Hatchery hygiene evaluation by microbiological examination

of hatchery samples
J. H. Kim and K. S. Kim1
College of Veterinary Medicine, Kyungpook National University, 1370 Sankyuk-dong, Buk-gu, Daegu,
702-701, Republic of Korea
hatchers where the bacterial contamination of the air
was low, bacterial counts were high, measuring over 100
cfu/16 cm2. Salmonella was mainly isolated from the
hatcher rooms, chick counting room, and the related
equipment and facilities but not from the areas used
for the earlier processing step such as the egg receiving
room, egg sorting room, setter rooms, and candlingtransfer room. The Salmonella serotype that was most
frequently isolated from the hatchery was Salmonella
Senftenberg. The other occasional Salmonella serotypes
such as Salmonella Schwarzengrund, Salmonella Madelia, Salmonella Montevideo, and Salmonella Enteritidis were isolated. The experimental group receiving
formaldehyde by constant rate infusion during hatching
had a significantly superior inhibitory effect on aerosol
bacterial count 4 h before hatching as compared with
the group receiving formaldehyde into a basin and the
negative control group (P < 0.05).

Key words: hatchery, contamination, Salmonella, formaldehyde, constant rate infusion

2010 Poultry Science 89:13891398
doi:10.3382/ps.2010-00661

INTRODUCTION
A hatchery plays the important role of collecting
hatching eggs from the breeder farm and selling newly
hatched chicks to a commercial poultry farm. However,
the environment of a hatchery can be a source of contamination that involves a variety of microorganisms
that can cause diseases in a poultry farm (Sheldon and
Brake, 1991; Scott and Swetnam, 1993). Furthermore,
a contaminated hatchery not only transmits diseases to
the poultry farm but also causes significant economic
losses for the poultry industry (Funk and Irwin, 1955;
Harry and Gordon, 1966). Therefore, Hazard Analysis
and Critical Control Point (HACCP) was recently applied for improvement of poultry farm hygiene; moreover, the importance of hatchery hygiene, egg hygiene,
2010 Poultry Science Association Inc.
Received January 21, 2010.
Accepted April 5, 2010.
1 Corresponding author: kimkiseuk@knu.ac.kr

and hygiene of equipment and facilities was realized all

over the world (Jordan et al., 2001).
Hatchery hygiene begins with the health of parent
stock. This is also related to biosecurity measures,
which include disinfecting and cleaning of the farm and
the avoidance of risk factors that can cause harm to
the hatching eggs even before they reach the hatchery.
Furthermore, standards for the evaluation of hatchery
hygiene have been set to measure overall contamination
by aerobic bacteria; the coliform and fungi contamination of eggs, fluff, air, and equipment; and facility surfaces involved in the processing steps from the egg sorting room to chick counting room (Nichols and Leaver,
1967; Jordan et al., 2001).
Microorganisms found on or in a few hatching eggs
can be easily distributed throughout the hatcher by
air movements during hatching and thus contaminate
all other chicks in the hatcher (Chute and Gershman,
1978; Sheldon and Brake, 1991). In particular, bacterial
contamination increases as the hatch nears completion
(Magwood, 1964a). Therefore, some researchers con-

1389

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ABSTRACT This study was conducted to investigate

the bacterial contamination of air and the surface of
equipment and facilities in hatchery. In addition, the
inhibitory effects of formaldehyde application methods on aerosol bacterial counts in the hatchers were
also investigated. In the operating hatchers, the contamination of air by aerobic bacteria, coliform, and
fungi was high, measuring over 300 cfu/63.6 cm2. In
the egg sorting room, contamination was moderate,
whereas in the remaining sampling sites such as the
setter room, candling-transfer room, and chick counting room, contamination was minimal, measuring less
than 10 cfu/63.6 cm2 for aerobic bacteria, 5 cfu/63.6
cm2 for coliform, and 2 cfu/63.6 cm2 for fungi. The bacterial contamination on the surface of the equipment
and facilities showed similar tendencies with that of
air. However, on the surfaces of the equipment and facilities in the hatcher room corridors and nonoperating

1390

Kim and Kim

HACCP application for hatcheries was not made and

a systematic investigation and evaluation of hatchery
hygiene have not been completed.
The purpose of this study was to investigate the contamination of the air and the surfaces of equipment and
facilities in a hatchery from August 2005 to September
2006 by aerobic bacteria, coliform, fungi, and Salmonella. In addition, the inhibitory effects of formaldehyde
application methods on aerosol bacterial counts in the
hatcher were also investigated.

