Download The Bacterial Flora of the Trachea, Liver, Spleen and Heart

Pertanika 10(3), 289 - 293 (1987)
The Bacterial Flora of the Trachea, Liver, Spleen and
Heart Blood of Chicken
M. SHAH-MAJID and H. JAH '
Department of Veterinary Pathology and Microbiology,
Faculty of Veterinary Medicine and Animal Science,
Universiti Pertanian Malaysia,
43400 Serdang, Selangor Darul Ehsan, Malaysia.
Key words: Chicken organs; bacterial flora.
ABSTRAK
Sejumlah 184 trakea, 147 hati, 150 limpa dan 148jantung daripada ayamyangsihat danyang
mati telah disampel dan flora bakteria dikenalpasti. Bakteria yang paling biasa dikultur daripada
organ-organ ayam yang sihat adalah Staphylococcus spp., Streptococcus spp., dan Escherichia coli.
Sementara itu Escherichia coli, Aeromonas spp., Staphylococcus spp., Proteus spp., dan Citrobacter
spp., pula adalah bakteria yang biasa didapati daripada ayam yang mati. Pasteurella haemolytica
telah didapati dalam bilangan yang kecil di dalam trakea, hati dan limpa ayam yang sihat, serta
dalam hati, limpa dan jantung ayam yang mati.
ABSTRACT
A total of 184 tracheas, 147 livers, 150 spleens and 148 hearts from apparently healthy and
dead chickens were sampled and the bacterial flora was determined. The bacteria most commonly
cultured from these four organs of healthy birds were Staphylococcus spp., Streptococcus spp., and
Escherichia coli. Escherichia coli, Aeromonas spp., Staphylococcus spp., Proteus spp., and Citrobacter spp., were the common isolates from dead chickens. Pasteurella haemolytica was present in
small numbers in the trachea, liver and spleen of healthy chickens and liver, spleen and heart of dead
chickens.
INTRODUCTION
Little information is available on the bacterial
flora of the trachea, liver, spleen and heartblood
of apparently healthy chickens and those that
were dead because of unknown aetiology or
disease manifestation. Price et al. (1957),
Smibert et al. (1958) and Bisgaard ^1977)
stated that the tracheal flora of healthy chickens
consisted primarily of gram positive microorganisms. A shift to a predominately gram
negative flora in chickens with respiratory
disease has been reported by Biddle and Cover
(1957), Price et al. (1957) and Bisgaard (1977).
Mushin et al (1980) postulated that Pasteurella
haemolytica is a member of the normal flora of
the respiratory tract of chickens and is also involved in pathological lesions (Bisgaard, 1977;
Mushin et ai, 1980). Hacking and Pettit (1974)
isolated P. haemolytica from liver and heart
blood of pullets and layers with high mortality
rate and decline in egg production. Addo and
Mohan (1985) isolated atypical P. haemolytica
Type A from nodular necrotic liver and heartblood of domestic fowl. Recently Shah-Majid et
al. (1986) isolated P. haemolytica from five
spleens with white foci from 3 week-old chickens
that were on Newcastle disease vaccination trial.
The purpose of this study was to determine
Part of junior author's Doctor of Veterinary Medicine Animal Industry Project.
M. SHAH MAJID AND H. JAH
the bacterial flora of the trachea, liver, spleen
and heart blood of apparently healthy and dead
chickens in the Universiti Pertanian Malaysia
(UPM) Poultry Unit and from a private poultry
farm.'Since Shah-Majid et al. (1986) reported
the presence of P. haemolytica in the spleen of
chickens, it was decided to record the incidence
of this organisms in these four organs of healthy
and dead birds. The Pasteurella haemolytica
described in this paper are those organisms that
are gram negative slender long rods. Piechulla et
ai (1985) suggested that these organisms do not
belong to the Genus Actinobacillus as suggested
by Mraz in 1969 but probably a new genus be
created for them.
MATERIALS AND METHODS
Organ samples of healthy chickens were obtained from the Universiti Pertanian Malaysia
(UPM) commercial unit. A total of 100 tracheas,
64 livers, 64 spleens and 63 hearts were randomly
selected from these healthy chickens. These
chickens were slaughtered by the 'Halal' method
and all organs were sampled within one to two
hours after slaughter. Organ samples from dead
chickens were collected from a private poultry
farm at Dengkil, Selangor, UPM commercial
unit and from the post mortem room of the
Faculty of Veterinary Medicine and Animal
Science, UPM, Serdang. A total of 84 tracheas,
83 livers, 86 spleens and 85 hearts were collected
from these dead chickens. At necropsy all dead
birds were examined grossly. The age group of
the dead chickens varied from one to sevenweek-old while the healthy chickens were sevenweek-old.
