Pak. J. Agri., Agril. Engg., Vet. Sci.

, 2021, 37 (2): 132-138

ISSN: 1023-1072 (Print), ISSN: 2663-7863 (Online)
https://doi.org/10.47432/2021.37.2.9

AVIAN ESCHERICHIA COLI INFECTION IN MEAT-TYPE

CHICKEN: PREVALENCE AND PATHOLOGY
*1 2 3 1 1
Q. A. Shah , N. M. Soomro , J. M. Shah , M. A. Hussain and S. F. Syed
1
Faculty of Veterinary and Animal Sciences, Lasbela University of Agriculture, Water and
Marine Sciences, Uthal, Balochistan, Pakistan
2
Faculty of Veterinary Medicine, University of Poonch, Rawala Kot Azad Jammu and Kashmir
3
Shaheed Benazir Bhutto University of Veterinary and Animal Sciences, Sakrand, Pakistan

ABSTRACT
Avian colibacillosis is considered the major bacterial disease in the poultry industry worldwide. This
study was conducted to determine prevalence and pathology by avian pathogenic Escherichia coli
(APEC) in broiler chicken. Various organs of affected broilers were checked for the E. coli infection.
One hundred and forty commercial poultry farms were divided into seven age groups (A, B, C, D, E, F,
st nd rd th th th th
and G). Group A (1 week), B (2 week), C (3 week), D (4 week), E (5 week), F (6 week), G (7 to
th
9 week). Investigations were conducted from day 1 until the marketing of birds based on clinical
findings. A total of 2491 (23.71% of sick birds) dead birds were observed positive for E. coli infection.
Three forms of infection were observed i.e. omphalitis, colisepticaemia, and colibacillosis at the rate of
1.55, 17 and 56.65%, respectively. Omphalitis was confirmed by microbial culture and revealed in only
groups A and B with 76.24 and 23.76 respectively. While, Colisepticaemia was observed at the rate of
3.11, 6.74, 20.18, 29.63, 19.16, and 22.18 in groups B, C, D, E, F, and G respectively. Whereas
Colibacillosis was noticed as 0.075, 3.35, 5.99, 19.20, 26.70, 16.11 and 28.55% in groups A, B, C, D,
E, F, and G respectively. Petechiation of heart, kidney, and gizzard besides hemorrhagic enteritis,
hemorrhagic tracheitis, and necrotic foci on the liver and caseous exudates in air sacs were the prime
gross lesions noticed in colibacillosis. Microscopically, no lesion was observed in serosa, muscularis
externa, and submucosa. However, loss of epithelial tissue and breaches in the mucosal layer of the
small intestine were observed. Furthermore, characteristic pink and grayish colonies were observed in
E. coli cultures on McConkey’s and Blood agar respectively.

Keywords: broiler, Escherichia coli, infection, pathology, prevalence

INTRODUCTION1 can combat the shortage of protein requirement

The poultry industry, as a source of animal by the cheapest available source of animal
proteins for a balanced human diet, occupies an protein through filling the gap of protein
important place in the sphere of food production. availability as the poultry have a rapid growth
The results of this study indicated that rate and low generation gap as compared to
colibacillosis is highly prevalent in the study other livestock.
area; these observations raise serious concern Avian Pathogenic Escherichia coli (E. coli)
and prompt attention and thus a huge economic cause severe economic losses in the animal and
could be reduced significantly and problematic poultry industry by increasing morbidity and
aspects (i.e. under feeding and malnutrition) are mortality rate as well as reducing the production
directly related to the health of the nation. of the animals. It causes a variety of infections
Pakistan commercial poultry is an important and through pan systemic involvement of various
significant contributor in livestock share in GDP vital systems of the body. Amongst, colibacillosis
that has added1809 (000 tons) of meat, and 21, is associated with respiratory and intestinal
285 millions eggs in FY 2021 as well as a major infections; it causes extra intestinal disease in
source of employment to almost 8 millions humans and mammals. Infections progress to
throughout the country (GoP, 2021). Producing many other organs as salpingitis, peritonitis,
more eggs and meat by growing more poultry cellulitis, perihepatitis, polyserositis, septicemia,
and death (Dho-Moulin, 1999; Barnes 2003;
* Jeong et al., 2012; Awad et al., 2016).
Corresponding author: [email protected]

© 2021, Sindh Agriculture University, Tandojam (http://pjaaevs.sau.edu.pk/index.php/ojs)