MATERIALS AND METHODS

Location and Time of Examination
Bacterial contamination of the air and the surfaces
of equipment and facilities for some 30 sites, which included an egg sorting room, a setter room, an egg candling room, a hatcher room, and a chick counting room
in a certain hatchery located in the Gyeongsangbuk-Do
province of Korea was examined 5 times from August
2005 to September 2006. The first examination was
conducted in August 2005 immediately after a cleaning and disinfection of the hatchery after the removal
of chicks from the site. The second investigation was
performed before cleaning and disinfection in October
2005. The third, fourth, and fifth examinations were
conducted 4 h after the chicks were removed and the
site was cleaned, disinfected, and the facilities were dry
in December 2005, April 2006, and September 2006,
respectively.

Examination for Contamination of Air

and Surface of Equipment and Facilities
in Hatchery
For each of the 20 to 24 sites that were suspected
of having a high bacterial contamination of the air
and the surface of equipment and facilities, 4 subsites
evenly arranged in each site were designated along each
step of the entire hatchery from the egg receiving room
to the chick counting room. For the air samples, plate
count agar (Difco, Detroit, MI), Endo agar (Difco), and
Sabouraud dextrose agar (Difco) were used for aerobic bacteria, coliform, and fungi, respectively. Three
agar plates were prepared that had been exposed for
10 min (compared with 1 min in operating hatcher).
In addition, a Rodac agar plate (TPC: KM0808, Han-il
Komed, Gyeonggi-do, Korea) and another Rodac agar
plate (ECC: KM0802, Han-il Komed) were prepared for
the surfaces of the equipment and facilities and were
used for aerobic bacteria and coliform, respectively.
Two agar plates were stamped to the surface of the
equipment and facilities. After being transported to the
laboratory, each plate was incubated at 37C for 24 to
48 h, and then a count of the colonies on the plate surface was done, yielding colony-forming units/63.6 cm2
and colony-forming units/16 cm2, respectively.

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ducted a study on the bacterial contamination of fluff

which presented these contaminated microorganisms
for evaluating the hygienic status in hatchery (Nichols
and Leaver, 1967; Chen et al., 2002). However, Magwood (1964a) studied the microbial contamination of
the air in hatcheries and compared the results with
those obtained from the microbiological examination of
fluff. He concluded that both tests measured the level
of contamination present in the environment but that
air sampling was particularly useful because it demonstrated the wide ranges in microbial count. Furthermore
Magwood and Marr (1964) found it necessary to investigate the contamination of the surface inside hatcheries. The efficacy of hatchery sanitation programs may
be increased by measuring and comparing the bacterial
contamination of the air and the surface inside hatcheries (Magwood, 1964b).
There have been several reports on damage to chicken
owing to insufficient hatchery hygiene. Chute and Gershman (1978) reported that the microorganisms that
were isolated from the air in a hatchery were found
in chickens with omphalitis. In addition, Devos et al.
(1966) suggested that the Escherichia coli infection of
chicks may have been associated with high levels of
Enterobacteriaceae in the fluff. Effective sanitation programs should be devised to reduce the bacterial contamination in the hatchery.
Formaldehyde has long been established as an effective disinfectant in commercial poultry hatcheries
(Beesley, 1980; Deeming, 1992; Steinlage et al., 2002).
Formaldehyde was used as a disinfectant in hatcheries
because of its ease of administration and effectiveness
against a wide variety of microorganisms (Sheldon and
Brake, 1991). However, the US Environmental Protection Agency had formaldehyde regulated under the
Toxic Substances Control Act because of its toxic effect
on humans (Sheldon and Brake, 1991). Also, these regulations include setting exposure limits and maintaining health records of exposed employees, along with the
provision of additional safety equipment and training
(Wilson and Mauldin, 1989). For these reasons, Samberg
and Meroz (1995) searched for alternative disinfectants;
however, a more effective alternative was not found.
Steinlage et al. (2002) presented an effective disinfection method, which administrated 37% formaldehyde
at a rate of 1 mL/h as constant rate infusion (CRI) by
using a Buretrol Intravenous Drip Set (Braun Medical
Inc., Bethlehem, PA) to reduce the potential exposure
of humans and chicks to formaldehyde and also to reduce bacterial contamination in hatchers.
To produce hygienic poultry meat, it is essential
to inhibit microorganisms from the breeder farm, the
hatchery, the broiler farm, and the chicken slaughter
house of an integrated broiler chicken operation. Hazard Analysis and Critical Control Point has been applied to chicken slaughter houses in Korea (Kim et al.,
2002. In addition, HACCP has been applied to poultry
farms including broiler farms and breeder farms since
2008 in Korea. However, an announcement about the