All samples were inoculated onto blood
agar and MacConkey agar. The tracheas were
swabbed and then inoculated onto these two
media. The livers, spleens and hearts were im-
TABLE1
Isolation of bacteria from various organs of healthy 7 week-old chickens
Organs
Trachea
Liver
Spleen
Heart blood
Staphylococcus spp.
95
40
26
30
Streptococcus spp.
60
23
7
25
Escherichia coli
42
18
1
10
Micrococcus spp.
22
-
Enterobacter spp.
12
Bacillus spp.
4
Citrobacter spp.
2
Pasteurella haemolytica
2
-
—
-
-
—
-
—
-
-
—
—
1
—
3
Klebsiella spp.
-
-
—
—
Aeronomas spp.
-
—
—
—
Salmonella spp.
-
—
—
—
Yersmia spp.
-
—
—
—
Edwardsiella spp.
-
—
—
Proteus spp.
-
—
—
—
Acinetobacter spp.
-
—
—
—
Pseudomonas spp.
-
—
_
_
Corynebacterium spp.
-
-
-
-
— No bacteria isolated
290
PERTANIKA VOL. 10 NO. 3, 1987
THE BACTERIAL FLORA OF THE TRACHEA, LIVER, SPLEEN AND HEART BLOOD OF CHICKEN
mersed briefly for three to four seconds in a
beaker of boiling water in order to reduce
surface contamination (Jang et al., 1976). The
interior portion of these organs was exposed and
the inner surface was swabbed and then inoculated onto the media mentioned.
All plates were incubated aerobically for 24
hours at 37°C. Plates with no visible growth were
reincubated for another 24 hours. Identification
of isolates was carried out according to the
method of Jang et al. (1976).
Statistical analysis was by chi-square.
RESULTS AND DISCUSSION
The aerobic bacterial flora isolated from apparently healthy and dead chickens is shown in
Table 1 and 2 respectively. Eight bacterial
species were isolated from healthy chickens while
17 species were isolated from dead chickens. The
dead chickens were those infected with Newcastle disease, chronic respiratory disease, coccidiosis, and those with yolk sac infection, congested liver and pulmonary oedema and with no
significant lesions based on gross examination.
The bacteria most commonly isolated from the
healthy chickens were Staphylococcus spp.,
Streptococcus spp. and Escherichia colt. Apart
from E. coli which is present in the trachea in
large numbers in this study, the tracheal flora of
healthy chickens consisted primarily of Gram
positive microorganisms. This is in accordance
with the findings of Price et al. (1957), Smibert
et al (1958) and Bisgaard (1977). Perhaps the
method of slaughter may play a significant role in
the contamination of the trachea with E. colt. P.
haemolytica was isolated in small numbers in
TABLE 2
Isolation of bacteria from various organs of dead chickens
Organs
Staphylococcus spp.
Streptococcus spp.
Escherichia colt
Micrococcus spp.
Trachea
Liver
Spleen
Heart blood
3 A (10) B
3(12)
7(10)
4(13)
l(-)
- (7)
17(51)
18 (48)
20 (62)
" (-)
22 (51)
- (5)
" (3)
7 «
- (2)
Enterobacter spp.
" (1)
- (6)
- (4)
Bacillus spp.
" (1)
5(10)
- (1)
1(6)
- (-)
2(7)
" (3)
- (2)
Citrobacter spp.
- (-)
2(5)
3(9)
" (3)
Klebsiella spp.
- (")
2(7)
1(2)
1(5)
- (4)
1(7)
Aeromonas spp.
2(14)
3(7)
3(11)
2(5)
Salmonella spp.
"
- (2)
- (3)
" (3)
Yersinia spp.
M-)
l(-)
-
(-)
-
(")
Edwardsiella spp.
~ (")
(-)
-
(-)
- (14)
l(-)
1(10)
-
Proteus spp.
- (1)
Pasteurella haemolytica
Acinetobacter spp.
(")
- (9)
~(7)
" (1)
Pseudomonas spp.
" (")
" (2)
- (3)
" (1)
"(1)
-
(-)
Corynebacterium spp.
- (")
- (1)
"
-
(-)
A
Number of bacteria isolated from 7 week-old chickens.
B
—
Number of bacteria isolated from 1 to 6 week-old rhickens
No bacteria isolated
PERTANIKA VOL. 10 NO. 3 t 1987
(-)
291
M. SHAH MAJID AND H. JAH
this study. These organisms were present in the
trachea of healthy birds although the number
isolated in this study is small when compared to
reports of Bisgaard (1977) and Mushin et al.
(1980). P. haemolytica was also present in the
liver and spleen of healthy and dead birds but
were not isolated from trachea of dead chickens
and heart blood of healthy chickens. There was
no significant difference between the prevalence
of P. haemolytica isolated from apparently
healthy chickens and dead chickens (P < 0.05).
