Prevalence of Escherichia coli in chicken meat and meat products in Egypt

Prevalence of Escherichia coli in chicken meat and meat products in Egypt

Mohamed M. Elsenduony¹, Lamiaa G. Moustafa¹, Madiha Salah Ibrahim^2*

¹Agriculture Research Center (ARC), Animal Health Research Institute, Reference Lab for Food Safety Analysis of Animal Origin, Alexandria Food Inspection Lab, Egypt

²Department of Microbiology, Faculty of Veterinary Medicine, Damanhour University, Egypt

Abstract

Meat from chicken source is considered one of the most common foods all over the world being easily digested, easily prepared, tasty nutritious food in addition for being economically reasonable.  However, it constitutes one of the highly nutritious media for many micro-organisms, especially Escherichia coli (E. coli).

Many steps in the production of raw chicken meat and meat products from slaughtering till reaching the consumers can be responsible for microbial contamination by foodborne bacteria. The current review covers the so far reported data on the incidence of E.coli in chicken meat and meat products in Egypt.

Introduction

Escherichia coli (E. coli) is a common gastro-intestinal bacteria in both animal and human and it can also found in water, soil and vegetation. Its presence in chicken meat products reflects fecal contamination of meat (Clarence et al. 2009).

1. coli has highly incidence rate in food manufacturing sector with noticeable severity on public health (Abu Elnaga et al. 2014). In addition, E. coli is mostly nonpathogenic, but some of them considered pathogenic and has the ability to induce a harm on animal and human health.

In humans, the pathogenic strains of E. coli have been ensnared in many cases of foodborne infections (Adzitey et al. 2020). Further, E. coli considered one of the most isolated bacteria from chicken carcass worldwide (Davis 2018). Poultry meat get contaminated at slaughtering during scalding, plucking, evisceration and chilling (Keener et al. 2004). Further, contamination during processing and handling by food handlers (Adeyanju and Ishola 2014). Therefore, strict hygienic practices should be adopted during primary processing of chicken carcasses to improve the bacteriological aspect of such products.

This review focused only on Escherichia coli isolated from chicken meat and chicken meat products, either raw or processed, through the so far available published data in Egypt. It covered the incidence of E. coli isolation, the prevalent serotypes, the antimicrobial susceptibility of the isolates, E. coli virulence and antimicrobial resistant genes. This is essential to update the knowledge on prevalence of E. coli in chicken meat to develop effective continuous monitoring plans to detect E. coli in human food-chain and for planning of effective strategies to control dissemination and spread of antibiotic resistant Escherichia coli, especially those with multi-drug resistant patterns, to humans.

Keywords: E. coli, MDR, chicken meat, Egypt

1. coli Prevalence in chicken meat and meat products

Ahmed (2004) detected Enteropathagenic Escherichia coli (E. coli) in 8%, 20% and 12% of luncheon, chicken nuggets and hot wings samples, respectively. While Ali et al. (2010) isolated E. coli from chicken frozen fillets and chicken frozen legs at 28 and 36%, respectively, Escherichia coli O157:H7 were accounted 4 % for each. Escherichia coli was isolated from examined chicken thigh and chicken breast samples by 16% and 12%, respectively (Saad et al. 2011). E. coli was detected in ready to eat meat (RTE) meat products at 10% total out of 120 examined samples of which represented as 10% (4/40) were from chicken luncheon (Awadallah et al. 2014). Further, Hassanin et al. (2014) examined chicken thigh and breast samples with the prevalence of Escherichia coli of 15% and 10%, respectively. Gwida et al. (2014) isolated Escherichia coli from chicken carcass from 16% of the tested samples. While in 2015 they recovered Escherichia coli from raw chicken meat at 35% (Gwida and El-Gohary 2015).Moreover, Saad et al. (2015) recorded that Escherichia coli was isolated from chicken hot wings, shawerma and nuggets by 5 %, 10 % and 25%, respectively from several hypermarkets at El-Gharbia and El- Dakahlyia, governorates.

Escherichia coli isolated from examined chicken thigh samples in 13.33 % (Ibrahim et al. 2015). Khalafalla et al. (2015) reported that E. coli was isolated from all examined.

Also, Escherichia coli O157:H7 was isolated from 0.8% of chicken carcass, while 2% of chicken meat was positive for non-O157:H7 STEC (Ahmed et al. 2017). Out of 90 investigated chicken samples; fresh and frozen, E. coli were isolated from 11.7 % from examined samples (Moawad et al. 2017).