1391

HATCHERY HYGIENE EVALUATION

Examination for Salmonella

Contamination in Hatchery

Comparison Test of Air Decontamination

in Hatcher by Formaldehyde Application
Methods During Hatching
This experiment was designed to look at alternative
methods of application of formaldehyde in the hatcher.
Formaldehyde was administrated 2 ways, the conventional method and the CRI method. In the conventional method, 37% formaldehyde was administrated
only once into a basin at the time of transfer, whereas
in the CRI method, the 37% formaldehyde was administrated at the same rate during hatching. The CRI was
attained with an Intravenous Infusion Set (Korea Vaccine, Seoul, Korea) falling into the basin.
The first trial consisted of 4 groups: the negative
control group (D), which did not receive 37% formaldehyde; the experimental group (A), which received
226.8 g of 37% formaldehyde through CRI at the time
of transfer, an additional dosage every 12 h, and a final
dose at 12 h before chick removal; group B, which received 56.7 g of 37% formaldehyde through CRI at the
time of transfer, at pipping, an additional dosage every

Statistical Analysis
A statistical analysis was conducted with SAS Version 9.1 (SAS Institute, 2003). One-way ANOVA and
a Tukey post-hoc test were used to compare the differences in the mean bacterial contamination of the 4 experimental groups after the use of various formaldehyde
application methods during hatching. The differences
were considered to be significant at P < 0.05.

RESULTS
Bacterial Contamination of Air Sample
in Hatchery
The results of air contamination by aerobic bacteria,
coliform, and fungi are presented in Table 1. On the
first and fifth sampling times, which collected samples
after cleaning and disinfection in a hatchery, aerobic
bacterial contamination in the operating hatchers was
high, measuring over 300 cfu/63.6 cm2 except for the
operating hatcher in the hatcher room-5 on the first
sampling time. In addition, in the operating hatchers,
coliform and fungi contamination was high, measuring
over 300 cfu/63.6 cm2 on the fifth sampling time. On
the second sampling time, which was conducted before
cleaning and disinfection, air contamination of aerobic
bacteria, coliform, and fungi in the candling-transfer
room, corridor in the hatcher room-2, and chick handling room was higher than the bacterial contamination of same sampling sites on the remaining sampling
times, which collected samples after cleaning and disinfection. In the egg sorting room, contamination by aerobic bacteria, coliform, and fungi was generally found
to be moderate, whereas in the other sampling sites,
it was minimal, measuring less than 10 cfu/63.6 cm2
for aerobic bacteria, 5 cfu/63.6 cm2 for coliform, and 2
cfu/63.6 cm2 for fungi.

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Twenty to 24 sites that were suspected of high Salmonella contamination were designated along the entire
hatchery at each of the processing steps from the egg receiving room to the chick counting room. By swabbing
the designated sites with a sterile cotton swab (Han-il
Komed), dust and debris from the surface of equipment and facilities were collected. After sterile transportation to the laboratory, each cotton swab in turn
was added to 10 mL of buffered peptone water (BPW,
Difco). After incubation of the samples in BPW at 37C
for 18 to 24 h, 0.1 mL of preenrichment BPW broth
was transferred to 10 mL of Rappaport-Vassiliadis R10
broth (Difco). After 24 to 48 h at 42C, the RappaportVassiliadis broth was streaked onto Salmonella-Shigella
agar (Difco), MacConkey agar (Difco), and Rambach
agar (Merck, Darmstadt, Germany) and incubated for
18 to 24 h at 37C. The suspicious colonies on each
plate were flooded with 1 drop of MUCAP reagent (Biolife, Milan, Italy) and after 3 to 5 min, the plates were
observed under a 366-nm Longwave UV lamp (UVP,
Upland, CA). The fluorescent colonies were presumed
to be Salmonella spp.
The presumptive colonies were selected and according to the method of Ewing (1986), the biochemical
tests were conducted. A serological test was performed
to react with Salmonella O antiserum (Difco). Colonies
showing typical agglutination by O antiserum were serotyped with Salmonella H antiserum (Difco) according
to Difco Laboratories (1977). In the case of a biphasic
organism, it was finally serotyped with a phase-changing test according to the method of Collins and Lyne
(1984).