P. haemolytica has been incriminated as a
primary or secondary pathogen in chickens and
has been isolated from the liver in cases of enlarged liver, severe peritonitis, salpingitis, respiratory disease and drop in egg production,
septicaemia and heart lesions (Harbourne, 1962;
Greenham and Hill, 1962; Hacking and Pettit,
1974; Addo and Mohan, 1985). Bisgaard (1977)
and Mushin et ai} (1980) postulated that P,
haemolytica, under conditions of stress or in
cases of extreme debilitation condition, may
acquire the role of an opportunist and participate in pathological processes.
Staphylococci and Streptococci were also
present in the liver, spleen and heartblood of
healthy chickens. Birds generally have no lymph
nodes, therefore, microorganisms reach the liver
(and spleen) via portal vene and lymph vessels.
The defense mechanisms located in the lymph
nodes in mammals are dispersed throughout the
tissues in birds. Consequently, microorganisms
enter the parenchymata to be destroyed forthwith. Staphylococcus spp. and Streptococcus
spp. were more frequently isolated from tracheas
of healthy chickens as compared to dead
chickens (P 0.05).
Gram negative microorganisms are the predominant bacteria isolated from all organs of the
dead birds and Salmonella spp., a known
primary pathogen was isolated from liver, spleen
and heart blood which indicates bacteremia.
Aeromonas spp. were frequently isolated in this
study. Shane and Gifford (1985) indicated that
Aeromonas hydrophila is widely distributed in
exotic and free living avian species and their
environment and maybe isolated from a variety
of organs or sites at post mortem. A hydrophila is
a typical organism of the water and is considered
a secondary invader. It needs stressors or other
292
diseases to infect the birds and the toxins they
may produce can be lethal.
It is concluded that our findings are similar
to those reported in the literature. The presence
of Staphylococcus spp., Streptococcus spp., P.
haemolytica, Aeromonas spp., in various organs
sampled needs further investigation.
REFERENCES
ADDO, P.B. and K. MOHAN. (1985): Atypical Pasteurella haemolytica type A from poultry. Avian Dis.
29:214-217.
BlDDLE, E.S. and M.S. COVER. (1957): The bacterial
flora of the respiratory tract of chickens affected
with chronic respiratory disease. Am.J. Vet. Res.
18:405-408.
B ISGA ARD, M. (1977): Incidence of Pasteur ella
haemolytica in the respiratory tract of apparently
healthy chickens and chickens with infectious
bronchitis. Characterization of 213 strains. Avian
Pathol. 6:285-292.
GREENHAM. L.W. and T.J. HILL. (1962): Observa-
tions on an avian strain of Pasteurella haemolytica. Vet. Rec. 74:861.
HACKING, W.C. and J.R, PETTIT. (1974): Pasteurella
haemolytica in pullets and laying hens. Avian
Dis. 18:483-486.
HARBOURNE, J.F. (1962): A haemolytic cocco-bacillus
recovered from poultry. Vet. Rec. 74: 640.
JANG, S.S., E.L. BIBERSTEIN and D.C. HIRSH. (1976):
A manual of Veterinary Clinical Bacteriology arid
Mycology, Veterinary Medical Teaching Hospital, School of Veterinary Medicine, University of
California, Davis.
MUSHIN, R., Y. WEISMAN and N. SINGER. (1980):
Pasteurella haemolytica found in the respiratory
tract of fowl, Avian Dis. 24: 1 6 2 - 168.
PIECHULLA. K., M. BISGAARD. H. GERLACH and W.
MANNHELM. (1985): Taxonomy of some recently
described avian Pasteurella /A ctinocillus-like
organisms as indicated by deoxyribonucleic acid
relatedness. Avian Pathol. 14: 281 - 3 1 1 .
PRICE, K.E., Z. ZOLLI, JR., W.B. HARDIE and M.J.
GALLIAN. (1957): Respiratory tract flora in CRD
and effect of antibiotics in the feed. Poultry Sci.
36:219-225.
SHAH-MAJID, M.,
I. AINI and A.
RAHMAN AZIZ.
(1986): Isolation of Pasteurella haemolytica from
the spleen of chickens. Pertanika. 9: 265 - 2 6 6 .
SHANE, S.M. and D.H. GIFFORD. (1985): Prevalence
and pathogenicity of A eromonas
Avian Dis. 29:681-689.
PERTANIKA VOL. 10 NO. 3,1987
hydrophila.
THE BACTERIAL FLORA OF THE TRACHEA, LIVER, SPLEEN AND HEART BLOOD OF CHICKEN
SMIBERT, R.M., H.M. D E VOLT and J.E. FABER, Jr.
159-166.
(1958): A study on the bacterial flora of the respiratory system of normal chickens. Poultry Sci. 37:
PERTANIKA VOL. 10 NO. 3, 1987
(Received 19January, 1987)
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