Veterinary Medicine
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Item Prevalence of Extended Spectrum Beta-Lactamases Producing Enterobacteriaceae in Sheep and Goats from Selected Markets in Ibadan, Oyo-State, Nigeria(Faculty of Basic Medical Scienecs, University of Ibadan, 2023) Adetunji, F. E.; Amosun, E. A.; Olatoye, I. O.; Ojo, O. E.Antìmicrobial misuse, unhygienic husbandry practices, dose interaction between humans and animai as obtained in livestock market facilitate thè emergence, dissemination and transmission of resistant Enterobacteriaceae. These organisms are responsible for various intestinal and extra-intestinal infections in human and animals. According to this report, thè prevalence of Extended Spectrum Beta-Lactamase (ESBL)-producing Enterobacteriaceae in sheep and goats from selected markets in Ibadan. Three hundred and four (304) samples were collected for a cross-sectional survey among thè sheep and goat markets in Ibadan. From thè same sheep and goats, 152 milk samples and 152 faeces samples were collected. In order to identify any antibiotic-resistant Enterobacteriaceae, these samples underwent bacteriological analysis. On MacConkey agar plates with 1 mg/L cefotaxime added, all samples were cultivated. Utilizing a biochemical test kit (Oxoid Microbact GNB 24E®), thè isolates were identified. ESBL products were evaluated utilizing a doublé disc diffusion test with discs impregnated with cefpodoxime and cefpodoxime-clavulanic acid. By using thè disc diffusion approach, antibiotic resistance was identified. Data was analysed using descriptive statistics. Eighty-eight ampicillin resistant Enterobacteriaceae strains were isolated from 304 samples collected and 23 (26.1%) of thè isolates were cefotaxime- resistant Enterobacteriaceae isolates. Only 9(10.2%) were confirmed phenotypic ESBL-producers and they were all from faeces. AH ESBL -producing Enterobacteriaceae were E. coli strain and showed 100% resistance to tetracycline and ceftazidime, 77.8% resistance to amoxicillin and sulphamethoxazole but susceptible to gentamycin and ciprofioxacin in this investigation. The ESBL-producing E. coli isolates showed different antibiotic resistance pattems. In this study, 100% of ESBL-producing E. coli were multidrug- resistant, showing resistance to at least three separate classes of antibiotics. Public awareness of thè significance of stringent hygiene in animai husbandry needs to be raised in light of thè possible threat that thè existence of multidrug resistant ESBL-producing E. coli in small ruminants poses to public healthItem Phenotypic Detection of Extended Spectrum Beta-Lactamase Production in Clinical and Non-Clinical Isolates of Klebsiella species of Animal Origins in Abeokuta, Nigeria(Nigerian Veterinary Medical Association, 2022) Amosun, E. A.; Ojo, O. E.Production of extended-spectrum (3-lactamases (ESBLs) confer resistance to P-lactam antibiotics. This study investigated thè presence of ESBL-producing Klebsiella spp in clinical and non-clinical samples froni different animai species in Abeokuta, Nigeria. The species of Klebsiella were determined by biochemical characterization (Oxoid Microbact GNB 24E®), while phenotypic ESBL-production was confirmed by using thè cefpodoxime and cefpodoxime/clavulanic acid combination disc kit. Ninety-five Klebsiella isolates were investigated in this study. Fifty-five (57.9%) of thè isolates were obtained ffom thè faeces of apparently healthy animals while 40 (42.1%) were fram clinical samples. The Klebsiella isolates were identified as follows: K. oxytoca (34.7%), K. pneumoniae (26.3%), K. ozaenae (18.9%), K. terrigena (13.7%), K. rhinooscleromatis (5.3%), K planticola (1.1%). Eight (8.4%) out ofthe 95 isolates of Klebsiella spp were identified as ESBL-producers. These included four isolates of Klebsiella pneumoniae, three of Klebsiella oxytoca and one of Klebsiella ozaenae. Five out of thè eight ESBL-producing isolates were ffom clinical samples while three isolates were ffom thè faeces of apparently healthy animals. The ESBL- producing isolates were 100% resistant to ampicillin, cefotaxime, ceftazidime, cefpodoxime, streptpmycin, tetracycline, compound sulfonamide, trimethoprim/sulphomethoxazole, and nalidixic acid. Apparently healthy carriers and sick animals can serve as sources of transmission