4 h, and a final dose at 4 h before chick removal; and

group C, which received 283.5 g of 37% formaldehyde
only once into a basin at the time of transfer.
The second trial consisted of 4 groups: the negative
control group (D) and the experimental groups (A and
C) were used with the same conditions as those of the
first trial. A new experimental group (E) was added,
which received 226.8 g of 37% formaldehyde through
CRI at the time of transfer, an additional dosage every
12 h, and a final dose of 283.5 g at 12 h before chick
removal.
To evaluate bacterial contamination of the air sample,
3 sites evenly arranged in each hatcher were sampled
every 4 h during transfer to chick removal. Plate count
agar prepared to evaluate the aerobic bacteria was exposed for 1 min. After transportation to the laboratory,
the plates were incubated at 37C for 48 h and then the
colonies on the plate surfaces were counted and found
to yield colony-forming units/63.6 cm2.

1392

Kim and Kim

Bacterial Contamination on Surface

of Equipment and Facilities in Hatchery

Salmonella Contamination in Hatchery

Salmonella contamination results from the hatchery are presented in Table 3. After the investigation of
the hatchery, the results revealed that Salmonella contamination was the highest at 50.0% in samples from
the second sampling time, which were collected before
cleaning and disinfection. The samples from the first,
third, fourth, and fifth sampling times, which were collected after cleaning and disinfection, had contamination levels that were 4.2 to 31.8% lower.
Salmonella was primarily isolated from the hatcher
rooms, chick counting room, and the related equipment
and facilities but not from earlier processing rooms
such as the egg receiving room, egg sorting room, setter
rooms, and candling-transfer room.
The Salmonella serotype isolated most frequently
in the hatchery was Salmonella Senftenberg. Other
occasional Salmonella serotypes included Salmonella
Schwarzengrund, Salmonella Madelia, Salmonella Montevideo, and Salmonella Enteritidis. On only one occasion in the third sampling time, 2 Salmonella serotypes,
Salmonella Enteritidis and Salmonella Senftenberg,
were detected in the store house for hatchery waste.

Comparison of Air Decontamination

in Hatcher by 37% Formaldehyde
Application Methods During Hatching
The results from the comparison of the air decontamination in the hatcher after 37% formaldehyde application methods during hatching by 2 trials are presented
in Figure 1. In the first trial (Figure 1a), the 2 experimental groups (A and B), which received 37% formaldehyde through CRI during hatching, and experimental
group C, which received 37% formaldehyde only once
into a basin at the time of transfer, had a significantly

DISCUSSION
Hatchery contamination by microorganisms acts as
an important medium in the spread of diseases in the
poultry farm including the parent stock farm (Funk
and Irwin, 1955; Harry and Gordon, 1966; Sheldon
and Brake, 1991; Scott and Swetnam, 1993). When
the bacterial contamination in a hatchery is high, the
decrease of growth rate and the increase of mortality
occur (Wright et al., 1959). To prevent hatchery contamination, a great interest has been taken in the aspects of hatchery hygiene, which include hatching eggs,
the equipment and facilities, and the vehicles used for
transporting hatching eggs and chicks (Jordan et al.,
2001). Stringent hygiene in the hatchery can minimize
contamination and maximize chick quality. When regular hatchery hygiene has been carried out, there has
been an impressive improvement in the sanitary conditions in hatcheries (Samberg and Meroz, 1995). New
equipment and facilities do not necessarily result in
good hygiene, but a good hygienic program is the best
way to improve hatchery hygiene (Chen et al., 2002).
Hatchery hygiene was evaluated by measuring the
bacterial contamination of air and the surface of equipment and facilities. By measuring the bacterial con-

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Contamination on the surface of the equipment and

facilities by aerobic bacteria and coliform is presented
in Table 2. In the corridor of the hatcher room and the
operating and nonoperating hatchers, aerobic bacteria
was high, measuring over 100 cfu/16 cm2 during all
sampling times regardless of a cleaning and disinfection
in a hatchery except for the corridor in the hatcher
room-2 on the first sampling time and nonoperating
hatcher in the hatcher room-3 and hatcher room-5 on
the second sampling time. In addition, in the some operating and nonoperating hatchers, coliform was high,
measuring over 100 cfu/16 cm2 on the second and fifth
sampling times. In the chick handling room, separating room, and chick counting room, contamination was
generally moderate, whereas in the remaining sampling
sites, it was minimal, measuring about 10 cfu/16 cm2
for aerobic bacteria and 5 cfu/16 cm2 for coliform.

