Prevalence, antibiotic resistance and genotypes of Campylobacter jejuni and Campylobacter coli isolated from chickens in Irbid governorate, Jordan

doi:10.1016/j.ijfoodmicro.2020.108656

International Journal of Food Microbiology

Volume 327, 16 August 2020, 108656

https://doi.org/10.1016/j.ijfoodmicro.2020.108656 Get rights and content

Highlights

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Prevalence rate of Campylobacter in chickens was 31.6%.
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C. coli was isolated more from chickens from informal than formal slaughterhouses.
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All tested isolates were multidrug resistant to 5 or more antimicrobials.
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Chicken isolates shared similarities at phenotyping and genetic levels.

Abstract

Campylobacter is the world's leading cause of bacterial gastroenteritis, causing nearly 9 million cases of food poisoning in Europe every year. Poultry is considered the main source of Campylobacter infection to humans. The objectives of the study were to determine occurrence of C. jejuni and C. coli in chickens, the antimicrobial resistance, genotypes, and relatedness of the isolates. A total of 177 chicken samples obtained from informal butcher shops (fresh), formal poultry slaughterhouses (refrigerated) and retail market (frozen) were analyzed. Isolation of Campylobacter spp. was conducted according to the ISO 10272-2006 method. Multiplex PCR was used for confirmation and identification of the isolates. The disk diffusion method was used to determine the antimicrobial resistance of the isolates and multilocus sequence typing was used for genotyping. The proportion of samples with Campylobacter spp. was 31.6% among all chicken samples (fresh and refrigerated 47.5%, frozen 0%) C. coli was isolated from 42.4% of chicken samples obtained from butcher shops and from 18.6% of samples obtained in formal slaughterhouses. C. jejuni was isolated from 17.0% of samples obtained in butcher shops and formal slaughterhouses. Campylobacter spp. was not isolated in frozen chicken samples. All tested isolates showed resistance toward ciprofloxacin and susceptibility toward imipenem and all of the isolates were multidrug resistant toward 5 or more antimicrobials. Three sequence types were identified among 10 C. coli isolates and seven sequence types were identified among 10 C. jejuni isolates. Among sequence types, chicken isolates shared similarities of both phenotypic and genetic levels.

Introduction

Campylobacteriosis is a disease caused by members of Campylobacter sp. In low and middle income countries (LMICs) infection in toddlers is mainly caused by C. jejuni (Oberhelman and Taylor, 2000), although C. coli, C. fetus and C. upsaliensis can also infect humans. Ingestion of only 500 cells may cause acute diarrhea to humans (Whiley et al., 2013).

All poultry species can carry Campylobacter; however, chickens pose a greater risk to humans due to the frequency of consumption (Humphrey et al., 2007). Consumption of raw or undercooked chicken or cross-contaminated from raw chicken can result in clinical cases (Nadeau et al., 2002). There is always a potential risk of contamination of chicken meat with Campylobacter, if hygienic measures are not applied properly during processing (Osaili et al., 2012). In Jordan, butcher shops (informal, small-scale chicken-slaughterhouses that sell directly to consumers) are still operating, which is the case in many other LMICs. These informal slaughtering facilities are characterized by poor hygiene, inadequate facilities, lack of cold, clean water and inspection during processing. As a result, they are likely to pose a higher risk for consumers (Carron et al., 2017; Cook et al., 2017). Data on Campylobacter infection in poultry in Jordan are available. A study carried out in 2012 found Campylobacter jejuni in around 40% of the studied broiler farms (Osaili et al., 2012). Al-Natour et al. (2018) investigate the occurrence of Campylobacter in 35-layer farms in Northern Jordan and found C. jejuni in 40% of chicken cloacae. More recently, a longitudinal study on a single semi-commercial poultry farm, suggests that there may be differences in the transmission dynamics of Campylobacter in this type of farms (which are an important source of poultry for the Jordanian population) and those observed in poultry farms in high-income countries, with potentially an earlier introduction of the pathogen into the flock, but slower within-flock transmission (Neves et al., 2019). Data on the presence of Campylobacter in poultry products in Jordan are limited, but evidence from other countries strongly suggests that Campylobacter is likely to be present in retail poultry in Jordan. A review conducted in 2009 suggests that in most countries, in which studies have been conducted, a majority of poultry products on retail are contaminated with Campylobacter (Suzuki and Yamamoto, 2009). According to the European Food Safety Agency (EFSA) in 2011, in Europe 8 out of 10 chickens were contaminated with Campylobacter sp. (EFSA, 2011). The frequency of Campylobacter in frozen samples is not common and freezing is suggested as a way for decontaminating slaughtered birds (Ilida and Faridah, 2012),

Macrolides, such as erythromycin and azithromycin, are the treatment of choice for Campylobacter, alternatively, fluoroquinolones (ciprofloxacin) and tetracycline can also be used (Osaili and Alaboudi, 2017; Siddiqui et al., 2015a, Siddiqui et al., 2015b). In severe cases such as bacteremia, aminoglycosides (gentamicin) are also used (Alfredson and Korolik, 2007; Corcoran et al., 2006; Kurinčič et al., 2007). Nevertheless, Campylobacter resistant strains have increased, probably as a result of the increased use and misuse of antibiotics in poultry farms (Silva et al., 2011), with strains being particularly resistant to chloramphenicol, tetracycline, macrolides, and fluoroquinolones (Cody et al., 2010; EFSA, 2011; Silva et al., 2011).

