0ml of A.V.M. to 25.0ml with water. Stand the flasks for

30 minutes to take the full colour. The solution was read against the

reagent blank at 440nm. The standards read before and after the

samples and take the average of the reading. Read the sample

concentration off the standard curve.

Calculation:

%P = sample concentration x dilution x 100

sample wt. (g) 106

100/106 is the factor used to convert mg/g p to %p.

3.5.2 Determinations of Iron

9.8 ml D.W. was added to the 0.2 sample in a centrifuge tube, then

added 0.2 ml of saturated potassium persulphate and 1 ml 25% sodium

thiocyanate. Added 5 ml of N amyl alcohole and shake the stoppered-

centrifuge tube vigorously. Centrifuge for 5 minutes. At 2000 rpm. Take

the supernant and compared the colours at 480.

For Fe standard, added 10 ml 65% perchlorid acid to 50 mg Fe in digestion

flask and boil until dissolved. Cooled and diluted to 100 ml with D.W. 2

ml of this standard in 100 ml D.W. used for working standard.

Calculation:

Iron = T x 50 = mg/ 100ml

S
T= Reading, S= sample

3.5.3 Copper Extraction

9.8 ml D.W. was added to the 0.2 sample in a centrifuge tube.

Transfer the contents in a bottle, added 5 ml of copper reagent to the bottle,

then shake the contents thoroughly until a yellow material to the bottom of

the bottle. Aspirate this material and transferred to a clean test tube and

read it at 435. For Cu stock standard prepared 50 mg Cu in 10 ml Conc.

Nitric Acid and diluted to 100 ml D.W. 1 ml stock standard in 100 ml

D.W. used for working Cu standard.

Calculation:

Copper = T x 50 = mg/ 100ml

S
3.5.4 Determination of Calcium

Calcium standard stock solution A:

Calcium carbonate dried at 103C for one hour, cooled in desiccator.

Weigh accurately 2.497 g into weighing boat. Transferred to 1litre

volumetric flask with about 100 ml deionised water. Whilst swirling,

slowly added 60 ml in hydrochloric acid. When carbonate has dissolved

make up to mark with deionised water.

Calcium intermediate stock solution B:

50 ml of stock A pipette to 1000 ml volumetric flask. Diluted to

mark with deinoised water.

Prepared an HCl extract solution, then diluted ash extract solution within

the range of 10-100 ml. Prepared standard solution by pipette 10.0 ml of

stock solution B (50 ug ml) to 50.0 ml volumetric flask, diluted to mark

with deionised water: contains 10.0ug ml. Prepared suitable standard

solution ( diluted 5-50 (+5ml/La3+ soln.) ) 1.0 ug ml. Prepared a blnk

solution (La3+) 5ml La3+ 50ml. Set up the atomic absorption

spectrophotometer according to instruction manual using the 422.7 nm

line. Adjust fuel to obtain the highest stable reading whilst aspiration a

standard solution. Sprayed deionised water into the flame and zero the

instrument. Sprayed standard solution (zeroing with a lanthanum mark)

then samples (zeroing with a ranged mark). Washed with deionised water

and take a zero reading between each. Run standards between every 5 or 6

samples solutions and after running samples.

Calculation:
% calcium = ug ml (from graph)x 100 (ash dilution) x dilution x 100
_____________________________________
sample wt. (g)
3. 5. 5 Determination of Sodium and Potassium

In test tube diluted 0.1 ml of the sample (serum) in 9.9 ml D.W.

Na stock standard:

58.45 g sodium chloride in 1L D.W.

working Na standard:

High standard: 8.0 ml of stock standard in 1Litre D.W.

Low standard: 7.0 ml of stock standard in 1litre D.W.

Calculation:

Na = T/S x140 (mg/L)

K stock standard:

7.45 g potassium chloride in 1L D.W.

working K standard:

High standard: 7.0 ml of stock standard complete to 1litre D.W.

Low standard: 5.0 ml of stock standard complete to 1litre D.W.

Calculation:

K= T/S x5 (mg/L)

3.6 Colour Measurement

The colour of samples was determined using a Hunter Lab

Tristimulus Colorimeter Model D 25 M.2 which was calibrated against

Hunter standard tile (c2-20219 (L = 91.93, a = -0.8 and b = -1.0). Hunter

lightness (L), redness (a) and yellowness (b) were recorded.

3.7 Oxidative Rancidity

The oxidative rancidity of fresh and treated samples were determind

using 2- thiobarbituric acid (TBA) method as described by Hoyland and

Taylor (1989). The method employed was as follows:

A - Preparation of Samples

To a 3g of fish sample 50 ml of distilled water were added and mixed,

the mixture was homogenized in a micro-blender jar for 5 minutes.

The homogenized sample was transferred to Kjeldhal flask and the

blender jar was washed with 47.5 ml of distilled water to remove all

the remaining of the homogenized sample. After addition of 2.5 ml of

4N HCl, the Kjeldhal flask was put on distillation apparatus and

heated at high temperature after the addition of anti-foaming granules.

A 50ml was collected by distillation.

B- TBA Preparation

To 0.2884 g of TBA 100 ml of glacial acid was added and heated

till TBA dissolved. 5 ml of this mixture was added to 5ml of the

samples.

C- Blank Prepration

The blank solution was prepared from 5 ml of distilled water plus 5

ml TBA solution.
D- Final Preparation of the Sample Solution

The flask containing sample solution (5 ml of the sample +5 ml of

TBA) was covered with aluminum foil, of a water bath for 35 minutes,

cooled for 10 minutes and the reading of oxidative rancidity was taken

using a spectrophotometer at the wave- length of 538 nm.

Calculation

7.8
Oxidative Rancidity (mg/ml) = Spectrophotometric Reading

sample weight (g)

3.8 Organoleptic Analysis

The final products of the processed sample of different

methods were assessed subjectively for product

acceptability. The samples of salted fermented fish treatment

were rated for colour, texture, odour and juiciness. Sensory

attributes were evaluated using four scale points, very

acceptable, acceptable, slightly acceptable and unacceptable.

3.9 Microbial Groups Studies

3.9.1 Total Viable Bacterial Count

 Cruckshank, (1975) methods is used to count the total viable

bacteria. The inoculum is deposited as drops from a calibrated dropping

pipette. Each drop, 0.02 ml in volume, is allowed to fall from a height of

2.5 cm onto the medium, where it spread over an area of 1.5-2 cm

diameter. Each of six plates receives one drop of each dilution in separate

numbered sectors. Counts are made in the drop areas showing the largest

number of colonies without confluence (up to 20 or more), the mean of

the six counts gives the viable count per 0.02 ml of the dilution.

3.9.2 Isolation and Identification of the Colonies Culture

Colonies to be identified were picked from the Nutrient Agar (N.A)
and Blood Agar (B.A). plated and purfied. Standared methods were used
for microscopic examination and biochemical tests, according to
Cruckshank, (1975)
3.10 Statistical Analysis
The data obtained were analyzed as a completely randomized
design (ANOVA one and two way) and the means were tested for
significance using Duncan Multiple range test as described by Statistical
Package for Social Science (SPSS Softword (Vearsion 11)).
 CHAPTER IV

RESULTS

The results derived from the examination of tables and other

illustration included in this section are given in the following:

4.1 Biochemical composition

The chemical composition tests were carried out to determine
protein (C.P), crude fat (C.F), ether extract (E.E), ash, dry matter
(D.M), moisture content (Moist.) and some minerals (P, Ca, Fe, Na,
K, and Cu) values for fresh and treated samples of Kawara (Alestes
sp.) and Debs (Labeo sp.) during three seasons.

The biochemical composition of the fresh samples of both

species (Kawara and Debs) over the three seasons are comparable in
both contents and values (Table 1).
The values obtained in Table 1 change in varying degrees when
subjected to salting at varying concentration during short and
prolonged times and seasons (Tables 2 to 7).

Two-way analysis of variance (appendices 13-16) was

performed to verify the significance of the effect of salt, sp., days and
their interaction on crude protein, crude fiber, ether extract, ash, dry
matter, moisture content and some minerals during different seasons.
From these appendices tables the following can be pointed out:
 1- Varying degree of significance (P 0.05 P0.00) were reported
with respect to correlated models, salt concentration, days,
type.
2- Significantly (P< 0.00) correlated interaction was found
between types and days in some correlated parameters
(appendices table 22 and 30).

4.2 General magnitude change between fresh and treated samples

The general magnitude of change between fresh and treated

material can be illustrated by a summary table (Table 8) for both species

(Kawara and Debs). Through correlation of the status of level of the

parameter as attached with the varying salt concentrations.

Table No. (1): Chemical composition of fresh samples during all season of experiments.

Parameters

Seasons Species
C.P C.F E.E Ash D.M Moist P Ca Fe Na k Cu

5.03
Kw 17.45b 1.05 1.63 1.05 29.60 70.41b 1.35 8.20b 56.50 81.00 5.53
5.03
Ds 18.37 1.19 1.47 0.67 21.74 77.90 1.57 8.57 55.33b 176.57b 5.10
Summer 0.20
SE .4583 7.16 8.33 0.19 3.92 3.74 0.10 0.18 0.58 2.21 0.21
NS
Sig. * NS NS NS NS * NS * * * NS
5.33
Kw 19.47 1.31 1.73 1.76 31.42 86.97 1.57 8.47 58.67 186.90 5.63
5.17
Ds 18.90 1.53 1.60 1.42 25.13 74.87 1.73 8.77 57.67b 181.33b 5.50
Autumn 8.33
SE 0.28 0.11 6.66 0.17 3.14 6.05 8.33 0.15 0.50 2.78 6.66
NS
Sig. * NS NS NS NS NS NS * * * NS
K 20.57 1.40 1.53 1.40 33.62 66.63b 1.53 5.63 56.67b 183.80b 5.57 5.33
D 19.77 1.47 1.37 1.60 26.10 72.70 1.60 8.67 66.00 187.67 5.60 5.30
Winter SE 0.40 3.33 8.33 0.10 3.76 3.03 3.33 1.51 4.66 1.93 1.66 1.66
Sig. * NS NS NS NS * NS NS * * NS NS

In this and subsequent tables means within the same column followed by different superscript are significantly
different (p<0.05).
* Significant at 5% level
NS not significant.
Legend:
C.P. = Crude protein Ca = Calcium k= potassium
C.F. = Crude fat Fe = iron Kw= Kawara
E.E. =Ether extract Na =Sodium Ds=Debs
D.M.=Dry matter Cu= Cobalt
P = phosphorus SE= Standard error

57
 Table No. (2): The effect of Salt concentration and time on C.P, C.F, E.E, Ash, D.M and Moist. of Kawara & Debs during summer.
C.P C.F E.E Ash D.M Moisture
Days

SP.

___Salt con._____ ___Salt con._____ ___Salt con._____ ___Salt con._____ ___Salt con._____ ___Salt con._____

20% 25% 30% SE Sig.

20% 25% 30% SE Sig. 20% 25% 30% SE Sig 20% 25% 30% SE Sig. 20% 25% 30% SE Sig. 20% 25% 30% SE Sig

4D Kw 18.67 19.27 18.53 0.23 NS 0.77 0.60 0.50 7.77 NS 1.60 1.37 1.10 0.44 NS 10.45 11.42 11.27 0.30 NS 41.66 41.61 46.33 1.56 NS 58.34 58.39 54.00 1.45 NS

Ds 17.5 17.40 16.33 0.39 NS 0.65 0.67 0.63 9.62 NS 1.30 1.23 1.33 2.94 NS 11.57 15.67 11.02 1.47 NS 35.26 38.67 37.29 0.99 NS 64.74 61.30 62.77 0.99 NS

8D Kw 18.33 19.13 18.03 0.33 NS 0.70 0.57 0.47 6.75 NS 1.47 1.30 1.00 0.14 NS 10.5 11.39 11.72 0.36 NS 44.87 42.59 39.97 1.41 NS 58.46 57.41 60.03 0.76 NS

Ds 15.77 15.57 17.87 0.73 NS 0.50 0.76 0.50 8.89 NS 1.26 1.23 1.10 5.09 NS 14.47 14.30 12.11 0.76 NS 41.95 38.83 41.19 0.94 NS 58.04 61.26 58.81 0.97 NS

12D Kw 18.2 18.37 17.70 0.20 NS 0.80 0.63 0.53 7.78 NS 1.63 1.48 1.20 0.13 NS 13.07 11.42 11.55 0.53 NS 40.54 41.79 41.20 0.36 NS 59.46 58.14 58.79 0.37 NS

Ds 15.70 15.93 15.12 0.23 NS 0.46 0.36 0.33 4.00 NS 1.03 1.00 0.9 4.00 NS 11.29 13.43 12.66 0.62 NS 57.56 54.78 56.41 0.80 NS 42.44 45.21 43.58 0.80 NS

1M Kw 15.33 16.76 17.62 0.66 NS 0.40 0.50 0.56 4.84 NS 1.43 1.13 0.90 0.15 NS 13.87 13.65 13.01 0.25 NS 36.15 34.97 42.93 2.48 NS 60.84 67.06 57.06 2.91 NS

Ds - 14.63 13.50 0.14 NS - 0.43 0.40 0.12 NS - 1.36 1.13 0.11 NS S 13.63 13.25 0.19 NS S 38.27 32.38 2.94 NS S 61.72 67.61 2.94 NS
2M Kw 15.40 16.08 16.25 0.26 NS 0.63 0.40 1.36 8.38 NS 1.20 1.23 1.03 5.55 NS 11.87 14.10 12.67 0.65 NS 42.51 45.21 43.58 0.78 NS 57.56 54.78 56.41 0.80 NS

Ds - 11.92 11.25 0.33 NS - 0.30 0.20 500 NS - 0.9 0.43 0.23 NS S 12.54 12.90 0.18 NS S 41.81 42.81 9.83 NS S 58.22 57.72 0.25 NS
3M Kw 15.70 15.93 15.12 0.23 NS 0.46 0.36 0.33 400 NS 1.03 1.00 0.90 400 NS 11.29 3.43 12.66 0.62 NS 57.56 54.78 56.41 0.80 NS 42.44 45.21 43.58 0.80 NS

Ds - 11.23 11.50 0.13 NS - 0.26 0.23 1.66 NS - 0.73 0.63 5.00 NS S 12.55 12.06 0.24 NS S 52.28 58.41 3.06 NS S 47.72 41.58 3.06 NS
4M Kw 14.67 16.34 14.58 0.57 NS 0.33 0.26 0.23 2.94 NS 0.76 0.86 0.70 4.84 NS 11.48c 13.77 12.84 0.66 * 54.24 53.90 55.24 0.40 NS 45.76 46.10 44.75 0.40 NS

Ds - 11.03 11.10 3.50 NS - 0.16 0.13 1.66 NS - 0.46 0.36 5.00 NS S 13.70 13.01 0.34 * S 58.66 61.64 1.48 NS S 41.33 38.38 1.47 NS
5M Kw - 14.62 14.49 6.50 NS - 0.16 0.13 1.66 NS - 0.50 0.53 .66 NS S 12.39 12.51 6.16 NS S 65.73 65.53 0.10 NS S 34.29 31.44 1.42 NS

Ds - 10.45 10.41 2.00 NS - 0.16 0.18 0.11 NS - 0.23 0.13 5.00 NS S 12.18 12.15 1.50 NS S 66.93 67.51 6.20 NS S 33.06 32.48 0.29 NS

6M Kw - 13.74 13.36 0.19 NS - 0.20b 0.13c 3.33a * - 0.33 0.13 0.10 NS S 11.66 11.60 3.00 NS S 72.34 74.84 1.25 NS S 27.65 25.15 1.25 NS

Ds - - 10.07 - NS - - 0.15 - * - - 0.23 - NS S S S 11.82 NS S S 70.31 - NS S S 29.86 - NS

58
Table No. (3): The effect of Salt concentration and time on P, Ca, Fe, Na, K, and Cu. of Kawara & Deds during summer.
P Ca Fe Na K Cu

___Salt con._____ ___Salt con._____ ___Salt con._____ ___Salt con._____ ___Salt con._____ ___Salt con._____
Species
Days

20% 25% 30% SE Sig. 20% 25% 30% SE Sig 20% 25% 30% SE Sig. 20% 25% 30% SE Sig. 20% 25% 30% SE Sig 20% 25% 30% SE Sig.