Abd Elzaher et al. (2018) isolated E. coli from 80.35% the internal surfaces and 78.57 % of external surfaces of the examined chicken. The incidence of E. coli isolation was higher from the carcass internal surfaces than the carcass external surface. While Ibrahim et al. (2018) reported incidence of E. coli were 13.3%, 16.6% and 23.3%, in samples of luncheon, nuggets, and shawerma, correspondingly.  In addition, Elbayoumi et al. (2018) reported that E. coli was isolated from breast and thigh samples by 14.3 and 20%, correspondingly. On the other hand, Atia (2018) Escherichia coli was isolated from 8 and 16 % of the examined breast and thigh samples, and Elshora (2019) reported that E. coli was isolated from breast samples by 8.57%.

Elsisy (2019) said that E. coli was isolated by 30 and 45 % from examined thigh and breast samples correspondingly. On the other side, E. coli could be detected in 2(6.66%) of examined nuggets samples (Gaafar et al. 2019).

Moustafa et al. (2019) recorded that the isolation of Escherichia coli from breast and thigh muscles of chicken carcasses at 40 and 48%, respectively. Similarly, Abo Elmagd et al. (2019) stated that out of 600 isolates obtained from 100 examined chicken meat, only 434 isolates were confirmed biochemically as Escherichia coli at a prevalence of 72.3 % (434/600), while when based on colonial morphology they reported an incidence of 100%.

Salem et al. (2019) reported high incidence of Escherichia coli in chicken fajitas (40%) followed by shishtawook (32%). It was evident from their results that fajitas were the most contaminated ones, followed by Shishtawook. They attributed this to more handling, spices and vegetables added during preparation which may be the source of the contaminating microorganisms.

Out of 100 examined samples, only 4 % were positive for Escherichia coli O157:H7, while 11% of the isolates were other species rather than E. coli O157. E. coli was detected in 6% and 9% breast and thigh samples, respectively (Shaltout et al. 2020).  Also, Abdelkarim et al. (2020) surveyed a total of 403 chicken carcass samples. The data showed a 66.3% prevalence of Escherichia coli. Arakeeb (2020) reported that E. coli was isolated from 62.5 and 42.8 % of breast and thigh samples, respectively. While, Escherichia coli was isolated from 4% raw chicken samples and 12% of chicken ready for consumption samples (Abd El-fatah et al. 2020).

1. coli were isolated from 88 % of chicken’s meat by (Shawish et al. 2021).

Furthermore, Escherichia coli O157:H7 was isolated in 1.1% of collected samples from different location in Egypt. The incidence of Escherichia coli O157:H7 was 1.2% from examined beef, raw and cooked chicken samples. Through these samples, Escherichia coli O157:H7 isolated mostly from cooked chicken samples by 1.9%. Regarding examined samples sources, Hurghada and Luxor Governorate showed the highest incidence of E. coli O157:H7 in 8.3% and 6.3%, respectively; however, the incidence in Cairo and Alexandria Governorates was 3.3 and 5.0 %. Lastly, there was no any E. coli O157:H7 isolate reported from either Aswan or Sharm El Sheikh Governorates (Refaay et al. 2022).

Coliform percentages in poultry meat product samples (n=25) was 76%, 88%, 100%, 100% in chicken pane, luncheon, burger and popcorn, respectively (Saad et al. 2022).

Elsenduony et al. (2022) found that all examined chicken breast samples were free from E. coli from all farms, while the incidence of E. coli were 16.6 % and 25 % from examined chicken thigh samples collected from two farms, correspondingly. While, all samples from breast were free from E. coli from all farms.

The prevalence of E. coli in chicken product samples was 50% (n=30); it was isolated from 65% (13) of nuggets, 35% (7) of luncheon and 50% (10) of pane samples (Morshdy et al. 2023)

Drawing a conclusion based on the comparison of the obtained results among all the above studies will not be consistent due to the vast differences in the sample size, type, collection and storage conditions. However, these data indicate, either high, moderate or low percentages of E. coli isolation from chicken or its products that contamination does exist from production till reaching the consumer.

Prevalent E. coli serotypes among chicken meat and chicken meat products

Serotyping is one of the methods used for identifying strains for epidemiological purposes. Identification of E. coli by detection of surface antigens using antibodies considered one of traditional technique; flagellar H-antigens, capsular K-antigens and O- polysaccharide antigens. Currently, there are 53 H-types besides 186 different Escherichia coli O-groups (Fratamico et al. 2016). Further, O-serogroups are related to the virulence factor profile of each strain.

