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Production of extended-spectrum β-lactamases (ESBLs) producing E. coli in animals and different methods of identifications from Ado Ekiti, Ekiti State, Nigeria, were investigated. Three hundred and fifty fecal samples, collected from apparently healthy cattle and pigs, were cultured and identified following standard procedures. ESBL phenotypic detection was carried out using combination disc test, double disc synergism test, and ESBL brilliance agar screening. Molecular detection of TEM, SHV, and CTX-M genes was carried out using standard molecular method. One hundred and fourteen E. coli isolates were recovered from the 350 samples processed, out of which 72 (63.2%) isolates were positive for ESBLs with multiple resistance to the antibiotics used. Eighty-one (71%) isolates were positive for ESBL by combination disc test, 90 (78.9%) were positive for double disc synergism test, and 93 (81.6%) were positive for ESBL brilliance agar. TEM and CTX-M genes were detected in 48 (42.1%) and 51 (44.7%) isolates, respectively. SHV gene was not detected in any of the isolates while TEM and CTX-M were detected in 33 (28.9%) isolates. This study showed high resistance of E. coli to antibiotics, particularly to the third generation cephalosporins. Regular monitoring and regulated use of antibiotics in livestock should be encouraged. [ABSTRACT FROM AUTHOR]

Background: Antimicrobial resistance (AMR) is an increasing global health concern reducing options for therapy of infections and also for perioperative prophylaxis. Many Enterobacteriaceae cannot be treated anymore with third generation cephalosporins (3GC) due to the production of certain 3GC hydrolysing enzymes (extended spectrum beta-lactamases, ESBLs). The role of animals as carriers and vectors of multi-resistant bacteria in different geographical regions is poorly understood. Therefore, we investigated the occurrence and molecular characteristics of ESBL-producing Escherichia coli (E. coli) in wild birds and slaughtered cattle in Ibadan, Nigeria. Cattle faecal samples (n = 250) and wild bird pooled faecal samples (cattle egrets, Bubulcus ibis, n = 28; white-faced whistling duck, Dendrocygna viduata, n = 24) were collected and cultured on cefotaxime-eosin methylene blue agar. Antimicrobial susceptibility was determined by agar diffusion assays and all 3GC resistant isolates were genotypically characterised for AMR genes, virulence associated genes (VAGs) and serotypes using DNA microarray-based assays., Results: All 3GC resistant isolates were E. coli: cattle (n = 53), egrets (n = 87) and whistling duck (n = 4); cultured from 32/250 (12.8%), 26/28 (92.9%), 2/24(8.3%), cattle, egrets and whistling duck faecal samples, respectively. blaCTX-M gene family was prevalent; blaCTX-M15 (83.3%) predominated over blaCTX-M9 (11.8%). All were susceptible to carbapenems. The majority of isolates were resistant to at least one of the other tested antimicrobials; multidrug resistance was highest in the isolates recovered from egrets. The isolates harboured diverse repositories of other AMR genes (including strB and sul2), integrons (predominantly class 1) and VAGs. The isolates recovered from egrets harboured more AMR genes; eight were unique to these isolates including tetG, gepA, and floR. The prevalent VAGs included hemL and iss; while 14 (including sepA) were unique to certain animal isolates. E. coli serotypes O9:H9, O9:H30 and O9:H4 predominated. An identical phenotypic microarray profile was detected in three isolates from egrets and cattle, indicative of a clonal relationship amongst these isolates., Conclusion: Wild birds and cattle harbour diverse ESBL-producing E. coli populations with potential of inter-species dissemination and virulence. Recommended guidelines to balance public health and habitat conservation should be implemented with continuous surveillance.

