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Abstract

Escherichia coli is one of the common microbial flora of poultry gut. Most of E. coli isolates are nonpath-ogenic but are considered to be an indicator of fecal contamination in food industry. A study was carried-out on the prevalence, incidence, isolation and antibiogram of E. coli from table eggs. A total of 100 table eggs were collected from various locations of district Peshawar, Pakistan and divided into three parts viz., the egg-yolk, egg-white and eggshell. These were cultured on different media and identified organism was subjected to antibiogram study using the disk diffusion method. The overall prevalence of E. coli was found as 37%. While, incidence was recorded as 15% in egg-shells, 12% in egg-whites and 10% in egg-yolks. It was concluded that the table eggs were contaminated with E. coli and higher incidence of E. coli was recorded in eggshells as compared to other components of the eggs. The antibiotics ciprofloxacin and enrofloxacin were recorded highly active against E. coli.
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INTRODUCTION
Poultry farming is widely adopted in Pakistan and al-
most every farmstead keeps some poultry mainly for
consumption and cash sales. e science and technolo-
gy have contributed widely for the expansion of poultry
industry and a number of strategies have been adopted
to modulate the quality of poultry products (Abel et al.,
2014). In Pakistan, there are about 25000 poultry farms,
providing employment and income for livelihood of fteen
thousand people. In the country, there are 400 hatcheries,
150 feed mills, 8.5 million broiler breeders, 0.428 million
layer breeders and their feed consumption is 5.51 million
metric tons per year (Anonyms, 2011; FAO, 2011).
E. coli are one of the common microbial ora of gut of
farm animals, poultry and human being. Most of E. coli
isolates are harmless, however, some strains are pathogenic
and may cause serious food poisoning in human beings
(Begum et al., 2014). A recent survey about prevalence of
virulence E. coli based on Congo red binding ability have
indicated more than 90% isolates as pathogenic (Yadav et
al., 2014). In past two decades, severe outbreaks with gas-
trointestinal symptoms have been occurred by food borne
pathogenic E. coli, particularly 0157:H7 (Armstrong et al.,
1996). E. coliand its related species are named as “enteric
bacteria”; because they mostly live in the intestinal tracts
of human and other animal species (Minnock et al., 2000).
About 10 to 15% of intestinal coliforms are opportunistic
and pathogenic serotypes and cause a variety of lesions in
immuno-compromised hosts including poultry (Daini et
al., 2008; Mailk et al., 2013); and may cause omphalitis,
yolk sac infection, cellulitis, colibacillosis and swollen head
syndrome (Gross, 1994).
Table eggs are the primary source of protein in human diet.
ese are used in a number of traditional Pakistani dishes
from decades. However, the recent studies have declared
Research Article
Abstract | Escherichia coli is one of the common microbial ora of poultry gut. Most of E. coli isolates are nonpath-
ogenic but are considered to be an indicator of fecal contamination in food industry. A study was carried-out on the
prevalence, incidence, isolation and antibiogram of E. coli from table eggs. A total of 100 table eggs were collected from
various locations of district Peshawar, Pakistan and divided into three parts viz., the egg-yolk, egg-white and eggshell.
ese were cultured on dierent media and identied organism was subjected to antibiogram study using the disk
diusion method. e overall prevalence of E. coli was found as 37%. While, incidence was recorded as 15% in egg-
shells, 12% in egg-whites and 10% in egg-yolks. It was concluded that the table eggs were contaminated with E. coli
and higher incidence of E. coli was recorded in eggshells as compared to other components of the eggs. e antibiotics
ciprooxacin and enrooxacin were recorded highly active against E. coli.
Keywords | Escherichia coli, Egg-shell, Egg-yolk, Prevalence, Egg-white, Antibiogram, Peshawar
AurAngzeb KhAn1*, rAhmAtullAh rind1, muhAmmAd ShoAib1, ASghAr Ali KAmboh1, gulfAm
Ali mughAl2, ShAKeel AhmAd lAKho1, KAnwAr KumAr mAlhi1, Ali rAzA nizAmAni3, AdnAn
YouSAf4
Isolation, Identication and Antibiogram of Escherichia coli from
Table Eggs
Editor | Sanjay Kumar Singh, Indian Veterinary Research Institute, Izatnagar 243122, Bareilly (UP), India.
Received | October 13, 2015; Revised | November 03, 2015; Accepted | November 06, 2015; Published | December 12, 2015
*Correspondence | Muhammad Shoaib, Department of Veterinary Microbiology, Sindh Agriculture University, Tandojam, Pakistan; Email: vet.socialray@gmail.
com
Citation | Khan A, Rind R, Shoaib M, Kamboh AA, Mughal GA, Lakho SA, Malhi KK, Nizamani AR, Yousaf A (2016). Isolation, identication and antibio-
gram of Escherichia coli from table eggs. J. Anim. Health Prod. 4(1): 1-5.
DOI | http://dx.doi.org/10.14737/journal.jahp/2016/4.1.1.5
ISSN | 2308–2801
Copyright © 2016 Khan et al. is is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distri-
bution, and reproduction in any medium, provided the original work is properly cited.
1Department of Veterinary Microbiology; 2Department of Livestock Management; 3Department of Veterinary Pa-
thology; 4Faculty of Animal Husbandry and Veterinary Sciences, Sindh Agriculture University, Tandojam, Pakistan.
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that enteric bacteria like Salmonella, E. coli, Listeria, etc.,
could contaminate these eggs and may cause egg-borne
diseases (Adesiyun et al., 2006; Adesiyun et al., 2007).
Some global epidemics have also been linked with egg
consumption and known to cause egg-borne pathogens
present in poultry eggs and their contents (CDC, 1990;
Rocourt et al., 2003). Food poisoning associated with egg-
borne pathogens may cause severe morbidity or mortali-
ty with diarrhea, vomiting, nausea and abdominal cramps
(Mitchell, 2005). e present investigation was therefore,
designed to study the prevalence and incidence of E. coli in
table eggs sold in retail market of district Peshawar. More-
over, the antibiogram study of isolated E. coli from poultry
eggs was also carried out to investigate the susceptibility
pattern of various antibiotics.