of ESBL-producing Klebsiella spp to other animals and humans.Item Phenotypic Detection of Extended Spectrum Beta-Lactamase Production in Clinical and Non-Clinical Isolates of Klebsiella species of Animal Origins in Abeokuta, Nigeria(Nigerian Veterinary Medical Association, 2022) Amosun, E. A.; Ojo, O. E.Production of extended-spectrum (3-lactamases (ESBLs) confer resistance to P-lactam antibiotics. This study investigated thè presence of ESBL-producing Klebsiella spp in clinical and non-clinical samples froni different animai species in Abeokuta, Nigeria. The species of Klebsiella were determined by biochemical characterization (Oxoid Microbact GNB 24E®), while phenotypic ESBL-production was confirmed by using thè cefpodoxime and cefpodoxime/clavulanic acid combination disc kit. Ninety-five Klebsiella isolates were investigated in this study. Fifty-five (57.9%) of thè isolates were obtained ffom thè faeces of apparently healthy animals while 40 (42.1%) were fram clinical samples. The Klebsiella isolates were identified as follows: K. oxytoca (34.7%), K. pneumoniae (26.3%), K. ozaenae (18.9%), K. terrigena (13.7%), K. rhinooscleromatis (5.3%), K planticola (1.1%). Eight (8.4%) out ofthe 95 isolates of Klebsiella spp were identified as ESBL-producers. These included four isolates of Klebsiella pneumoniae, three of Klebsiella oxytoca and one of Klebsiella ozaenae. Five out of thè eight ESBL-producing isolates were ffom clinical samples while three isolates were ffom thè faeces of apparently healthy animals. The ESBL- producing isolates were 100% resistant to ampicillin, cefotaxime, ceftazidime, cefpodoxime, streptpmycin, tetracycline, compound sulfonamide, trimethoprim/sulphomethoxazole, and nalidixic acid. Apparently healthy carriers and sick animals can serve as sources of transmission of ESBL-producing Klebsiella spp to other animals and humans.Item Multidrug resistant Enterohaemorrhagic Esherichia coli serogroups in the faeces of hunted wildlife, Abeokuta, Nigeria(Istituto Zooprofilattico Sperimentale dell'Abruzzo e del Molise "G. Caporale Teramo, Italy, 2022) Ojo, O. E.; Amosun, E. A.; Opebiyi, O. O.; Oyekunle, M. A.; Dipeolu, M. A.; Otesile, E. B.Wildlife plays significant roles in the dissemination and zoonotic transmission of pathogens. The enterohaemorrhagic Escherichia coli (EHEC) are associated with complicated cases of food-borne illnesses. This study investigated the presence of EHEC serogroups (026, 045, 0103, 0145, 091, 0111, 0128, 0121 and 0157) in wildlife species: cane rats (Thryonomys swinderianus), royal antelope (Neotraguspygmaeus), African giant rats (Cricetomysgambianus) and \Naterbuck(Kobusellipsiprymnus). EHEC and non-EHEC isolates from these wildlife sources were tested for susceptibility to antimicrobial agents. Overall, 127 (83.0 %) out of 153 samples yielded E. coli. Nine (5.9%) samples were positive for EHEC belonging to three serogroups as follows: 026 (n = 2), 0111 (n = 2) and 0103 (n = 5). The EHEC isolates were from cane rats (n = 6) and royal antelope (n = 3) and possessed virulence-associated genes stx, (77.8%), stx2 (100.0%), eaeA (100.0%) and hlyA (100.0%). Overall, 127 £ coli isolates showed resistance to ampicillin (99.2%), ceftiofur (90.6%), tetracycline (90.0%), cephalexin (87.4%), cefotaxime (50.4%), streptomycin 42.5%, ceftazidime (41.7%), nalidixic acid (37.0%), ciprofloxacin (43.6%), amoxicillin/davulanic acid (32.3%), gentamicin (27.6%), sulphamethoxazole/ trimethoprim (25.2%), norfloxacin (17.3%) and chloramphenicol (11.0%). The role of wildlife in the dissemination and transmission of antimicrobial resistant and zoonotic bacteria should not be neglected for effective preventive and control strategies.Item Investigation of Extended Spectrum Beta-Lactamase Producing Escherichia coli and other Cefotaxime-Resistant Bacteria in Cow Milk in Nigeria(Israel Veterinary Medical Association (IVMA), Raanana, Israel, 2021) Amosun, E. A.; Ogunbadewa, A. J.; Ojo, O. E.; Akinade, S. A."Production of extended spectrum fl-lactamases (ESBLs) confers resistance to cefotaxime as well as many third and fourth generation cephalosporins in bacteria. ESBL-producing E. coli strains have been implicated in refractory infections in humans and dairy cows in Nigeria. The present study investigated thè presence of cefotaxime-resistant