superior inhibitory effect on aerosol bacterial counts 12

h after transfer than those of the negative control group
(D) (P < 0.05). The 2 experimental groups (A and
B) also had a significantly superior inhibitory effect on
aerosol bacterial counts 4 h before the hatching than
group C. Between the 2 experimental groups (A and
B), experimental group A, which received 226.8 g of
37% formaldehyde through CRI at the time of transfer,
an additional dosage every 12 h, and a final dose at 12
h before chick removal, had significantly lower aerosol
bacterial levels 4 h before the hatching as compared
with group B, which received 56.7 g of 37% formaldehyde through CRI at the time of transfer, at pipping,
an additional dosage every 4 h, and a final dose at 4 h
before chick removal (P < 0.05).
In the second trial (Figure 1b), the 2 experimental
groups (A and E) received 37% formaldehyde through
CRI during hatching and group C had a significantly
superior inhibitory effect on aerosol bacterial counts 24
h after transfer than the negative control group (D) (P
< 0.05). The 2 experimental groups (A and E) had a
significantly superior inhibitory effect on aerosol bacterial counts 4 h before hatching than group C. There
was no significant difference in the level of bacterial
contamination between experimental group A, which
received 226.8 g of 37% formaldehyde through CRI at
the time of transfer, an additional dosage every 12 h,
and a final dose at 12 h before chick removal, and group
E, which received 226.8 g of 37% formaldehyde through
CRI at the time of transfer, an additional dosage every
12 h, and a final dose of 283.5 g at 12 h before chick
removal.

>3002

1.0
2.8
0

>3002

2.0
0.5
1.0

1.0

0.3

43.5

10.0

2.0

25.5
28.0

0.5
0.8

2.8
0.5
1.0
0
0

0.8

0.8

4.0

0.8

22.82

3.0
2.5
3.5
0.5
11.0
8.0
3.8
0.5

Second

0.3
1.0
1.3
0.5
14.8
6.0
6.3
0.5

First

0.5
0.5
0.5

1.0
12.0
4.5

0.8

2.04

8.02

2.3

0.5
1.3

1.5

2.3

0.5
2.5
1.5
0.5
1.3
1.3
0.8
2.3

Third

9.8
0.3
1.3

4.3
13.0
3.5
1.8
4.8

0.52

0.84

10.84

3.3

1.0
1.8

1.5

8.0

7.0
5.0
3.3
6.3
48.8
3.0
5.8
1.3

Fourth

0.5
1.0
1.3

2.8
13.0
0.5
0.8
1.3

4.34

>3002

>3002

4.0

3.0
0.5

3.0

5.5

1.7
1.5
3.0
2.3
17.8
6.8
5.0
0

Fifth

1.0
0
1.3

0.3
0
0.3
0.3
0

6.52

11.52

36.02

0.3
0.5

2.5

0.8
0.8
0.5
0.8
11.5
6.0
8.0
0

First

3.0
1.8
0.3

21.0

12.0

0.5

0.3

2.0
33.8

1.5

1.0
2.5
2.0
0.8
3.0
1.7
1.5
1.0

Second

0.3
0.5
0

0
4.7
2.5

04

7.72

0.3

0
0.8

0.3

0.3
0.3
0.5
0.3
0.3
0
0.3
0

Third

Coliform

1.5
0
3.0

2.5
9.0
0.8
2.8
1.0

02

0.34

1.84

0.5

1.0
0.8

0.3

2.0

1.0
0.3
0.3
2.3
14.5
0.5
1.3
0.8

Fourth

Sampling time1

57.82
13.52

>3002
>3002

2.3
0.8
1.3

0.5

2.8

0.3
0.8
0.3

0.5
1.3

1.5
0

0.5
0.3
0.5
0.5
0.8

4.8

0.3

0.5
0.8
0.3
0.3
0.8

0.5

0.8

113.52

1.0
1.5
1.0
10.8
10.3
3.3
7.0
0.8

0
0.3
1.0
0.5
12.3
0.5
1.5
0.5

2.84

First

Fifth

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1.3
2.0
5.3

62.3

8.3

1.0

0.8

1.8
24.8

1.0

1.0

2.8
1.3
2.0
0.5
9.0
1.0
0
0.5

Second

0.3
0
0

0.8
3.7
2.5

0.3

04

02

0
0

0
0
0.3
0
0
0
0.3
2.0

Third

Fungi

0.3
0
0

1.5
4.5
0.3
0.8
0.3

02

04

2.84

0.3
2.0

1.5

1.5

0
0.7
0.3
0
1.5
0.5
0
0.5

Fourth

0
1.3
0

2.3
2.3
0
0
0.5

0.54

>3002

>3002

2.3

0.3
0

0.3

0.3
0.3
1.0
0
0.8
0.8
0.5
0.3

Fifth

1First = samples were collected immediately after cleaning and disinfection after the removal of chicks from the hatchery in August 2005; second = samples were collected before cleaning and disinfection in October 2005; third, fourth, and fifth = samples were collected 4 h after cleaning and disinfection, by which time the facilities were dry, after the removal of chicks from the hatchery in January
2006, April 2006, and September 2006, respectively.
2Operating hatcher (eggs are incubated).
3Not tested.
4Nonoperating hatcher (after a cleaning and disinfection after the removal of chicks).