MATERIALS AND METHODS
StudY deSign
A total of one hundred poultry table eggs were collected
randomly from dierent markets existed in various local-
ities of district Peshawar, Khyber Pakhtunkhwa, Pakistan.
Eggs were collected from four dierent localities (n= 25
from each locality) i.e., Bacha Khan Chowk, Karkhano
Road, Nahaqi and Palossi Markets of Peshawar. Although,
the eggs were kept at room temperature at sale outlets, so it
was ensured that these should not be older than 24 hours.
Moreover, the eggs with visible fecal shell contamination
were not taken as samples. e collected eggs were trans-
ported to laboratory under cold chain and were kept in
refrigerator at 4°C until they were processed for microbial
contamination.
lAborAtorY ProcedureS
For the isolation of E. coli, table eggs were processed ac-
cording to procedure described by Adesiyun et al., (2006).
In brief, using aseptic conditions one sterile swab mois-
tened in normal saline (0.9% NaCl w/v) was applied to the
surface of each egg. It was dipped in 1ml saline in universal
bottle to form a representative egg shell sample. For egg-
yolk and egg-white samples, the eggs were immersed in
75% ethanol for 5 minutes and then pointed end of each
egg was disinfected on Bunsen burner ame for 5-10 sec-
onds. en, a small hole was made on the shell surface and
the egg-yolk and egg-white were emptied separately into
the sterilized polythene bags. e contents were blended
manually. e resultant mixtures and egg shell samples
were used for bacteriological culture as described earlier
(Nazia et al., 2015). e isolated E. coli were then subjected
towards dierent biochemical and sugar fermentation tests
for species conrmation like starch test, lipid hydrolysis
test, casein hydrolysis test, gelatin hydrolysis, carbohydrate
fermentation test, triple sugar iron test, which were based
on their capability to breakdown complex molecules in to
simpler nutritional elements.
In-vItro SuScePtibilitY of E. coli to
AntimicrobiAlS
e antibiotics (Difco, Michigan, USA) used during the
study were amoxicillin (10ug), colistin (10ug), gentamycin
(10ug), enrooxacin (05ug), kanamycin (10ug), ciproox-
acin (05ug), noroxacin (10ug), tetracycline (30ug) and
doxycycline (30ug). All the E. coli isolates of table eggs
were investigated for their in-vitro susceptibility pattern to
various antimicrobial agents using disk diusion method as
described by Bauer et al. (1966). In brief, the Muller Hin-
ton agar (Difco, Michigan, USA) was prepared, dispensed
in Petri dishes and surface was dried by incubating at 37oC
for 30 minutes. e isolated colonies were selected and sus-
pended evenly in 4ml sterile normal saline solution (0.9%
NaCl w/v; pH: 7.0). A sterile cotton swab was dipped into
the suspension and culture was smeared on the surface of
Muller Hinton agar in such a way that all agar surfaces
would be covered evenly with the bacterial suspension. e
plates were then placed in incubator for 30 minutes to get
dried. e antibiotic discs were placed on the agar surface
with the disc disperser and slightly pressed with sterile
forceps to keep it adhere to the surface. e plates were
then closed, wrapped in polythene bag, inverted in such
a way that medium and discs would be in upward portion
and placed in incubator for 24 hours at 37oC. e zones of
inhibition were observed as a clear area, free from growth
around the discs. Clear zones of inhibition made against
organism by the antibiotics were recorded in mm from the
centre of disc of zone with the observed annotations.
dAtA AnAlYSiS
All the experimental results were calculated and presented
in percentage format using the Excel Spreadsheets.
RESULTS
Of the total 100 table eggs examined, the overall prevalence
of E. coli was recorded as 37.00%, while 63.00% eggs were
found free from E. coli contamination (Table 1). Of the 25
eggs examined from Bacha Khan Chowk, the prevalence
of the E. coli species was noted in 40.00% eggs. Similarly,
25 eggs acquired from Karkhano Road market, the prev-
alence was observed as 48.00%. When the same number
eggs were examined from Nahaqi market, the prevalence
of E. coli was recorded as 32.00% in eggs. Whereas 25 eggs
collected from Palossi market showed the prevalence of E.
coli as 28.00% (Table 2).
