Professional Documents
Culture Documents
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).
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
1389
1390
1391
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.
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).
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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-
>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
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)
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
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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Kim and Kim
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
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
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1396
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.
1397
1398
REFERENCES