Isolation and antimicrobial susceptibility pattern of Escherichia Coli from laying chicken and egg in Jimma Town, Ethiopia

Food borne diseases are one of the main important issues in the world. Escherichia coli are considered as the most prevalent food borne pathogen. A cross sectional study was conducted from January 6- 2018 to September 10-2019 on cloacae swab; farm and market’s egg at Jimma town to isolate, identify and determine antimicrobial susceptibility patterns. Result Out of 415 total samples, 156 (37.59%) were identified as E. coli from farm eggshell 21/83 (25.30%), farm egg content 12/83 (14.46%), cloacae swab 42/83 (50.60%) and market egg shell 45/83 (54.23%). In the current study antimicrobial susceptibility test of E. coli isolates from different sample types revealed varying degree of susceptibility to antimicrobial. Isolated E. coli was highly susceptible to Ciprofloxacin (100%), Gentamicine (100%), Streptomycin ( 96.4% ) , sulfonamides (94%), Kanamycin (91%), Chloramphenicol ( 89.5% ) , tetraccine (78.9%) , Trimethoprim (75%) these are considered appropriate for empirical treatment of E. coli in the study area. Moreover, resistance of isolates with 100%, 75% 50%, 25%, 1.6% and 0.5% was developed to ampicillin, neomycin, cefoxitin oxytetracycline , Streptomycin and tetraccine respectively. The presence of E.coli within egg/and chicken can pose serious public health problems. The appropriate hygienic practices of eggs at both farms and markets should be under taken. The present study was conducted to isolate, identify and determine antimicrobial susceptibility patterns of E. coli from farm egg, cloacae swab and market egg samples in Jimma town. Food borne diseases is one of the important issues worldwide. E. coli is considered the most prevalent food borne pathogen that has gained increased attention in recent years. In the present the prevalence of E. coli from egg is in line with the study of Germini al., 2009. But this finding is lower than some reports from previous works The prevalence of E. coli from cloacae in this study is in line with the privies 15]. In contrast the finding of this prevalence in the present study is higher than the study of [16, 22, 23, 24 in which E. coli was isolated from cloacae samples taken from poultry farms. In another study, [25, 26] E. coli was isolated from faeces of poultry sample at different farms in Nigeria that may shows contamination of cloacae of chicken. Observed variation in prevalence among studies could be attributed to differences in sampling and isolation procedures, used culture, fecal contact to egg, hygienic condition of poultry house, study design and season variation in which samples are collected against the antimicrobial discs tested. The isolated bacteria were susceptible to most of drugs for the treatment and multi drug resistance occurred in 25% of isolates for in vitro tested in this


Introduction
Poultry feeds are food materials used in raising poultry birds. Poultry feeds are referred to as complete feeds as they are designed to contain all the nutritional materials in the form of meat and egg production these have significant importance in humane food [1]. Advances in science and technology have widely contributed for the expansion of the poultry industry and a number of strategies have been adopted to modulate the quality of poultry products like egg and meat [2].
Escherichia coli (E. coli) are major factors for declining of food quality including food originated from poultry due to poultry and poultry products being a primary source of bacterial infection. It has most often been associated with the consumption of contaminated foods of animal origin, such as poultry, swine, dairy products and eggs [3].
Most of E. coli isolates are harmless; however, some strains are pathogenic and may cause serious food poisoning in human beings [4]. In the past two decades, severe outbreaks with gas-trointestinal symptoms have occurred by food borne pathogenic E. coli, particularly 0157:H7 [5]. 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 [6]. Moreover E.coli is one of foods borne bacteria and resistant to one or more antimicrobial drugs used in medicine and agriculture. Antibiotic resistance in E. coli is of particular concern because it is the most common gram-negative pathogen in humans [7]. Some global epidemics have also been linked with egg consumption and known to cause egg-borne pathogens present in poultry eggs and their contents. Food poisoning are associated with egg-borne pathogens may cause severe morbidity or mortality period with diarrhea, vomiting, nausea and abdominal cramps [4].
In the previous study, which carried out the determination of isolation and antimicrobial drug resistance patterns of E. coli isolates and estimates, the level of the pathogen was connected at eastern Ethiopia on poultry farm of cloacae swabs sample and in different studies were conducted on the prevalence of E.coli from both markets and farm egg [8]. There is very little information regarding the E.coli profile from egg and cloacae of poultry and drug susceptibility patterns particularly in Jimma Town, south western Ethiopia. Therefore, this study was aimed to isolate E.coli from cloacae swabs, mark and farm egg and, asses the antimicrobial susceptibility patterns of the isolates.

Description of the Study Area
The study was conducted on market egg, farm and cloacae swabs of laying chicken in Jimma town, Oromia regional state, Southwestern Ethiopia.