bacteria including ESBL-producing E. coli in thè milk of apparendy healthy cows from smallholder dairy herds. A total of 168 non-duplicate milk samples were collected from 34 catde herds in three locai government areas of Oyo State, Nigeria. Cefotaxime-resistant bacteria were isolated from milk samples by selective culture on MacConkey agar supplemented with cefotaxime (lmg/L). Isolates were identified by biochemical tests. Phenotypic ESBL production was determined using cefpodoxime/ cefpodoxime-clavulanic acid combinatìon discs. Presence of ESBL genes (Mttem, blasm and blacvx} and phylogenetic groups (A, Bl, B2, C, D, E, and F) of E. coli were determined by polymerase chain reaction assay. Susceptibility to other antimicrobials was carried out using thè Kirby-Bauer disk diffusion method. Cefotaxime-resistant bacteria were detected in milk samples of in 40 (23.8%) out of 168 individuai cows. At least one sample from 19 (55.9%) out of 34 herds yielded a cefotaxime-resistant isolate. The isolates were identified as Escherichia coli (9/168; 5.4%), Enterohacter amnigenus (8/168; 4.8%) and Pseudomonas aeruginosa (23/168; 13.7%). Five (3.0%) of all 168 samples were positive for phenotypic ESBL-producing bacteria. All thè ESBL-producing bacteria were E. coli and possessed blacrx-u which is of thè èlacvx-M-is gene variant. Two isolates also possessed thè blcrcem gene. ESBL-producing E. coli isolates belonged to phylogenetic groups A (n=3) and Bl (n=2). Overall, thè cefotaxime-resistant bacteria showed 100.0% resistance to ampicillin and cefotaxime, 90.0% to tetracycline, 72.2% to amoxicillin/clavulanic acid, 47.5% to streptomycin, 40.0% to sulphamethoxazole/trimethoprim, 37.5% to chloramphenicol, 32.5% to ceftazidime and 10.0% to each of ciprofloxacin and gentamicin. Keywords: ESBL Resistance Genes; Cow Milk; Cefotaxime-Resistance; Escherichia coli.Item Antimicrobial use and detection of cefotaxime-resistant Enterobacteriaceae in thè pig production chain, Ogun State, Nigeria(Vigot Freres. Paris, France, 2020) Ojo, O. E.; Akinyemi, M. I.; Amosun, E. A.; Hassan, J. O.; Dipeolu, M. A.Cefotaxime belongs to thè third-generation cephalosporin group of antimicro- bials, which are classified as criticai for thè treatment of infections in humans. The upsurge in thè incidence of cefotaxime-resistant (C-R) bacteria from animai sources is of global public health importance. This study investigated thè pres- ence of C-R Enterobacteriaceae in thè pig production chain in Ogun State, Nigeria, and examined C-R isolates for production of extended-spectrum P-lactamase (ESBL). The knowledge, attitude and practices of pig farmers regarding antimicrobial usage were also investigated. C-R bacteria were detected in 54 (17.8%) out of 303 samples. C-R isolates were identified as Escherichia coli (n = 22), Klebsiella spp. (n = 17), Enterobacter aerogenes (n = 10) and Citrobacter freundii (n = 5). The organisms were present in feces from on-farm pigs (15/109; 13.7%), fresh pork at slaughterhouses (19/40; 47.5%), frozen pork at retail shops (7/28; 25.0%), cutting surfaces of butchers' tools (7/52; 13.5%), and abattoir effluent water (6/41; 14.6%). No C-R bacteria were detected in ready-to-eat pork. Three isolates of Es. coli and one of K. pneumoniae were ESBL-producers and pos- sessed WaCTX_M_15 ESBL gene variant. ESBL-producing Es. coli belonged to phylo- genetic group A. All C-R isolates were resistant to more than three antimicrobi- als from different classes of antimicrobials. Tetracycline, ampicillin, amoxicillin, ciprofloxacin and enrofloxacin were among thè commonly used antimicrobials in pig production, whereas cephalosporins were rarely used. Farmers knew that pigs could serve as reservoirs of pathogenic bacteria transmissible to humans. However, they were not aware that thè use of antimicrobials in pig production could lead to thè development and proliferation of antimicrobial-resistant bacteria in pigs. Efforts should be made to improve awareness among farmers on thè roles of antimicrobial use in thè emergence and dissemination of antimicrobial- resistant bacteria in animai production.Item Prevalence and antimicrobial resistance profiles of bacteria isolated from the urine of dogs in Ibadan, Nigeria(Faculty of Veterinary Medicine, University of Ibadan, Ibadan, Nigeria, 