4D K 1.6 1.47 1.27 9.69 NS 8.62 8.20 7.93 8.25 NS 54.33 49.67 47.67 1.98 NS 274.33 403.00 560.00 82.60 NS 5.47 5.17 5.1 0.11 NS 4.07 4.37 4.43 0.11 NS

D 1.33 1.23 1.10 6.75 NS 7.90 7.87 7.57 0.11 NS 52.33 57.67 59.33 2.11 NS 375.33 542.67 723.33 100.48 NS 8.57 6.43 7.83 0.44 NS 4.70 4.46 4.20 0.14 NS
8D K 1.57 1.33 1.17 0.12 NS 8.50 8.07 7.80 0.20 NS 53.00 48.33 45.67 2.14 NS 345.33c 501.67b 638.33 84.64 * 5.33 5.10 5.03 9.09 NS 3.90 4.23 4.33 0.13 NS

D 1.16 1.17 1.03 4.44 NS 7.76 7.63 7.56 5.87 NS 54.67 59.67 61.66 2.08 NS 359.66c 556.67b 773.00 119.36 * 6.20 6.46 6.26 8.01 NS 4.60 4.30 4.03 0.16 NS
12D K 1.30 1.20b 1.03b 7.77 * 7.75 7.43b 7.40b 0.1 * 52.50 44.67 42.33 3.07 NS 335.00c 506.00b 680.67 99.78 * 5.23 4.96 4.83 0.11 NS 3.96 4.13 4.33 0.10 NS

D 1.10b 1.12 0.90b 6.66 * 7.10 6.83b 6.80b 0.10 * 43.00 39.00 38.00 1.52 NS 220.00c 416.00b 566.66 100.22 * 4.43 4.13 4.03 0.12 NS 3.90 3.60 3.53 0.11 NS
1M K 1.37 1.27 1.00 0.10 NS 7.23 7.37 7.53 8.58 NS 51.33 59.00 63.00 3.42 NS 327.33 521.00 720.00 113.35 NS 6.03 5.90 5.50 0.16 NS 4.53 4.20 4.03 0.14 *

D S 1.36 1.26 5.00 NS S 7.10 7.16 3.33 NS S 50.66 54.66 2.00 NS S 319.00 515.00 98.00 NS S 6.23 6.11 5.00 NS S 4.30 4.10 1.45 *
2M K 1.26 1.36 1.10 7.77 NS 7.50 7.16 7.13 0.11 NS 49.00 41.00 39.00 3.05 NS 302.66c 496.00b 673.00 106.94* 4.93 4.63 4.84 3.33 NS 3.76 3.93 4.30 0.15 *

D S 1.03 0.83 0.10 NS S 7.10 7.20 5.00 NS S 51.66 57.00 2.66 NS S 479.33b 675.66 98.16* S 5.60 5.20 0.20 NS S 4.06 4.03 1.66 *
3M K 1.10 1.11 0.90 6.66 NS 7.13 6.83 6.80 0.10 NS 43.00 39.00 38.00 1.52 NS 220.0c 410.00b 566.66 100.22* 4.43 4.13 4.03 0.12 NS 3.90 3.60 3.53 0.11 NS

D S 0.76 0.70 3.33 NS S 6.86 7.1 0.13 NS S 44.33 49.66 2.66 NS S 399.00b 600.00 100.50* S 5.76 4.46 0.65 NS S 3.76 3.60 8.33 NS
4M K 0.93 0.96 0.83 4.00 NS 7.06 6.83 6.53 0.15 NS 40.00 36.00 36.33 1.28 NS 202.22c 397.66b 549.33 100.33* 4.06 3.90 3.70 0.10 NS 3.73 3.33 3.23 0.15 NS

D S 0.53 0.46 3.33 NS S 6.50 6.30 8.3 NS S 38.00 41.33 1.66 NS S 310.33b 565.00 127.33* S 4.23 4.40 8.33 NS S 3.13 3.10 1.66 NS
5M K S 0.66 0.62 0.01 NS S 6.10 5.92 6.66 NS S 30.00 27.00 1.33 NS S 351.33b 508.33 78.50* S 3.46 3.13 0.16 NS S 3.13 3.23 5.00 NS

D S 0.40 0.33 3.33 NS S 6.16 6.12 0.11 NS S 30.66 28.61 1.10 NS S 264.33b 501.00 118.33* S 4.10 3.83 0.13 NS S 2.80 2.56 0.12 NS
6M K S 0.56 0.46 5.00 NS S 5.80 5.60 0.10 NS S 25.00 26.00 0.50 NS S 334.66 493.00 79.16 NS S 3.10 3.03 3.33 NS S 2.70 2.63 3.33 NS

D S S 0.16 - NS S S 5.73 - NS S S 21.33 - NS S S 237.66 - NS S S 3.40 - NS S S 1.90 - NS

k=Kawara D=Day
D=Debs M=Month

59
Table No. (4): The effect of Salt concentration and time in C.P, C.F, E.E, Ash, D.M and Moist. Of Kawara & Debs SP. during autumn.

C.P C.F E.E Ash D.M Moistuse

___Salt con._____ ___Salt con._____ ___Salt con._____ ___Salt con._____ ___Salt con._____
___Salt con._____
species
Days

20% 25% 30% SE sig.

20% 25% 30% SE Sig.
20% 25% 30% SE Sig 20% 25% 30% SE Sig. 20% 25% 30% SE Sig. 20% 25% 30% SE Sig
4D K 19.00 18.63 18.20 0.23 NS 0.93a 0.90ab 0.80ab 4.00 * 1.63 1.50 1.20 0.12 NS 10.77 10.77 11.72 11.65 NS 40.10 40.73 40.93 0.25 NS 59.90 59.26 59.06 0.25 NS

D 18.03 18.70 18.53 0.20 NS 1.00a 0.90ab 0.80ab 5.77 * 1.43 1.30 1.20 6.75 NS 11.30 12.20 13.06 0.50 NS 33.65 34.81 37.05 0.99 NS 66.34 63.89 62.94 1.01 NS
8D K 18.73 18.36 17.93 0.23 NS 0.83 0.83 0.70 4.44 NS 1.53 1.36 1.20 9.62 NS 10.83 11.83 11.82 0.33 NS 41.10 42.53 42.10 0.42 NS 58.88 57.46 57.90 0.41 NS

D 18.43 18.63 18.33 8.81 NS 0.80 0.70 0.73 2.94 NS 1.33 1.23 1.06 7.77 NS 11.40 12.36 13.23 0.52 NS 39.77 38.52 40.15 0.49 NS 60.22 61.44 59.84 0.48 NS
12D K 18.73 18.36 17.96 0.22 NS 0.63 0.66 0.60 1.92 NS 1.40 1.30 1.10 8.81 NS 11.72 11.94 12.24 0.14 NS 41.10 41.00 41.99 0.31 NS 58.90 58.99 58.00 0.31 NS

D 18.30 18.46 18.16 8.67 NS 0.86 0.80 0.96 4.84 NS 1.06 1.10 0.96 4.00 NS 11.65 12.78 13.21 0.46 NS 40.10 37.84 40.58 0.84 NS 59.89 60.07 59.41 0.19 NS
1M K - 17.90 17.73 8.33 NS - 0.56 0.53 1.66 NS - 1.20 1.06 6.66 NS - 12.72 12.18 0.26 NS - 60.83 59.05 0.88 NS - 39.17 40.91 0.87 NS

D - 18.23 18.06 8.33 NS - 0.76 0.73 1.66 NS - 1.00 1.03 1.66 NS - 13.53 13.56 1.66 NS - 63.55 60.58 1.48 NS - 36.45 39.41 1.48 NS
2M K - 7.22 17.20 0.12 NS - 0.46 0.33 6.66 NS - 1.03 0.96 3.33 NS - 12.35 12.66 0.15 NS - 65.51 63.07 1.22 NS - 34.48 36.92 1.22 NS

D - 18.00 17.83 8.33 NS - 0.66 0.73 3.33 NS - 1.03 1.03 0.00 NS - 13.80 14.00 1.00 NS - 63.38 63.16 0.10 NS - 36.62 36.83 0.10 NS
3M K - 16.50 16.03 0.23 NS - 0.35 0.33 1.22 NS - 0.80a 0.70ab 5.00 * - 13.26 13.33 3.66 NS - 71.25 74.24 1.49 * - 28.74 26.16 1.29 *

D - 17.73 17.56 8.33 NS - 0.63 0.53 5.00 NS - 0.83a 0.70ab 6.66 * - 14.06 14.00 3.33 NS - 71.13 74.04 1.45 * - 28.86 25.95 1.45 *
4M K - 16.10 16.00 5.00 NS - 0.30 0.23 3.33 NS - 0.63 0.60 1.66 NS - 13.78 13.59 9.50 NS - 76.00 77.09 0.54 NS - 24.00 22.90 0.54 NS

D - - 16.90 - NS - - 0.43 - NS - - 0.56 - NS - - 14.16 - NS - - 79.31 - NS - - 20.68 - NS

5M K - - 16.33 - * - - 0.16 - * - - 0.40 - * - - 13.03 - NS - - 82.00 - * - - 14.25 - *

D - - 16.70 - * - - 0.30 - * - - 0.43 - * - - 14.30 - NS - - 83.08 - * - - 16.91 - *

6M K - - 16.23 - * - - 0.13 - * - - 0.30 - * - - 13.03 - NS - - 85.74 - * - - 14.25 - *

D - - 16.30 - * - - 0.26 - * - - 0.23 - * - - 15.63 - NS - - 87.15 - * - - 13.12 - *

60
 Table No (5): The effect of Salt concentration and time on P, Ca, Fe, Na, K, and Cu. of Kawara & Debs during autumn.

P Ca Fe Na K Cu

___Salt con._____ ___Salt con._____ ___Salt con._____ ___Salt con._____ ___Salt con._____ ___Salt con._____
Species
Days

20% 25% 30% SE Sig. 20% 25% 30% SE Sig 20% 25% 30% SE 20% 25% 30% SE Sig. 20% 25% 30% SE Sig 20% 25% 30% SE Sig.
Sig.
4D K 1.70 1.60 1.53 4.84 NS 8.76 8.63 8.56 5.87 NS 57.00a 54.66b 48.66c 2.48 * 391.66 499.00 580.00 54.54 NS 5.63 5.50 5.16 0.13 NS 4.36 4.43 4.40 1.92 NS

D 1.36 1.23 1.16 5.87 NS 8.10 7.96 7.86 6.75 NS 57.66c 56.33b 51.00c 2.03 * 383.00 549.66 726.66 99.22 NS 6.46 6.40 6.40 2.22 NS 4.53 4.43 4.30 6.75 NS
8D K 1.56 1.40 1.36 6.18 NS 8.60 8.43 8.33 7.77 NS 51.33 49.00 45.66 1.64 NS 375.66 466.66 541.66 47.99 NS 5.36 5.33 5.03 0.10 NS 4.13 4.16 4.16 1.11 NS

D 1.23 1.10 1.03 5.87 NS 8.03 7.90 8.13 6.75 NS 55.33 53.66 49.00 1.89 NS 376.66 536.66 713.33 97.22 NS 6.30 6.20 6.16 4.00 NS 4.43 4.30 4.13 8.67 NS
12D K 1.40 1.20 1.20 6.66 NS 8.30 8.20 8.13 4.84 NS 49.00 45.66 46.00 1.05 NS 358.00 456.33 524.66 48.37 NS 5.40 5.30 4.83 0.17 NS 4.03 4.00 4.03 1.11 NS

D 1.20 1.16 1.06 4.00 NS 8.03 8.00 7.90 4.00 NS 52.33 51.66 48.33 1.23 NS 362.33 523.33 699.33 97.31 NS 6.13 6.03 5.96 4.84 NS 4.30 4.23 4.20 2.94 NS
1M K - 167 1.10 3.33 NS - 8.33 7.96 3.33 NS - 44.33 45.66 0.66 NS - 326.67 504.66 89.00 NS - 5.33 4.80 0.11 NS - 3.00 3.73 3.33 NS

D - 0.96 1.03 3.33 NS - 7.90 7.96 3.33 NS - 53.00 51.33 0.83 NS - 486.66 615.00 64.16 NS - 5.96 5.70 0.13 NS - 3.93 3.96 1.66 NS
2M K - 1.03 0.96 3.33 NS - 7.73 7.63 5.00 NS - 42.00 44.33 1.16 NS - 307.66 494.00 93.16 NS - 4.60 4.63 1.63 NS - 3.60 3.65 1.70 NS

D - 0.90 0.93 0.11 NS - 7.90 7.70 0.10 NS - 49.00 46.33 1.33 NS - 463.33 637.66 87.16 NS - 5.50 5.46 1.66 NS - 3.73 3.70 1.66 NS
3M K - 0.80 0.76 1.66 NS - 7.56 7.43 6.66 NS - 35.66 32.66 1.50 NS - 352.00 481.66 64.83 NS - 4.20 4.03 8.33 NS - 3.23 3.13 5.00 NS

D - 0.80 0.60 0.10 NS - 7.36 7.10 0.13 NS - 40.00 38.66 0.66 NS - 413.33 583.33 85.00 NS - 5.03 4.63 0.20 NS - 3.43 3.40 1.66 NS
4M K - 0.70 0.63 3.33 NS - 7.30 7.16 6.66 NS - 31.33 29.00 1.16 NS - 283.33 455.00 85.83 NS - 4.03 3.80 0.11 NS - 2.96 3.10 6.66 NS

D - - 0.40 - NS - - 6.83 - NS - - 32.00 - NS - - 510.66 - NS - - 4.50 - NS - - 2.96 - NS

5M K - - 0.36 - NS - - 6.83 - NS - - 21.66 - NS - - 402.66 - NS - - 3.26 - NS - - 2.60 - NS

D - - 0.266 - NS - - 6.60 - NS - - 29.33 - NS - - 490.33 - NS - - 4.00 - NS - - 2.73 - NS

6M K - - 0.23 - * - - 6.50 - * - - 20.33 - NS - - 384.33 - * - - 3.03 - NS - - 2.43 - *

D - - 0.23 - * - - 6.10 - * - - 28.66 - NS - - 411.33 - * - - 3.53 - NS - - 2.50 - *

 Table No. (6): The effect of Salt concentration and time on C.P, C.F, E.E, Ash, D.M and Moist. of Kawara & Debs
during winter.
C.P C.F E.E Ash D.M Moistuse

___Salt con._____ ___Salt con._____ ___Salt con._____ ___Salt con._____ ___Salt con._____
Species
Days

___Salt con._____

20% 25% 30% SE Sig. 20% 25% 30% SE Sig 20% 25% 30% SE Sig. 20% 25% 30% SE Sig. 20% 25% 30% SE Sig 20% 25% 30% SE Sig.