This work is licensed under a Creative Commons Attribution 4.0 International License
 Pak. J. Agri., Agril. Engg., Vet. Sci., 2021, 37 (2)

th
Various studies on Epidemiological and 42 days in group F (6 week) and from day 43
prevalence of E. coli infection described have up to marketing in group G, respectively.
been reported earlier in calves, pigs, ducks and
turkeys. These findings revealed the prevalence Gross pathological observations
of positive strains. These investigators found this During the visits, a postmortem of dead birds
infection as the most significant bacterial was conducted, the lesions observed were
disease in poultry worldwide (Harel et al., 1991; recorded and positive samples were collected
Dozois et al., 1992, Dozois et al., 1997; Stordeur and preserved in 10% formol saline solution and
et al., 2002). E. coli, the predominant organism brought to the Post Graduate Pathological
isolated from number of lesions in various avian Laboratory of the Department.
studies showed reduced weight gain, poor feed Histopathological observations
conversion, down grading of carcasses and To determine the histopathological changes,
condemnation at slaughter, infection of soft collected samples were processed with
tissues as well initiating cellulitis particularly in ascending grades of ethyl alcohol (starting from
the broiler (Baracho et al., 2006). 30% to absolute) for dehydration. Then the
Clinically E. coli infection is the most sample was cleared in xylene and embedded in
devastating disease that it consequences huge paraffin wax. After embedding the tissues were
mortality, culling, treatment costs; infected birds cut in 4-5µ section size. Ribbons were placed in
show dissemination of infection. Several the hot water bath to get rid of the wrinkle
researchers have revealed purulent formation. These sections were mounted on
inflammation of avian lungs, presence of slides and stained with Hemotoxyline and Eosin.
apoptotic cells. They also found dullness and These slides were then examined for
depression with an elevation of temperature, in histopathological changes.
appetence, diarrhea, cyanosis, salpingitis and
peritonitis in layers, infection of the reproductive Media prepared for isolation of E. coli
tract, perihepatitis, pericarditis, damaged Commercially available McConkey’s agar was
lymphatic organs and immune function.( Barnes, prepared by mixing 05 grams of McConkey’s
et al., 2008; Horn et al., 2012; Landman et al., agar with 100 ml-distilled water in conical flask.
2013; Pors et al., 2014; Sun et al., 2015; Jia et A homogenous mixture was obtained by gently
al., 2017). Since, Avian E. coli infections are mixing and heating on the hot plate. The mixture
ubiquitous and widely spread in Pakistan and was cooled at the room temperature, pH was
cause heavy economic losses to the poultry adjusted at 7.4, (by adding NaOH and NaHCO3),
farmers. Thus, this study aimed to determine the autoclaved at 15 lbs. pressure at 121°C for 15
prevalence and pathology produced by avian minutes, after autoclaving, the medium was
pathogenic Escherichia coli in different age cooled at 50°C and then l5ml was distributed
groups of broiler in the surroundings of Karachi, into sterilized petri dishes.
Pakistan. Blood agar base
MATERIALS AND METHODS Commercially available Blood Agar media was
Prevalence of infection in broiler chicken used for the culture of E. coli. Media consisted
The investigations were carried out to record the Lab-Lemco Powder l0 gms, Peptone l0 gms,
prevalence of E. coli infections in different age NaclO 5 gms, and Agar l5 gms. Forty grams of
groups of broilers at different commercial poultry blood agar base medium was dehydrated in l000
farms in surroundings of Karachi, Pakistan. To ml of distilled water and after mixing the pH of
observe the pathogenesis in different age medium was adjusted at 7.3. Then the medium
groups of chicks, 140 commercial broiler poultry was autoclaved for 15mm. at 121°C under 15
farms were visited; the disease was investigated lbs pressure. After cooling at 50°C about 10%
from day first up to the marketing. Total number fresh defibrinated sheep blood was added and
of farms (140) was divided into seven groups (A, subsequently distributed into petridishes. After
B, C, D, E, F and G), each group having twenty solidification the plates were incubated for 24
broiler poultry farms (comprising 1000-7000 hrs at 37°C for the sterility of the medium. The
chicks). Group-A represents the age group from sterile blood agar plates were used for primary
st
day 1 to day 7 (1 week), from 08 to 14 days in isolation of E. coli.
nd
group B (2 week), from 15 to 21 days in group Culture of sample
rd th
C (3 week), 22 to 28 days in group D (4 The samples from infected yolk and liver with E.
th
week), 29 to 35 days in group E (5 week), 36 to coli were collected from different chicks and

