Isolation, Identification and Antimicrobial Resistance Profile of Staphylococcus Aureus in Cockroaches (Periplaneta Americana)
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Journal of Advanced Veterinary and Animal Research ISSN 2311-7710 (Electronic) http://doi.org/10.5455/javar. 2016.c153 September 2016 A periodical of the Network for the Veterinarians of Bangladesh (BDvetNET) Vol 3 No 3, Pages 221-228. Original Article Isolation, identification and antimicrobial resistance profile of Staphylococcus aureus in Cockroaches (Periplaneta americana) Ariful Islam, Aurjun Deb Nath, Kamrul Islam, Shariful Islam, Shovon Chakma, Muhammad Belal Hossain, Abdullah Al-Faruq and Mohammad Mahmudul Hassan • Received: May 28, 2016 • Revised: June 27, 2016 • Accepted: June 28, 2016 • Published Online: June 29, 2016 AFFILIATIONS ABSTRACT Ariful Islam Objective: The study was conducted to determine the prevalence of Staphylococcus Shariful Islam aureus in cockroaches (Periplaneta americana), and to assess the antimicrobial EcoHealth Alliance, New York, NY, resistance profiles of the isolated bacteria. USA. Materials and methods: A total of 150 cockroaches (P. americana) were & randomly captured from three households and four restaurants in Chittagong City Institute of Epidemiology, Disease Control Corporation, Bangladesh during July to December 2014. The cockroaches were and Research (IEDCR), Mohakhali- transported to the bacteriology laboratory at the Poultry Research and Training 1212, Bangladesh. Centre (PRTC), Chittagong Veterinary and Animal Sciences University. The Aurjun Deb Nath isolation and identification of Staphylococcus spp. from the external surface wash and Abdullah Al Faruq gut homogenates by pooling cockroaches were done by following conventional Mohammad Mahmudul bacteriological examinations followed by biochemical characterization. The Hassan antibiotic susceptibility profiles of the isolates were determined using disc Faculty of Veterinary Medicine, Chittagong diffusion method. Veterinary and Animal Sciences Results: In this study, the overall prevalence of S. aureus was 38% (n=57/150). University, Khulshi, Chittagong 4225, Higher prevalence of Staphylococcus spp. was observed among the cockroaches from Bangladesh. restaurant (49.3%; n=37/75) as compared to those of households (26.7%; Kamrul Islam n=20/75) having a significant difference (P<0.05). Highest level of resistance by Shovon Chakma the Staphylococcus spp. was found to Penicillin (68%) followed by Erythromycin Institute of Epidemiology, Disease Control (60%), Oxacillin (46%) and Clindamycin (31%). On the other hand, the and Research (IEDCR), Mohakhali- Staphylococci isolates were highly sensitive to Cephalothin (84%) and Kanamycin 1212, Bangladesh. (65%). Conclusion: The rational use of antibiotics needs to be adopted in both human Muhammad Belal Hossain and animal medicine practices to prevent the emergence of drug resistant International Centre for Diarrhoeal Disease Research, Bangladesh (icddr,b), Mohakhali- Staphylococcus spp. 1212, Bangladesh. CORRESPONDENCE KEYWORDS Mohammad Mahmudul Hassan Antibiogram, Cockroach, External washing, Gut content, Prevalence, Resistance Faculty of Veterinary Medicine, Chittagong Veterinary and Animal Sciences University, Khulshi, Chittagong 4225, Bangladesh E-mail: [email protected] How to cite: Islam A, Nath AD, Islam K, Islam S, Chakma S, Hossain MB, Al-Faruq A, Hassan MM (2016). Isolation, identification and antimicrobial resistance profile of Staphylococcus aureus in Cockroaches (Periplaneta americana). Journal of Advanced Veterinary and Animal Research, 3(3): 221- 228. http://bdvets.org/javar/ Islam et al./ J. Adv. Vet. Anim. Res., 3 (3): 221-228, September 2016 221 INTRODUCTION human or veterinary medicine, and for the production of food-animal (Bager and Helmuth, 2001; Anderson et al., Cockroach is one of the most common pests in urban 2003; Schroeder et al., 2004). These resistant microbes environmental settings that are associated intimately to may act as a potential source in spreading antimicrobial the food which carries and spread antimicrobial resistant resistance gene to human pathogens (Schroeder et al., bacterium frequently (Bennett and Owens, 1986). 