e results regarding the incidence of E. coli in dierent
components of eggs has been summarized in Table 3. Of
the 100 egg shells examined, the incidence was recorded as
15.00%. Similarly, among 100 egg-whites, the incidence of
E. coli was noted as 12.00%, whereas within 100 egg-yolks
E. coli was detected in 10.00% egg-yolks.
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Table 1: e overall prevalence of Escherichia coli in table eggs collected from retail markets of Peshawar
Total No. of eggs examined No. of eggs positive % of eggs positive No. of eggs negative % of eggs negative
100 37 37 63 63
Table 2: e number and percentage prevalence of Escherichia coli in table eggs collected from dierent localities of
Peshawar
S. No Name of area Total No. of eggs
examined
Total No. of
positive eggs
% of positive
eggs
Total No. of
negative eggs
% of negative
eggs
1 Bacha Khan Chowk 25 10 40 15 60
2 Karkhano Road 25 12 48 13 52
3 Nahaqi 25 8 32 17 68
4 Palossi 25 7 28 18 72
Table 3: e number and percentage incidence of Escherichia coli in dierent components of table eggs
Egg com-
ponents
Total No. of egg com-
ponents examined
Number of posi-
tive components
Percentage of posi-
tive components
Number of nega-
tive components
Percentage of nega-
tive components
Egg-yolk 100 10 10.00 90 90.00
Egg-white 100 12 12.00 88 88.00
Egg-shell 100 15 15.00 85 85.00
Table 4: Antibiogram results of Escherichia coli isolates of
table eggs
Antibiotic
discs used
Zone around
discs
Indication
of sensitivity
Degree of
sensitivity
Amoxicillin
(10ug)
2 mm + Weakly sen-
sitive
Colistin
(10ug)
8 mm +++ Quite sensitive
Gentamycin
(10ug)
10 mm +++ Quite sensitive
Enrooxacin
(05ug)
14 mm ++++ Highly sensi-
tive
Kanamycin
(10ug)
2 mm + Weakly sen-
sitive
Ciprooxacin
(05ug)
14mm ++++ Highly sensi-
tive
Noroxacin
(10ug)
4 mm ++ Moderately
sensitive
Tetracycline
(30ug)
0 mm - Not sensitive
Doxycycline
(30ug)
3 mm ++ Moderately
sensitive
-: Absence of clear zones around disc; +: clear zone up to 2mm; ++:
clear zone with >2-5 mm; +++: clear zone with >5-10 mm; ++++:
clear zone with >10-15mm
During present experiments, nine dierent antibiotics
were tested to demonstrate the in-vitro susceptibility of
E. coli isolates recognized from the table eggs and results
were given in Table 4. e antibiotics ciprooxacin and
enrooxacin were recorded as highly active against E. coli
isolates and inhibited its growth, while antibiotics colistin
and gentamycin were recorded as quite active against E.
coli. Whereas, drugs noroxacin and doxycycline showed
moderate sensitivity against E. coli, as these drugs inhib-
ited the growth of the organisms and showed small zones
of inhibition (>2-5mm) around the discs. Furthermore,
the antibiotics amoxicillin and kanamycin were marked as
weakly active against the organism. However, the antibi-
otic tetracycline failed to inhibit the growth of bacterial
organism on agar plate and was recorded as completely re-
sistant against E. coli.
DISCUSSION
It has been estimated that many nutrient substances found
in table eggs create an excellent environment for the
growth and development of potential spoilage or infectious
microorganisms. Present study has demonstrated an over-
all 37.00% E. coli contamination in table eggs. is nd-
ing is in agreement with a study conducted by Adesiyun et
al. (2006) in Trinidad. e researchers reported a 71/184
(38.6%) table eggs positive for enteric microbes including
E. coli, Salmonella, etc. Likewise another Polish study re-
ported a 40.30% bacterial contamination in table eggs with
E. coli as most dominant contaminant (Stępień-Pyśniak,
2010).
We got 28-48% contamination of bacterial organism in
table eggs collected from dierent localities of Peshawar.
However, another study reported the 36.3 to 69.6% con-
tamination in poultry eggs collected from dierent points
i.e., supermarket, mall and farm (Adesiyun et al., 2006).
ese dierences might be due to dierence in man-
agement, handling and hygienic conditions used at farm
and/or sale outlets. e poultry eggs can get contamina-
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tion either horizontally (through the shell) or vertically
(trans-ovarial), and could serve a potential source of path-
ogens participating in the etiology of foodborne diseas-
es (Stępień-Pyśniak, 2010). Indar et al. (1998) reported
trans-ovarial transmission of Salmonella spp. in table eggs
collected from commercial poultry farms in Trinidad.
Although, egg-yolk contains maternal immunoglobulin
IgG (also called IgY), but its level could be inuenced by
various factors like, functional quality of immunological
system and/or antibiotics exposure to fowl (Tokarzewski,
2002). Moreover the quantitative contamination of eggs
depends upon bacterial load in the environment where
eggs laid and/or handled (Stępień-Pyśniak, 2010).
e results of the present study indicated the bacterial
contamination level as 15, 12 and 10% on eggshells, egg-
whites and egg-yolk respectively. Adesiyun et al. (2006)
reported in their investigation the contamination level as
19% and 13% in eggshell and egg contents respectively. It
has been suggested that temperature, and/or storage con-
ditions provided to the eggs at retail outlets signicantly
impact the bacterial load of eggs without aecting the bac-
terial prevalence (Suresh et al., 2005). In consistent with
this study, Stępień-Pyśniak (2010) also reported a high
contamination level of eggshells as compared to other in-
ternal contents. is is probably due to exposure of egg-
shell with the environment.
e results about in-vitro susceptibility of E. coli isolat-
ed from table eggs of poultry birds recorded during pres-
ent investigation were in line to the ndings reported by
previous studies. Like, Akond et al. (2009) isolated and
identied E. coli from poultry sources of dierent poul-
try markets and sensitivity to antimicrobials was recorded
as 86, 80, 60, 36, 30 and 26% to noroxacin, gentamicin
and chloramphenicol, neomycin, tetracycline, streptomy-
cin and ampicillin, respectively. Raji et al. (2007) observed
ciprooxacin as highly active (85-100%) antibiotic against
E. coli isolates. Adesiyun et al. (2007) studied the resist-
ance of bacterial species to seven antimicrobial agents us-
ing the Disc Diusion Method. An overall, 131 bacterial
isolates of E. coli and Enterobacteriaceae were tested, and
125 (95.4%) exhibited resistance to one or more antimi-
crobial agents. e high resistance was recorded against
streptomycin (90.1%), tetracycline (51.9%) and kanamy-
cin (30.5%).
In present investigation we have found 3/9 (33.33%) an-
timicrobial agents as resistant or weakly sensitive to E. coli
isolated from table eggs. is nding is in agreement with
a study conducted by Musgrove et al. (2006). e study in-
dicated that most (73.2%) of E. coli isolated from eggshells
were susceptible to all antimicrobial agents. Moreover, the
E. coli isolates showed 29.9, 6.2 and 3.1% resistance to tet-
racycline, streptomycin and gentamicin respectively. Simi-
lar results were also reported by Ansari et al. (2014).