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cultured by sterilized platinum loop into the indicated higher infarction of E. coli in birds of
th
McConkey’s nutrient agar and blood agar and the 5 week and subsequently decreased in
incubated for 24 hours at 37°C for characteristic older birds as indicated in (Table 2).
and morphological observations. The study was
conducted at Central Veterinary Research and Omphalitis
Diagnostic Laboratory, Tandojam. The maximum rate of infection was observed in
group A with 76.24% and minimum in group B
RESULTS with 23.76% whereas, omphalitis was not seen
The systematic investigations were carried out in other groups (Table 3). The gross lesions
to record the prevalence of E. coli infections in observed were panophthalmitis, unabsorbed
the boiler at commercial poultry farms in the yolk, congestion of the small intestine, and
surroundings of Karachi. The period of studies necrotic foci in the liver (Figure 3A and 3B).
lasted five months. During the period, 140
commercial poultry broiler farms were visited to Colisepticaemia
confirm the disease. The maximum rate of infection 29.63% was
observed in group E whereas, minimum of
Prevalence of E. coli infections 3.11% in group B while no infection was
During the study period, the prevalence of E. coli observed in group A. However, the rate of
infections was recorded at various broiler farms. infection in other groups was as 22.18, 20.18,
The highest rate of prevalence was found in 18.63 and 6.74% were seen in group G,D,F and
group E (33.2%), followed by group G (27.7%), C respectively (Table 3).
whereas, the prevalence of E. coli infection was The gross pathological lesions about
observed as 14.6%, 13.6%, 17.7%, 27.5%, and colisepticaemia were observed in certain
26.9% in groups A, B, C, D and F respectively visceral organs such as the liver, heart, and air
(Table1). sacs.The most common lesions were fibrinous
perihepatitis, hepatomegaly with pale-colored
Table 1. Prevalence of E. coli in different groups necrotic foci, fibrinous pericarditis along clotted
Groups Total No. of No. of Sick No. of Positive
birds reared Birds (%) Birds (%) blood present in the heart, the air sacs were
st
A (1 Week) 12500 996 (7.69 %) 146 (14.65) found inflamed due to presence of caseous
B (2nd Week) 15000 1499 (9.99%) 204 (13.60)
C (3rd Week) 15700 1587 (10.10%) 282 (17.76 exudates.
D (4th Week) 14000 1620 (11.57%) 447 (27.59
E (5th Week) 12000 1675 (13.95%) 557 (33.23
F (7th Week) 14600 1490 (10.20%) 401 (26.91 Colibacillosis
G (8th Week) 17000 1637 (9.62%) 454 (27.73)
Grand Total 100800 10504 (10.42%) 2491 (23.71)
The maximum rate of infection was observed in
group G as 28.55% whereas, the minimum was
Gross pathology 0.075% in group A. However, the rate of
Gross pathological lesions varied at different infection was 26.70, 19.20, 16.11, 5.99, and
broiler farms comprising the birds of various age 3.35 in group E, D, F, and B respectively as
groups. The lesions observed in different organs shown in (Table 3).

Table 2. Specific lesions were observed in various age groups infected with E. coli
Form of Infection Specific Lesions observed Groups Total
A B C D E F G
1.Obphalitis and its Unabsorbed yolk, congested blood 125 15 - - - - - 140
forms 1.1 Mushy vessels, and panophthalmitis
1.2 Naval ill Emaciation 13 28 - - - - - 41
2.Colisepticaemia 2.1 Fibrinous perihepatitis - - 38 112 291 139 168 748
2.2 Fibrinous Pericarditis - - 33 72 68 74 88 335
2.3 Air Sacculitis - 62 63 217 230 148 185 905
3. Colibacillosis 3.1 Enteritis 5 33 59 225 323 202 294 1141
3.2 Necrotic foci on liver - 39 71 228 192 173 231 934
3.3 Hemorrhages on heart - 20 23 139 179 43 83 487
3.4 Tracheitis - 50 73 207 297 171 327 1125
3.5 Nephritis - 50 74 160 280 185 331 1080
3.6 Osteodystrophy - - 35 161 233 155 332 916
3.7 Petechiation of Gizzard - 30 62 152 265 138 293 940
4. E. coli and other 4.1 H.P.S - 51 3 55 223 202 162 696
infections 4.2 N.D. - - 60 80 - 49 - 189
4.3 I.B.D. - - - 106 120 20 80 326
4.4 Coccidiosis - - 105 50 32 35 100 322
4.5 Water belly (Ascites) - - 13 - - 27 72 112
4.6 C.R.D 3 70 92 276 323 194 296 1254
Grant Total 146 448 804 2240 3056 1955 3042 11691

Group A represents 1st week, B 2nd Week, C 3rd Week, D 4th Week, E 5th Week, F 6th Week, and G7th up to marketing.