2004). Cockroaches are largely found in multi-family dwellings, and may act as a carrier of several microorganism The isolation and identification of cockroach associated affecting public health (Wood et al., 1981). Both in with Staphylococcus spp. and their antimicrobial resistance restaurants and households, cockroaches are commonly profile are of public health importance and imperative to found in dark rooms, kitchens, bathrooms and food the scientific community to take appropriate strategies storage rooms, and they have the ability to move from and could provide information on the supplementary one part of a building to another and even in dark light emergence of drug resistance. Therefore, the present (Rivault, 1989). The cockroaches come in contact with research was aimed to asses the prevalence and garbage, feces, stored food, sewage, and biological wastes antibiogram profile of S. aureus isolated from cockroaches (Rivault et al., 1993). Close association to the human in restaurants and households. dwellings, contact with environmental wastages might have influence in the emergence and spread of anti- MATERIALS AND METHODS microbial resistant Staphylococcus aureus from cockroaches to human and vice versa. Cockroach collection and identification: Three household and four restaurants of Chittagong City Emergence and rapid spread of antimicrobial resistance Corporation (CCC) were randomly selected where large bugs particularly S. aureus has become a grave global number of people had their foods every day. The concern. Multidrug resistant (MDR) Staphylococcus poses a households had at least six members and the restaurants growing problem for human health and has been consi- represented medium level eating centers which attended dered as horrifying public health threat (Ahaduzzaman et around three hundred and fifty customers per day. al., 2014). The dismaying bug is commonly found within Sampled cockroaches were collected using sterile screw- tissues, skin surfaces or in foods contaminated by capped 250-mL jars and sterile hand-gloves (Paul et al., exposure to infected human beings or animals. Close 1992), and were transported to the “Poultry Research and interface between human, animal, and environmental Training Centre” (PRTC), Chittagong Veterinary and components both biological and physical, the threat of Animal Sciences University (CVASU) for bacteriological pathogenic Staphylococci is continuously surging up analyses. The insect identification has been confirmed in towards a complex situation (Tachbele et al., 2006). accordance with Lane and Crosskey (1993). Staphylococcus spp. is recognized as a foremost cause of food poisoning, wound infections, skin infections, Estimation of sample size: The required sample size infections of internal organ, and once resistant S. aureus was based on the prevalence of 13% (n=24/187) S. aureus emerges in such infections, it will be very difficult to among isolated bacterial pathogens in conserved continue treatment regimens, because bacterium may cockroaches of hospitals and dwelling of Northeast acquire resistance against commonly used drugs. Foods Algeria (Tine et al., 2014). However, we are assuming that are usually contaminated by the different pathogens due cockroaches might show 10% Staphylococcal prevalence to unhygienic handling of food results. Close contact of in the study population. Based on this assumption, we human and animal is one of the possible sources of food estimated that 150 cockroaches would be sampled for our contamination, and spreading of pathogen to foods or study (population size 100000, confidence limits 5%, utensils through rodents and pests such as cockroaches. design effect 1.0 and clusters 1). The sample size was calculated using windows version of the Epi InfoTM Around the globe, many researchers reported that while 7.1.5.0 software. living close to human dwellings cockroaches play vital roles in transmitting pathogens like bacteria, virus, Processing of cockroaches to isolate Staphylococcus helminthes and protozoan (Agbodaze and Owusu, 1989; aureus: Captured cockroaches humanely killed using Fotedar et al., 1991; Cloarec et al., 1992; Pai et al., 2003), chloroform soaked cotton in a screw capped sterile jar. and some of isolated bacterial pathogens were resistant to Body surface of cockroaches were washed with various antimicrobials (Fathpour et al., 2003). Different physiological saline after vortexing for 2 min and taken as types of food sources contain resistant bacteria that are a homogenate sample. Before gastrointestinal tract (GIT) resistant to one or more antimicrobials that are used in dissection, each cockroach was decontaminated with 95% Islam et al./ J. Adv. Vet. Anim. Res., 3 (3): 221-228, September 2016 222 Table 1. Antibiotics used, concentrations and zone diameter interpretative standards based on CLSI (2007) Antimicrobial Antibiotics Concentration Zone diameter (mm) R I S Aminoglycosides Kanamycin 30 µg ≤ 13 14-17 ≥ 18 Macrolides Erythromycin 15 µg ≤ 13 14-22 ≥ 23 Cephems Cephalothin 30 µg ≤ 14 15-17 ≥ 18 Lincosamides Clindamycin 2 µg ≤ 14 15-20 ≥ 21 Penicillins Penicillin 10 unit ≤ 28 - ≥ 29 Penicillins Oxacillin 1 µg ≤ 10 11-12 ≥ 13 R=Resistant, I=intermediately resistant, S=Sensitive ethanol, and the residue of ethanol was removed with procedure was used on Mueller-Hinton (MH) agar (Bauer saline solution. The gut was dissected aseptically using et al., 1966), and a 0.5 McFarland standard was prepared sterile needles and washed