CONCLUSIONS
It could be concluded from present investigation that table
eggs sold in retail market of district Peshawar contained
E. coli, hence may pose a health hazard to human beings
if consumed improperly cooked or raw eggs. Eggshells
contained more bacterial contaminants as compared to
egg contents. Antimicrobial agent tetracycline was found
completely resistant to E. coli isolates, whereas, amoxicillin
and kanamycin were observed as weakly sensitive. ere is
a need to educate the people to adopt signicant hygienic
measures in handling of table eggs and should not be con-
sumed inadequately cooked eggs or egg products.
ACKNOWLEDGEMENTS
e author is thankful to the honourable Supervisor, Dr.
Rahmatullah Rind, Professor, Department of Veterinary
Microbiology, Faculty of Animal Husbandry and Veter-
inary Sciences, Sindh Agriculture University, Tandojam,
for his courteous professional pieces of advice, untiring
guidance, full support, motivation, fruitful suggestions and
encouragement during the whole period of study and also
thankful to the respectable and sincere teacher Dr. Asghar
Ali Kamboh, Assistant professor, Department of Veteri-
nary Microbiology SAU Tandojam for his technical facili-
tation and all others for their co-operation during research
work.
CONFLICT OF INTEREST
ere is no conict of interest.
AUTHORS’ CONTRIBUTION
Mr. Aurangzeb was the main researcher, Dr. Rahmatullah
Rind was his supervisor, Dr. Asghar Ali Kamboh revised
the article, Muhammad Shoaib did all the correspondence,
Gulfam Ali Mughal, Shakeel Ahmad Lakho, Kanwar Ku-
mar Malhi, Ali Raza Nizamani and Adnan Yousaf con-
tributions in statistics, and other activities related to the
research.
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... rodents play, further field and laboratory research is needed (Khan et al 2016). As a preventative step against the probable entry of viruses and pathogens, it is advised that poultry buildings be rodent-proof and the immediate environs are unappealing to rodents. ...
... A shrew population in a poultry farm may pose a high risk of bacterial contamination in poultry products. Shrew populations can be eliminated from poultry farms through extensive live trapping and rodent proofing, which reduces the risk of bacterial infection to poultry birds and products (Khan et al 2016). ...
Article
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Rodents can be abundant around poultry houses, share their habitat with waterfowl and can readily enter poultry houses. Infective bacteria have been found in the feces, urine, and blood of Eurasian Shrew (Sorex Araneus) that live in poultry farms. 22 shrews were caught, and their guts were examined for harmful germs. In feces, Escherichia coli and Proteus species were found in 59.09% and 18.18% of cases, respectively. Salmonella spp. were found in 72.72% of feces and urine samples. Rodents can be abundant around poultry houses, share their habitat with waterfowl and can readily enter poultry houses. Different viruses and bacteria can replicate in rodents without adaptation, resulting in high viral titres in lungs and nasal turbinates, virus presence in nasal washes and saliva, and transmission to naïve contact animals. Therefore, active viruses shedding by infected rodents may play a role in transmission to poultry. Making poultry houses rodent-proof and the immediate surroundings unattractive for rodents are recommended as preventive measures against possible any viruses and bacteria introduction. It is determined that common shrews serve as a reservoir for dangerous microorganisms, which can be avoided using effective shrew-eradication measures on poultry farms.
... Spring had the highest prevalence of E.coli infection 32%, followed by winter 26.15%, autumn 13.16%, and summer 12.50%. Our findings are comparable to those of (Khan et al., 2016) and (Javed et al. 1991), who found an E.coli infection rate of 11.74% in the layer. According to (Yunus et al., 2009), E.coli infections and coryza outbreaks were equally distributed throughout the year in broilers, but more than half of all chronic respiratory disease (CRD) cases were seen from April to June, and half of coryza and one-third of CRD outbreaks were seen in layers from April to June. ...
Article
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The purpose of this study was to document the frequency of respiratory diseases in broiler and layer flocks in the Rawalpindi District of Pakistan from July 2020 to June 2021. Infectious coryza, colibacillosis, and CRD were identified in 48, 30, and 22 of 240 broiler flocks, respectively. Infectious coryza, colibacillosis, and CRD were reported to be prevalent in 20%, 12.50%, and 9.17% of the population, respectively. Infectious coryza, collibacillosis, and CRD were discovered in 41, 34 and 28 of 160 layer flocks, respectively. Infectious coryza, colibacillosis, and CRD were reported to be prevalent in layer at 25.63%, 21.25%, and 17.50%, respectively. Respiratory diseases were observed throughout the year, but were more common during the winter months. A total of n = 201 samples (liver, heart, and lungs) were cultured during the investigation, with n = 89 proving to be positive for E.coli isolates. Lincomycin, norfloxacin, and neomycin were the most effective antibiotics against E.coli infections, while oxytetracyclin, doxycyclin, and colistin had the least effective zones of growth suppression. Prevalence, respiratory diseases, and poultry are some of the terms used in this study.
... Brucellosis in cattle can cause mastitis, orchitis, abortion, arthritis, and infertility (Nicoletti et al., 2012). In several places of the world, the disease has been identified as endemic (Khan et al., 2016). However, with good management, it has been decreased and eradicated in certain developed countries (Hungerford et al., 2012). ...
Preprint
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The prevalence of Brucella abortus in cattle was determined using n = 800 samples (n = 400 serum and n = 400 blood samples) collected from organised dairy farms in various areas (Chakri, Taxila,) and the Rawalpindi Main City. Serological techniques such as the Milk Ring Test (MRT), Serum Agglutination Test (SAT), and Rose Bengal Plate Test were used to diagnose Brucella abortus (RBPT). Brucella abortus was discovered in 5.38 % of cattle in Rawalpindi research areas. Using serological tests, the prevalence of Brucella abortus in local breeds was determined to be 6.5 %, 4.75 %, and 4.75 %, respectively. In the Holstein Friesian breed, the prevalence was found to be 4.25 %, 2.25 %, and 1.75 %, respectively. In general, Brucella abortus RBPT (3.25 %), MRT (1.75 %), and SAT (1.75 %) were found in cattle aged 1 to 5 years. The bacterium was shown to be prevalent in cattle aged 6-9 years (7.5 %), MRT (5.25 %), and SAT (4.75 %). In Rawalpindi, Brucella abortus was found in serum and milk samples from cattle. Cattle in the older age groups were shown to be more sensitive to Brucella abortus than those in the younger age groups. Brucellosis was more common in local cattle than in Holstein Friesian cattle.
