ORIGINAL_ARTICLE This study was conducted to isolate generic extended-spectrum cephalosporin (ESC)-resistant and extended-spectrum β-lactamase (ESBL)-producing enterobacteria from horses in Nigeria, and to determine the antibacterial resistance profile. Rectal swabs were collected from 155, systematic randomly selected, apparently-healthy horses. Isolation of ESC-resistant enterobacteria was done using Mac Conkey agar with ceftazidime. ESBL production was assessed by combination disc method. Resistance of the isolates was determined using disc diffusion method. Out of 155 samples, 5.2% gave positive growth. From these, 11 ESC-resistant enterobacteria comprising of 36.4% E. coli, 36.4% Salmonella spp. and 27.2% Proteus spp., were obtained. From 11 isolates, 45.5% consisting of all the 4 E. coli and 1 Proteus isolates, were ESBL-producers, these were recovered from 4 (2.6%) out of the 155 horses sampled. Resistance of the E. coli isolates was 25% to aztreonam (AZT), 75% to amoxicillin-clavulanic acid (AMC), gentamicin (GEN), perfloxacin (PEF), and sulphamethoxazoletrimethoprim (SXT-TRI), 50% to ofloxacin (OFL) and 100% to ampicillin (AMP), ceftazidime (CTZ), cefotaxime (CTX), chloramphenicol (CHL), streptomycin (STR), tetracycline (TET), sparfloxacin (SPA), ciprofloxacin (CIP), norfloxacin (NOR) and enrofloxacin (ENR). Resistance of the Salmonella isolates was 50% to PEF and 100% to CTZ, CTX, AMP, AZT, AMC, CHL, GEN, STR, TET, SPA, CIP, OFL, NOR and ENR. Resistance of the Proteus isolates was 25% to AMC, CHL, STR, TET, SPA and NOR, and 100% to CTZ, CTX, AZT and AMP. Resistance of the isolates to more than 3 classes of antibacterial agents tested was 75% for Proteus and 100% for E. coli and Salmonella, respectively. This study showed that horses in Nigeria are potential reservoirs and disseminators of ESC-resistant and ESBL-producing Enterobacteriaceae. https://macvetrev.mk 2018-10-15T09:00:00 123 132 10.2478/macvetrev-2018-0015 antibiogram extended-spectrum cephalosporin-resistant extended-spectrum β-lactamase-producing Enterobactericeae equine Madubuike Umunna Anyanwu madubuike.anyanwu@unn.edu.ng false 1 Department of Veterinary Pathology and Microbiology, University of Nigeria, 400001 Nsukka, Enugu State Nigeria LEAD_AUTHOR Ifeoma Chinyere Ugwu false 2 Department of Veterinary Pathology and Microbiology, University of Nigeria, 400001 Nsukka, Enugu State Nigeria AUTHOR Collins Uchenna Onah false 3 Department of Veterinary Pathology and Microbiology, University of Nigeria, 400001 Nsukka, Enugu State Nigeria AUTHOR Huber, H., Zweifel, C., Wittenbrink, M.M., Roger, S. (2013). ESBL-producing uropathogenic Escherichia coli isolated from dogs and cats in Switzerland. Vet Microbiol. 162, 992-996. PMid:23177909 1 10.1016/j.vetmic.2012.10.029 Apostolakos, I., Franz, E., van Hoek, A.H.A.M., Florijn, A., Veenman, C., Sloet-van Oldruitenborgh-Oosterbaan, M.M., Dierikx, C., van Duijkeren, E. (2017). Occurrence and molecular characteristics of ESBL/AmpC-producing Escherichia coli in faecal samples from horses in an equine clinic. J Antimicrob Chemother. 72 (7):1915-1921. PMid:28333298 2 10.1093/jac/dkx072 World Health Organization (WHO) (2017). Global priority list of antibiotic-resistant bacteria to guide research, discovery, and development of new antibiotics http://www.who.int/medicines/publications/WHO-PPL-Short_Summary_25Feb-ET_NM_WHO.pdf 3 Dierikx, C.M., van Duijkeren, E., Schoormans, A.H., van Essen-Zandbergen, A., Veldman, K., Kant A., Huijsdens XW., van der Zwaluw K., Wagenaar JA., Mevius DJ. (2012). Occurrence and characteristics of extended-spectrum-β-lactamase- and AmpC-producing clinical isolates derived from companion animals and horses. J Antimicrob Chemother. 