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Table 3. Rate of infections observed in various forms of E. coli in different age groups
Form of Specific Groups with Percentage No. of birds
Infection Lesions A % B % C % D % E % F % G % examined
(Omphalitis and observed
its form)
Types Unabsorbed 125 76.24 15 23.76 - - - - - - - - - 181
Mushy yolk,
congested
blood
vessels and
panophthalmitis
Naval ill infection Emaciation 13 28 - - - - -

-
Colisepticaemia 2.1 Fibrinous - - - 3.11 38 6.74 112 20.18 291 29.63 139 18.63 168 22.18 1988
Perihepatitis
2.2 Fibrinous - - 33 72 68 74 88
Pericarditis
2.3 Air - 62 63 217 230 148 185
Sacculitis
Colibacillosis 3.1 Enteritis 5 0.075 33 3.35 59 5.99 225 19.20 323 26.70 202 16.11 294 28.55 6623
3.2 Necrotic - 39 71 228 192 173 231
foci on liver
3.3 - 20 23 139 179 43 83
Hemorrhages
on heart
3.4 Tracheitis - 50 73 207 297 171 327
3.5 Nephritis - 50 74 160 280 185 331
3.6 - - 35 161 233 155 332
Osteodystrophy
3.7 - 30 62 152 265 138 293
Petechiation of
Gizzard
Group A represents 1st week, B 2nd Week, C 3rd Week, D 4th Week, E 5th Week, F 6th Week, and G7th up to marketing.

Figure 3. Various pathological conditions caused by E. coli in broiler chicken. Figure 3A shows panophthalmitis
in day-old chick. Figure 3B shows classical postmortem signs for omphalitis due to E. coli. Figure 3C and D,
Shows hemorrhagic tracheitis and enteritis respectively, severe hemorrhages can be observed. Figure 3E and F,
represent cultural characteristics of E. coli in the broiler on blood agar and McConkey’s agar. Figure 3G
represents the morphology of Escherichia coli after successful culture and Figure 3H shows disruption of villus
due to E. coli in a histological study (H and E, magnification 40X).

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The gross pathological lesions varied in infection rate was recorded as 14.6%, 13.6%,
different visceral organs in the case of 17.7%, 27.5%, 33.2%, 26.9% and 27.7% in
colibacillosis which included necrotic foci and group A,B,C,D,E,F and G, respectively.
hemorrhages in the liver, hemorrhagic patches The gross pathological lesions varied in
in both the heart and gizzard. The kidney was different organs of various age groups.
also inflamed and in some cases showed Omphalitis was observed as 76.2% in group A
necrotic lesions in the peripheral region. Gall and 23.7% in group. Colisepticaemia as 3.1%,
bladder was enlarged and distended in some of 6.7%, 21.1%, 18.1%, and 22.1% in birds of
the cases, whereas in some cases, the lesions group B, C, D, E, F, and G, while in group A,
of C.R.D were associated (Table 3) with Colisepticaemia was not observed. Whereas
hemorrhagic tracheitis (Figure 3C), hemorrhagic colibacillosis was found in all groups as 0.07% in
enteritis (Figure 3D) along with the lesions of A,3.3% in B, 5.9% in C,19.2%, in D, 26.7% in
mixed infections (Table 3). In general, E,16.1% in F, and 28.5% in G groups of birds.
colibacillosis with 56.65% was noticed as The specific macroscopic lesions observed in
maximum, whereas, colisepticaemia with 17% the case of omphalitis included unabsorbed yolk
was observed as a minimum. While other material (sac) congested intestinal mucosa
associated diseases with 24.80% and omphalitis along with panophthalmitis whereas in the case
nd rd
with 1.55% were 2 and 3 week respectively of colisepticaemia there were fibrinous
(Table 4). pericarditis, perihepatitis, and airsaculitis. While,
in the case of colibacillosis, gross lesions
Table 4. Rate of infection in various types of E. coli along observed were petechiation in the heart, kidney
with mixed infection
and gizzard besides enteritis tracheitis and
Disease No. of Lesions Percentage necrotic foci in liver and caseous exudate in
observed air sacs. Histopathological lesions revealed
Omphalitis 181 1.55
congestion, degenerative changes in the
Colisepticaemia 1,988 17.00
Colibacillosis 6,623 56.65 intestine, and leukocytic infiltration in liver, heart
Escherichia coli and 2,899 24.80 and kidneys with sometimes necrotic foci
other infections especially in the heart. Based on the observed
Grand Total 11,691 100 lesions, the form of colibacillosis in the current
study could be a hepatic form of colibacillosis.
Histopathological observations Histopathological lesions in the intestine, lungs
There was no any histopathological lesion and liver, were also identified by previous
observed in serosa, musclaris externa and sub- studies (Gosh et al., 2006; Talha et al., 2001;
mucosa but the lesions in the mucosa revealed Riaz et al., 2016).
loss of epithelial tissue along with desquamation Colibacillosis is a contagious disease of
(Figure 3H), leukocytic infiltration of necrotic foci chicken caused by E. coli (Barnes et al., 2008),
were found in liver, heart, and kidney. it is classified as one of the major causes of an
increase in morbidity and mortality rate in
Cultural and morphological characteristics of commercial farms, consequently leading to large
Escherichia coli economic losses to the poultry farming through
The morphological study was conducted by its association with other diversified diseases, as
culturing the specimens on different media a primary or secondary pathogen (McPeake et
revealed either singly or paired colonies with al., 2005; Tonu et al., 2012; Mbanga and
pinkish, small, circular, smooth, translucent, and Nyararai 2015).
convex colonies on McConkey’s agar and grey- Avian colibacillosis noticed to be a major
white (Figure 3E), glistening circular with convex infectious disease in all age birds, and has
colonies on blood agar (Figure 3F). various local and systemic symptoms. The
current study agrees with previous findings, in
DISCUSSION our finding various lesions were found in the
One hundred forty commercial broiler farms liver, intestine and lungs. Died chicks showed
were surveyed to record the prevalence of E. postmortem signs for omphalitis intestinal
coli infections in the surroundings of Karachi hemorrhagic with tracheitis and enteritis
Pakistan. The studies were carried out to record respectively with severe hemorrhages observed.
the infection of E. coli in different age groups of The poor flock performance and managements
birds from day one up to the marketing. The practices correlated with increased early