Outbreaks of Salmonellosis remain a major public health problem globally. This study determined the diversity and antibiotic resistance gene profile of Salmonella enterica serovars isolated from humans and food animals. Using standard methods, Salmonella spp. were isolated from fecal samples, profiled for antimicrobial susceptibility and resistance genes. Seventy-one Salmonella isolates were recovered from both humans and food animals comprising cattle, sheep, and chicken. Forty-four serovars were identified, with dominant Salmonella Budapest (31.8%). Rare serovars were present in chicken ( S . Alfort, S . Wichita, S . Linton, S . Ealing, and S . Ebrie) and humans ( S . Mowanjum, S . Huettwillen, S . Limete, and S . Chagoua). Sixty-eight percent of isolates were sensitive to all test antibiotics, while the highest rate of resistance was to nalidixic acid (16.9%; n  = 12), followed by ciprofloxacin (11.3%; n  = 8) and tetracycline (9.9%; n  = 8). Five isolates (7%) were multidrug-resistant and antimicrobial resistance genes coding resistance to tetracycline ( tetA ), beta-lactam ( bla TEM ), and quinolone/fluoroquinolone ( qnrB and qnrS ) were detected. Evolutionary analysis of gyrA gene sequences of human and food animal Salmonella isolates revealed variations but are evolutionarily interconnected. Isolates were grouped into four clades with S . Budapest isolate from cattle clustering with S . Budapest isolated from chicken, whereas S . Essen isolated from sheep and chicken was grouped into a clade. Diverse S. enterica serovars with high antibiotic resistance profile are ubiquitous in food animals; hence, there is a need for surveillance and prudent use of antibiotics in human and veterinary medicine.

Zoonotic transmission of mycobacteria between humans and other animal species is an important aspect of the epidemiology of tuberculosis (TB) in regions of the world where the burden of the disease in humans and other animals is high. This is particularly important in areas in which sociocultural factors increase the possibility for interspecies transmission of different mycobacteria. Carrying out a review of the published literature involving laboratory-confirmed TB cases (by culture) and/or biochemical and molecular identification, we described the presence of Mycobacterium bovis and M. tuberculosis infections in humans and animals from 1975 to 2014. Overall, 1693 isolates of M. tuberculosis complex and other mycobacteria were identified and reported, of which 1131 represented M. tuberculosis , 286 represented M. bovis , 71 represented M. africanum , and 205 represented other mycobacteria. Importantly, 1.3% (15/1131) of the M. tuberculosis isolates reported were identified in cattle, while 8.0% (23/286) of the M. bovis isolates reported were isolated from humans. We suggest that representative sampling of TB cases in both hosts, studied by molecular identification tools, will help significantly in deciphering ongoing transmission between animals and humans in both directions and will enhance TB control in Nigeria.

Infections caused by non-typhoidal Salmonella (NTS) are common around the world, with high morbidity and mortality rates recorded annually. Salmonella serovars harbor plasmids of various sizes which may play roles in antibiotic resistance and virulence. The aim of this study was to profile and determine the role of plasmids in ciprofloxacin resistance and virulence of Salmonella serovars. Using alkaline lysis method 25 NTS serovars from food animals and humans were assayed for plasmids. Isolates ability to resist healthy human serum, bind congo red, produce hemolysin and susceptibility to ciprofloxacin before and after plasmid curing were evaluated. Mobility of plasmids was determined by conjugation. Fifteen (60%) of the 25 Salmonella serovars harbored plasmids with sizes ranging from 0.4 to 38.4 kb. S . Budapest serovars harbored 5-9 plasmids, while S . Essen and S . Mura had six plasmids each. S . Chomedey and a S . Budapest serovar were sensitive to ciprofloxacin after plasmid curing while other serovars remained resistant to ciprofloxacin after plasmid curing. All Salmonella isolates had the ability to withstand human serum before and after plasmid curing, however, some serovars lost their ability to bind congo red after plasmid curing. All Salmonella isolates that initially displayed hemolysin activity retained their ability after curing. Thirteen (86.7%) of the 15 serovars that harbored plasmids conjugatively transferred their plasmids to E. coli K-12 (DH5α). Having Salmonella serovars that harbor transferrable plasmids in the food chain can drive antibiotic resistance and enhanced virulence of otherwise less virulence strains.