67(6):1368-1374. PMid:22382469 4 10.1093/jac/dks049 Olusa, T.A.O., Adegunwa, A.K., Aderonmu, A.A., Adeyefa, C.A.O. (2010). Serologic evidence of equine H7 influenza virus in polo horses in Nigeria. Sci World J. 5(2):17-19. 5 10.4314/swj.v5i2.61509 Turaki, U.A., Kumsha, H.A., Biu, A.A., Bokko, P.B. (2014). Prevalence of piroplasmosis amongst local horses in Northeastern Nigeria. IOSR J Agric Vet Sci. 7(12):4-7. 6 Ehizibolo, D.O., Kamani, J., Eizibolo, P.O., Egwu, K.O., Dogo, G.I., Salami-Shinaba, J.O. (2012). Prevalence and significance of parasites of horses in some states of Northern Nigeria. J Equine Sci. 23(1):1-4. PMid:24833991 PMCid:PMC4013976 7 10.1294/jes.23.1 Ardo, B., Abubakar, D.M. (2016). Seroprevalence of horse (Equus caballus) brucellosis on the Mambilla plateau of Taraba State, Nigeria. J Equine Sci. 27(1):1-6. PMid:27073329 PMCid:PMC4828245 8 10.1294/jes.27.1 Sule, WF, Oluwayelu, D.O., Adedokun, R.A.M., Rufai, N., McCracken, F., Mansfield, K.L., Johnson, N. (2015). High seroprevelance of West Nile Virus antibodies observed in horses from Southwestern Nigeria. Vec Borne Zoonot Dis. 15(3):218-220. PMid:25793479 PMCid:PMC4369928 9 10.1089/vbz.2014.1706 Agina, O.A., Ihedioha, J.I., Anyanwu, M.U., Ngwu, M.I. (2016). Clinicopathological and microbiological findings associated with wounds in Nigerian horses. Comp Clin Pathol. 26 (1):181-188. 10 Agina, O.A., Ihedioha, J.I. (2017). Occurrence of wounds in Nigerian horses. J Appl Anim Welf. Sci. PMid:28696771 11 10.1080/1088∁.2017.1343149 Alonso, C.A., Zarazaga, M., Ben Sallem, R., Jouini, A., Ben Slama, K., Torres, C. (2017). Antibiotic resistance in Escherichia coli in animal husbandry:the African perspective. Lett Appl Microbiol. 64, 318-334. PMid:28208218 12 10.1111/lam.12724 Ugwu, I.C., Anyanwu, M.U., Ugwu, C.C., Ugwuanyi, O.W. (2015). Prevalence and antibiogram of generic extended-spectrum β-lactam-resistant enterobacteria in healthy pigs. Not Sci Biol. 7(3):273-280. 13 10.15835/nsb.7.3.9616 Huijbers, P.M., de Kraker, M., Graat, E.A., van Hoek, A.H., van Santen, M.G., de Jong, M.C., van Duijkeren, E., de Greeff, S.C. (2013). Prevalence of extended-spectrum β-lactamase-producing Enterobacteriaceae in humans living in municipalities with high and low broiler density. Clin Microbiol Infect. 19(6):E256-59. PMid:23397953 14 10.1111/1469-0691.12150 Maddox, T.W., Clegg, P.D., Williams, N.J., Pinchbeck, G.L. (2015). Antimicrobial resistance in bacteria from horses:Epidemiology of antimicrobial resistance. Equine Vet J. 47(6):756-765. PMid:26084443 15 10.1111/evj.12471 Woerther, P., Burdet, C., Chachaty, E., Andremont, A. (2013). Trends in human fecal carriage of extended-spectrum-β-lactamases in the community:toward the globalization of CTX-M. Clin Microbiol Rev. 26(4):744-758. PMid:24092853 PMCid:PMC3811232 16 10.1128/CMR.00023-13 Dolejska, M., Duskova, E., Rybarikova, J., Janoszowska, D., Roubalova, E., Dibdakova, K., Maceckova, G., Kohoutova, L., Literak, I., Smola, J., Cizek, A. (2011). Plasmids carrying blaCTX-M-1 and qnr genes in Escherichia coli isolates from an equine clinic and a horseback riding centre. J Antimicrob Chemother 66(4):757-764. PMid:21393204 17 10.1093/jac/dkq500 Ewers, C., Stamm, I., Pfeifer, Y., Wieler, L.H., Kopp, P.A., et al. (2014). Clonal spread of highly successful ST15-CTX-M-15 Klebsiella pneumoniae in companion animals and horses. J Antimicrob Chemother 69(10):2676-2680. PMid:24974381 18 10.1093/jac/dku217 Faleke, O.O., Jolayemi, K.O., Igoh, Y.O., Jibril, A.H., Ayedun, J.O. (2017). Salmonella species on meat contact surfaces and processing water in Sokoto main market and abattoir, Nigeria. Mac Vet Rev. 40(1):59-65. 