4D K 20.03 19.90 19.40 0.19 NS 1.40 1.16 1.26 6.75 NS 2.03 1.90 1.83 5.87 NS 10.40 10.10 10.40 0.10 NS 27.97 27.95 20.79 2.39 NS 71.39 72.04 79.19 2.49 NS

D 19.90 19.56 19.66 9.87 NS 1.53 1.50 1.43 2.94 NS 1.66 1.53 1.50 5.09 NS 10.46 10.43 10.63 6.18 NS 29.44 32.27 33.24 1.13 NS 70.55 67.73 66.76 1.13 NS
8D K 19.83 19.73 19.33 0.15 NS 1.30 1.10 1.20 5.77 NS 1.86 1.70 1.70 5.55 NS 10.46 10.20 10.46 8.88 NS 26.78 27.00 20.80 2.02 NS 73.22 73.02 79.19 2.02 NS

D 18.90 18.63 18.80 7.77 NS 1.43 1.33 1.36 2.94 NS 1.43 1.40 1.36 1.92 NS 10.23 10.86 11.10 0.25 NS 28.10 32.64 32.26 1.45 NS 71.89 67.36 67.74 1.45 NS
12D K 19.46 19.20 19.16 9.49 NS 1.20 1.20 1.30 3.33 NS 1.73 1.63 1.60 4.00 NS 10.93 10.80 10.46 0.13 NS 26.52 27.22 21.14 1.91 NS 73.53 72.77 78.88 1.92 NS

D 18.36 18.26 18.06 8.81 NS 1.36 1.30 1.20 4.84 NS 1.40 1.33 1.33 2.22 NS 10.83 11.20 11.26 0.13 NS 29.00 32.70 32.76 1.24 NS 71.00 67.29 67.23 1.24 NS
1M K 19.17 18.96 19.06 5.77 NS 0.76 0.70 0.76 2.22 NS 1.53 1.43 1.40 4.00 NS 11.76 12.06 12.16 0.12 NS 74.76 73.08 79.01 1.76 NS 25.23 26.91 20.95 1.77 NS

D 18.36 18.00 18.06 0.11 NS 1.13 1.16 1.10 1.92 NS 1.20 1.23 1.30 2.94 NS 11.23 11.60 11.60 0.12 NS 29.91 35.10 35.04 1.71 NS 70.08 64.89 64.95 1.71 NS
2M K 19.06 19.03 18.53 0.17 NS 0.53 0.50 0.40 4.00 NS 1.40 1.16 1.13 8.38 NS 12.45 12.47 12.49 9.68 NS 79.38 76.57 80.79 1.23 NS 20.61 23.42 19.21 1.23 NS

D 17.80 17.60 17.66 5.87 NS 0.86 1.00 0.90 4.00 NS 1.00 0.96 0.90 2.94 NS 11.70 12.16 12.20 0.16 NS 75.93 70.25 70.10 1.91 NS 24.07 29.74 29.89 1.91 NS
3M K - 18.20 18.26 18.26 NS - 0.36 0.30 4.33 NS - 0.90 1.00 5.00 NS - 13.35 13.60 0.12 NS - 78.51 81.01 1.25 NS - 21.48 18.98 1.25 NS

D - 17.03 17.16 6.66 NS - 0.90 0.86 1.66 NS - 0.83 0.86 1.66 NS - 12.73 12.93 0.10 NS - 71.77 71.11 0.32 NS - 28.23 28.88 0.32 NS
4M K - 17.76 17.70 3.33 NS - 0.33 0.31 0.12 NS - 0.73 0.83 5.00 NS - 13.31 13.56 0.12 NS - 80.87 82.13 0.63 * - 19.12 17.86 0.63 *

D - 16.93 16.83 5.00 NS - 0.76 0.70 3.33 NS - 0.70 0.63 3.33 NS - 13.30 13.43 6.500 NS - 80.44 81.95 0.75 * - 19.55 18.05 0.75 *
5M K - 17.03 17.16 6.66 NS - 0.26 0.25 0.13 NS - 0.50 0.46 1.66 NS - 13.83 13.66 8.33 NS - 79.29 83.93 2.31 NS - 20.70 16.06 2.31 NS

D - 16.10 16.23 6.66 NS - 0.46 0.36 5.00 NS - 0.30 0.36 3.33 NS - 13.30 13.56 0.13 NS - 83.14 85.96 1.41 NS - 16.86 14.03 1.41 NS
6M K 16.20 16.16 1.66 NS - 0.16 0.23 3.33 NS - 0.36 0.43 3.33 NS - 14.16 14.00 8.33 NS - 84.46 85.16 0.35 NS - 15.53 14.83 0.35 NS

D - 14.56 13.96 0.30 NS - 0.23 0.33 0.13 NS - 0.20 0.16 1.66 NS - 9.93 14.96 2.51 NS - 86.03 86.23 0.41 NS - 13.96 13.99 0.21 NS

1
 Table No. (7): The effect of Salt concentration and time on P, Ca, Fe, Na, K, and Cu. of Kawara & Debs during
Winter.
P Ca Fe Na K Cu

___Salt con._____ ___Salt con._____ ___Salt con._____ ___Salt con._____ ___Salt con._____ ___Salt con._____
Species
Days

20% 25% 30% SE Sig. 20% 25% 30% SE Sig 20% 25% 30% SE Sig. 20% 25% 30% SE Sig. 20% 25% 30% SE Sig 20% 25% 30% SE Sig.

K 1.73 1.70 1.70 1.11 NS 8.83 8.66 8.76 4.84 NS 61.66 62.33 60.66 0.48 NS 360.00c 507.66b 664.33a 87.86 * 5.70 5.50 5.50 6.66 NS 4.93 4.90 4.83 2.94 NS
4D
D 1.50 1.43 1.40 2.94 NS 8.16 8.13 8.03 4.00 NS 66.33 63.33 61.66 1.36 NS 386.66c 580.00b 764.00a 108.9 * 6.43 6.26 6.23 6.18 NS 4.63 4.50 4.33 8.67 NS
K 1.63 1.56 1.70 3.84 NS 8.73 8.70 8.73 1.11 NS 61.33 61.00 59.66 0.50 NS 355.00c 505.33b 662.00a 88.62 * 5.46 5.30 5.20 7.77 NS 4.86 4.70 4.70 5.55 NS
8D
D 1.40 1.36 1.36 1.11 NS 8.00 8.26 7.90 0.10 NS 64.33 61.66 60.00 1.26 NS 378.66c 577.66b 758.66a 109.73 * 6.23 6.13 6.06 4.84 NS 4.36 4.40 4.20 6.18 NS
K 1.43 1.40 1.30 4.00 NS 8.40 8.16 8.30 6.75 NS 57.33 56.33 56.00 0.40 NS 322.66c 503.66b 652.66a 95.41 * 5.30 5.20 5.16 4.00 NS 4.60 4.46 4.43 5.09 NS
12D
D 1.30 1.23 1.26 1.92 NS 8.06 8.10 8.60 0.17 NS 61.00 59.00 57.66 0.96 NS 361.66c 528.00b 750.00a 112.48* 5.90 5.83 6.00 4.84 NS 4.03 3.93 3.96 2.94 NS
K 1.30 1.33 1.40 2.94 NS 8.10 8.13 8.03 2.94 NS 52.66 51.00 50.33 0.69 NS 305.00c 495.66b 610.00a 88.96 * 5.00 4.93 4.96 1.92 NS 4.30 4.20 4.20 3.33 NS
1M
D 1.23 1.20 1.10 4.00 NS 8.10 8.13 7.83 9.49 NS 59.00 57.33 55.66 0.96 NS 357.00c 531.33b 728.33a 107.26 * 5.80 5.73 5.70 2.94 NS 4.00 3.86 3.86 4.44 NS
K 1.10 1.20 1.03 4.84 NS 8.10 8.26 7.90 0.10 NS 49.00 48.33 47.00 0.58 NS 294.00c 500.00b 604.00 a 91.08 * 4.76 4.70 4.73 1.92 NS 4.03 3.90 4.03 4.44 NS
2M
D 1.06 1.03 1.03 1.11 NS 7.70 7.53 7.56 5.09 NS 52.33 53.66 53.66 0.44 NS 351.33c 524.33b 706.33a 102.49 * 5.46 5.43 5.40 1.92 NS 3.40 3.50 3.36 4.00 NS
K - 1.03 0.93 5.00 NS - 7.83 7.70 6.66 NS - 43.33 42.66 0.33 NS - 483.33b 649.66a 83.16 * - 4.40 4.40 0.00 NS - 3.60 3.73 6.66 NS
3M
D - 0.90 0.83 3.33 NS - 7.16 7.03 6.66 NS - 50.00 50.12 0.22 NS - 504.33b 690.00a 92.83 * - 4.96 5.06 5.00 NS - 3.16 3.33 8.33 NS
K - 0.83 0.70 6.66 NS - 7.76 7.56 0.10 NS - 38.00 40.00 1.00 NS - 434.00 508.00 37.00 NS - 4.03 4.00 1.66 NS - 3.33 3.60 0.13 NS
4M
D - 0.73 0.70 1.66 NS - 6.93 6.86 3.33 NS - 47.66 46.33 0.66 NS - 559.66 662.00 51.16 NS - 4.73 4.70 1.66 NS - 3.03 3.16 6.66 NS
K - 0.43 0.46 1.66 NS - 7.53 7.26 0.13 NS - 34.66 31.66 1.50 NS - 410.66 490.00 39.66 NS - 3.43 3.42 0.12 NS - 3.16 3.40 0.11 NS
5M
D - 0.50 0.40 5.00 NS - 6.43 6.73 0.15 NS - 38.00 38.03 0.11 NS - 406.66 608.66 101.00 NS - 4.10 4.23 6.66 NS - 2.80 3.00 0.10 NS

K - 0.36 0.43 3.33 NS - 7.50 7.23 0.13 NS - 33.66 31.00 1.33 NS - 396.00 475.66 39.83 NS - 3.06 3.33 0.13 NS - 3.03 3.30 0.13 NS
6M
D - 0.43 0.36 3.33 NS - 6.36 6.96 0.3 0 NS - 35.66 36.00 0.16 NS - 392.00 588.33 98.16 NS - 4.17 4.36 9.50 NS - 2.70 2.83 6.66 NS

2
 Table 8 The magnitude of change between fresh and treated materials during seasons

Parameters
Season __________________
Sampl
es C.P C.F E.E Ash DM Moist. P Ca Fe K Cu
F.K. 17.45 1.05 1.63 1.05 29.60 70.41 1.35 8.20 56.50 81.00 5.53
T.K. 16.53 0.49 1.15 13.51 73.59 26.40 1.24 7.38 51.78 5.18 4.25
Summer
F.D. 18.37 1.19 1.47 0.67 21.74 77.90 1.57 8.57 55.33 176.57 5.10
T.D. 14.07 0.41 1.25 13.44 70.13 29.86 1.31 7.15 54.33 6.17 4.2
F.K. 58.6
T.K. 19.47 1.31 1.73 1.76 31.42 86.97 1.57 8.47 186.90 5.63
17.81 0.55 1.13 12.45 59.94 40.04 1.39 8.15 7 5.07 3.37
Autumn
45.0
F.D. 57.6
T.D. 18.90 1.53 1.60 1.42 25.13 74.87 1.73 8.77 181.33 5.50
18.15 0.74 1.02 13.55 62.07 37.93 1.00 7.93 7 5.83 3.95
52.17
F.K. 20.57 1.40 1.53 1.40 33.62 66.63 1.53 5.63 56.67 183.80 5.57
T.K. 19.06 0.74 1.45 11.99 75.62 24.36 1.34 8.09 51.33 4.96 4.23
Winter
F.D. 19.77 1.47 1.37 1.60 26.10 72.70 1.60 8.67 66.00 187.67 5.60
T.D. 18.14 1.13 1.24 11.48 33.35 66.64 1.18 8.02 57.33 5.74 3.91

F.K.= Fresh Kawara

F.D.=Fresh Debs
T.K.= Treated Kawara
T.D.= Treated Debs

3
4.3 Effect of the quality of the treated samples

The given tables 2-7 can be illustrated by a number of observation

such as:

1- The findings obtained during the first 4, 8, and 12 days show a

significant variations in the enlisted parameters in terms of time

and per species, but in generals times the quality is kept within

comparable levels.

2- During the first levels before gradual deterioration starts toward the

end of the sixth month which marks the onset for further losses in

quality.

This fact can be further illustrated by the fate of crude protein of

both species during summer, autumn and winter seasons as in Figs 1-

3.Further findings attached to oxidative rancidity, yellowness are

given in Figs 4-7.

Oxidative rancidity, lightness, redness and yellowness were tested

for statistical significant by two-way analysis of variance. The effect

of days, type of fish, salt and their interaction on these physical

parameters were found significantly (P<0.05) in corrected model, type

and salt (appendices table 17, 18, 25 and 26).

 Correlated interaction between type and days was only found

significantly (P<0.05) during summer and winter seasons in some

physical parameters (appendices tables 18 and 26).

1
Fig. (1) Crude protein of Kawara and Debs during summer season
(during the first 12 days (D) and subsequent 6 month (M)).

25
20
15

%10
5
0
1M

2M

2M

3M

4M

4M

5M

6M
4

8
12

12

Time
Kawara sp. Debs sp.

Fig. (2) Crude protein of Kawara and Debs during autumn season

25
20
15
%
10
5
0
4
4
8
12
12

M
M
M
M
M
M
M
M
1
2
2
3
4
4
5
6

Time
Kawara sp. Debs sp.

2
Fig. (3) Crude protein of Kawara and Debs during winter season

25
20
15
10
%5
0
4

1M

2M

2M

3M

4M

5M

6M
12

12

Kawara sp. Debs sp. Time

Fig. (4) Oxidative Rancidity of Kawara during seasons

4
3
2
%1

0
1M

2M
2M

3M

4M
4M

5M

6M
4

4
8
12
12

Time
Kawara sp. in summer Kawara sp. in autumn
Kawara sp. in winter

3
Fig. (5) Oxidative Rancidity of Debs during seasons

6
5
4
%3
2
1
0

1M

2M

3M

4M

5M
4

8
12

12
Time
Debs sp. in summer Debs sp.in autuomn
Debs sp. in winter

Fig. (6) Yellowness of Kawara during seasons

20

15

10
%
5

0
12

12
4

1M

2M

2M

3M

4M

4M

5M

6M

Kawara sp. in summer Tim

Kawara sp. in autumn
Kawara sp. in winter

4
 Fig. (7) Yellowness of Debs during seasons

20

15

10

% 5
0
Time
4

8
12

12

1M

2M

3M

4M

5M

Debs sp. in summer Debs in autumn

Debs sp. in winter

5
 Table 9 Summary of levels of chemical parameters obtained at
different salt concentrations for the two species in the different
seasons (tables 2-7)

Debs
Seasons Kawara

High levels are obtained at 20% High levels are obtained at 20%
concentration for C.F, E.E, Ash, P, concentration for D.M, Moisture, Ca, K
Summer and Cu.
Ca, Fe and K.
At 25% for C.P and 30% for Moisture, D.M, At 25% for C.F, Ash, P and Fe
Cu and Na At 30% for C.P, E.E and Na

High levels are obtained at 20% High level are obtained at 20%
Autumn concentration for C.P, C.F, Ash, moisture, concentration for D.M, Moisture, Ash, P,
P, Ca, Fe, k &Cu
At 25% for E.E & Cu At 25% for C.P
At 30% for C.F & Na At 30% for C.F, E.E, Ca, Na.