The isolated strains of E. coli by Ahmed (2004) were serologically identified as 055: K59 0111: K58 and 0124: K72 in addition to untypable isolates.

1. coli O119: K69 and O55: K59 were isolated from breast and thigh, respectively. Thigh and breast O119:K69 and O125:K70, thigh only O26:K60 and breast only O55:K59 and O124:K72 (Saad et al. 2011).

Hassanin et al. ( 2014) serologically identified Escherichia coli as O55:K59 (enteropathogenic; EPEC), O124:K72 and O125:K70 (enterotoxigenic; ETEC). Furthermore, the incidence of Enteropathogenic E. coli which serologically identified (O119:K69 and O55:K59) was 4%, Enterotoxogenic E. coli (O125:K70) was 4%, Enterohaemorrhagic E. coli (O26:K60) was 4% from the breast only and Enteroinvasive E. coli (O124:K72) was 4% from thigh only.

Further, E. coli isolated from chicken luncheon were identified as O55:K59 (3 isolates) and O111:K58 (1 isolate). They were categorized as enteropathogenic E. coli (EPEC) and enterohaemorrhagic Escherichia coli - Shiga toxigenic Escherichia coli , respectively (Hassanin et al. (2014).

Incidence of EPEC and STEC in chicken meat may be related to possible cross contamination during food handling and processing or may be during the evisceration process, mainly due to the rupture of the intestine at preparation (Awadallah et al. 2014).

Further, 5% of each of the following serotypes were detected in chicken nuggets: O119:H6, O86, O125:H21, O124, O26. Also, O55:H7 and O26 were detected in 5% of the chicken shawarma samples. In addition, O111:H4 was the only serotype in chicken hot wings at 5% as well (Saad et al. 2015). Further, serological identification of E. coli isolates by Gwida and El-Gohary (2015) revealed the presence of EHEC; O26:H11and O103:H2, EPEC; O128:H2 and O111:H2 and ETEC O78.

Non-O157:H7 E. coli comprised O26:H11 (2), O111:H8 (3), O113:H21 (1) and O55:H7 (1) (Ahmed et al. 2017). Further, Twenty‐one E. coli isolates from chicken meat samples were serotyped as O1 (2), O18 (3), O20 (1), O78 (1), O119 (1), and O146 (1) (Moawad et al. 2017).

As reported by Ibrahim et al. (2018), 4 out of 30 chicken luncheon samples were positive for E. coli (13.3%), one of them (25%) identified as Enterotoxogenic E. coli (O128:H2),while two of them (50%) identified as Enteropathogenic E. coli (O91:H21 and O78) and other isolates (25%) were Enteroinvasive E. coli (O124). Moreover, 5 out of 30 s chicken nuggets samples (16.6%) confirmed to be contaminated by Escherichia coli, one of them (20%) identified as Enterohaemorrhagic E. coli (O26:H11), four of them (80%) were Enteropathogenic E. coli (O1:H7 (2), O44:H18(1) and O78(1), also, 7 out of 30 half cooked chicken shawerma samples were E. coli (23.3%), the isolates identified as Enteropathogenic E. coli (O91:H21 (1), O127:H6 (1), O55:H7 (1), O44:H18 (1), O153:H2 (2), and O78 (2).

Salem et al. (2019) reported that they could isolate different serotypes of E. coli from shishtawook beside chicken fajitas, as O113:H4 (4%), O111:H2 (4%), O127:H6 (4%) ,O26:H11 (4%) and O103 (4%), while O111:H2 (4%), O119:H6 (8%), O113:H4 (4%), O26:H11 (4%), O127:H6 (4%) ,O91:H21 (4%) and O124 (4%) were isolated from fajitas.

Serological identification by Gaafar et al. (2019) of isolated E. coli revealed detection of O78 and O1:H7 (EPEC) in chicken nuggets samples at 6.66%.

While the incidence of other serotypes were O127:H6 (4%), O114:H21 (2%), O26 (2%) and O126 (3%) (Shaltout et al., 2020). Moreover, Shawish et al. (2021) reported the most common E. coli serotypes were (O128, O86) by 4.5% ; (O26, O 121) by 9%,(O146) by 11.4% ; (O 111 and O91) by 13.6 %;(O119) by 18.2 % and (O45) by 15.9 % .

15 E. coli isolates from popcorn, burger, luncheon and fresh pane and serotyped as O142 (20%), O128:H2 (6.66%), O114 (13.33%) and O124 (13.33%) (Saad et al. 2022).