... The feed samples were homogenized in peptone water in the beaker as described previously (Gyang et al., 2019) by thoroughly mixing 1 g of each sample in 9 ml of peptone water. In case of table eggs, yolks were treated as described previously (Khan et al., 2016). Briefly, the eggs were soaked in 75% alcohol for 5 min before being sterilized for 5-10 s on a hot flame. ...
Research
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The current study provides information on Bacillus spp. contamination along with present status in commercially available poultry and animal feeds as well as animal-derived products in Bangladesh. The research has been conducted to determine if animal feed and its components are a source of Bacillus spp. contamination in feed and food chain. Out of 180 different feeds, milk, egg, and human stool samples, 218 Bacillus spp. were isolated and identified by cultural morphology, microscopic, biochemical, and molecular characteristics where B. cereus, B. subtilis, B. amyloliquefaciens, B. licheniformis, B. thuringiensis, B. megaterium, and B. coagulans accounted for 51, 22, 9.1, 5.9, 5, 3.6, and 2.2%, respectively. Regarding the enumeration of total viable count and total Bacillus count, correspondingly 67 and 39% samples were found to be contaminated with above 10,000 CFU/g, while highest contamination was 85 and 75% in broiler feed, respectively. The total number of bacteria above the regulatory limits in commercially available feeds indicates a poor compliance with regulation and abuse administration in the Bangladeshi market. Moreover, a hospital-based survey showed that food-borne Bacillus spp. contributed to 4.5% human diarrhea cases and 25% food contamination associated with vegetables, rice, RTE food, milk, and egg, accounting for 46, 34, 14, 4, and 2%, respectively. B. cereus was the dominant isolate correspondingly accounting for 56 and 51% egg and milk contamination followed by B. amyloliquefaciens (32%) and B. thuringiensis (12%) in egg and B. subtilis (25%), B. amyloliquefaciens (12%), B. thuringiensis (6.4%), and B. coagulans (3.2%) in milk, respectively. Toxin gene profiling of Bacillus spp. revealed that B. cereus constituted a principal part of virulence, while B. thuringiensis, B. licheniformis, B. megaterium, B. coagulans, and B. subtilis showed genetic diversity and B. amyloliquefaciens had not carried any toxin gene. Detection rate of enterotoxin genes (nheA, nheB, nheC, cytK, hblA, hblC, hblD, and entFM) showed that 55% isolates carried nheABC genes, 80% entFM, and 71% cytK, whereas only 33% of the isolates contained hblACD gene clusters. These virulence genes were posing a threat to human health due to Frontiers in Microbiology | www.frontiersin.org 1 January 2022 | Volume 12 | Article 783103 Haque et al. Bacillus spp. Contamination and Toxigenicity spread across the food and feed chain. Finally, our findings support the hypothesis that B. cereus might contribute to clinical diarrhea, gizzard erosion, and lung infection in duck and poultry, and that it contaminates animal-derived foods resulting in toxicity and antibacterial resistance to humans. Therefore, maximal tolerance limits of Bacillus spp. and their potential risks to the animal industry are urgently needed to clarify. Moreover, Bacillus spp.-induced toxin residual must be altered for human health via food chain transmission.
... All the hatchable eggs were graded using the MOBA 9A egg grading machine based on their quality and weight. Only oval-shaped and good-quality eggs were chosen, with the poor shell, break, bloody stained, and elongated eggs being excluded (Khan et al., 2016). During the whole investigation, the egg room temperature and humidity were 75 0F and 65 0F, respectively, with fresh air at a rate of 2 CFM/1000 eggs. ...
... All the hatchable eggs were graded using the MOBA 9A egg grading machine based on their quality and weight. Only oval-shaped and good-quality eggs were chosen, with the poor shell, break, bloody stained, and elongated eggs being excluded (Khan et al., 2016). During the whole investigation, the egg room temperature and humidity were 75 0F and 65 0F, respectively, with fresh air at a rate of 2 CFM/1000 eggs. ...
Article
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Hatchery productivity is determined by hatchability as well as the quantity of high-quality day old chicks (DOC) produced. The hatchery industry has been looking for innovative ways to improve incubation con�ditions, such as using different incubation MS or SS machines. The goal of this study was to evaluate findings as of two incubator systems—MS and SS—for all hatchery criteria such as broiler performance, hatchability and chicks quality. Chicks Performance hatch window, Hatchability and water loss vari�ables were all subjected to a completely randomized experimental design with two treatments (MS and SS). A 2 x 2 factorial configuration was used to study performance characteristics (incubator type x chick sex). MS-incubated eggs lost more weight between incubation and transfer (P<0.05). Hatchabil�ity of eggs incubated in SS was maximum (P<0.05), and the hatch window of chicks in SS was longer (P<0.05). Embryo diagnostic demonstrated increased end mortality (P<0.05) and greater ratios of living and dead piped and broken eggs (P<0.05) for embryos incubated in MS. SS chicks had higher physical quality (P<0.05). For performance results, there was no interface (P > 0.05) between the investigated parameters. Broiler performance is influenced by incubator type. SS In terms of absorption of yolk ages, incubator chicks outperformed MS chicks (P > 0.05), and male broilers outperformed female broilers (P<0.05). Although performance characteristics were unaffected by incubation type, the SS incubation system demonstrated to be superior to the MS in addressing during embryo development, with maxi�mum hatchability and quality chicks.