2019) Amosun, E. A.; Amusa, A. O.; Ojo, O. E.Bacteria exist as part of thè norma! flora of thè urogenital tract of animals. However, some of them may cause severe opportunistic infections in immuno-suppressed host requiring antimicrobial chemotherapy. Antìmicrobial resistance may lead to therapy fatture and protracted tllnesses. This study investigated thè diverse reservoirs of antimicrobial resistant bacteria in thè urogenital tract of dogs in lbadan. A total of 114 urine samples from male (42) and female (72) dogs were examined for thè presence of bacteria by inoculation onto cultural media. Bacteria identification was based on cultural, microscopie and biochemical characteristics. Bacteria isolates were tested for susceptibility to amoxicillin, amoxillin/ clavulanic acid, cefixime, ceftazidime, cefuroxime, ciprofloxacin, ampicillin/cloxacillm, erythromycin, gentamicin, nitrofurantoin, ofloxacin, perfloxacin, ceftriaxone, streptomycin, trimethoprim/sulfamethozoJeand cefuroxime. Overall, 184 bacteria isolates were identified from thè 64 (56.14%) of thè 114 urine samples examined. The remaining 47 (41.2%) of 114 samples did notyield significant bacterial growth on thè various bacteriological media. The bacteria detected in thè urine samples inciuded Escherìchia coli (59); Staphylococcvs aureus (40), Slreptococci spp (32), Klebsiella spp (22), Pseudomonas aemgìnosa (12), coagulase negative staphylococcus spp (7), Enterobacter spp (7) and Proteus mirabilis (5). The organisms showed varying degrees of susceptibility to antimicrobial agents. Many of thè organisms demonstrated resistance to at least three antimicrobials from different classes. Forty-twò resistance groups were observed among thè Gram-positive bacterial isolates while eleven resistance groups were observed among thè Gram-negative isolates. ‘APX-CRX-AM-CET-S-SXT-E-PEF-CN’ and ‘CAZ-CPR-NITAUG-OFL-CXM-GENV CRX ’ were thè predom inant resistant pattems for Gram-positive and Gram-negative isolates respectively. This study revealed a diversity and high level of multidrug resistance bacteria in thè urogenital traets of dogs. These bacteria maybe important as primary or opportunistic aetiological agents of infection in die affected host with thè possibility of zoonotic transmission to human companions.Item Antibiotic Resistance Profiling of Escherichia coli O157:H7 from Guinea fowl and Chicken in Ibadan North Local Government area, Nigeria(Faculty of Veterinary Medicine, University of Ibadan, Ibadan, Nigeria, 2017) Amosun, E. A.; Ayantoke, I. A.; Agba, J. U.; Ojo, O. E.Escherichia coli Ol 57 is a major zoonotic disease of thè poultry industry. 315 cloacae samples (177 chickens and 138 guinea fowls) were screened for enterohaemorrhagic E. coli (EHEC 0157). The samples were pre-enriched with tryptic soy broth, incubated over- night at 37°C for 18 hours, sub-cultured on MacConkey agar and incubated at 37°C for 24 hours. Rose pink colonies on MacConkey agar (putative E. coli) were selected for bio- chemical test. The susceptibility of identified E. coli isolates to antimicrobial agents was determined by thè standard Kirby-Bauer disk diffusion method on Mueller-Hinton agar. Confirmed E. coli were sub-cultured on Sorbitol MacConkey agar piate (SMAC) and incubated at 37°C for 24hours. Colourless or pale colonies (non-sorbitolfermenters) were further tested. Isolates that produced colourless colonies and agglutinatimi with E. coli 0157 latex test were tested with E. coli HI antiserum by a slide agglutination test as well as other identified E. coli isolates irrespective of their sorbitol fermentatimi reactions were identified serologically using a latex agglutination test kit. Of thè 315 samples, 9.5% were positive for EHEC Ol 57 including 7.3% and 12.3% of thè chickens and thè guinea fowls, respectively. The differences in thè two species were not statistica!ly (P<0.05) significant The resistance of thè EHEC 0157 to antim icrobials were; amoxicillin/clavulanic acid 66.67%, nitrofurantoin 60.00%, gentamicin 56.67%, trimethoprim/sulphamethoxazole 50.00%, tet- racycline 50.00%, pefloxacin43.33%, amoxicillin 33.33 %, ciprofloxacin 33.33 %,ofloxacin 30.00%, and ceftriaxone 20.00%. E. coli OJ57:H7 isolated from guinea fowls were all susceptible to amoxicillin and ciprofloxacin while EHEC 0157:H7 from chickens showed 100% resistance