High levels are obtained at 20% High levels are obtained at 20%
concentration for CP, DM, Moisture, CF, concentration for D.M, Moisture, Ash, P, Fe,
EE, P, Ca, K & Cu. K & cu.
Winter
At 25% for Fe At 30% for C.P, C.F, E.E, Ca & Na.
At 30% for Ash & Na

6
4.4 Total viable bacterial counts

The total viable bacterial counts of fresh sample throughout

selected structures and whole fish indicate differences between the two

species over the three seasons (Table 10). In fresh samples, 16 species of

bacteria were found during the three seasons (Table 11). Staphylococcus

gallinarum was found in all organs in debs sp. during autumn, while

E.coli found during winter season (Table 11).

In the treated material the occurrence of bacteria is closely correlated

with the salt concentration on the one hand and both preliminary

treatment during the first 12 days as well as the prolonged phase lasting

six month (Tables 11,12,13).

There are some bacterial species found during the three seasons of study:

- In summer season Staphylococcus lentus was the dominant bacteria

found in kawara sp. during the first days of treatment at 20% salt

concentration, while Staphylococcus xylosus found at 25% and

30% salt concentration during (Table 12).

- For debs sp. Staphylococcus Caseolyticus and Staphylococcus

auricularis were found at 25% and 30% respectively (Table 12).

- In autumn season Staphylococcus gallinarum and Staphylococcus

Saprophyticus were the dominant bacteria found at different salt

concentration (Table 13).

7
 - In winter season Stomatococcus, Staphylococcus gallinarum,

Staphylococcus xylosus and Staphylococcus Saprophyticus were

found during the first days of treatment at different salt

concentration for two species (Table 14).

Total viable bacterial count was subjected to two-way analysis of

variance for statistical significance. The effect of salt, type and their

interaction found significantly (P<0.05) during summer and autumn

seasons (appendices tables 19 and 27). Significantly (P<0.05) correlated

interaction was found between salt and days during winter season

(appendix table 34).

8
Table No. (10): Total Viable Bacteria count of fresh samples during
all season of experiments.

Parameters
Seasons Species
Gill Gut Muscle Skin Whole
K 1900.00 2850.00 70.33 1860.00 1516.67
D 3176.67 3813.33 756.67 2756.67 990.00
Summer
t 3.98 6.92 1.21 5.15 4.76
Sig. 0.15 0.09 0.44 0.12 0.13
K 8633.33 8000.00 3533.33 28666.67 6766.67
D 2000.00 4700.00 396.67 33666.67 560.00
Autumn
t 1.60 3.85 1.25 12.47 1.18
Sig. 0.36 0.16 0.43 0.51 0.44
K 363.33 470.00 16.33 983.33 2283.33
D 280.00 17.67 1540.00 3513.33 4336.67
Winter
t 7.72 1.08 1.02 1.78 3.22
Sig. 0.08 0.48 0.49 0.33 0.19

9
Table No. (11): Bacteria species found in fresh samples during different seasons

Seasons Kawara Debs

Gill Gut Muscle Skin whole Gill Gut Muscle Skin whole
Summer 1+3 16 4 7 1+4 4+1 5 4+12 4+14 4+1

Autumn 4+6+1 6+1 4+6+1 4+6 4 4 4+6 4 4+13 4

Winter 5 10 7 9+11 10+7 5+7 5 5+7 5+7 5+7

1=Stomatococcus 8=Enterobactor agglomenerus 15= Micrococcus variaus

2= Staphylococcus
3= Staphylococcus. capitis
4= Staphylococcus gallinarum
5=Escherichia coli
6= Corynebacterium
7= Staphylococcus caprae
9= Enterobacter cloacae - = Spoilage
10 = Enterobacter agglomenerus
11= Staphylococcus auricularis
12= Staphylococcus equorum
13= Staphylococcus schleifor
14= Staphylococcus caseolyticus
 Table No. (12): Bacterial found in treated samples in summer season

Kawara
Debs
Season Days
20% 25% 30%
20% 25% 30%
4D 17 + 3 + 18 17 + 16 + 18 17 + 23 4+ 6 4+ 11 17 + 11
8D 18 + 27 17 23 17 + 18 + 20 20+ 11 20+ 17 + 21
12 D 18 + 17 17 + 21 12 + 21 4+ 17 + 20 18 + 21 18 + 11
1M 4 + 33 17 + 21 12+ 21 22 + 12 21 21
Summer 2M 21 17 + 21 12+ 21 - 21 21 + 11
3M 4 21 4 + 21 - 21 11
4M 6 19 4 - 19+ 4 + 21 21
5M - 19 19+ 4 - 21 4+ 11
6M - 19 19+ 4 - 21 4+ 11

17= Staphelococcus xylosus 23= Staphelococcus sciuri

18= Staphelococcus lentus
19= Staphelococcus saprophyticus
20= Micrococcus.lylae
21= Staphelococcus caseolyticus
22= Staphelococcus cohnii

1
 Table No. (13): Bacterial found in treated samples in autumn season

Kawara Debs
______________________________________
Da ________________________________
Season
ys 20% 25%
20% 25% 30%
30%
4D 4 C+S 4 4 4 4
8D 4+ 23 + 19 4+ 23 + 19 4+ 23 i + 19 4+ 23 + 19 4+ 23 + 19 4+ 23 + 19
12 D 4+ 23 + 19 4+ 23 + 19 4+ 19 4+ 23 + 19 4+ 23 + 19 4+ 23 + 19
1M 4+ 12 + 19 4+ 12 + 19 4+ 12 + 19 - 4+ 12 + 19 12 + 19
Autum
2M 19 4+ 19 - - 12 + 19 19
n 3M - Nill S - 24 25
4M - Nill Nill - 24 25
5M - 12 + 23 12 + 7 - 12 12E + 23
6M - - S+ 7 - - -

24 = Micrococcus lutes
25 = Aerococcus

2
 Table No. (14): Bacterial found in treated samples in winter season

Day Kawara Debs

Season
s ________________________________ ______________________________________
20% 25% 30% 20% 25% 30%
4D 1 + 20 + 17 + 4 1 + 20l + 17 1+ 20 + 17 1 + 20 + 17 + 4 1 + 20 + 17 + 4 1+ 20l + 17 + 4
8D 1+ 20 + 17 + 4 1+ 20 + 17 + 4 1+ 20 + 17 + 4 1 + 20 + 17 + 4 1 + 20 + 17 + 4 1 + 20 + 17 + 4
12 D 4 + 12 + 23 4 + 12 + 7 4 + 12 + 7 4 + 12 + 7 4 + 12 + 7 4 + 12 + 7 + 23
1M 12 + 7 12 + 7 121 + 7 12 + 7 + 23 12 + 7 + 23 7 + 23
Autumn 2M 24 + 12 1 + 24 + 12 1 + 24 + 12 1 + 26+ 12 + 7 26 + 12 6+ 12
3M - 4 12 - 12 12
4M - 12 12 - 6 25+ 12
5M - Nill 6 - Nill 12
6M - 26+ 12 26+ 12 - 6 + 12 6 + 12

M= Micrococcus

3
 CHAPTER V

DISCUSSION

It is quite pertinent to state that the specific objectives

forwarded in the introduction of the thesis comprise the main issues to

be tackled in the discussion. These objectives were geared to target

the progressive promotion of the traditional treatments so far

practiced, in the direction of adoption of scientific approaches the

determination of the magnitude of losses in quality of fish resulting

from spoilage, the call for adoption of standards and specification

currently applied domestically, regionally, and internationally

wherever appropriate in order to furnish the practitioners with definite

recommendations pertaining to high level preservation.

As regards the promotion of traditional methods, these are

presently partially or completely abandoned through a number of

practices including the sorting out of fish species proposed for

treatments by keeping records of the biochemical composition and

nutritive values of their flesh; determination of the treatments that

suits each; determination of the quality of salt to be used and methods

of application, the calculation of salt concentration that secures the

appropriate preservation level; the onset of spoilage and the fate of the
end products and the subsequent requirements of packaging,

transportation and marketing.

The specifications and standards comprise the proposed

recommendation that can be forwarded to the practitioners related to

freshly treated samples; the impact of salting on flesh quality and the

recommended doses till being availed to the consumer over a

prolonged period of time.

The results so obtained, through carried out on two selected

species while some others have been studied on other species, draw

the attention to a significant fact that encourages extension of

investigation to a wide range of Sudanese fishes as long as there is a

consistency in treatments, unified methodologies and reliable

evaluation.

1
 CHAPTER VI

CONCLUSIONS

The conclusions obtained from this study are outlined in the following:

1- The research has addressed the magnitude of losses resulting from

spoilage that are likely to occur during the post-harvest treatment

by provisions that cut them down to the minimum level through the

adoption of salting and fermentation methods of preservation.

2- The findings so obtained have targeted the progressive promotion

of the traditional treatment in the direction of the following

scientific approaches based on specific parameters such as the salt

quality, salt concentration, fermentation time and impact on the

overall nutritive value.

3- The need for the adoption of standards and specifications at

domestic, regional and international levels is both advocated and

emphasized in order to secure successful competition and

marketing for the products.

4- The findings attached to the selected fish species Debs (Labeo sp.)

and Kawara (Alestes sp.) are indicative of the possibility of wide

application amongst a wide range of fish if practitioners wherever

and whenever so wish to work using the appropriate methods.

2
5- The specific findings are summarized as following:

(a) The biochemical composition of fresh samples change in

varying degrees when treated with different salt

concentrations. (Crude protein of fresh Kawara and Deds

was 19.16 and 19.01%, these percentages was changed in

treated samples to 16.53, 17.81,19.06 for Kawara and

14.07,18.15, 18.14 for Debs during summer, autumn and

winter seasons respectively.)

(b) There is a significant change in nutritive values between

fresh and treated samples. (The caloric content of protein of

fresh Kawara and Debs was 78.56 and 77.94 (4.1 cal per

mg), these calories changed to 67.77 and 57.69 for Kawara

and Debs respectively, when treated with salt during summer

season.)

(c) There are correlations between parameters and different salt

concentrations for the two species in different seasons. (High

level of crude fat obtained at 25%, 30% and 30% for Debs

during summer, autumn and winter seasons respectively,

where the highest level of C.F for Kawara obtained at 20%

during all seasons)

3
 (d) The quality of treated samples keeps up for one month

before the start of deterioration.

(e) Appropriate storage keeps up to 6 month for kawara during

all season, and to 4 month for Debs in summer and autumn,

and to 6 month in winter season.

(f) There are differences in the total viable bacterial count

between two samples of fresh fish during three seasons.

(g) The occurrence of bacteria is closely correlated with the salt

concentration. In summer season the total viable bacterial

count was 756.67 and 11.33 for Kawara at 20 and 30% salt

concentration, where it was 43666.67 and 5466.66 for Debs

at 20 and 30% respectively during 8 days of treatment.

Generally the total viable bacterial count increase with salt

concentration increased during all seasons.

6- Recommendation:

(a) Adoption of practice throughout the fisheries

(b) Circulation of the practice to involve other Sudanese fish

(c) Adoption of standard and specification with respect to salt

quality, concentration and impact on products (based on

panel test).

4
 CHAPTER VII

REFERANCES

A. O. A. C (1990). Official Methods of Analysis, 13th ed. Association of

Official Analytical Chemists, Washington D. C., U.S.A. 1094
pp.

Abdalla, M. T. (1989). Microbiology and biochemistry of fessiekh

fermentation. M. Sc. Thesis, University of Khartoum.

Abu Gideiri, Y. B., Ali, M. E. and Mahamoud, Z. N. (2004). Review of

research on the Nile bulti, Oreocheomis Niloticus (Trewavas)
in Sudan. 43-44 pp.

Abu Gideiri, Omima, O. (2001). Some biochemical and microbiological

aspects of fesseikh industry in Sudan. B. SC. (Hons.)
Dissertation, Department of Zoology, Faculty of Science,
University of Khartoum, Sudan.

Adam, H. M. (1996). Body weight characterstics and physical

composition of some fish species frome lake Nubia. M. Sc.
Thesis, University of Khatoum.

Agab, M. A and Beshir, R. B. (1987). Traditionally salted fermented fish

Fasseikh. Sudan Journal of Food Science and Technology. .
17:

Akamatsu, M. (1959). Bacteriological studies on the spoilage of fish

sausage. 1. Number of bacteria present in the meat of fish
sasuage on the market. Jap. Soc. Sci. fish. Bull.25: 545-548.

Akanda, G. R. and Oladosu, H. O (1988). Production and acceptability

trials of solar dried, minced and salted fish cakes from
Miscellaneous fish species. In: FAO, fisheries report. 1990,
No. 400: 231-238.

5
Allen, J. and Angelo, St. (1991). Lipid oxidation in: Food agriculture and
food chemistry. 111 American Chemical Society (202 nd:
1991) New York. IV series.

Ali, M. E., Babiker, S. A. and Tibin, A. (1996). Body charcterstics, yield

indices and proximate chemical composition of commercial
fish species of Lake Nubia. FAO 6th Consultation on Fish
Technology in Africa. Kisumu, Kenya. 27-30.

Amano, K. (1962). The influence of fermentation on the nutrative value

of fish with special reference to fermented fish products of
south-east Asia. In: Fish Nutrition, ed., E. Heen and R.
Kreuzer, Fishing News (Books), London.180. cited by Mackie,
I. M.; Hardy, R. and Hobbs, G. (1971).FAO. Fisheries report
No.100, Fermented Fish Products.

Anon. (1970). Refrigeration of fish. Part 3. US Department of Interior,

fishery leaflet No. 429.

Anonymous, N. (1982). Report on Thai Traditional Fermented Food

Research Project, National Research Council of Thailand.

Applied Microbiology. 29, 106.

Beatty, S. A. and Fougere, H. (1957). The processing of dried-salted fish.

Bull. No.112, Fish. Res. Bd. Can.

Beddows, C. G. (1985). Fermented fish and fish products: In

Microbiology of Fermented Food. B. Wood, (ed.), J. B. El
Sevier Science Puplishing, New York,2, 1-40.

Beddows, C. G.; Ismail, M. and Steinkraus, K. H. (1976). The use of

bromelain in the hydrolysis of mackerel and the investigation
of fermented fish aromas. Journal of Food Technology.11: 379-
388.

6
Beddows, C. G.; Ardeshir, A. G. and Wan, J. D. (1979). Biochemical
changes occuring during the manfacture of Budu. Journal
Science of Food and Agriculture. 30:1097-103.

Bhumiratana, A. (1980). Traditional fermented foods in Thailand. In:

Proceeding of the Conference on Oriental Fermented Foods,
FIRDI, Taiwan, 58-70.

Boleman, S. J. ; Boleman, S. L. ; Miller, R. K., Taylor, J. F. ; Cross, H.

R. ; Wheeler, T. L ; Koolmaraie, M. ; Shachelford, S. D. ;
Miller, M. F. ; West, R. L. ; Johnson, D. D. and Stavell, J.W.
(1997). Consumer evaluation of beef of known categories of
tenderness. Journal of Animal Science. 75 : 1521-1524.

Boez, L. and Guillerm, J. (1930). Le facteur microbine dan la fabriction

de la saumare indochinoise (nuoc-mam).C.R.Acad. Sci. 190 :
534-535.