... Each species has a favorite host [23]. Brucella pathogen phenotype isolates have been known to have host specificity in the past [9]. However, due to mixed farming, sharing of the same pasture by small and large ruminants, mixed livestock shelters, the presence of reservoir hosts in a farm, and uncontrolled animal movements, Brucella species may cross infect their non-preferred hosts [25]. ...
Article
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A molecular research was carried out in the areas of Lahore in order to confirm the existence of B. abortus antigen in Caprine and ovine by utilizing molecular techniques in this study. Small ruminants can be infected with B. abortus (the causative agent of bovine brucellosis), which complicates brucellosis control efforts because most brucellosis control programmes rely on immunological testing rather than genomic testing to determine the specific species circulating in ruminants. Now at this investigation, n = 1270 goat serum samples and n = 770 sheep serum samples were collected, respectively. After already being tested with the Rose Bengal test, all positive specimens were examined to the real-time PCR technique. RBT confirmed brucellosis prevalence of 21.43±0.37% and 18.11±0.12 in caprine and ovine respectively. Out of 230 positive goat samples, real-time PCR found B. abortus in 150 samples (65.21±0.51) in sheep and goats and 118 samples (71.51±0.21%) out of 165 seropositive sheep samples. Brucella abortus infection in small ruminants could be caused by a combination of factors including mixed farming of small and large ruminants, sharing of the same pasture, and the presence of reservoir hosts on a farm, all of which could be risk factors for Brucella species cross-infection in non-species hosts. It has been determined that B. abortus is the causative agent of caprine and ovine brucellosis in the country of Pakistan. Results of this study can be utilised to develop successful brucellosis eradication and control strategies in small ruminants, which can be applied to other animals.
... The highest prevalence in the Kazla market was probably due to the poor hygienic practice in the market. This finding agrees with the previous study where the prevalence of E. coli was reported as 37.0% in table eggs [29]. In the present study, Salmonella spp. in table egg from the selective market was 30%. ...
Article
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Chicken eggs are a major component of people’s diets, with an average yearly consumption of approximately 103 eggs per person in Bangladesh. Eggs act as an important carrier of food-borne pathogen worldwide. The study was conducted to identify the prevalence and antibiotic resistance pattern of E. coli, Salmonella spp., and Staphylococcus spp., in eggs isolated from farms and different markets of Rajshahi, Bangladesh. A total of 60 eggs were collected randomly between April to December 2019. The isolation and identification of bacterial pathogen was done in accordance with standard procedures. The bacterial isolates were subjected to antibiotic susceptibility testing against seven commonly used antibiotics using Kirby-Bauer disk diffusion method. An overall prevalence of E. coli, Salmonella spp., and Staphylococcus spp. were found to be 35.0%, 28.33%, and 23.33%, respectively. E. coli were found highly resistant to penicillin (100%), tetracycline (80.95%), ampicillin (100%), and erythromycin (85.71%) and were sensitive to amoxicillin (71.42%), ciprofloxacin (85.71%), and gentamicin (95.23%). Salmonella spp. was highly resistant to penicillin (100%), erythromycin (82.35%) and tetracycline (82.35%), and was sensitive to gentamicin (94.11%), amoxicillin (76.47%) and ciprofloxacin (70.58%). Staphylococcus spp. was resistant to penicillin (100%), erythromycin (78.57%), tetracycline (85.71%), amoxicillin (100%), and ampicillin (100%) but sensitive to ciprofloxacin (85.71%), and gentamicin (92.85%). The higher prevalence of multidrug-resistant (MDR) bacteria can easily enter the food chain, which poses a public health threat.
... Both pathogens penetrate the egg through the pores of the eggshell during incubation. Salmonella spp. is one of the bacteria with the highest presence in the eggshells (Spickler et al., 2011), Escherichia coli is another pathogenic bacteria that causes embryonic death if it gets through the pores of the eggshell (Khan et al., 2016). Microscopic fungi such as Aspergillus fumigatus, Fusarium culmorum and Fusarium equiseti penetrate the egg and consequently, embryos become contaminated with spores and mycelium and die (Tomczyk et al., 2018). ...
Article
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Objective: To describe the factors that affect the incubation of fertile eggs of Creole hens (Gallus gallus domesticus) at Mezquital Valley, Hidalgo, Mexico. Design/Methodology/Approach: 448 eggs of Creole hens were incubated in a Casser model 7622 incubator, in which the number of infertile and fertile eggs was evaluated; within the number of fertile eggs, the number of dead embryos was determined; due to dehydration or damage caused by fungi and bacteria. From the live hatched chickens, those that died due to omphalitis were recorded. Results: 272 of 448 incubated eggs were fertile; 119 of them hatched and 153 were dead embryos. While 176 were infertile. Among the dead embryos, 84 of dehydration, 69 due to fungal and bacterial infection and 11 live hatched chicks due to omphalitis. Limitations/implications: The literature review found little information on factors affecting the hatching of fertile Creole chicken eggs in backyard flocks. Findings/conclusions: Infection by fungi and bacteria increase embryonic mortality, while omphalitis increases mortality during the first days of life of the backyard Creole chickens. High storage temperature in the nest or incubator causes embryo dehydration.
... Milk samples were cultured using a conventional technique as described by Khan et al. [29]. Plates were then incubated for 24 h at 37°C, wherein growth of bacteria was observed after every colony was separated using the specific media as follows: Staph-110 for Staphylococcus, blood agar for Streptococcus, and eosin methylene blue agar for E. coli. ...