Dressing room
Main office
Main corridor
Egg receiving room
Egg sorting room
Storage room for egg
Presetter room
Corridor in the setter
room-2
Setter in the setter
room-1
Setter in the setter
room-3
Candling-transfer room
Corridor in the hatcher
room-2
Corridor in the hatcher
room-5
Hatcher in the hatcher
room-1
Hatcher in the hatcher
room-3
Hatcher in the hatcher
room-5
Chick handling room
Separating room
Chick counting room
Chick storage room
Egg box and egg store
room
Setter tray store room
Hatcher tray store room
Chick box store room

Sampling site

Aerobic bacteria

Table 1. Bacterial contamination of air samples by process steps in hatchery (cfu/63.6 cm2)

HATCHERY HYGIENE EVALUATION

1393

0.3
3.0
5.8
0
0.3
0.3
0
0.5
0.3
0.8
2.5
91.0
>100
>1002
>1002
>1002
19.5
3.5
31.8
0
0
1.8
1.0
2.8

Dressing room
Main office
Main corridor
Egg receiving room
Egg sorting room
Storage room for egg
Presetter room
Corridor in the setter room-2
Setter in the setter room-1
Setter in the setter room-3
Candling-transfer room
Corridor in the hatcher room-2
Corridor in the hatcher room-5
Hatcher in the hatcher room-1
Hatcher in the hatcher room-3
Hatcher in the hatcher room-5
Chick handling room
Separating room
Chick counting room
Chick storage room
Egg box and egg store room
Setter tray store room
Hatcher tray store room
Chick box store room

0.3
1.8
3.0
5.0
0.3
5.3
15.8
0.5
3.3
0.3
1.8
>100
>100
>1002
53.82
25.82
21.5

19.5
3.3
1.8
3.5
4.8
43.5

Second
0.5
5.8
3.5
0.3
0
0.3
2.3
0
0
1.0
1.5
>100
>100
>1002
>1002
4
0
12.3
0.5
0
1.0
0.8
0.5
3.8

Third
0.3
3.8
2.8
3.0
3.3
9.3
0.5
0.8
1.3
0.3
1.5
>100
>100
>1003
>1003
>1002
13.0
11.5
13.0
1.5
2.3
5.0
1.5
3.8

Fourth
0.8
11.0
3.3
1.3
1.3
2.8
0.8
5.0
16.3
0
1.3
>100
>100
>1003
>1003
>1002
2.0
>100
6.3
15.3
12.3
0.3
0
15.3

Fifth
0
0
0
0
0
0
0
0
0
0
0
0.8
0.8
4.82
0.32
6.32
0
0
0
0
0
0
0
0

First

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Third
0.5
1.0
1.0
0.3
0.3
0
0.5
0
0
0
0.3
1.5
12.3
4.52
38.02

0
2.5
0
0
0.3
0.8
0.3
0.3

Second
0.3
0
1.3
0
0
0
0
0
0
0.3
0
10.8
0
2.02
>1002
1.02
7.3

5.8
6.0
0
0.3
1.0
0.8

Coliform

1.0
0
0
0.8
0.3
0.5
0.8
0
0
0.3
0.3
11.3
0.3
49.03
0.33
02
0.3
0.3
10.0
1.3
0
0.8
0
0.5

Fourth

0
0.8
0
0
0.5
0.5
0.8
0
4.8
0
0.3
1.0
3.8
>1003
>1003
12.02
0
>100
0
4.5
0
0
0
0

Fifth

1First = samples were collected immediately after cleaning and disinfection after the removal of chicks from the hatchery in August 2005; second = samples were collected before cleaning and disinfection in October 2005; third, fourth, and fifth = samples were collected 4 h after cleaning and disinfection, by which time the facilities were dry, after the removal of chicks from the hatchery in January
2006, April 2006, and September 2006, respectively.
2Nonoperating hatcher (after a cleaning and disinfection after the removal of chicks).
3Operating hatcher (eggs are incubated).
4Not tested.