Elsenduony et al. (2022) recorded that by the examination of chicken thigh samples from farm no.4; 8.33% and 16.7% of samples were positive for E. coli which serotyped as O2: H6 (EPEC) and O91: H21 (EHEC), correspondingly. While E. coli O78 and E. coli O44: H18 were isolated from the examined farm no.5 thigh samples with an incidence rate of 8.33% for each serotype.

Elbayoumi et al. (2018) isolate different E. coli serotypes (O78 and O26:H11) from breast samples with incidence of 8.6% and 25.71%, correspondingly .While, from thigh muscle samples , the isolated E. coli serotypes were O78 , O91:H21 and O2: H6,  with incidence  of 5.71% for each, correspondingly. Edris et al. (2015) said that E. coli O78 was isolated from chicken thigh samples and E. coli O26, O78 from breast samples.

Such result may well be a small or huge diverse comparing to others because of the huge scale of Escherichia coli serological typing.  Phenotypic serotyping methods for differentiating E. coli considered not necessarily accurate. Thus, molecular serotyping and subtyping methods for E. coli allow more accuracy rather than other methods in addition to the advancement in investigation of outbreaks of foodborne disease and tracing-back the main sources.

Prevalence of E. coli virulence genes among chicken meat and its products

Analysis of different genes associated with virulence in E. coli isolates was performed in some not all of the studies. Further, limited number of genes was analyzed per study and there was inconsistency in the detected genes.

Saad et al. (2011) reported that elt gene was detected by molecular examination of isolated E. coli (O55: K59 and O119: K69) from examined thigh and breast chicken samples.

Hassanin, et al. (2014) failed to detect stx1 and stx2 genes in any of the analyzed isolates.

PCR analysis by Awadallah et al. (2014) of E. coli serotypes showed the presence of Stx1 gene and Stx2 gene in O111:K58 isolates. The stx1 gene was not detected in all E. coli O157:H7 isolates or other serotypes rather than O157:H7, while stx2 was detected in 100% of the E. coli O157:H7, it was detected in 60% of other serotypes rather than O157:H7. In addition, eaeA and hlyA were detected in E. coli O157:H7 and non- O157:H7 STEC at 100% and 60%, respectively (Ahmed et al. 2017). Abd Elzaher et al. (2018) showed that 33.3% (2/6) of E. coli isolates from chicken carcasses were positive for the eaeA gene.

fliC gene of E. coli is considered as an indicator for E. coli O157:H7 (Carey et al. 2009). PCR investigation by Shaltout et al., (2020) revealed that fliC gene was detected in all E. coli O157 isolates.

1. coli isolates ECKFGF5 and ECDMGF6 were negative for stx1 and stx2 genes. While, stx2 were detected in E. coli ECGHBF1 and ECMFLF4 strains (29%) (Abdelkarim et al. 2020). Moreover, Refaay et al. (2022) reported that after examination of E. coli O157:H7 isolates, the result showed that they harbored eae gene and stx1 gene was not detected. While, only one isolate harbored stx2 gene beside eae gene, but was not from chicken meat .

Antimicrobial susceptibility among E. coli from chicken meat and chicken meat products

Antibiotics have and are still extensively used in poultry farms as therapy and as growth promotors. This greatly inflated the emergence of antimicrobial resistance in food borne pathogens with its hazards on human health and consequent treatment failure. Especially, antimicrobial resistant E. coli is considered one of the major challenges in both humans and animals.

Antibiotic susceptibility was mostly carried out using the disc diffusion method, but automated Vitek2 analysis was used by some researchers. Different studies used different antibiotics, however, most of the antibiotic categories were covered. In addition, not all the studies investigated the antibiotic susceptibility of their isolates.

Ali et al. (2010) isolated E. coli from poultry products that showed 100% resistance to Cephalexin, Erythromycin and Penicillin G, while 100% sensitivity to Amikacin, Ciprofloxacin and Gentamicin.

Gwida and El-Gohary (2015) showed that raw chicken meat was 100% resistant to Ampicilin, Cefoxitin and Tetracycline, while variably resistant to other tested antibiotics.

Moawad et al. (2017) reported that E. coli isolates showed resistance to Tetracycline, Ampicillin, Streptomycin, Trimethoprim/sulphamethoxazole and Amoxicillin–clavulanic acid with 80, 80, 60, 66.7 and 66.7%, respectively. Eight and seven isolates were susceptible to Ciprofloxacin, Enrofloxacin and Ceftriaxone correspondingly, and all tested isolates were susceptible to Colistin.