Article
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Background and Aim: Mastitis has been identified as the most prevalent and economically imperative disease among dairy animals. Thus, understanding its common bacterial pathogens and risk factors is necessary to improve udder health at herd, region, or country level. However, scientific research on caprine mastitis, especially on Beetal breed, has remained to be insufficient in Pakistan. Therefore, this study aimed to evaluate the epidemiology and antibiogram assay of common mastitis-causing bacterial agents, that is, Staphylococcus, Streptococcus, and Escherichia coli, in dairy goats. Materials and Methods: In total, 500 Beetal goats, irrespective of age and those that were not treated with any kind of antimicrobial agents during the past 120 h, were screened using California Mastitis Test in Pattoki, Kasur District, whereas epidemiological factors were recorded. The milk samples of mastitic goats were then collected and processed using standard methods. Each sample was primarily cultured on nutrient agar. Using a specific medium, each bacterial colony was separated using several streak methods. Six antibiotic disks belonging to different antibiotic groups were used for antibiogram profiling of bacterial isolates. Chi-square test was used to assess the association of baseline characteristics and mastitis occurrence. Meanwhile, multivariable logistic regression (p
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The study was conducted to investigate the effect of different stocking densities and quantitative feeding regimens on starting broiler chicks. A total of one hundred and sixty two (162) Marshall strain chicks having average initial weight of 44.23g at day old were randomly distributed and used for the trials. The treatments under stocking densities were D1 (0.25m2/bird), D2 (0.17m2/bird) and D3 (0.13m2/bird) while under the quantitative feeding regimen were 0% feeding restriction (F1), 15% feed restriction (F2) and 30% feed restriction (F3). All were replicated three times making a total of 27 observations and randomly distributed into a 3 × 3 factorial arrangement in a completely randomized design. The growth performance, economic cost, nutrient digestibility and haematological characteristics were measured, data obtained were subjected to one way analysis of variance. Results showed that stocking density had no significant effect (p>0.05) on the daily feed intake and the body weight gain. The body weight gain decreased with increasing stocking density from 20.53g in D1 to 19.25g in D2. The best feed conversion ratio (FCR) value of 3.05 was recorded in D3 compared to 3.20 recorded in D2. Stocking density had no effect on the dry matter digestibility, crude protein digestibility, crude fibre digestibility and ether extract digestibility (p>0.05). The stocking density however had an effect (p < 0.05) on the haematological parameters like red blood cell (RBC), mean cell volume, mean cell haemoglobin concentration, blood glucose and blood corticosterone levels. The blood cholesterol was significantly highest in D1 (94.28mG/dl) compared to the lowest value of 88.17mG/dl in D3. Stocking density had no significant effect on the cost of feed per kg weight gain. The quantitative feed restriction significantly decreased both the daily feed intake and weight gain. A lower daily weight gain of 16.40g in F3 compared to 22.95g in F1. A better FCR value was recorded in the restricted groups compared to the F1. Birds in F3 recorded a marginally better digestibility coefficient when compared to F1 and F2 respectively. The packed cell volume and RBC was significantly lower in feed restricted birds compared to F1 but the white blood cells were significantly highest in F3 than in F1 and F2. The blood glucose and cholesterol level was significantly highest in F1 than F2 and F3 birds. The heterophil: lymphocyte ratio (H: L) was significantly highest in F3 than in F1 and F2 respectively. Feed restricted birds (F2 and F3) had significantly (p < 0.05) lower cost of feed intake/ bird (N5.32 and N6.15, respectively) compared to N7.64 in F1 birds. Also, the cost of feed/kg weight gain was significantly lowest in F3 at N317.20 compared to N334.53 in F1. It was concluded that that birds in D3 and F3 had the best FCR and a better digestibility coefficient though higher haematological values were recorded in the D1 and F1 compared to other treatments. Birds in F3 had lower haematological profiles and reduced costs of production per bird due to the quantitative feed restriction.
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Neonatal calf diarrhea (NCD) is a common disease affecting the newborn calf and the most critical period is in the first few days following birth of the calf which is also known as calf scours. Keeping animals in close confinement where the opportunity for transmission of causative agents of NCD. The diarrhea and other clinical signs seen with the disease are caused by the interaction of any of several possible infectious causes. This study was carried out to isolate, identify and detect the antimicrobial resistant profile of E. coli and Salmonella from diarrheic calves. A total of one hundred and twenty five fecal specimens were collected directly from the rectum of diarrheic calves. Of the samples collected 35 (25%) and 11 (8.8%) was found positive for E. coli and Salmonella respectively. Antimicrobial resistance of these two isolate was found against Amoxycillin and Tetracycline whereas a high sensitivity was found towards Ciprofloxacin, Levofloxacillin, Azithromycin and Cefotaxime. Serotyping was done by using specific antisera to identify variants of the somatic (O) and flagellar (H) antigens. Cultural and biochemical features also reveal the presence of pathogens in the diarrheic calves.
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This study was designed to determine the isolation rate, serovars and biochemical profiles of e. coli from cases of colibacillosis and dead-in-shell embryos in Zaria-Northern Nigeria. The isolation rate of e. coli from hatcheries studied were 4.67% and 7.50% from farms of Simtu Agricultural Company and National Animal Production Research Institute (NAPRI) Shika Zaria, Nigeria respectively. Twenty e. coli isolates from clinical cases of colibacillosis were also used for this study. The Simtu farm e. coli isolates showed 97.5% motility, while isolates from both NAPRI and clinical colibacillosis cases were 100% motile. The results of carbohydrate fermentation are variable without specific character, except for e. coli isolates from clinical cases of colibaccillosis that showed 100% fermentation especially for lactose, ducitol, rhamnose and xylose. The major serovars recorded from clinical cases of colibacillosis were serovars O8:K50 and O9:K30. Serovars from the dead-in-shell embryos were O78:K80, O8:K50, O9:K30, and O26:K60. Untypable isolates made up the greatest percentage of serogroup of e. coli studied. The antibiotic susceptibility testing indicated that many of the isolates were resistant to more than one antibiotic. Ciprofloxacin was the antibiotic to which majority of isolates were sensitive (85% of the clinical cases and 100% of both the Simtu and the NAPRI farms' isolates). It is concluded that other methods for controlling e. coli should be evaluated, so that the emergence of resistant isolates be limited and the cost involved in prophylactic and therapeutic treatment programs be reduced.