19 10.1515/macvetrev-2017-0011 Johns, I.C., Adams, E.L. (2015). Trends in antimicrobial resistance in equine bacterial isolates:1999-2012. Vet Rec. PMid:25628448 20 10.1136/vr.102708 World Health Organization (WHO) (2014). Antimicrobial resistance:global report on surveillance. World Health Organization 21 Schmiedel, J., Falgenhauer, L., Domann, E., Bauerfeind, R., Prenger-Berninghoff, E., et al. (2014). Multiresistant extended-spectrum β-lactamase-producing Enterobacteriaceae from humans, companion animals and horses in central Hesse, Germany. BMC Microbiol. 14, 187. PMid:25014994 PMCid:PMC4105247 22 10.1186/1471-2180-14-187 Vo, A.T., van Duijkeren, E., Fluit, A.C., Gaastra, W. (2007). Characteristics of extended-spectrum cephalosporin-resistant Escherichia coli and Klebsiella pneumoniae isolates from horses. Veterinary Microbiol. 124(3-4):248-255. PMid:17521833 23 10.1016/j.vetmic.2007.04.027 Smet, A., Boyen, F., Flahou, B., Doublet, B., Praud, K., Martens, A., Butaye, P., Cloeckaert, A., Haesebrouck, F. (2012). Emergence of CTX-M-2-producing Escherichia coli in diseased horses:evidence of genetic exchanges of bla (CTX-M-2) linked to ISCR1. J Antimicrob Chemother. 67(5):1289-1291. PMid:2232⇀ 24 10.1093/jac/dks016 Ewers, C., Bethe, A., Stamm, I., Grobbel, M., Kopp, PA, et al. (2014). CTX-M-15-D-ST648 Escherichia coli from companion animals and horses:another pandemic clone combining multiresistance and extraintestinal virulence?J Antimicrob Chemother. 69(5):1224-1230. PMid:24398338 25 10.1093/jac/dkt516 van Spijk, J.N, Schmitt, S., Fürst, A.E., Schoster, A. (2016). A retrospective analysis of antimicrobial resistance in bacterial pathogens in an equine hospital (2012-2015). Schweiz Arch Tierheilkd. 158(6):433-442. PMid:27504838 26 10.17236/sat00069 Maddox, T.W., Clegg, P.D., Diggle, P.J., Wedley, A.L., Dawson, S., Pinchbeck, G.L., Williams, N.J. (2012). Cross-sectional study of antimicrobial-resistant bacteria in horses. Part 1:Prevalence of antimicrobial-resistant Escherichia coli and methicillin-resistant Staphylococcus aureus. Equine Vet J. 44(3):289-296. PMid:21848534 27 10.1111/j.2042-3306.2011.00441.x Maddox, T.W., Pinchbeck, G.L., Clegg, P.D., Wedley, A.L., Dawson, S., Williams, N.J. (2012). Cross-sectional study of antimicrobial-resistant bacteria in horses. Part 2:Risk factors for faecal carriage of antimicrobial-resistant Escherichia coli in horses. Equine Vet J. 44(3):297-303. PMid:2184⅘ 28 10.1111/j.2042-3306.2011.00440.x Chah, K.F., Oboegbulem, S.I. (2007). Extended spectrum beta-lactamase production among ampicillin-resistant Escherichia coli strains from chicken in Enugu State, Nigeria. Braz J Microbiol. 38, 681-686. 29 10.1590/S1517-83822007000400018 Duru, C., Nwanegbo, E., Adikwu, M., Ejikeugwu, C., Esimone, C. (2013). Extended-spectrum beta-lactamase-producing Escherichia coli strains of poultry origin in Owerri, Nigeria. World J Med Sci. 8(4):349-354. 30 Eze, E., Nwakeze, E., Oji, A., Ejikeugwu, C., Iroha, I. (2013). Microbiological investigation of Escherichia coli isolates from cloacal and feacal swabs of broiler chickens for extended- spectrum beta-lactamase (ESBL) enzymes. J Pharm Biol Sci. 7(5):96-99. 