Currently, Multilocus Sequence Typing (MLST) is the leading and most discriminative method for genotyping (Duarte et al., 2016). It implies the use of housekeeping genes (6–7 genes), which may represent the genome for MLST (Dingle et al., 2001). Relying on the fact that the proteins, which encode these housekeeping genes, evolve slowly, MLST will provide data for accurate phylogenic estimation, typing and strain relatedness (Maiden, 2006). Therefore, the aims of the study were to determine i) the occurrence of C. jejuni and C. coli in chickens at informal butcher shops (fresh), formal poultry slaughterhouses (refrigerated) and retail market (frozen) and ii) the antimicrobial resistance, genotypes and relatedness of the isolates.

Section snippets

Sample collection

The following formula (Krousel-Wood et al., 2006) was used to determine sample size:

n = (1.96)²PQ/d², whereas:

n = sample size required.

1.96 z value at α-error = 0.05

P = prevalence of the disease.

Q = 1 – P.

d = tolerated margin of error.

The reported prevalence rate from previous local studies for chickens was 34.4%, Accordingly the number of samples was determined as:

n = ((1.96)² × 0.344 X (1–0.344))/(0.07)²

n = 177.

Over the period January to May 2017, 177 fresh, refrigerated (not more than

Occurrence of Campylobacter in chicken

Out of 177 chicken samples 64 samples (36.2%), had morphology compatible with that of Campylobacter reference strains. Biochemical tests identified C. coli in 40 samples and C. jejuni in 18. Final confirmation of the isolates was carried out using PCR. The results confirmed that 36 of the isolates were C. coli and the remaining 20 isolates were C. jejuni. The frequency of Campylobacter spp. among the studied samples of local chicken from butcher shops (fresh), local chicken from formal

Discussion

Chicken meat has been identified as the main source of human Campylobacter infection and studies aiming at attributing the source of human infection or the relatedness of isolates are conducted (Nadeau et al., 2002; Ravel et al., 2017). The occurrence of Campylobacter among chickens sold in Jordan included in this study (31.6% overall among fresh, refrigerated and frozen) is comparable to results from different countries such as Italy 34% (Stella et al., 2017), China 45% (Zhu et al., 2017),

Acknowledgment

This work was part of the OIE Veterinary Education Twinning Programme between The Royal Veterinary College and the Jordan University of Science and Technology funded by the World Organization for Animal Health (OIE) and partly funded by Deanship of research at Jordan University Science and Technology (grant number 365/2017).

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Prevalence, antibiotic resistance and genotypes of Campylobacter jejuni and Campylobacter coli isolated from chickens in Irbid governorate, Jordan

Highlights

Abstract

Introduction

Section snippets

Sample collection

Occurrence of Campylobacter in chicken

Discussion

Acknowledgment

Food Control

Prev. Vet. Med.

Lancet

Int. J. Antimicrob. Agents

J. Microbiol. Methods

Int. J. Food Microbiol.

Food Control

Int. J. Food Microbiol.

Int. J. Food Microbiol.

Food Control

J. Food Prot.

Mol. Cell. Probes

Food Control

J. Food Prot.

Int. J. Food Microbiol.

Asian Pac J Trop Med

Asian Pac J Trop Med

Clin. Microbiol. Infect.

Food Microbiol.

Food Control

In vitro susceptibility of Campylobacter jejuni from Kuwait to tigecycline & other antimicrobial agents

Ind. J. Med. Res.

Antibiotic resistance and resistance mechanisms in Campylobacter jejuni and Campylobacter coli

FEMS Microbiol. Lett.

Resistance of Campylobacter jejuni isolated from layer farms in northern Jordan using microbroth dilution and disc diffusion techniques

J. Food Sci.

Prevalence and risk factors associated with Campylobacter among layer farms

Vet. Sci. Dev.

Improvement of modified charcoal-cefoperazone-deoxycholate agar by supplementation with a high concentration of polymyxin B for detection of Campylobacter jejuni and C. coli in chicken carcass rinses

Appl. Environ. Microbiol.

Working conditions and public health risks in slaughterhouses in western Kenya

BMC Public Health