Pathogenic Escherichia coli can be released with the wastes coming from slaughterhouses into the environment, where they can persist. We investigated the presence of diarrheagenic E. coli in specimens taken at an abattoir located in the Zaria region, Nigeria, in samples of water from the river Koreye, where the effluent from the abattoir spills in, and vegetable specimens taken at a nearby farm. All the isolated E. coli were assayed for the production of Shiga toxins (Stx) by using the Ridascreen verotoxin Immunoassay and by PCR amplification of genes associated with the diarrheagenic E. coli. Three strains from the rectal content of two slaughtered animals and a cabbage were positive for the presence of the Stx-coding genes. Additionally we have isolated one Enteroaggregative E. coli (EAggEC) from the abattoir effluent and two Subtilase-producing E. coli from the slaughterhouse's effluent and a sample of carrots. Our results provide evidence that pathogenic E. coli can contaminate the environment as a result of the discharge into the environment of untreated abattoir effluent, representing a reservoir for STEC and other diarrheagenic E. coli favouring their spread to crops.

Introduction: Tuberculosis remains a global public health problem. In 2011, tuberculosis incidence was 133 per 100,000 in Nigeria. In Nigeria, little is known about the factors associated with tuberculosis, especially in the northern part and only few studies have characterized the Mycobacterium species that cause tuberculosis infection in humans. This study determined factors associated with tuberculosis and identified Mycobacterium species causing human tuberculosis in North-West, Nigeria., Methods: We conducted a hospital based case control study between April and July 2010 in Zaria. Cases were newly diagnosed sputum smear-positive tuberculosis patients >15 years while controls were patients >15 years attending the hospital for other reasons but were negative for tuber-culosis. We used a structured questionnaire to obtain information on demographics, knowledge of transmission of tuberculosis, and exposure to some factors. We preformed descriptive, bivariate and backward elimination logistic regression. Sputa from cases were analyzed by multiplex polymerase chain reaction (PCR) based on genomic regions of difference., Results: The mean ages of the cases and controls were 36, standard deviation (SD) 9.0 and 36, SD 9.7 respectively. Only 10 (9.8%) and nine (8.8%) of cases and controls respectively had a good knowledge of the transmission of tuberculosis. Contact with a tuberculosis patient (adjusted odds ratio (AOR) 12.3, 95% confidence interval (CI) 5.2-28.8), consumption of unpasteurized milk (AOR 6.4, CI 2.4-17.2), keeping pets (AOR 5.6, CI 2.3-13.7), associating closely with cattle (AOR 5.6, CI 1.3-6.8), and overcrowding (AOR 4.8, CI 1.8-13.1) were significantly associated with tuberculosis. Of the 102 sputa analyzed, 91 (89%) were M. tuberculosis, 8 (7.8%) were M africanum., Conclusion: We identified possible opportunities for intervention to limit the spread of tuberculosis. We recommend that the Nigeria tuberculosis control program consider some of these factors as a way to mitigate the spread of tuberculosis in Nigeria.

Introduction: Bovine tuberculosis (BTB) is widespread yet poorly controlled in Nigeria hence posing a public health threat. This study determined the prevalence of Mycobacterium tuberculosis complex (MTC) and factors associated with MTC among slaughtered cattle at Jos South Abattoir in Plateau State, Nigeria., Methods: We conducted a cross sectional study in which we collected 168 lung samples systematically from 485 slaughtered cattle from May-June, 2012, and tested for acid fast bacilli (AFB) using Ziehl-Neelsen test and a duplex polymerase chain reaction technique (PCR) for MTC detection. Data on cattle socio-demographic characteristics and risk factors for zoonotic BTB infection was obtained and analyzed using Epi info version 3.5.3 to determine frequency, proportions, and prevalence odds ratios. Multiple logistic regression was done at 95% Confidence Interval (CI)., Results: The mean age of the cattle was 5.6 ± 1.3 years and (108) 64.3% were females. Majority were indigenous White Fulani breed of cattle (58.5%) and about half (54.8%) were slightly emaciated. Prevalence of MTB complex was 21.4% by AFB test and 16.7% by duplex PCR. Of 33 (19.6%) lungs with lesions, 27 (81.8%) were positive for AFB; while of 135 (80.4%) lungs without lesions, 9 (6.7%) were positive for AFB. Lungs with lesions were 52 times more likely to test positive to AFB test compared to tissues without lesions (AOR=52.3; 95% CI: 16.4-191.8)., Conclusion: The presence of MTC in cattle signifies its potential risk to public health. Presence of lesions on lungs is a reliable indicator of MTC infection that meat inspectors should look out for.