First

Sampling site

Aerobic bacteria

Sampling time1

Table 2. Bacterial contamination on the surface of equipment and facilities by process steps in hatchery (cfu/16 cm2)

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Egg receiving room

Egg sorting room (conveyor)
Egg sorting room (inhalant)
Setter room 2 (indoor exhaust pipe)
Setter room (dome)
Setter room indoor and outdoor
Setter tray cleaning and store room
Outdoor ventilating outlet of setter room
Candling-transfer room (conveyor)
Candling-transfer room (inhalant)
Hatcher room 3 (indoor exhaust pipe)
Hatcher room 3 (outdoor ceiling)
Hatcher room (indoor and outdoor)
Air inlet of hatcher room
Hatcher room-outdoor ceiling (operating)
Hatcher tray cleaning and store room
Hatcher tray before cleaning
Hatcher tray after cleaning
Outdoor ventilating outlet of hatcher room
Vaccination equipment
Chick box receiving room
Chick box before cleaning
Chick box after cleaning
Store house of hatchery waste

Chick counting room (disterk)

Chick counting room (separator)
Chick counting room (conveyor)
Total (%)

Salmonella Senftenberg
Salmonella Senftenberg
Salmonella Senftenberg
11/22 (50.0)

Salmonella Senftenberg
Salmonella Senftenberg
Salmonella Senftenberg

Salmonella Senftenberg
Salmonella Senftenberg

Salmonella Montevideo
Salmonella Schwarzengrund

Salmonella Madelia

Second

Salmonella Senftenberg
Salmonella Senftenberg

Salmonella Senftenberg

Salmonella Senftenberg

Salmonella Senftenberg

Salmonella Enteritidis,
Salmonella Senftenberg

Salmonella Senftenberg

7/22 (31.8)

Third

Sampling time1

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Salmonella Senftenberg

4/23 (17.4)

1/24 (4.2)

Salmonella Senftenberg

Salmonella Senftenberg

Salmonella Senftenberg

Salmonella Senftenberg

Fifth

Fourth

1First = samples were collected immediately after cleaning and disinfection after the removal of chicks from the hatchery in August 2005; second = samples were collected before cleaning and disinfection in October 2005; third, fourth, and fifth = samples were collected 4 h after cleaning and disinfection, by which time the facilities were dry, after the removal of chicks from the hatchery in January
2006, April 2006, and September 2006, respectively.
2Not tested.
3No. isolated/no. tested.

4/203 (20.0)

Salmonella Senftenberg
Salmonella Senftenberg
Salmonella Senftenberg

Salmonella Senftenberg

First

Sampling site

Table 3. Salmonella contamination by process steps in hatchery

HATCHERY HYGIENE EVALUATION

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Figure 1. The changes of aerosol bacterial counts in hatcher by 37% formaldehyde application methods during hatching (a: first trial, b: second
trial). CRI = constant rate infusion; TNTC = too numerous to count.

HATCHERY HYGIENE EVALUATION

entire hatchery from the egg sorting room to the chick

counting room. It was determined that the main contamination source of Salmonella in the hatchery was
the hatcher room and the related equipment and facilities. The Salmonella that was isolated from poultry and
its environment in Korea has 43 different serotypes.
Salmonella Blockley, Salmonella Enteritidis, Salmonella
Newport, and Salmonella Hilingdon were mainly isolated in poultry meat (Kim and Tak, 1998; Lee et al.,
1999; Sung et al., 2002) and Salmonella Typhimurium
and Salmonella Enteritidis were isolated from the poultry environment (Woo et al., 2000). The cecal contents
of 1-d-old chicks in a hatchery were detected to have
Salmonella Typhimurim (Oh and Choi, 1994). Byrd
et al. (1999) reported that 11 different serotypes were
isolated from the hatchery and most of the serotypes
were Salmonella Heidelberg and Salmonella Kentucky
in the United States. However, the Salmonella serotype
that was isolated most frequently in the hatchery was
Salmonella Senftenberg in Korea. It was also found
that Salmonella serotypes have different distributions
nationally and regionally. Salmonella Senftenberg was
announced as a cause of gastroenteritis in patients who
ate food that was cross-contaminated by turkey meat
in the United States (LEcuyer et al., 1996) and in
children who ate cereal contaminated with Salmonella
Senftenberg in the United Kingdom (Rushdy et al.,
1998). Salmonella Senftenberg in the hatchery can be
transmitted to the final product, which is poultry meat,
and subsequently cause gastroenteritis. Consequently,
Salmonella contamination in hatcheries should be minimized and further investigation is needed to identify
the transmission routes of Salmonella Senftenberg from
the hatchery to slaughter house.
The reduction of bacterial contamination in the
hatcher is very important because the hatcher is a major source of contamination within the hatchery. Effective cleaning and disinfection programs are vital in
the hatchery. Many different disinfectants and disinfection methods are used to reduce contamination in
a hatchery; these include selecting disinfectants after
testing for bacterial tolerance to the disinfectants with
the bacteria that were mainly isolated from the hatchery, administrating hydrogen peroxide during the incubation period, administrating 37% formaldehyde in
the hatcher, and fumigating the inside of the hatcher
with formaldehyde after hatching (Nichols and Leaver, 1967; Hodgetts et al., 1976; Samberg and Meroz,
1995; Sander and Wilson, 1999; Steinlage et al., 2002).
However, a more effective disinfectant than formaldehyde was not reported before this study. The most effective method was the use of 37% formaldehyde as a
CRI to reduce bacterial contamination in the hatcher
(Steinlage et al., 2002). In this study, the hatcher that
received 37% formaldehyde as a CRI during hatching
had a significantly superior inhibitory effect on aerosol
bacterial counts 4 h before hatching than the hatcher
that received 37% formaldehyde once into a basin at
the time of transfer.