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The aim of this study was to analyse the qualitative composition of Gram-negative microbes, mainly of the family Enterobacteriaceae, including pathogenic bacteria such as Salmonella, in the albumens and yolks and on the shells of hens' eggs, depending on their source and on the temperature and duration of their storage. A total of 375 table eggs were studied, from a large-scale poultry farm, a small-scale poultry farm and a supermarket. Each group was divided into 5 subgroups according to the temperature and duration of their storage during the study. Two serotypes of bacteria of the genus Salmonella were identified: S. Enteritidis and S. Arizonae. Strains of Salmonella spp. were also isolated. Apart from Salmonella and Escherichia coli, among the most frequently isolated bacteria of the family Enterobacteriaceae were Enterobacter spp., Klebsiella spp. and Citrobacter freundii. Qualitative analysis of the bacterial microflora of the eggs also showed the presence of other Gram negative bacteria, including Acinetobacter spp., Pseudomonas spp., Tatumella ptyseos, Providencia stuartii, Serratia liquefaciens, Flavimonas oryzihabitans, Vibrio metschnikovii, Leclercia adecarboxylata, Kluyvera spp., Rahnella aquatilis, Proteus mirabilis, and Achromobacter spp. The study demonstrated that the conditions applied, i.e., the temperature and duration of storage, did not significantly influence the prevalence of particular species of Gram-negative bacteria in the eggs. However, based on the analysis of contamination of eggs with Salmonella depending on their source, it can be concluded that the system in which the hens are housed affects the risk of contamination of eggs with these pathogens.
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A total of sixty-one resistant stains of Escherichia coli from 105 clinical specimens obtained from different body sites at the University College Hospital, Ibadan were subjected to antimicrobial susceptibility testing and plasmid profiling. Among the various classes of antibiotic tested, high resistance was found with Amoxicillin (90.16%), followed by tetracycline (88.52%) and co-trimoxazole (78.69%) while Nitrofurantoin and Ofloxacin being the most potent with (90.16%) and (54.10%) sensitivity. All the strains that were resistant to any antimicrobial agents were also resistant to amoxicillin. Plasmids ranging in molecular sizes from 0.12kb to 23.1kb were extracted from 40 (65.57%) of these isolates and grouped into seven plasmid profiles.Transformation experiment revealed that 40% of the resistant strains carried a common R – plasmid of size 23.1kb. Plasmid determined resistance to Amoxicillin was identified.
Article
The prevalence of potential spoilage microorganisms on the shells and in the egg contents of table eggs sold in Trinidad was determined. Table eggs samples were obtained from 23 poultry layer farms, 14 shopping malls and 102 supermarkets across the country. Each farm was visited twice approximately one month apart and 25 pooled eggs constituted a composite sample. Shopping malls were each visited twice usually one month apart while supermarkets were each visited once over a 4-month period. For both mall and supermarkets, six pooled eggs constituted a composite sample. Swabs of egg shells and pooled yolk and albumen (egg content) were tested for selected bacteria using standard methods. The resistance of bacteria to seven antimicrobial agents was detected using the disc diffusion method. Of a total of 184 composite eggs (shells, yolk/albumen or both) sampled, 71 (38.6%) samples were positive for enteric microbes, other than E. coli, Salmonella, Campylobacter spp. and Listeria spp. Enterobacter spp. and Klebsiella spp. were isolated from 15 (8.2%) and 14 (7.6%), respectively, of pooled egg shells alone and from 6 (3.3%) and 3 (1.6%), respectively, of egg content samples alone. Prevalence of enteric bacteria in egg contents was generally higher than found on egg shells with faeces/blood or cracks compared with those without, but the differences were not significant (P>0.05; X2). The microbial load of egg content was not significantly affected by type of housing of laying birds, source of feeds, use of medicated feeds and temperatures at which eggs were kept at sale outlets. Of a total of 131 bacterial isolates tested, 125 (95.4%) exhibited resistance to one or more antimicrobial agents and resistance was high to streptomycin (90.1%), tetracycline (51.9%) and kanamycin (30.5%). Failure to properly handle or heat table eggs sold in Trinidad poses a potential health hazard to consumers because of their poor microbial quality and high frequency of resistance to antimicrobial agent.
Article
The antimicrobial sensitivity of Salmonella spp. and Escherichia coli isolated from the shells and contents of table eggs sampled from sale outlets in Trinidad was determined using the disc diffusion method. The phage types of S. Enteritidis isolates, the phenotypic characteristics of E. coli isolates and the presence of O157 strain were also investigated. Of a total of 74 isolates of Salmonella tested, 17 (22.9%) exhibited resistance to one or more of the seven antimicrobial agents used compared with 104 (88.1%) of 118 E. coli isolates. The difference was statistically significant (P < 0.05; X2). For both microorganisms, resistance was relatively high to streptomycin (54.2%) and tetracycline (35.9%) but low to gentamicin (11.5%) and sulphamethoxazole/trimethoprim (9.4%). Only 1 (1.4%) isolate of Salmonella was multi-resistant while 55 (46.6%) of E. coli isolates were resistant to three or more antimicrobial agents. The frequency of resistance to antimicrobial agents amongst both bacteria was not significantly (P > 0.05; X2) affected by the location of isolation on the egg (shell or content) or source of eggs (farms, shopping malls or other retailers). Eight (19.5%) of 41 S. Enteritidis isolates tested were resistant compared to 4 (26.7%) of 15 isolates of S. Ohio. All S. Enteritidis isolates belonged to phage type 1 (PT1) and all E. coli isolates were non-haemolytic, non-mucoid, sorbitol fermenters and non-O157 strains. It was concluded that the relatively high resistance amongst the bacteria tested could pose therapeutic problems in consumers, particularly in egg-borne salmonellosis or colibacillosis.