Botta, J. R.; Kennedy, K. and Squires, B. E. (1987a). Effect of methods of

catching and time of seasons on composition of atlantic cod
(Gadus morhua). Journal of Food Science. 52: 922-927.

Botta, J. R.; Bonnell, G. Squieres, B.E (1987b). Effect of methods of

catching and time of season on sensory quality of fresh raw
Atlantic cod (Gadus morhua). Journals of Food Science. 52:
928-931.

Chakraborty, P. K. (1978). Technological development of artificial and

solar drying of fish in India. Proc. IPFC, 18(3):322-329.

Christine, P. M.; Lone, G.; Mette, M. and Peter, L. M. (2001).

Fermentation and microflora of plaa-som, a Thai fermented fish

7
 product prepared with different salt concentrations.
International Journal of Food Microbiology. 73: 61-70.

Chrystall, B. B.; Culioli, J.; Demeyer, D.;, Honikel, K. O.; Moller, A. J.;
Purslow, P.; Swagele, F.; Shorthose, R. and Uytterhaegen, L.
(1994). Recommendation of refrence methods for assessment
of meat tenderness. Proc. 40th ICOMST the Hague, the
Netherlands, 6:1-6.

Cole, R. C. (1963). Fish curing industry of Aden. Report of Tropical

Products Institute. (22/63).

Clucas, I. J. and Ward, A. R. (1996). Post-Harvest fisheries development:

A Guide to handling, preservation, processing and quality.
Natural Resources Institute (NRI). U. K.

Compbeel-platt, G. (1987). Fermented foods of the world: A Dictionary

and Guide. Butter-worths, London.

Crisan, E. V. and Sands, A. (1975). The microbiology of four fermented

fish sauces. Applied Microbiology. 29, 106.

Crukkshank ,R.(1975). Medical Microbiology. Vol 2. p307, (ed.) Dugwd,

J. P., Marmiom, B.P, Swain, R.H. and livingstome, Ch.
Academic press. London.

Crutchfield, J. A. (1959). Report in fish markting in Uganda. FAO

Fisheries Technical Paper No. 160.

Del Rosario, R.R. (1980). Traditional filipino fermented foods. In:

Proceedings of the conference on oriental fermented foods,
FIRDI, Taiwan, 71-87.

Desrosier, N. W., and Desrosier, J. N. (1977). Technology of food

preservation, 4th edn.AVI, Westport, Can. 558p.

8
Dirar, H.A. (1993). Indigenous fermented food of the Sudan. Fish product
Fesseikh P 347-366. Faculty of Agriculture, University of
Khartoum. Khartoum. Sudan.

Disney, J. G. (1974). Fish handling and processing at Gwadur, Pakistan

tropical product institute. London- cited by FAO, (1981). The
prevention of losses in cured fish. FAO fisheries Technical
papers, 219, 87.

Doe, P. E., Ahamed, M.; Muslemuddin, M. and Sachithanathan, K.

(1977). A polythene drier for improved sun drying of fish. Food
Technology. 29(11): 437-441.

Doe, P. E.; Curran, C. A. and Poulter, R. J. (1983). Determination of the

water activity and shelf-life of dried fish products. Journal of
Food Technology.17. 202-208.

Dyer, W. J (1974). Stability of minced fish, an important factor in

standard formulation. In second Technical Seminar on
Mechanical Recovery and Utilization of fish flesh. Washington,
DC: National Fisheries Institute.

Eddy, B. P. (1958). Microbiology of fish and meat curing brines. 11.

International Symposium. Food Microbiology proc., London.
AVI publishing Co., Westport, Can.

El Tom, A.M. (1989). Microbiology and Bichemistry of Fessiekh

fermentation. M. Sc. Thesis, Unversity of Khatoum, Sudan.

El-Sebahy, L. A. and Metwalli, M. S. (1988).Changes in some chemical

charactersistics and lipid composition of salted fermented Bouri
fish muscle (Mugil cephalus). Food chemistry. 31: 41-50.

9
Evelyn, T. P. T., and Mc Dermott, L. A. (1961). Bacteriological studies
of fresh water fish. 1. Isolation of aerobic bacteria from several
species of Ontario fish. Canadian Journal of Microbiology.7:,
375-382.

FAO (1962). Fish in Nutrition (eds E.Heen and R. Kreuzer), Fishing

News (Books). London.

FAO (1970). Smoke curing of fish. FAO-Fisheries Report No. (88),43.

FAO (1981). The preservation of losses in cured fish. Technical Paper

No. 219.

FAO (1984). Year-book of Fishery Statistics. Vol.59. Food and

Agriculture Organization of the United Nations, 1984.

FAO (1986). The production of fish meal and oil. Fisheries Technical
Paper.142.

FAO (1989). Solar drying of fish and paddy. FAO, Environment and
Energy Paper, (10).

FAO (1990). Characterization of traditional smoked-dried fish in Nigeria.

A case study, NORFICO, Consultancy report. In: CIFA
Technical paper No. 19. Fish Technology. FAO, 21-22 October
1990.Cairo, Egypt.

FAO. (1992). Fermented fish in Africa. A study on proceesing, marketing

and composition. FAO Fisheries Technical paper. No. 329, 57.

Fey, M. S. and Regenstein, J. M. (1982). Extending shelf life of fresh fish

wet red hake and salmon using Co2-O2 modified atmosphere
and potassium sorbate ice at 0-1C. Journal of Food Fcience. 47:
1048-1054.

Frazier, W. G. (1958). Food Microbiology. 3rd ed. Mcgaraw. Hill Book

Company, London.

10
Gracey, F. H. (1986). The preservation of meat. Thorntons meat hygine.
7th. Ed.. the English language book society and Baciliers
Tindall, London.

Green, B. E (1969). Lipid oxidation and pigment changes in raw beef.

Journal of Food Science. 34: 110.

Hall, R. J. (1989). Application of biotechnology to traditional

fermentation.. In: Biotechnology and the food industry. P. L.
Rogers and G. H. Fleet (eds.). Gordon and Breach Science
Publishers. New York. 241-277

Halleck, F. E., Ball, C.O. and Stier, E. F. (1958). Factors affecting the
quality of prepackaged meat. IV.Microbilogical studies.B.
Effect of package characteristics and of atmospheric pressure in
package upon bacterial flora of meat. Food Technology. 12:
301-306.

Hamed, M. G.; Elias, N. A.; El-Wakeil, F. A. and Foda, I. O. (1973).

Studies on the microorganisms in salted Mullet fish
Feseeiekh. Journal of Food Science. 1: 1-16.

Hamm, R. (1994). Influence of pH on the protein net charge in the

myofibrillar system. Rec. Meat Conf. Proc.47: 5-9.

Hansen, L. T.; Rontved, S.D.; and Huss, H. H. (1998). Microbiology

quality and shelf life of cod-smoked salmon from three
different processing plants. Food Microbiology. 15: 137-150.

Harris, P. and Tall, J. (1994). Substrate specificity of mackerel flesh

lipoxygenase. In: Rancidity in foods. J.C. Allen and R.J.
Hamilton, (eds). (1994). Chapman and Hall plishing.London,3:
257-271.

11
Hassan, Y. M.; Abdel-Al, A.; Awad, A.T. and Awad, A. A. (1972). An
introduction to agricultural research carried out in Egypt (1900-
1970). Food and dairy industries. The National Center for
Information and Documintation, Egypt.

Hess, E. (1950). Bacterial fish spoilage and its control.Food

Technology.4: 477-480.

Hillman, H. (1983). Kitchen Science [ Boston: Houghton Mifflin] B.H.

himelbloom, J.E. Routledge and S.L. biede (1983) Colour
changes in blue carbs (Callinectes sapidus) during cooking.
Jornal of Food Science 48: 652-653.

Howgate, P. (1979). Food microscopy. Vol. 2, 343-389, edited by J.G.

Vaughan. Acadamic Press, London.

Hoyland, D. V. and Taylor, A. J (1989). Prepration and storage stability

of dried-salted mutton. International Journal of Food Sceince
and Technology. 2186: 13-15.

Hultin, H. O.; Mcdonald, R. E. and Kelleher, S. D. (1982). Lipid

oxidation in fish muscle microsomes. In Chemistry and
Biochemistry of Marine Food Products, (eds R. E. Martine; G.
J. Flick; C. E. Hebart and D. R. Ward), AVI puplishing,
Westport, Ct, USA, Chap.1. 1.

Huss, H. H and Valimarson, G. (1990). Micorobiology of salted fish.

FAO Technical Paper. 10(1), 190.

Idris, Z. O. (1981). Studies on morphmetric, parametric and meristic

counts of the family Characid. B. Sc. (Hons.) Dissertation,
Department of Zoology, Faculty of Science, University of
Khartoum, Sudan.

12
Igene,J. O.; Pearson, A. M.; Merkel, K. A. and Coleman, T. H.(1979).
Effect of frozen storge time, cooking and holding temperature
upon extractable lipids and TBA values of beef and chicken.
Journal of Animal Science. 49-701.

ILO (1982). Small-scale processing of fish Ist(ed). ILO-Geneva-

Switzerland.

ILO. International Labour organization (1986). Solar drying practical

methods of food preservation (Ist ed.) 1986 Ilo, Genva, Swiss.

ILO (1989). Small-scale processing of fish. Technical

Momorandum.No.3:13-41. Geneva.

Ingram, M. and Kitchell, A. (1967). Labrotory observation Effect of

common salt on Dermestes Maculates infesting dried fish. FAO
fisheries report No. 329.

Ishag, N. A. (1998). Quality of traditionally produced dehydrated beef.

M. Sc. Thesis, University of Khatoum.

James, D. G., and Krone, W. (1977). Development in fish utilization. In:

Proceedings of the conference on the handling, processing and
marketing of tropical fish. Held in London, 5-9 July 1976.
London, Tropical Products Institute 467-472.

Jan, P., Nedyalka, Y. and Micheal, G. (2001). Antioxidants in Food.

Woodheat puplishing limited. pp. 421-450. England.

Jane, A. B., Jean, A. C., Kropt, D. H. and Tucker, T. J. (1987). Flavour,

colour and other characterstics of beef Longissimus muscle
heated to seven internal temperatures between 55C and 85C.
Journal of Food Science.52(3), 533.

13
Jauegui Carlos, A. and Baker, C. (1980). Discolouration problems in
mechanically deboned fish. Journal of Food Science. 45: 1068-
1069.

John, H. B.(1994). Food Colour and Appearance. Ist (ed.), 284-318.

Blackie Academic and Professional, London.

Johnson, W.A. (1994). Freezing and refrigerated storage in Fisheries.

FAO, Fisheries Technical Paper No.340. FSCL. Food science
Laboratory.

Josephson, D. B.; Lindsay, R. and Olafsdotter, G. (1986). Measurement

of volatile aroma constituents as a means of following sensory
deterioration of fresh fish and fishery products. In Seafood
Quality Determination, D.E. Kramer and J. Liston (eds.),
Elsevier, New York, pp. 27-47.

Judge, M. D., Aberle, E.; Forrest, J.; Hedrick, H. and Market, R. (1990).
Principle of meat science. (2nd ed.) Kendall Hunt. Iwoa. USA.

Kenedy, L. and Wood, C. D (1983). The use of solar dryers to reduce

losses of sun-dried fish wet seasons in Malawi. Report of
Tropical Producrs Institute.(TPI).

King, F.J. (1973). Improving the supply of minced bloch for the fish stick
trace, A progress Report. Marine Fisheries Review, 38(8) 26.

Kofi, M. E (1992). Fermented fish in Africa, FAO Fisheries Technical

Paper 329.

Kramlich, W. E (1975). Processed meat detroiration. In: Processed meats

John morel and Company. Chicago. 111 inois 2 nd , U.S.A.

Krempt, A. (1929). Traitement industriel des produits de la peche.

Rapport sr la fon ctionnement du service oceonographique des
peches Iindochine .13:36-43.

14
Kropf, D. H (1980). Effect of retail display conditions on meat colour.
Proceed. Annual Recip. Meat conference. (RMC) of the
American Meat Science.Assoc.,33:15-32.

Lobuza, T. P. (1971). Kinetics of lipid oxidation in foods. CRC Crit. Rev.

food technology. 356.

Lands,W.E.M.(1986). Fish and human Health. Academic Press, ,pp.1-

170.

Lawrie, K. A. (1990). Meat science. 5 th. ed. Program Press, Oxsford. UK.

Levandiov, L. P. (1958). Quantitative characterstics of fish after salting.

Israel program of scientific Translation (PST) Catalogue, No.
109: 39-45.

Locker, R.H., and Hagyard, C.J. (1963). A cold shortening effect in beef
muscle. J.Sci.Food.Agric. 14, 787.

Maga, J. A. (1987). The flavour chemistry of smoke. Food Reviews

International. 3, 139-183.

Mahmoud, Z. N. (1977). Studies on meat qulality of some common Nile

fish. M. Sc. Thesis, Department of Zoology, Faculty of
Science, University of Khartoum. Sudan.

Margy, W. (1992). Starter cultures in traditional fermented meats. In:

A pplications of Biotchnology to traditional fermented food,

report of an Ad Hoc panel of the Board on Scince and
Technology for International Development. (1992). National
Academy Press. Washington. 128-159.

Martin, R. E. (1976). Mechanicaly-deboned fish flesh. Food Technology.

30 (9): 64-68.

15
Medynski, A.; Pospiech, E.; and Kniat, R. (2000). Effect of various
concentrations of lactic acid and sodium chloride on selected
physico-chemical meat traits. Meat Science. 55: 285-290.

Mmdlewa, T. C. (1990). Sun dried salted fish as a mean of reducing post-

harvest losses in Botswana. CIFA technical paper No.19.
Proceeding of Symposium on Post-harvest Fish Technology.
FAO, 21-22, October, 1990. Clucas and Ward(1996) Cairo,
Egypt.

Moy, J. H. (1977). Solar dehydration Vs-radiation preservation of foods.

In-Proc. Conf. Fd. Agric. Malysia 2000, Kuala lumpur.

Narayanaswarny, D.; Narasimha Rao, C.V. and Ggovindan, T. K. (1980).

Pentration of sodium chloride during prolonged salting of fish.
Fish Technology, 17(1): 63-64.

Ofstad, R.; Kidman, S.; Myklebust, R.; Olsen, R. L, and Hermansson, A.

M.(1995). Liquid-holding capacity and structural changes in
comminuted salmon (Salmo salar) muscle as influenced by pH,
salt and temperature. Lebemsmittel Wissenschaft und
Technologie.28: 329-339.

Olympia, M.; Ono, H.; Shinmyo, A. and Takano, M. (1992). Lactic acid
bacteria in a fermented fishery product, Burong bangus.
Journal of Fermentation and Bioengenering. 73: 193-197.

Owens, J. D. and Mendoza, L. S. (1985). Enzymatically hydrolysed and

bacterially fermented fishery products. Journal of Food
Technology. 20:273-293.

Pearson, A. M., and Gillett, T. A (1996). Procesed meat (3ed ed.).

Champman and Hall, New York.

16
Peter, F. (1997). Traditional Food. Intermediated technology publication.
103-105.Southampton Row, London.

Peter, S.F., and Allan,W. (1984). Principle of Fermentation Technology.

Pergamon Press. Oxford. UK. 1-10.