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mortality rates in broiler forms, Histopathological 2008. Colibacillosis (Eds.). Diseases of

observations in the present study had similarities Poultry. Iowa State Press, Ames, pp. 691-
to observations earlier (Barnes et al., 2008; Riaz 732.
et al., 2016). Dho-Moulin, M. and J. M. Fairbrother. 1999.
In conclusion, the results of this study Avian pathogenic Escherichia coli (APEC).
confirm the prevalence of avian pathogenic E. Veterinary Research, 30 (2): 299-316.
coli in the field, based on morphological and Dozois, C. M., J. M. Fairbrother, J. O. S. E. E.
histological lesions and cultural findings. The Harel and M. A. R. C. Bosse. 1992. Pap-
economic and poultry health burden of this and pil-related DNA sequences and other
disease emphasizes the effective surveillance virulence determinants associated with
and control measures to reduce economic Escherichia coli isolated from septicemic
losses inflicted to the farmer due to this hazard. chickens and turkeys. Infection and
Immunity, 60 (7): 2648-2656.
Novelty statement Dozois, C. M., S. Clément, C. Desautels, E.
We report that no such data have been Oswald and J. M. Fairbrother. 1997.
published earlier and this is first ever report on Expression of P, S, and F1C adhesions by
natural infection of Avian Escherichia coli in Cytotoxic necrotizing factor 1-producing
Pakistan. This data will prove to be complete Escherichia coli from septicemic and
data on Avian Escherichia coli in meat-type diarrheic pigs. FEMS Microbiology Letters,
chicken from day 1, upto marketing. 152 (2): 307-312.
Ghosh, R. C., S. D. Hirpurkar and P. R.
AUTHOR’S CONTRIBUTION Suryawanshi. 2006. Concurrent
Q. A. Shah: Executed the experiment, data colibacillosis and infectious bursal disease in
collection, analysis and overall manuscript write- broiler chicks. Indian Veterinary Journal, 83
up (9): 1019-1020.
N. M. Soomro: Helped in the development of GoP, 2021. Economic Survey of Pakistan
experiment and generation of the data (Ministry of Finance, Government of
J. M. Shah: Provided technical support in data Pakistan).
and sample collection from the study area Harel, F., Daigle, S. Maiti, C. Desautels, A.
M. A. Hussain: Helped in data collection Labigne and J. M. Fairbrother. 1991.
S. F. Sayed: Helped in data collection and Occurrence of pap-, sfa-, and afa-related
culture of E. coli (Prevalence) sequences among F165-positive
Escherichia coli from diseased animals,
CONFLICT OF INTEREST FEMS Microbiology Letters, 82 (2): 177-182.
There is conflict of interest among authors. Horn, F., A. M. R. Corrêa, N. L. Barbieri, S.
Glodde, K. D. Weyrauch and B. Kaspers.
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(Received: September 29, 2021; Accepted: October 28, 2021)

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