31 Ojo, O.E., Schwarz, S., Michael, G.B. (2016). Detection and characterization of extended-spectrum β-lactamase-producing Escherichia coli from chicken production chains in Nigeria. Vet Microbiol. 194, 62-68. PMid:27157499 32 10.1016/j.vetmic.2016.04.022 Abubakar, M.B., Salihu, M.D., Aliyu, R.M., Bello, A., Tukur, H., Shuaibu, A.B. (2016). Occurrence and antimicrobial resistance of ESBL-producing Escherichia coli in indigenous chickens and retailed table-eggs in Sokoto metropolis, Nigeria. Schol J Biol Sci. 5(2):56-60 33 Olowe, O.A., Adewumi, O., Odewale, G., Ojurongbe, O., Adefioye, O.J. (2015). Phenotypic and molecular characterization of extended-spectrum beta-lactamase producing Escherichia coli obtained from animal fecal samples in Ado Ekiti, Nigeria. J Env Publ Health 34 Ogefere, H.O., Agbe, S.O., Ibadin, E.E. (2017). Detection of extended-spectrum beta-lactamases among Gram-negative bacilli recovered from cattle faeces in Benin city, Nigeria. Not Sci Biol. 9(2):177-181. 35 10.15835/nsb9210005 Anyanwu, M.U., Ugwu, I.C., Ezekwelu, M.O., Okoroafor, U.N. (2017). Prevalence and antibiogram of generic extended β-lactam-resistant enterobacteria in healthy dogs. Not Sci Biol. 9(1):22-33. 36 10.15835/nsb919940 Cheesebrough, M. (2000). District laboratory practice in tropical countries Part 2. Cambridge University Press;Cambridge, pp. 63-70. 37 Clinical and Laboratory Standards Institute (CLSI) (2014). Performance standards for antimicrobial susceptibility testing;twenty-fourth informational supplement, M100-S24 34(1):61-188. 38 Clinical and Laboratory Standards Institute (CLSI) (2017). Performance standards for antimicrobial susceptibility testing, 27th Edition M100. 39 Walther, B., Lübke-Becker, A., Stamm, I., Gehlen, H., Barton, A.K., Janssen, T., Wieler, L.H., Guenther, S. (2014). Suspected nosocomial infections with multi-drug resistant E. coli, including extended-spectrum beta-lactamase (ESBL)-producing strains, in an equine clinic. Berl Munch Tierarztl Wochenschr. 127(11-12):421-427. PMid:25−51 40 Hagget, E.F. (2014). Antimicrobial use in foal:do we need to change how we think?Equine Vet J. 46, 137-138. PMid:24548374 41 10.1111/evj.12178 Ahmed, M.O., Clegg, P.D., Williams, N.J., Baptiste, K.E., Bennett, M. (2010). Antimicrobial resistance in equine faecal Escherichia coli isolates from North West England. Ann Clin Microbiol Antimicrob. 9, 12. PMid:20374640 PMCid:PMC2867969 42 10.1186/1476-0711-9-12 Schaufler, K., Bethe, A., Lubke-Becker, A., Ewers, C., Kohn, B., Wieler, L.H., Guenther, S. (2015). Putative connection between zoonotic multiresistant extended-spectrum beta-lactamase (ESBL)-producing Escherichia coli in dog feces from a veterinary campus and clinical isolates from dogs. Infect Ecol Epidemiol. 5, 25334. PMid:25656467 PMCid:PMC4318939 43 10.3402/iee.v5.25334 Damborg, P., Marskar, P., Baptiste, K.E., Guardabassi, L. (2012). Faecal shedding of CTX-M-producing Escherichia coli in horses receiving broad-spectrum antimicrobial prophylaxis after hospital admission. Vet Microbiol. 154(3-4):298-304. PMid:21820821 44 10.1016/j.vetmic.2011.07.005 Beceiro, A., Maharjan, S., Gaulton, T., Doumith, M., Soares, N.C., Dhanji, H., et al. (2011). False extended-spectrum {beta}-lactamase phenotype in clinical isolates of Escherichia coli associated with increased expression of OXA-1 or TEM-1 penicillinases and loss of porins. J Antimicrob Chemother. 66, 2006-2010. PMid:21742679 45 10.1093/jac/dkr265