Abd Elzaher et al. (2018) showed that E. coli isolates were 100% resistance to Penicillin, Trimethoprim/Sulfamethoxazole. While were sensitive to Cefotaxime (35.92%) followed by Chloramphenicol (37.75%), Rifampicin (44.25%), Norfloxacin (50.27%), Ofloxacin (61.77%), Doxycycline (76.97%), Cephradine (93.4%), Amoxicillin (93.7%) and Tobramycin (100%),

The incidence of multidrug-resistant (MDR) strains in chicken meat cuts-up samples was found to be 81%. 3 out of 262 isolates were sensitive to all antibiotic agents, while 212 out of 262 isolates were multi drug resistance. The resistance rates were lowest for Gentamicin (7.2%) and Chloramphenicol (12.6%). On other hand, it was highest for Vancomycin (99%) and Tetracycline (96%). Other antibiotic showed ranged resistance pattern from 24-73% (Abdelkarim et al. 2020).

Abd El-fatah et al. (2020) showed 100% resistance of E. coli isolates to Clindamycin and Rifampin and 78.9% resistance to Ciprofloxacin.

Shawish et al. (2021) revealed a strong resistance of E. coli isolates against Tetracycline (33.3%), Ampicillin (26.6%), Kanamycin and Trimethoprim-Sulphamethoxazole (20%), Cefotaxime (12%), Chloramphenicol and Ciprofloxacin (8%). Whereas; the lowest degree of resistance was noticed against Streptomycin (6.6%), Ceftazidime (5.3%) and Nalidixic acid (4%).

Antibiotic resistance pattern using Vitek 2 by Elsenduony et al. (2022) revealed that all E. coli isolates were Extended Spectrum Beta-Lactamase ( ESBL) negative and showed resistance to Trimethoprim/ Sulfamethoxazole, Piperacillin, Ticarcillin, and Ciprofloxacin but were sensitive to other antibiotics used.

Refaay et al. (2022) reported that 100 % of E. coli O157:H7 isolates showed resistance to Cefixime, Ciprofloxacin, Ampicillin and Cotrimoxazole. 85.7% showed resistance to Pipracillin-Tazopactam, Piperacillin and 57.1% showed resistance to Gentamicin. However, the lowest resistance rate (14.3%) was observed against Aztreonam . Furthermore, the isolates showed high sensitivity to Meropenem (71.4%) and Amikacin (85.7%). E. coli O157:H7 isolates showed multidrug resistance.

Generally, MDR was common among all the E. coli isolates in the above-mentioned studies with diverse patterns of antibiotic resistance profiles among the isolates that were derived from different sources and/or regions. Monitoring antibiotic usage in poultry production is essential to control the development and spread of MDR bacteria.

Antimicrobial resistance genes harbored by E. coli from chicken meat and chicken meat products

Very few analyses of the antibiotic resistance genes were performed by the papers reviewed here, by using RT PCR, resistance-associated genes were detected during molecular examination of E. coli isolates isolated from chicken meat. Five out of ten isolates harbored bla_TEM (1 and 104), while bla_CTX-M (1 and 14) was positive in four isolates. Three isolates carried qnrB and qnrA while bla_OXA-1 was detected in three isolates. Genes qnrS ,bla_CMY and bla_SHV were not detected in any isolates. mcr-1 gene and bla_{OXA-23 gene} were not identified in E. coli isolates (Moawad et al. 2017). Abd Elzaher et al. (2018) showed that acc6-Ib-cr genes is detected in 33.3% (2/6) of E. coli isolates from chicken carcasses, where qnrA gene was not detected in all examined isolates. Further, all E. coli isolates obtained from chicken samples contained bla_TEM gene but not bla_CTX gene (Abd El-fatah et al. 2020).

Conclusion

In Egypt, great economic losses to the poultry industry occurred due to Avian pathogenic E. coli (APEC) affecting the affordable protein supply to a wide population, while extra intestinal enteropathogenic E. coli (ExPEC) causes serious conditions in human. Despite the existence of defining virulence genes shared between APEC and ExPEC of humans, it is not fully ruled out whether APEC is the main source of the ExEPEC infection in human, especially through the food chain suggesting APEC zoonosis. Thus, understanding the prevalence and the phenotypic as well as the genotypic characteristics of E. coli isolated from food would greatly unravel the role of poultry and poultry products in the spread of ExPEC among humans.

Further, monitoring the antibiotic usage in poultry industry is essential to control the spread of antimicrobial resistant bacteria. Moreover, applying strict hygienic conditions during all the production steps of poultry meat processing is essential to control foodborne infections in human.