Introduction: The main objective of the study was the molecular characterization of extended spectrum β-lactamases (ESBL) in Escherichia coli isolates collected from human and bovine samples in Oyo state, Nigeria., Methodology: Between August 2010-2011, 114 E. coli isolates were collected from hospitals (n = 57) and bovine (n = 57). PCR and sequencing were used for identification of ESBLs, upstream sequences, plasmid-mediated quinolone resistance (PMQR) genes and class 1 integrons. Plasmid incompatibility groups were identified among ESBL-positive isolates by PCR. Genetic relatdness was assessed by rep-PCR and MLST. Transfer of ESBL determinants to the recipient strain E. coli J53 was performed by broth mating assays., Results: CTX-M15 was the unique ESBL found in eight human isolates. Six CTX-M-15 producers also carry the aac(6')-lb-cr gene and/or qnrB gene, and class 1 integrons. FIA, FIB, H11, H12, F, Y and K were the plasmid replicon types found. CTX-M-15 and PMQR determinants were transferred by conjugation in two E. coli assigned by MLST to ST131 and ST2695, a new allele., Conclusions: The study highlights the dissemination hability of CTX-M-15 associated with PMQR, and the presence of class 1 integrons, able to capture additional genes, justifying the urgent need of antimicrobial resistance surveillance in Nigeria.

Setting: Bovine tuberculosis (TB) is endemic in the cattle population in Nigeria. Livestock workers are at risk of Mycobacterium bovis infection and unaware of their health status., Objective: To determine the occurrence of pulmonary M. bovis infection among livestock workers., Design: A cross-sectional study of livestock traders was conducted for TB through screening of sputum samples using a simple random sampling method coupled with oral interview on the assumption of sub-clinical pulmonary TB infection. Specimens were cultured, and the isolates analysed using molecular typing techniques., Results: Overall, 10% (7/70) of the livestock traders had a positive culture indicative of M. tuberculosis complex, which were differentiated into M. bovis (n = 2) and M. tuberculosis (n = 5) using deletion typing. Further spoligotyping analyses of the M. bovis and two available M. tuberculosis isolates classified the strains as SB1432 and SB09444 and LAM_10 CAM and T1 using respectively www.mbovis.org and spotclust databases. Prolonged cough and >3 years in the livestock trade were risk factors for infection., Conclusion: We confirm that there is undetected pulmonary M. bovis infection among livestock traders in Nigeria. Further studies on the role of occupationally exposed workers in the transmission of M. bovis infection to the larger community are required.

The characterisation of 24 lactic acid bacteria (LAB) isolates from Nigerian traditional fermented dairy foods, including some cow's intestine isolates, was conducted in order to select isolates for potential use as probiotics. LAB isolates were identified by partial sequencing the 16S rRNA gene as belonging to the species Lactobacillus paracasei, Lactobacillus brevis and mainly Weissella confusa. At the end of a characterisation process, 2 L. paracasei and 2 W. confusa isolates were selected, and their resistance to a simulated gastrointestinal digestion and their ability to adhere to eukaryotic cell lines were assessed. The survival to the simulated gastrointestinal passage was higher when bacterial suspensions were made in skimmed milk (2.0±0.8 log units reduction) or at the simulated gastric juice pH 3 (2.7±0.9 log units reduction) than at pH 2.0 (5.5±0.7 log units reduction). Adhesion of LAB to both intestinal and vaginal epithelial models was comparable or higher than that of the reference Lactobacillus rhamnosus GG. However, some of the isolates increased the adhesion of the pathogen Escherichia coli LMG2092 to HT-29 and HeLa monolayers. Overall, isolates L. paracasei UI14 and W. confusa UI7 are good candidates for further studying potential benefits that support their use as probiotics. This is one of the few articles reporting the characterisation and the probiotic potential of Weissella, although more studies are needed in order to establish their safety for potential probiotic applications., (Copyright © 2011 Elsevier B.V. All rights reserved.)

We determined the prevalence of bovine tuberculosis in Jigawa State, northwestern Nigeria using the comparative intradermal tuberculin test in 22 herds consisting of 922 cattle with overall and individual prevalence of 1.08% (10/922) and 45.45% (10/22) obtained, respectively.