Poulter, R. G (1980). Final report of work at Institute of Fish Technology

1978-1980, Clombo, Syri-Lanka, London, Tropical Products
Institute, Cited from Abu Gideiri, Y.B.; Ali, M.E. and
Mohamoud, Z.N (1999). Post- harvest treatment of the Nile
Bulti Oreochromis niloticus. Department of Zoology. Reserch
Series. No.1. University of Khartoum.

Reid, D. S.(1973). Water activity concepts in intermediate moisture

foods. In intermediate moisture foods, R. Davies, G.G. Birch
and K.J. Parker (eds), Applied Science, London, pp. 54-65.

Reiss, U. and Tappel, A.L. (1973). In: Food, Journal, 8, 199-202.

Rock-Well, G. E., and Ebertz, E. G. (1924). How salt preserves:.

Infection Diseases. 35:573.

Rosenthal, A. j (1999). Flesh foods and their Analogues.In: Food texture

measurment and perception, M.L. Greaser and A.M. Pearson
(eds). Aspen publisher, Inc.Marylan.p.228-233.

Ruiter, A. (1995). Fish and fishery products. Composition, nutritive

properties and stability. CAB international. Oxon, U.K.

Sadjeli, M. C.; Ahamed, J.; Hasjmi, A., Arbain and Sukendar. (1974).
Laporan penelitian kwalitas ikan kering yang diawet dengan
garam swasta dan yang diawet danger garam pemerintah. P.N.
garam cabang niaga kal.sel/teng. Dan fakultas perikanan unlam.
Indonesia.

17
Saisithi, P. (1987). Traditional fermented fish products with special
reference to Thai Products. Asean Food Journal. 3: 3-10.

Saisithi, P.; Kasemsarn, B.; Liston, J. and Dollar, A. M. (1966).

Micorobiology and chemistry of fermented fish. Journal of
Food Science. 31: 105-110.

Samson, R. A, Kooij, J. A. and Boer, E. D. (1987). Microbiological

quality of commercial tempe in the Netherlands. Journal of
Food Protection. 50: 92-94.

Sanders, T. A. B (1994). Nutritional aspect of rancidity. In: Rancidity in

food. 3rd . J.C. Allen and R.J.Hamilton. Blackie Acadamic and
Professional. UK. 128-139.

Scott, W. J. (1957). Water relations of food spoilage microorganisms.

Advantage food research, 7, 83. Acadamic Press Inc.,
Publishers. New York. USA.

Shahidi, F. (1992). Prevention of lipid oxidation in muscle foods by

nitrite and nitrite-free compositions. In lipid oxidation in food,
ed. A.J.St. Angelo, ACS Symposium series 500. American
chemical Society. Washington, pp. 161-182.

Shahidi, F.; Chong, X. and Dunajski, E. (1994). Freshness quality of harp

seal (Phoca groen landica) meat. Journal of Agriculture and
Food Chemistry.42: 868-872.

Shahine, A. B. (1956). Chemical composition of salted fermented fish

(Fessiekh) at various stages of fermentation. M. Sc. Cairo
University.

Sharp, J. G (1957). Deterioration of dehydrated meat during storage. J.

Sci. Food. Agri. 8:14-21.

18
Smith, G.; Hole, M. and Hanson,W. H. (1990). Assessment of lipid
oxidation in Indonesian salted-dried marine catfish (Arius
thalassinus). J. Sci. Food. Agri. 51: 193-205.

Sojofn, F. ; Margret, S. S., Hannes, H. ; Ole, T. and Jean, L. V. (2000).

Effects of different salting and smoking processes on the
microstructurer, the texture and yeild of Atlantic salmon
(Salmo salar) fillets. Food Research International. 33: 847-855.

Spanier, A, M., Miller, J. A and Bland, J. M. (1992). Lipid oxidation:

Effects on meat proteins. In lipid oxidation in food. A.J.St.
Angelo (eds). ACS Symposium series 500, American Chemical
Society. Washingon, pp. 161-182 pp.

Sundhaghul, M.; Daengsubha, W. and Suyanandana, P. (1975).

Thailands traditional fermented food products: a brief
description. Thailand journal of Agricultural Science.8: 205-
219.

Tanasumpawat, S.; Hashimoto, Y.; Ezaki, T.; kozaki, M. and Komagata,

K. (1991). Identification of Staphyococcus Carnosus strains
from fermented fish and soy sauce mach. Journal of Genetic
and Applied microbiology. 37: 479-494.

Tanasupawat, S.; Hashimoto, Y.; Ezaki, T.; Kozki, M. and Komagata, K.

(1992). Staphylococcus Piscifermentans sp. Nov., from
fermented fish in Thailand. International Journal of
Bacteriology. 42: 577-581.

Tarr, H. A. (1965). The maillard reaction in flesh foods. Food

Technology.15-19.

Terrel, R.N. (1983). The water relation of food born bacteria pathogens.
A Review. Journal of Milk and Food Technology. 36 (5).

19
The Fertilizer and Feeding Stuff Regulation (1976). H. M. S.O., 49 High
Holborn, London WCIV 6 HB.

Tressler, D. K. (1950). Marine products of commerce, New York,

Reinhold. 782p.

Toury, J.; Ware, A.; Gorgi, R. and Cros, J. (1970). Fish in the diet in
Senegal, quantitative and qualitative aspects, methods of
preservation. Food Nutrition.8: 6-13.

Um, M. N. and Lee, C. H. (1996). Isolation and Identification of

staphylococcus sp. From Korean fermented fish products.
Journal of Microbiology and Biotechnology. 6: 340-346.

Van Arsdel, W. B.; Copley, M. j. and Morgan, A. I. (1973). Food

dehydration, practices and applications. The AVI publishing
Company, Inc. New York. USA.

Wang, H. L. and Hesseltine, C. W. (1981). Use of microbial culture:

Legume and cereal products. Food Technology. 33(1): 79-83.

Wasson, D.H., Reppond, K.D., Kandianis, T.M. (1991). Antioxidants to

preserve rock-fish colour. Journal of food science. 56, 1564-
1566.

Watanabe, K. (1982). Fish handling and processing in tropical Africa. In:

proceedings of the FAO expert consultation on fish technology
in Africa, Cosablanca, Morocco, pp. 7-11

Waterman, J. J (1976). The production of dried fish. FAO Fisheries

Technical Paper, No 160, 52.

Younathan, M. T. and Watt, B. M. (1959). Lipid oxidation in food. Fish

Technology. 24: 728-734.

20
Zakhia, N., and Cua, J.L. (1991). A percu surla quality du tilapia
seche et commercialise au Mali. FAO Fisheries Repot (400).
176-181.

Zapata, J. F., Ledward, D. A and Lawrie, R. A (1990). Prepration and

storage stability of dried salted Mutton. Meat Science, 27:109.

21
APPENDICES

22
 Appendix (1) The effect of Species of fishes and time on C.P., C.F., E.E., Ash, Moist., and D.M. of Kawara &
Debs during Summer.
C.P C.F E.E Ash D.M Moist.
Species
Days

Mean SE Sig. Mean SE Sig. mean SE Sig. mean SE Sig. mean SE Sig. mean SE Sig.

4D K 18.82 0.23 * 0.8 7.78 NS 1.5 0.14 NS 10.81 0.30 NS 40.15 1.56 * 59.85 1.45 *

D 17.1b 0.39 * 0.65 9.62 NS 1.29 2.94 NS 12.76 1.47 NS 37.07 0.99 * 62.95 0.99 *
8D K 18.50 0.33 NS 0.57 6.75 NS 1.25 0.13 NS 11.20 0.36 NS 42.47 1.41 * 58.63 0.76 *

D 16.40b 0.74 NS 0.59 8.88 NS 1.20 5.09 NS 13.62 0.76 NS 40.65 0.94 * 59.37 0.97 *
12 K 18.08 0.20 * 0.65 7.77 NS 1.43 0.12 NS 12.01 0.53 NS 41.17 0.36 NS 58.79 0.37 NS
D
D 16.48b 0.55 * 0.41 4.84 NS 1.15 0.21 NS 13.35 0.14 NS 39.45 1.91 NS 60.66 1.84 NS
1M K 16.57 0.66 NS 0.48 4.84 NS 1.15 0.15 NS 13.50 0.25 NS 38.02 2.48 NS 61.65 2.91 NS

D 14.06b 0.56 NS 0.43 0.00 NS 1.25 0.11 NS 13.44 0.19 NS 35.33 2.94 NS 64.67 2.94 NS
2M K 15.91 0..20 * 0.46 8.38 NS 0.14 5.55 NS 12.88 0.65 NS 43.77 0.78 NS 56.25 0.80 NS

D 11.58b 0.33 * 0.25 5.00 NS 0.66 0.23 NS 12.72 0.18 NS 41.91 9.83 NS 57.97 0.25 NS
3M K 15.58 0.23 * 0.38 4.00 NS 0.97 4.00 * 12.46 0.62 NS 56.25 0.80 NS 43.74 0.80 NS

D 11.36b 0.13 * 0.25b 1.66 NS 0.68b 5.00 * 12.30 0.24 NS 55.34 3.06 NS 44.65 3.06 NS

4M K 15.19 0.57 * 0.27 2.94 * 0.77 4.84 * 12.70 0.66 NS 45.46 0.40 * 45.53 0.40 *

D 11.06b 3.50 * 0.15 b 1.66 * 0.41b 5.00 * 13.35 0.34 NS 60.15 1.48 * 39.85 1.47 *
5M K 14.55 6.50 * 0.15 1.66 NS 0.15 1.66 * 12.45 6.16 * 65.63 0.10 * 32.86 1.42 NS

D 10.43b 2.00 * 0.16 0.00 NS 0.18 5.00 * 12.16 1.50 * 67.22 0.29 * 32.77 0.29 NS

6M K 13.55 0.19 NS 0.16 3.33 NS 0.33 0.00 NS 11.63 3.00 NS 73.59 1.25 NS 26.40 1.25 NS

D 10.07 - NS 0.13 - NS 0.23 - NS 11.82 - NS 70.31 - NS 29.68 - NS

100
Appendix (2) The effect of Species of fishes and time on P, Ca, Fe, Na, K, and Cu of Kawara & Debs during
summer.
P Ca Fe Na K Cu
Species
Days

Mean SE Sig. Mean SE Sig. Mean SE Sig. Mean SE Sig. Mean SE Sig. Mean SE Sig.
.

4D K 1.44 9.69 NS 8.60 0.19 NS 55.0 1.98 NS 412.44 8.26 NS 5.24 0.11 * 4.29 0.11 NS

D 1.22 6.76 NS 7.78 0.11 NS 56.44 2.11 NS 547.11 100.48 NS 6.94 0.45 * 4.46 0.14 NS
8D K 1.35 0.11 NS 8.12 0.20 NS 49.00b 2.10 * 495.11 84.64 NS 5.1 9.09 * 4.1 0.13 NS

D 1.12 4.44 NS 7.65 5.87 NS 58.66 2.18 * 563.11 119.36 NS 6.31 8.01 * 4.31 0.16 NS
12D K 1.17 7.77 NS 7.54 0.10 NS 46.50b 3.07 * 507.22 99.78 NS 5.01 0.11 * 4.14 0.10 NS

D 1.11 8.67 NS 7.46 0.10 NS 58.33 1.53 * 542.44 104.59 NS 6.58 5.87 * 4.28 0.16 NS
1M K 1.21 0.10 NS 7.37 8.58 NS 57.77 3.42 NS 522.77 113.53 NS 5.81 0.16 NS 4.25 0.14 NS

D 1.31 5.00 NS 7.13 3.33 NS 52.66 2.00 NS 417.00 98.00 NS 6.18 5.00 NS 4.20 0.10 NS
2M K 1.24 7.77 NS 7.26 0.11 NS 43.00 3.05 NS 490.55 106.94 NS 4.90 3.33 * 4.00 0.15 NS

D 0.93 0.10 NS 7.15 5.00 NS 54.33 2.66 NS 577.50 98.16 NS 5.40 0.20 * 4.05 1.66 NS
3M K 1.03 0.11 * 6.93 0.10 NS 40.00 1.52 NS 398.88 100.22 NS 4.20 0.12 NS 3.67 0.11 NS

D 0.73b 3.33 * 7.00 0.13 NS 47.00 2.66 NS 499.50 100.50 NS 5.11 0.65 NS 3.68 8.33 NS
4M K 0.91 0.19 * 6.81 0.15 NS 37.44 1.28 NS 383.22 100.33 NS 3.88 0.10 NS 3.43 0.15 NS

D 0.50b 0.82 * 6.41 8.33 NS 39.66 1.66 NS 437.66 127.33 NS 4.31 8.33 NS 3.11 1.66 NS

5M K 0.66 0.00 * 6.03 6.66 NS 28.66 1.33 NS 429.83 78.50 NS 3.30 0.16 NS 3.18 5.00 NS

D 0.36b 3.33 * 16.16 0.00 NS 29.66 1.00 NS 382.66 118.33 NS 3.96 0.13 NS 2.68 0.11 NS

6M K 0.51 5.00 NS 5.70 0.10 NS 25.50 0.50 NS 413.83 79.16 NS 3.06 3.33 NS 2.66 3.33 NS

D 0.16 - NS 5.73 - NS 21.33 - NS 237.66 - NS 3.40 - NS 1.90 - NS

101
Appendix (3) The effect of Species of fishes and time on C.P., C.F., E.E., Ash, Moist., and D.M. of Kawara &
Debs during autumn.
Days C.P C.F E.E Ash D.M Moist.
SP.