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tamination of air in a hatchery using an air sampler

(TDL slit sampler model B with vacuum supplied by
a Gast air pump model 0321), which on calibration
passed 28.88 to 30.30 L per minute when operated in
the range of 26.67 to 35.56 cm of vacuum, the bacterial
contamination was found to be high in the operating
hatcher and hatcher room (Magwood, 1964a). In addition, Nichols and Leaver (1967) reported that the fluff
on the hatching days, 58% of total bacteria, 9% of coliform, and 3% of fungi had high contamination over 106
cfu/g, respectively. In general, the bacterial contamination is very high in the operating hatcher. In the present
study, aerobic bacteria, coliform, and fungi of air had
high levels of contamination in the operating hatcher but had low levels of contamination in the hatcher
room. In particular, aerobic bacterial contamination in
the operating hatchers on the first and fifth sampling
times was very high, measuring over 300 cfu/63.6 cm2
except for the hatcher in hatcher room-5, in which the
usage frequency was relatively low compared with the
other hatcher rooms, on the first sampling time. A direct relationship has been established between airborne
microorganisms and the amount of contamination on
various hatchery surfaces (Magwood, 1964b). In this
study, the bacterial contamination on the surface of
the equipment and facilities showed similar tendencies
with that of air. The air in the corridors of the hatcher
room and the nonoperating hatcher had a low level of
aerobic bacterial contamination, whereas the level of
aerobic bacterial contamination on the surface of the
equipment and facilities was very high.
The bacterial contamination after fumigation was
much lower than before fumigation when the bacterial
contamination of the fluff in the hatcher was compared
(Nichols and Leaver, 1967). In addition, Sander and
Wilson (1999) reported that the bacterial contamination of the experimental group that was disinfected
with hydrogen peroxide was lower than the group that
was not disinfected. In the present study, the third,
fourth, and fifth experiments that were performed with
cleaning and disinfecting had a lower level of bacterial contamination than that of the second experiment
that was performed without cleaning or disinfecting the
hatchery. It was suggested that hatchery hygiene can
be greatly improved by cleaning and disinfecting the
hatchery machinery and environment.
Salmonellosis is one of the major foodborne causes
of gastroenteritis and is frequently associated with contaminated poultry meat (Bryan and Doyle, 1995). Salmonellosis also produces economic losses in the poultry
industry (Saif et al., 2003). Salmonella contamination
in a hatchery can produce poor-quality chicks, resulting in a decreased feed conversion rate, increased mortality, and poor flock uniformity. It was reported that
Salmonella was isolated from fluff in a few hatcheries
(Chen et al., 2002) and that 12.1% of the hatcher trays
were contaminated with Salmonella (Byrd et al., 1999).
However, the previous study only examined the fluff
and hatcher trays. The present study investigated the

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Kim and Kim

In conclusion, administration of formaldehyde by a

CRI as opposed to the conventional method currently
used in Korea of a single administration of 37% formaldehyde into a basin at the time of transfer had a
superior inhibitory effect on aerosol bacterial counts.
Therefore, widespread use of the CRI method can be
expected to have a positive effect on hatchery hygiene
and productivity. Further, this study provides useful
information on the application of HACCP in the hatcheries of Korea.

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