A total of 96 lactic acid bacteria (LAB) were isolated from African indigenous fermented products and cow's intestines to study their inhibitory capability against multi-drug-resistant uropathogens. Escherichia coli accounted for approximately 45% of isolated uropathogens, followed by Staphylococcus spp. (20%). The Gram negative uropathogens were highly resistant to quinolones, co-trimoxazole, teicoplanin and some beta-lactams, while the Staphylococcus spp. showed high resistance to aminoglycosides, beta-lactams and macrolides. Twenty-four LAB isolates were selected based on their antimicrobial activity against two uropathogenic Staphylococcus aureus strains and bacteriocin production. LAB strains showing antimicrobial activity were grouped into smaller groups through amplified ribosomal DNA restriction analysis (ARDRA). Representative strains were identified as Weissella spp., Enterococcus faecium, Lactococcus lactis and Lactobacillus brevis through sequencing of 16S rDNA. The Weissella spp. and L. brevis strains demonstrated remarkable inhibitory activity against seven strains of Gram negative uropathogens. Two strains of L. lactis produced a bacteriocin-like inhibitory substance active against Lactobacillus sakei. In this study, an unusual high rate of co-trimoxazole, quinolones and macrolides resistance among uropathogens from south west Nigeria was discovered. Based on their sensitivity to Weissella spp., there is a potential for using these LAB as a natural approach for the protection against the uropathogens assayed.

Two-hundred and forty-five strains of Clostridium perfringens isolated from the faeces of apparently healthy sheep and cattle and from their environments (paddock soils) in Kano and Kaduna States of Nigeria were studied. The isolates were examined by the toxin-antitoxin neutralisation tests performed intradermally in depilated albino guinea-pigs. One-hundred and twenty-seven (53.1%) of the isolates were type A, 17 (7.1%) were type B, 14 (4.9%) were type C, 44 (18.4%) were type D and 19 (7.9%) were type E. Eighteen (7.5%) were not typable while six were lost during storage. The significance of this distribution and the potential danger to animal health, especially as regards enterotoxaemias, cannot be over-emphasized.

Fourteen isolates of yersiniae belonging to three species isolated from cattle and pigs were tested for their sensitivity to 12 antimicrobial agents by the tube dilution technique. All the isolates were sensitive to gentamicin and 93.0%, 93.0%, 85.7% and 85.7% sensitive to tetracycline, nitrofurantoin, chloramphenicol and sulphamethoxazole-trimethoprim, respectively. There were no consistent differences in the rates and patterns of resistance based on source, species and serotypes of organisms.

Random bacteriological examination of 300 bile samples aseptically drawn from the gallbladders of adult cattle at slaughter revealed the presence of Escherichia coli in 30%, Enterobacter aerogenes in 14% and Klebsiella aerogenes in 56%. Bacteriological examination of another 116 faecal samples revealed Esch. coli in 60% and Ent. aerogenes in 40%, while parasitological examination revealed 50-100 strongyle ova per gram from pre-rain samples of faeces, 100-850 strongyle eggs per gram for faeces collected during the rains, and less than 50 strongyle eggs per gram for dry season faeces. No salmonellae were recovered from either the faeces or bile, and no Clostridium or any helminth of significant public health interest were recovered from faeces but it is still suggested that indirect consumption of bile and faeces from these animals may constitute a public health hazard.

A total of 252 rodents were trapped in the environs of Ahmadu Bello University, Zaria, over a two-year period, of which 221 were brown field rats (Arvicanthus niloticus). Only these were found positive (4.5%). Titers were detected against Leptospira interrogans, serovars australis, icterohaemorrhagiae, tarassovi and hardjo. All positive sera except one reacted against a single serovar. Eight isolates were obtained from field rats: five were serovar australis, one ballum and two unidentified. Five isolates were recovered from 74 bovine kidneys examined: one was serovar pyrogenes and four unidentified. One unidentified isolate failed to react with any of the 16 screening leptospira antisera and another was tentatively considered to be serovar ballum. Two isolates, one ballum and one unidentified, were recovered from the Kubani stream waters. No leptospira was isolated from piggery sewage effluents collected at Kano and Kaduna. It is suggested that serovar ballum be included in any screening battery for leptospirosis in man and animals in Nigeria.