Mean SE Sig. Mean SE Sig. Mean SE Sig. Mean SE Sig. Mean SE Sig. Mean SE Sig.
.
4D K 18.61 0.23 NS 0.87 4.00 NS 1.44 8.12 NS 11.38 0.30 NS 40.58 0.25 * 59.41 0.25 *

D 18.42 0.20 NS 0.90 5.77 NS 1.31 6.75 NS 12.18 0 .50 NS 35.17 0.99 * 64.39 1.01 *
8D K 18.34 0.23 NS 0.78 4.44 NS 1.36 9.62 NS 11.49 0.33 NS 41.95 0.42 * 58.00 0 .41 *
18.46 8.81 NS 39.48 0.49 *
D 0.74 2.94 NS 1.21 7.77 NS 12.33 0.52 NS 60.50 0.48 *
12D K 18.34 0.22 NS 0.63 b 1.92 * 1.26 8.81 NS 11.97 0.14 NS 41.36 0.31 NS 58.63 0.31 *
12.55 0 .46 NS
D 18.31 8.00 NS 0 .87a 4.84 * 1.04 4.00 NS 39.51 0.84 NS 59.79 0 .19 *
1M K 17.81 8.33 NS 0.55 b 1.66 * 1.13 6.66 NS 12.45 0.26 NS 59.94 0.88 NS 40.87 0.87 NS

D 18.15 8.33 NS 0.75a 1.66 * 1.01 1.66 NS 13.55 1.66 NS 62.06 1.48 NS 37.93 1.48 NS
2M K 17.20b 0.00 * 0.40 6.66 NS 1.00 3.33 NS 12.50b 0.15 * 64.29 1.22 NS 35.70 1.22 NS

D 17.91a 8.33 * 0.70 3.33 NS 1.03 0.00 NS 13.90a 1.00 * 63.27 0.10 NS 36.72 0.10 NS
3M K 16.26b 0.23 * 16.26 0.00 NS 0.75 5.00 NS 13.0b 3.66 * 72.74 1.49 NS 27.45 1.29 NS

D 17.65a 8.33 * 0.58 5.00 NS 0.76 6.66 NS 14.03a 3.33 * 72.58 1.45 NS 27.41 1.45 NS
4M K 16.0 5.00 NS 0.26 3.33 NS 0.61 1.66 NS 13.68 9.50 NS 76.54 0.54 NS 23.45 0.54 NS

D 16.90 - NS 0.43 - NS 0.56 - NS 14.16 - NS 79.31 - NS 20.68 - NS

5M K 16.33 0.18 NS 0.16 6.66 NS 0.40 1.66 NS 13.03 1.63 NS 82.00 0 .54 * 14.25 1.33 *

D 16.70 0.18 NS 0.30 6.66 NS 0.43 1.66 NS 14.30 1.63 NS 83.08 0.54 * 16.91 1.33 *

6M K 16.23 3.33 NS 0.13 6.66 NS 0.30 3.33 NS 13.03b 1.30 * 85.74 0.70 * 14.25 0.56 NS

D 16.30 3.33 NS 0.26 6.66 NS 0.23 3.33 NS 15.63a 1.30 * 87.15 0.70 * 13.12 0.56 NS

107
 Appendix (4). The effect of Species of fishes and time on P, Ca, Fe, Na, K, and Cu of Kawara & Debs during
P Ca Fe Na K Cu
Species
Days

Mean SE Sig. Mean SE Sig. Mean SE Sig. Mean SE Sig. Mean SE Sig. Mean SE Sig.
.
autumn.
4D K 1.61a 4.84 * 8.65 5.87 * 53.44 82.4 NS 490.22 54.54 NS 5.43b 0.13 * 4.40 1.92 NS

D 1.25b 5.87 * 7.97 6.75 * 55.00 2.03 NS 553.11 99.22 NS 6.42b 2.22 * 4.42 6.75 NS
8D K 1.44a 6.18 * 8.45 7.77 * 48.66 1.64 NS 461.33 47.99 NS 5.24 0.10 * 4.15 1.11 NS

D 1.12b 5.87 * 8.02 6.75 * 52.66 1.89 NS 542.22 97.22 NS 6.22 4.00 * 4.28 8.67 NS
12D K 1.26 6.66 NS 8.21a 4.84 * 46.88 1.05 NS 446.33 48.37 NS 5.17 0.17 * 4.00 1.11 *

D 1.14 4.00 NS 7.97 b 4.00 * 50.77 1.23 NS 528.33 97.31 NS 6.04 4.84 * 4.24 2.94 *
1M K 1.13 3.33 NS 8.00 3.33 NS 45.00b 0.66 * 415.66 89.00 NS 4.91 0 .11 * 3.76 3.33 *

D 1.00 3.33 NS 7.93 3.33 NS 52.16a 0.83 * 550.83 64.16 NS 5.83 0.13 * 3.93 0.00 *
2M K 1.00 3.33 NS 7.68 5.00 NS 43.16 1.16 NS 400.83 93.16 NS 4.60 0.00 * 3.60 0.00 *

D 0.90 0.00 NS 7.80 0.10 NS 47.66 1.33 NS 550.50 87.16 NS 5.48 1.66 * 3.71 1.66 *
3M K 0.78 1.66 NS 7.50 6.66 NS 34.16 1.50 NS 416.83 64.83 NS 4.11 8.33 NS 3.18 5.00 *

D 0.70 0.10 NS 7.23 0.13 NS 39.33 0.66 NS 498.33 85.00 NS 4.83 0.20 NS 3.41 1.66 *
4M K 0.66 3.33 NS 7.23 6.66 NS 30.16 1.16 NS 369.16 85.83 NS 3.91 0.11 NS 3.00 6.66 NS

D 0.40 - NS 6.83 - NS 32.00 - NS 510.66 - NS 4.50 - NS 2.96 - NS

5M K 0.36 5.00 NS 6.83 0.11 NS 21.66 3.83 NS 402.66 43.83 NS 3.26 0.36 NS 2.60 6.66 NS

D 0.26 5.00 NS 6.60 0.11 NS 29.33 3.83 NS 490.33 43.83 NS 4.00 0.36 NS 2.73 6.66 NS
6M K 0.23 0.00 NS 6.50 0.20 NS 20.33 4.16 NS 384.33b 13.50 * 3.03 0.25 NS 2.43 3.33 NS

D 0.23 0.00 NS 6.10 0.20 NS 28.66 4.16 NS 411.33a 411.33 * 3.53 0.25 NS 2.50 3.33 NS

108
 Appendix (5). The effect of Species of fishes and time on C.P., C.F., E.E., Ash, Moist., and D.M. of Kawara &
Debs during winter.
C.P C.F E.E Ash D.M Moist.
Species
Days

Mean SE Sig. Mean SE Sig. Mean SE Sig. Mean SE Sig. Mean SE Sig. Mean SE Sig.
.
4D K 19.77 0.19 NS 1.27a 6.75 * 1.92a 5.87 * 10.30 0.10 NS 25.57 2.39 NS 74.21 2.49 NS

D 19.71 9.87 NS 1.48b 2.94 * 1.56b 5.09 * 10.51 6.18 NS 31.65 1.13 NS 68.34 1.13 NS
8D K 19.63a 0.15 * 1.20b 5.77 * 1.75a 5.55 * 10.37 8.88 NS 24.86 2.02 NS 75.14 2.02 NS

D 18.77b 7.77 * 1.37a 2.94 * 1.40b 1.92 * 10.73 0.25 NS 31.00 1.45 NS 68.99 1.45 NS
12D K 19.27a 9.49 * 1.23a 3.33 NS 1.65a 4.00 * 10.73 0.13 NS 24.96 1.91 * 75.06 1.92 *

D 18.23a 8.81 * 1.28a 4.84 NS 1.35b 2.22 * 11.10 0.13 NS 31.48 1.24 * 68.51 1.24 *
1M K 19.06a 5.77 * 0.74b 2.22 * 1.45a 4.00 * 12.00a 0.12 * 75.62 1.76 * 24.36 1.77 *

D 18.14b 0.11 * 1.13a 1.92 * 1.24b 2.94 * 11.47b 0.12 * 33.35 1.71 * 66.64 1.71 *
2M K 18.87a 0.17 * 0.47b 4.00 * 1.23a 8.38 * 12.47a 9.68 * 78.91 1.23 * 21.08 1.23 *

D 17.68b 5.87 * 0.92a 4.00 * 0.95b 2.94 * 12.02b 0.16 * 72.09 1.91 * 27.90 1.91 *
3M K 18.23a 3.33 * 0.36b 0.00 * 0.95 5.00 NS 13.48 0.12 NS 79.76 1.25 * 20.23 1.25 *

D 17.10b 6.66 * 0.88a 1.66 * 0.85 1.66 NS 12.83 0.10 NS 71.44 0.32 * 28.55 0.32 *
4M K 17.73a 3.33 * 0.33b 0.00 * 0.78 5.00 NS 13.43 0.12 NS 81.50 0.63 NS 18.49 0.63 NS

D 16.88b 5.00 * 0.73a 3.33 * 0.66 3.33 NS 13.36 6.50 NS 81.19 0.75 NS 18.80 0.75 NS
5M K 17.10a 6.66 * 0.26 0.00 NS 0.48 1.66 NS 13.75 8.33 NS 81.61 2.31 NS 18.38 2.31 NS

D 16.16b 6.66 * .4167 5.00 NS 033 3.33 NS 13.43 0.13 NS 84.55 1.41 NS 15.44 1.41 NS
6M K 16.18a 1.66 * 0.20 3.33 NS 0.40a 3.33 * 14.08 8.33 NS 84.81 0.35 NS 15.18 0.35 NS

D 14.26b 0.30 * 0.23 0.00 NS 0.18b 1.66 * 12.45 2.51 NS 86.03 0.00 NS 13.96 0.00 NS

109
Appendix (6). The effect of Species of fishes and time on P, Ca, Fe, Na, K, and Cu of Kawara & Debs during
winter.

P Ca Fe Na K Cu

Days Sp.
Mean SE Sig. Mean SE Sig. Mean SE Sig. Mean SE Sig. Mean SE Sig. Mean SE Sig.
.
K 1.71a 1.11 * 8.75a 4.84 * 61.55 0.48 NS 510.66 87.86 NS 5.56 6.66 * 4.88 2.94 *
4D
D 1.44b 2.94 * 8.11b 4.00 * 63.77 1.36 NS 576.88 108.93 NS 6.31 6.18 * 4.48 8.67 *
K 1.63a 3.84 * 8.72a 1.11 * 60.66 050 NS 507.44 88.62 NS 5.32 7.77 * 4.75 5.55 *
8D
D 1.37b 1.11 * 8.05b 0.10 * 62.00 1.26 NS 571.66 109.73 NS 6.14 4.84 * 4.32 6.18 *
K 1.37 4.00 NS 8.28 6.75 NS 56.55 0.40 NS 493.00 95.41 NS 5.22 4.00 * 4.50 5.09 *
12D
D 1.26 1.92 NS 8.25 0.17 NS 59.22 0.96 NS 546.55 112.48 NS 5.91 4.84 * 3.97 2.94 *
K 1.34a 2.94 * 8.08 2.94 NS 51.33b 0.69 * 470.22 88.96 NS 4.96 1.92 * 4.23 3.33 *
1M
D 1.17b 4.00 * 8.02 9.49 NS 57.33a 0.96 * 538.88 107.26 NS 5.7444 2.94 * 3.91 4.44 *
K 1.11 4.84 NS 8.08 0.10 NS 48.11 0.58 NS 466.00 91.08 NS 4.73 1.92 NS 3.98 4.44 NS
2M
D 1.04 1.11 NS 7.60 5.09 NS 53.22 0.44 NS 527.33 102.49 NS 5.43 1.92 NS 3.42 4.00 NS
K 0.98 5.00 NS 7.76a 6.66 * 43.00b 0.33 * 566.50 83.16 NS 4.40 0.00 * 3.66 6.66 NS
3M
D 0.86 3.33 NS 7.10b 6.66 * 50.00a 0.00 * 597.16 92.83 NS 5.01 5.00 * 3.25 8.33 NS
K 0.76 6.66 NS 7.66a 0.10 * 39.00b 1.00 * 471.00 37.00 NS 4.01 1.66 * 3.46 0.13 NS
4M
D 0.72 1.66 NS 6.90b 3.33 * 47.00a 0.66 * 610.83 51.16 NS 4.71 1.66 * 3.10 6.66 NS
K 0.45 1.66 NS 7.40 0.13 NS 33.16 1.50 NS 450.33 39.66 NS 3.43 0.00 * 3.28 0.11 NS
5M
D 0.45 5.00 NS 6.58 0.15 NS 38.00 0.00 NS 507.66 101.00 NS 4.16 6.66 * 2.90 0.10 NS
K 0.40 3.33 NS 7.36 0.13 NS 32.33 1.33 NS 435.83 39.83 NS 3.20 0.13 * 3.16 0.13 NS
6M
D 0.40 3.33 NS 6.66 0.30 NS 35.83 0.16 NS 490.16 98.16 NS 4.27 9.50 * 2.7667 6.66 NS

110
 Appendix (7). The effect of Salt concentration and time on Physical analysis, pH, and total viable Bacteria count
of Kawara & Debs during autumn.
b a l Ran pH B.count

___Salt con._____ ___Salt con._____ ___Salt con._____ ___Salt con._____ ___Salt con._____ ___Salt con._____
Species
Days

20% 25% 30% SE Sig. 20% 25% 30% SE Sig 20% 25% 30% SE Sig. 20% 25% 30% SE Sig. 20% 25% 30% SE Sig 20% 25% 30% SE sig.

4D K 8.83 7.13 6.60 0.67 NS 4.93 3.40 2.86 0.61 NS 46.86 43.86 45.53 0.86 NS 1.05 1.15 2.03 0.31 NS 7.13 7.13 7.16 1.11 NS 12600.00 7566.66 590.00 3482.08 NS

D 6.83 8.26 8.36 0.49 NS 4.43 5.93 4.46 0.49 NS 41.73 42.96 45.36 1.06 NS 0.96 1.06 1.21 7.27 NS 7.16 7.13 7.13 1.1 NS 4800.00 1833.33 1566.66 1036.19 NS

8D K 7.93 7.50 7.40 0.16 NS 5.90 4.03 4.20 0.59 NS 45.43 44.46 43.33 0.60 NS 1.12 1.29 1.51 0.11 NS 7.10 7.06 7.06 1.11 NS 19600.00 1800.00 373.33 6184.83 NS

D 8.23 8.40 9.16 0.28 NS 6.70 6.50 4.53 0.69 NS 41.06 41.76 47.30 1.97 NS 1.43 1.71 2.22 0.23 NS 6.90 7.00 6.83 4.84 NS 1863.33 1433.33 753.33 323.12 NS
12D K 7.33b 8.03a 8.10a 0.24 * 4.00 4.13 3.63 0.14 NS 46.03 47.46 47.06 0.42 NS 2.30 2.85 3.18 0.25 NS 7.10 7.06 7.13 1.92 NS 16700.00 4056.66 27.00 5022.62 NS

D 7.50b 8.43a 8.10a 0.27 * 4.80 4.96 4.66 8.67 NS 44.16 45.83 46.76 0.76 NS 1.87 2.35 2.94 0.30 NS 6.83 6.63 6.80 6.18 NS 7066.66 423.33 200.33 2252.53 NS
1M K - 8.70 9.96 0.63 NS - 5.40 5.93 0.26 NS - 43.20 44.33 0.56 NS - 1.63 1.70 3.50 NS - 7.03 7.03 0.00 NS - 124.33 90.00 17.16 NS

D - 10.16 7.96 1.10 NS - 7.03 6.90 6.66 NS - 47.43 47.93 0.25 NS - 1.49 1.52 1.50 NS - 6.63 6.63 0.00 NS - 13.13 13.86 0.36 NS
2M K - 10.96 11.76 0.40 NS - 7.80 8.16 0.18 NS - 45.86 46.30 0.21 NS - 1.25 1.28 1.66 NS - 6.67 6.80 8.33 NS - 0.00 0.00 0.00 NS

D - 7.73 9.93 1.10 NS - 6.43 6.93 0.25 NS - 42.36 40.43 0.96 NS - 1.06 1.10 1.83 NS - 6.43 6.33 5.00 NS - 0.00 0.00 0.00 NS
3M K - 10.53 12.13 0.80 NS - 6.90 7.50 0.30 NS - 44.96 44.60 0.18 NS - 1.19 1.21 6.66 NS - 6.83 6.73 5.00 NS - 0.73 1.36 0.31 NS

D - 8.86 10.03 0.58 NS - 5.70 6.30 0.30 NS - 45.66 43.76 0.95 NS - 1.10 1.18 4.16 NS - 6.33 6.46 6.66 NS - 0.33 0.93 0.30 NS
4M K - 12.10 11.90 1.00 NS - 8.16 8.00 8.33 NS - 39.90 40.60 0.35 * - 1.23 1.22 1.66 NS - 6.73 6.46 0.13 NS - 1.10 4.83 1.86 NS