to tetracycline and pefloxacin. The presence of virulent multidrug resistali E. coli 0157 strains in thè faeces of guinea fowls and chickens showed thè dangers that handlers and consumers are exposed and thè implications of disseminatimi of thè pathogens to thè environment through faecal shedding.Item Evaluation of bacteria and antibiotic resistance profiling of Wara (White soft cheese) in Oyo- State, South West, Nigeria(Nigerian Veterinary Medical Association, 2017) Amosun, E. A.; Agbato, A. O.; Daodu, O. B.; Ojo, O. E.Fresh milk from healthy cows contains few bacteria but contamination during handling and processing can rapidly increase bacterial population and subsequently depreciate it quality. This study evaluated thè presence of bacteria and their antibiotic resistance profiles in white soft cheese (Wara) in three major producing locai Government areas in Oyo-State, Nigeria. The cheese samples were collected in sterile universal bottles immediately after production. Samples were immediately transported to thè laboratory for bacteria isolation, identification and antibiotic susceptibility test of thè isolates. A total of one hundred and sixty-five (165) bacterial isolates of ten different bacterial genera were identified from 102 cheese samples. The isolation rates of different species/genera were Lactobacillus spp (39.8%), E. coli (32.5%), Klebsiella spp (9%), Citrobacter spp (2.4%), Acinetobacter spp (3.6%), Pseudomonas spp (6.6%), Bordetella spp (0.6%), Proteus spp (1.2%), Staphylococcus spp (3.6%) and Strepiococcus agalactiae (0.6%). Results demonstrated substantial differences in their resistance pattems to various antibiotics. The isolates were tested against ten different antibiotics. The Gram negative isolates showed twenty-five resistance pattems while Gram positive isolates revealed fifty-four distinct resistance groups. In all, Gram positive isolates showed resistance to ceftriaxone (61.1%), gentamicin (51.4%), chloramphenicol (51.1%), amoxicillin (48.6%), ciprofloxacin (44.4%), cotrimoxazole (43.1%), perfloxacin (40.3%), streptomycin (40.3%) Erythromycin (31.9) and ofloxacin (30.6%) while Gram positive showed resistance to chloramphenicol (22.6%), trimethoprim /sulfamethoxazole (20.4%), amoxillin/clavulanic acid. (10.8%), streptomycin (9.7%), gentamicin (8.6%), sparfloxacin (6.5%), amoxicilin (5.4%), Perfloxacin (5.4%), ofloxacin (4.3%) and ciprofloxacin (3.2%). Multi-drug resistance to three or more antibiotics was observed in 69 isolates, 56 Gram positive and 13 Gram negative bacteria. This study confirmed that white soft cheese “wara” can be a source of bacteria with multidrug resistant phenotypes.Item Multidrug Resistant Enterohaemorrhagic Escherichia coli O157:H7 in Pigeons in Ibadan, Nigeria(African Union, InterAfrican Bureau for Animal Resources (AU-IBAR), 2017) Amosun, E. A.; Aweda, D. I.; Ojo, O. E.Pigeons are commonly seen around human dwellings and in city centres.The movement of these birds from place to place makes them a veritable vehicle for environmental dissemination of pathogens. Enterohaemorrhagic E coli (EHEC) OI57:H7 can cause severe and sometimes fatal gastroenteritis in humans.This study investigated the occurrence and antimicrobial susceptibility of EHEC OI57:H7 in the faeces. One hundred and twenty five cloacae samples (82 adult pigeons and 43 squabs) were collected from three different locations in Ibadan metropolis. Enterohaemorrhagic E coli Ol 57:H7 was isolated from samples on BCIG-SMAC following a selective pre-enrichment culture in modified Tryptic Soy Broth supplemented with novobiocin. Suspected non-sorbitol fermenting Ecoli isolates were serologically identified as serotype 0157:H7 using latex agglutination method. Enterohaemorrhagic E coli isolates were tested for susceptibility to antimicrobial agents by the Kirby Bauer disk diffusion method. Out of all 125 samples examined, 23(18.4%) were confirmed as EHEC 0I57.H7 serotype. Isolates showed resistance to nitrofuranton (100%), ceftriazone (100%), amoxicillin (91.3%), augmentin (73.9%), gentamycin (60.9%), cotrimozazole (60.9%), pefloxacin (47.8%) , tetracycline (34.8%), ciprofloxacin (30.4%) and ofloxacin (17.4%). This study showed that pigeons harbour multidrug resistant EHEC OI57:H7 and may contribute to environmental contamination through faecal shedding.