Study Design
A cross-sectional study was conducted from January, 2018 to November, 2018 to isolate, identify and characterize antimicrobial susceptibility patterns of E.coli species from market egg, farm egg and cloacae swabs of laying chicken in Jimma town. Kito Furdisa Development Enterprise, Emabet and Daniel poultry farm respectively. Eggs were collected in sterile bags as soon as egg laid using sterile glove and transported to the laboratory and cloacae swabs from these laying sample of eggs were collected and swabs were placed in sterile tube contain 10 ml peptone water. Eighty three egg sellers were selected randomly from Jimma town markets randomly and Single egg from each egg seller were collected in sterile bags randomly using sterile glove.
Cloacae swabs were incubated at 37 o C for 24 h as soon as samples were collected. Samples of egg were processed according to (Loongyai et at., 2011) in which external shell of eggs were swab with sterile cotton swabs dipped in sterile peptone broth and placed in 10 ml of peptone broth and subsequently incubated for 24 h at 37 o C. In order to collect the egg contents, eggs were surface sterilized by immersion in 75% alcohol for 2 min, air dried in a sterile chamber for 10 min, then cracked with a sterile knife. Each egg's content was mixed thoroughly and 1 ml of the mixed egg content was inoculated into 9 ml of peptone broth and incubated at 37C for 24 h.

Isolation and identification of E. coli
A loop from incubated peptone water of egg shells swabbed cloacae swabs and egg content were streaked on MacConkey agar and incubated for 24 hr at 37 °C. Colonies on MacConkey agar having bright pink-red via lactose fermentation were taken and streak on Eosin Methline Bleu (EMB) agar and plates were incubated for 24 hr at 37 °C. Then the plates were examined for the presence of colonies that may resemble E.coli according to the technique recommended by [9]. The organisms showing characteristic colony morphology of E. coli on EMB agar having black golden on the center of colonies were isolated (finger1). Isolated colonies were taken directly from the plate and transferred to nutrient agar for farther identification of E.coli with biochemical test.

Discussion
The present study was conducted to isolate, identify and determine antimicrobial susceptibility patterns of E. coli from farm egg, cloacae swab and market egg samples in Jimma town. Food borne diseases is one of the important issues worldwide. E. coli is considered the most prevalent food borne pathogen that has gained increased attention in recent years. In the present the prevalence of E. coli from egg is in line with the study of Germini et al., 2009. But this finding is lower than some reports from previous works (13). The prevalence of E. coli from cloacae in this study is in line with the privies studies [14,15]. In contrast the finding of this prevalence in the present study is higher than the study of [16,22,23,24 ] in which E. coli was isolated from cloacae samples taken from poultry farms. In another study, [25,26] [19]. This variation is probably attributed to the expression of resistant gene code by the pathogen which associated with emerging and re-emerging aspects of the isolates [21].

Conclusion 8
The present study shows a substantial presence of E. coli farm egg, cloacae swab and market egg samples in the town. In this study, the overall prevalence of E. coli that contaminates the market egg shell was higher that may show contamination is avail in the market than farm. Though most of the E.
coli isolates subjected to antimicrobial susceptibility tests show different degrees of resistance against the antimicrobial discs tested. The isolated bacteria were susceptible to most of the drugs used, those important for the treatment and multi drug resistance occurred in 25% of isolates for in vitro tested in this study.
Based on the above conclusion the following recommendations were forwarded: Poultry farm should be trained to ensure the hygienic practices of poultry farm. Hygiene measures must be implemented on the market egg contamination to prevent the contamination of egg.
In vitro drug sensitivity testing of coli should be performed so that proper treatments can be instituted for E. coli infected patients at animal and human health center.
Further study on molecular characterization of both coli O157:H7 and other shiga toxin producing E. coli strains should be conducted.

Limitation
The isolates were not molecularly characterized due to the lack of resources. All participants were informed about the aim of research. Additionally, written consent was obtained from Jimma town administration office.

Consent to Publication
Not applicable

Availability of Data and Material
The data sets developed and analyzed during the current study are available from the first author or from the corresponding authors upon request.

Competing Interests
The authors declare that they have no competing interest.

Funding
Jimma University College of Agriculture and Veterinary Medicine was sponsored for the research to design the study and collection, analyzing, and interpretation of the data and writing of the manuscript.
Author's Contribution D.T, E.S and M.A were participated in the conception of the research idea, Methodology, carried out the laboratory work, analyze the data and approve the manuscript. Figure 1 Red to Pink color on Mack Conckey and green metallic sheen on EMB from left to right  measuring antimicrobial disk diffusion susceptibility test for Escherichia coli isolates