D - - 14.33 - NS - - 7.63 - NS - - 40.56 - * - - 1.18 - NS - - 6.40 - NS - - 0.66 - NS

5M K - - 11.76 - * - - 8.33 - NS - - 40.50 - * - - 1.21 - * - - 6.36 - NS - - 2.13 - NS

D - - 11.70 - * - - 9.63 - NS - - 34.10 - * - - 1.18 - * - - 6.36 - NS - - 2.53 - NS

6M K - - 12.03 - * - - 9.00 - NS - - 37.90 - * - - 1.80 - NS - - 6.43 - NS - - 0.23 - NS

D - - 10.03 - * - - 12.20 - NS - - 30.66 - * - - 1.35 - NS - - 6.23 - NS - - 0.00 - NS

111
Appendix (8) Physical Analysis, and pH of fresh samples

Parameters
seasons Species
b a L Ran. pH
K 6.80 6.18 48.32 0.09 7.30
D 5.76 6.60 46.28b 0.10 7.43
Summer
SE 0.52 0.21 1.02 5.00 6.66
Sig. NS * * NS *
K 6.83 6.20 49.23 0.08 7.27
D 4.90 6.30 43.37 0.18 7.43
Autumn
SE 0.96 5.00 2.93 5.16 8.33
Sig. NS NS * NS *
K 5.07 6.43b 43.40b 0.09 7.10b
D 6.97 6.87 49.07 0.09 7.77
Winter
SE 0.95 0.21 2.83 1.66 0.33
Sig. NS * * NS *

Legend:

a = Redness

b= Yellowness

l= Lightness

112
 Appendix (9). The effect of salt concentration and time on Physical analysis, pH, and total viable Bacteria count
of Kawara & Debs during summer.

b a l Ran pH B.count

___Salt con._____ ___Salt con._____ ___Salt con._____ ___Salt con._____ ___Salt con._____ ___Salt con._____

20% 25% 30% SE Sig. 20% 25% 30% SE Sig. 20% 25% 30% SE Sig. 20% 25% 30% SE Sig. 20% 25% 30% SE Sig 20% 25% 30% SE Sig.
SP.
ays

4D K 9.73 9.33 10.83 0.45 NS 4.60 4.43 5.10 0.20 NS 48.00 48.20 50.13 0.68 NS 1.14 1.29 2.08 0.29 NS 7.13 7.16 7.13 1.11 NS 126.67 26.33 16.33 47.20 NS

D 10.60 11.01 9.79 0.36 NS 5.94 5.91 4.71 0.41 NS 46.57 49.66 47.27 0.93 NS 1.00 1.27 1.62 0.18 NS 7.33 7.26 7.30 1.92 NS 11400.00 11200.00 5100.00 2067.47 NS
8D K 10.76 9.67 8.67 0.60 NS 7.03 6.13 4.80 0.65 NS 51.70 50.67 50.30 0.42 NS 1.28 1.49 2.91 0.51 NS 7.13 7.10 7.11 1.11 NS 756.67 22.33 11.33 246.63 NS

D 11.81 11.51 10.87 0.27 NS 7.93 7.47 5.67 0.68 NS 45.36 46.50 48.37 0.87 NS 1.91 3.91 5.33 0.99 NS 7.20 7.16 7.23 1.92 NS 43666.67 1130.00 5466.66 135.22 NS
12D K 10.83 10.20 9.86 0.34 NS 6.96 6.53 6.26 0.20 NS 51.76 49.63 50.30 0.63 NS 2.33b 2.99 b 3.76 0.41 * 6.70 6.73 6.86 5.09 NS 0.00 21.00 14.66 6.21 NS

D 12.06 11.90 12.56 0.20 NS 4.73 4.06 4.30 0.19 NS 44.08 44.90 45.20 0.34 NS 1.47 1.32 b 1.45ab 4.57 * 6.80 6.68 6.82 2.22 NS 6.50 5.53 4.66 0.52 NS
1M K 10.03 10.90 11.03 0.31 NS 6.36 5.20 4.69 0.43 NS 45.76 45.40 45.32 0.12 NS 1.79 1.62 1.99 0.10 NS 7.20 6.90 6.20 2.94 NS 5.66 0.00 0.00 1.88 NS

D S 11.17 14.09 1.45 NS S 6.66 8.30 0.81 NS S 46.03 46.50 0.23 NS S 1.27 1.86 0.29 NS S 6.93 7.00 3.33 NS S 0.00 0.00 0.00 NS
2M K 12.10 11.66 12.26 0.23 NS 4.70 3.63 4.16 0.30 NS 43.96 44.76 45.00 0.31 NS 1.45b 1.37c 1.61 7.05 * 6.80 6.90 6.6 2.23 NS 24.33 0.00 0.00 8.11 NS

D S 13.87 12.86 0.50 NS S 5.67 5.50 0.41 NS S 47.22 47.42 1.00 NS S 1.23 1.27 2.00 * S 6.90 6.20 3.35 NS S 5.33 0.00 2. 00 NS
3M K 12.06 11.90 12.26 0.20 NS 4.73 4.06 4.30 0.19 NS 44.03 44.90 45.20 0.34 NS 1.47 1.32 1.45 4.57 NS 6.80 6.86 6.50 4.58 NS 6.50 5.53 4.66 2.22 NS

D S 12.86 13.86 0.50 NS S 7.97 8.60 0.31 NS S 41.81 44.74 1.43 NS S 1.18 1.15 1.50 NS S 6.86 6.70 8.33 NS S 41.33 6.33 17.50 NS
4M K 11.13 11.80 11.20 0.19 NS 7.70 7.30 7.03 0.19 NS 38.46 39.36 39.96 0.43 NS 1.43 1.19b 1.09c 0.10 * 6.72 6.80 6.71 4.44 NS 0.00 0.00 0.00 0.00 NS

D S 12.27 14.13 0.82 NS S 9.15 9.40 0.12 NS S 39.09 43.01 1.96 NS S 1.16 1.17 5.00 * S 6.70 6.20 5.02 NS S 16.33 71.33 27.50 NS
5M K S 13.26 13.23 0.11 NS S 10.66 9.93 0.36 NS S 39.40 39.80 0.23 NS S 1.45 1.38 3.50 NS S 6.70 6.66 5.00 NS S 0.00 0.00 0.00 NS

D S 12.26 14.06 0.90 NS S 9.89 10.47 0.29 NS S 36.12 42.52 3.20 NS S 1.11 1.28 8.33 NS S 6.60 6.43 8.33 NS S 5.00 15.66 5.33 NS
6M K S 13.06 13.10 1.66 NS S 10.80 11.73 0.46 NS S 39.80 39.83 1.66 NS S 1.56 1.50 3.00 NS S 6.43 6.52 5.01 NS S 0.00 0.00 0.00 NS

D S S 14.66 - NS S S 11.20 - NS S S 42.28 - NS S S 1.32 - NS S S 6.40 - NS S S 0.00 _ NS

113
Appendix (10). The effect of Species of fishes and time on P, Ca, Fe, Na, K, and Cu of Kawara & Debs during
autumn.

P Ca Fe Na K Cu
Species
Days

Mean SE Sig. Mean SE Sig. Mean SE Sig. Mean SE Sig. Mean SE Sig. Mean SE Sig.
.
4D K 1.61a 4.84 * 8.66 5.87 * 53.45 82.4 NS 490.23 54.54 NS 5.43b 0.13 * 4.41 1.92 NS

D 1.25b 5.87 * 7.97 6.75 * 55.00 2.03 NS 553.11 99.22 NS 6.42b 2.22 * 4.42 6.75 NS
8D K 1.44a 6.18 * 8.46 7.77 * 48.67 1.64 NS 461.34 47.99 NS 5.25 0.10 * 4.16 1.11 NS

D 1.12b 5.87 * 8.02 6.75 * 52.66 1.89 NS 542.22 97.22 NS 6.22 4.00 * 4.28 8.67 NS
12D K 1.27 6.66 NS 8.21a 4.84 * 46.89 1.05 NS 446.34 48.37 NS 5.18 0.17 * 4.00 1.11 *

D 1.14 4.00 NS 7.97 b 4.00 * 50.77 1.23 NS 528.33 97.31 NS 6.04 4.84 * 4.24 2.94 *
1M K 1.13 3.33 NS 8.00 3.33 NS 45.00b 0.66 * 415.66 89.00 NS 4.91 0 .11 * 3.76 3.33 *

D 1.00 3.33 NS 7.93 3.33 NS 52.16a 0.83 * 550.83 64.16 NS 5.83 0.13 * 3.93 0.00 *
2M K 1.00 3.33 NS 7.68 5.00 NS 43.16 1.16 NS 400.83 93.16 NS 4.60 0.00 * 3.60 0.00 *

D 0.90 0.00 NS 7.80 0.10 NS 47.66 1.33 NS 550.50 87.16 NS 5.48 1.66 * 3.71 1.66 *
3M K 0.79 1.66 NS 7.51 6.66 NS 34.17 1.50 NS 416.84 64.83 NS 4.12 8.33 NS 3.19 5.00 *

D 0.70 0.10 NS 7.23 0.13 NS 39.33 0.66 NS 498.33 85.00 NS 4.83 0.20 NS 3.41 1.66 *
4M K 0.67 3.33 NS 7.24 6.66 NS 30.17 1.16 NS 369.17 85.83 NS 3.92 0.11 NS 3.00 6.66 NS

D 0.40 - NS 6.83 - NS 32.00 - NS 510.66 - NS 4.50 - NS 2.96 - NS

5M K 0.36 5.00 NS 6.83 0.11 NS 21.66 3.83 NS 402.66 43.83 NS 3.26 0.36 NS 2.60 6.66 NS

D 0.26 5.00 NS 6.60 0.11 NS 29.33 3.83 NS 490.33 43.83 NS 4.00 0.36 NS 2.73 6.66 NS
6M K 0.23 0.00 NS 6.50 0.20 NS 20.33 4.16 NS 384.33b 13.50 * 3.03 0.25 NS 2.43 3.33 NS

D 0.23 0.00 NS 6.10 0.20 NS 28.66 4.16 NS 411.33a 411.33 * 3.53 0.25 NS 2.50 3.33 NS
Appendix (11): The effect of Species of fishes and time on Physical analysis, pH, and total viable Bacteria count
of Kwara & Debs during summer.
b a l Ran. pH B.count
Species
Days

Mean SE Sig. Mean SE Sig. Mean SE Sig. Mean SE Sig. Mean SE Sig. Mean SE Sig.
.
4D K 9.97 0.45 NS 4.71 0.20 NS 48.78 0.68 NS 1.50 0.29 NS 7.41 1.11 * 68.44 47.18 *

D 10.47 0.36 NS 5.52 0.41 NS 47.84 0.93 NS 1.29 0.18 NS 7.30 1.93 * 9233.3 2067.47 *
8D K 9.70 0.60 NS 5.98 0.64 NS 50.88 0.41 * 1.89b 0.51 * 7.12 1.11 NS 263.44 246.63 NS

D 11.39 0.27 NS 7.02 0.68 NS 46.74b 0.87 * 3.71 0.99 * 7.20 1.92 NS 16754.44 13514.22 NS
12D K 10.23 0.33 NS 6.58 0.20 NS 50.56 0.63 * 3.03 0.41 NS 6.76 5.09 * 11.88 6.21 NS

D 11.68 0.42 NS 7.08 0.53 NS 48.72b 0.13 * 2.68 0.47 NS 7.11 2.94 * 2299.22 2085.19 NS
1M K 10.65 0.31 NS 5.51 0.43 NS 45.52 0.12 NS 1.80 0.10 NS 6.94 2.94 NS 1.88 1.88 NS

D 12.63 1.45 NS 7.48 0.81 NS 46.26 0.23 NS 1.56 0.29 NS 6.8 3.33 NS 0.00 0.00 NS
2M K 12.07 0.23 NS 4.16 b 0.30 * 44.57b 0.31 * 1.48 7.05 NS 6.9 2.22 NS 8.11 8.11 NS

D 1337 0.50 NS 6.08 0.41 * 47.32 1.00 * 1.25 2.00 NS 6.5 3.33 NS 2.66 2.66 NS
3M K 12.17b 0.20 * 4.36 0.19 NS 44.71 0.34 NS 1.41 4.57 * 6.8 2.22 NS 5.56 0.52 NS

D 13.36 0.50 * 8.28 0.31 NS 43.31 1.43 NS 1.17b 1.50 * 6.6 8.33 NS 23.83 17.50 NS
4M K 11.44 0.19 NS 7.34b 0.19 * 39.81 0.43 NS 1.24 0.10 NS 6.70 4.44 NS 0.00 0.00 NS

D 13.30 0.82 NS 9.27 0.12 * 42.28 1.96 NS 1.16 5.00 NS 6.62 5.00 NS 43.83 27.50 NS
5M K 13.35 0.11 NS 10.30 0.36 NS 39.63 0.23 NS 1.41 3.50 NS 6.65 5.00 NS 0.00 0.00 NS

D 13.16 0.90 NS 10.18 0.29 NS 39.32 3.20 NS 1.19 8.33 NS 6.50 8.33 NS 10.33 5.33 NS
6M K 13.08b 1.66 * 11.26 0.46 NS 39.81b 1.66 * 1.53 3.00 NS 4.48 5.00 NS 0.00 0.00 Ns

D 14.66 - * 11.20 - NS 42.28 - * 1.32 - NS 6.40 - NS 0.00 - NS

Appendix (12). The effect of Species of fishes and time on Physical
analysis, pH, and total viable Bacteria count of Kawara & Debs during
winter.

b a l Rancidity
Species
Days

Mean SE Sig. Mean SE Sig. Mean SE Sig. Mean SE Sig. Mean

.
K 7.80 0.63 NS 4.12 0.49 NS 45.50 0.72 NS 1.14 7.96 NS 7.07
4D
D 7.12 0.49 NS 5.22 0.43 NS 44.56 0.88 NS 1.05 4.76 NS 7.33
K 7.65 0.59 NS 4.08 0.49 NS 45.52 0.18 NS 1.02 6.93 NS 7.01
8D
D 7.24 0.52 NS 5.30 0.52 NS 44.36 0.68 NS 1.01 4.69 NS 7.13
K 7.83 0.24 NS 4.06a 0.30 * 46.03 0.29 NS 2.07 5.16 NS 6.90
12D
D 7.60 0.31 NS 4.94a 0.12 * 44.62 0.56 NS 1.99 3.28 NS 6.75
K 9.02 9.49 NS 5.41b 0.21 * 45.26 0.48 NS 2.22 4.82 NS 6.80
1M
D 8.50 0.26 NS 6.78a 0.14 * 45.16 0.95 NS 2.12 3.13 NS 6.70
K 9.37 0.25 NS 6.08a 0.17 * 44.52 0.22 NS 2.27 4.44 NS 6.56
2M
D 8.96 5.77 NS 6.96a 3.84 * 43.85 0.41 NS 2.04 0.12 NS 6.62
K 9.80a 6.66 * 6.53 0.23 NS 42.03 0.33 NS 2.64 0.33 NS 6.61
3M
D 9.05b 5.00 * 7.01 1.66 NS 43.38 0.45 NS 2.18 5.00 NS 6.43
K 10.28 0.18 NS 7.46 0.43 NS 41.38b 0.25 * 2.37 0.16 NS 6.61
4M
D 9.56 3.33 NS 7.56 0.13 NS 42.60a 6.66 * 2.19 5.16 NS 6.38
K 17.91 0.15 NS 10.31 0.11 NS 37.76 0.23 NS 1.44 6.16 NS 6.45
5M
D 8.48 0.11 NS 10.58 0.18 NS 38.51 0.45 NS 1.46 3.16 NS 6.30
K 18.01b 5.00 * 10.68 1.66 NS 33.80 0.23 NS 0.95 3.00 NS 6.31
6M
D 18.65a 8.33 * 10.38 8.33 NS 32.33 0.30 NS 1.06 3.66 NS 6.30