Prevalence and Antimicrobial Susceptibility of Escherichia Coli O157 in Beef at Butcher Shops and Restaurants in Central Ethiopi
Total Page：16
File Type：pdf, Size：1020Kb
Beyi et al. BMC Microbiology (2017) 17:49 DOI 10.1186/s12866-017-0964-z RESEARCHARTICLE Open Access Prevalence and antimicrobial susceptibility of Escherichia coli O157 in beef at butcher shops and restaurants in central Ethiopia Ashenafi Feyisa Beyi1,2*, Akafete Teklu Fite1, Ephrem Tora1, Asdesach Tafese1, Tadele Genu1, Tamirat Kaba3, Tariku Jibat Beyene1,4, Takele Beyene1, Mesula Geloye Korsa1,5, Fanos Tadesse1, Lieven De Zutter6, Bruno Maria Goddeeris6,7 and Eric Cox6 Abstract Background: Ethiopia bears the largest burden of foodborne diseases in Africa, and diarrheal diseases are the second leading causes of premature deaths. Enterohemorrhagic Escherichia coli O157 causes an asymptomatic infection to severe diarrhea and/or hemolytic-uremic syndrome in humans. Methods: A total of 440 beef carcass and in-contact surface swabs from 55 butcher shops and 85 minced beef samples from 40 restaurants in central Ethiopia were collected and examined for the presence of E. coli O157. Standard microbiological methods were used to isolate and identify E. coli O157 and to characterize the antimicrobial resistance of the isolates. Results: E. coli O157 was detected in 4.5% carcass swabs (n = 5) and 3.6% cutting board swabs (n = 4) samples from butcher shops. E. coli O157 was not detected in any of the minced beef samples obtained from restaurants. All isolates (n = 9) were 100% susceptible to five drugs, but five isolates were resistant to amoxicillin, two isolates to streptomycin and three isolates to chloramphenicol. One isolate was resistant to two drugs and another to three drugs. Conclusions: The present study shows a low prevalence of E. coli O157 in beef sold at butcher shops. Nevertheless, given the low infective dose of this pathogen and the deep-rooted tradition of consuming raw or undercooked beef, the current prevalence should not be considered lightly from a public health perspective. Keywords: Antimicrobial susceptibility, Beef, Butcher shops, Escherichia coli O157, Minced beef, Restaurants Background consumption of contaminated and improperly cooked In the 21st century, foodborne diseases have become one minced beef and unpasteurized cow milk [4]. Butcher of the important issues all over the world [1]. Due to houses and restaurants are frequently incriminated as poor infrastructure and low level of awareness, this sources of E. coli O157 for human infections [5]. problem is worse in developing countries [2]. Major Ethiopia ranks second after Nigeria in the health burden pathogenic microorganisms that frequently have been of zoonotic diseases in Africa [6]. This country is located associated with foods of animal origin include Enterohe- in a sub-region that experiences the second highest food- morrhagic Escherichia coli O157. In humans this pathogen borne disease burden in the world, where E. coli O157 is causes asymptomatic infection to severe diarrhea and/or one of the leading causes of foodborne disease disability hemolytic-uremic syndrome (HUS) [3]. Human infections adjusted life years [7]. In Ethiopia, years of life lost due to with E. coli O157 have been mostly associated with the diarrheal diseases was 2.6 million in 2010, and diarrheal diseases are the second leading cause of premature death after lower respiratory infections [8]. Information about * Correspondence: [email protected] E. coli 1College of Veterinary Medicine and Agriculture, Addis Ababa University, human infections with O157 is limited in this Bishoftu, Ethiopia country, however, in a study conducted on 422 diarrheic 2 Department of Animal Sciences, University of Florida, Gainesville, FL, USA children under 5 years in northern part of Ethiopia, E. coli Full list of author information is available at the end of the article © The Author(s). 2017 Open Access This article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated. Beyi et al. BMC Microbiology (2017) 17:49 Page 2 of 6 O157:H7 was isolated from 59 (28.9%) of the children [9]. cities as sampling frames. When visiting each selected The habit of consuming raw and/or undercooked meat is butcher shop and restaurant, the purpose of the study one of the factors that exacerbate the transmission of was explained to the manager, and a letter from the foodborne pathogens including E. coli O157 in the coun- College of Veterinary Medicine and Agriculture Addis try. Sufficient heating of meat kills these organisms [4]; Ababa University demonstrated the approval and cred- however, in Ethiopia, most people prefer to eat raw or ibility of our work. From each butcher shop on average undercooked beef (locally called kitfo, dulet,andkurt). eight samples were collected on two separate days. A Treatment of Enterohemorrhagic E. coli infections total of 440 swab samples consisting of 110 samples with antibiotics may worsen the illness, presumably by from beef carcass, knife, cutting board, and butcher’s breaking up the bacteria with the release of more toxins hands at butcher shops, respectively, and 85 minced beef and increased toxin production [10]. However, early ad- (locally also called kitfo) samples from restaurants were ministration using some antimicrobials is effective [11]. collected (Table 1). Beef carcasses were swabbed follow- Unfortunately, inappropriate ways of antimicrobial uses ing the method described in ISO17604 (2003). A sterile have contributed to the increase in antimicrobial resist- cotton tipped swab (2X3cm) fitted with shaft, was first ance [12]. soaked in an approximately 10 ml of buffered peptone In Ethiopia, only few small-scale studies estimating the water (BPW; Oxoid, Hampshire, UK) and subsequently prevalence and/or assessing the antimicrobial sensitivity rubbed horizontally and vertically several times on the profile of enterohemorrhagic E. coli O157 in feces, skin carcass surface. After the rubbing process, the shaft was swabs and carcasses of sheep, goats, and cattle at abattoirs broken by pressing it against the inner wall of the test were conducted [13–16]. But studies at the level of con- tube leaving the cotton swab inside the test tube. The sumption particularly in butcher shops and restaurants is whole surface of the carcass was swabbed by one swab, lacking. Consequently, this study was designed to address and swabs from multiple carcasses from the same the information gap pertaining to the prevalence and anti- butcher shop were pooled together. Carcass in-contact biotic susceptibility profiles of E. coli O157 in beef carcass surfaces including knife, butcher’s hands and cutting and minced beef at butcher shops and restaurants respect- board swabs were obtained by swabbing them using a ively in central Ethiopia. cotton swab pre-moistened in BPW. The junction of the handle and blade of all knives in each butcher shop was Methods swabbed and pooled together. Similarly, both sides of Study design and sample collection the two hands of the butcher, and the whole surface of From December 2013 to April 2014, carcass and carcass the wooden cutting board were swabbed. The carcass in-contact surface swabs of 55 randomly selected swab, the butcher’s hands swab, the cutting board swab butcher shops, and minced beef samples of 40 randomly and the knife swab were kept separately. The minced selected restaurants were collected in four cities (Addis beef samples were taken from ready-to-eat undercooked Ababa, Bishoftu, Batu, and Holetta) in central Ethiopia. (n = 45) and raw minced beef (n = 40) at the selected These cities have municipal abattoirs which render restaurants. All samples were transported in cold boxes slaughter services to their respective dwellers. Butcher to the laboratory. The samples were stored at 4 to 7 °C shops and restaurants get their beef from cattle slaugh- and analyzed within 6 to 12 h as described in ISO tered in the abattoirs of their respective cities. Neverthe- 16654:2001. less, back yard or illegal slaughtering of animals is a common practice. The abattoirs are barely equipped Sample processing with necessary facilities, and shortage of clean water is Ninety ml of modified tryptone soy broth supplemented one of the chronic problems for most abattoirs in the with novobiocin (mTSB + N; Oxoid) were added to country. In addition, the sanitary condition of most 10 ml swab sample. Conversely, 25 g of each minced restaurants and butcher shops is poor; generally, such beef sample were collected in a Stomacher bag. After food catering firms are loosely monitored and regulated. adding 225 ml mTSB + N, each sample was homoge- In Ethiopia, fresh beef cuts are commonly purchased nized using a Stomacher 400 (Seward Medical, England) from butcher shops and are consumed at home or at the for two minutes and transferred into a sterile flask. After same butcher shop, either cooked or raw. In butcher’s incubation at 41.5 ± 0.5 °C for 24 h, all enrichment shops, a beef carcass is kept in open-air at environmen- broths were plated onto sorbitol MacConkey agar tal temperature (in Addis Ababa 7 to 25 °C daily (Oxoid) supplemented with 0.05 mg/l cefixime and temperature).