Extended spectrum and metallo-beta-lactamase Pseudomonas species from poultry and piggery waste

Beta-lactamase producing bacteria have become a public health burden due to antibiotics usage in livestock production. This study was carried out to detect extended spectrum beta-lactamase (ESBL) and metallo-beta-lactamase (MBL) producing Pseudomonas spp. from poultry droppings and piggery dung in Ibadan. Poultry droppings and piggery dung were collected from the University of Ibadan livestock farms while isolation of Pseudomonas spp. was done using Pseudomonas base agar supplemented with Pseudomonas C-N supplement and were conventionally characterized. Detection of ESBL and MBL producing isolates were by double disc synergy test and imipenem-EDTA combined disc test respectively. Antimicrobial susceptibility test was by disc diffusion method against trimethoprim (5 µg), amoxicillin/clavulanate (30 µg), cefotaxime (30 µg), ceftazidime (30 µg), cefepime (30 µg), aztreonam (30 µg), imipenem (10 µg), gentamicin (10 µg) and ciprofloxacin (10 µg). A total of 108 Pseudomonas spp. were isolated comprising 53.7% from poultry droppings and 46.3% from piggery dung. The isolates include P. aeruginosa (63.0%), P. putida (24.0%) and P. stutzeri (13.0%). While the ESBL producers were P. aeruginosa (10.2%) and P. stutzeri (1.9%), none of the isolates produced MBL. However, 63.6% the ESBL producers showed resistance to trimethoprim while 61.5% were multidrug resistant. The high prevalence of antibiotics resistance and multidrug resistant strains observed among the Pseudomonas spp. infer that poultry droppings and piggery dung can serves as a reservoir for growth and dissemination of clinically significant antibiotics resistance among bacterial species. DOI: http://dx.doi.org/10.5281/zenodo.348957

Pseudomonas species from cattle dung producing extended spectrum and metallo beta-lactamases

Indiscriminate use of antibiotics in livestock contributes to emergence of antimicrobial resistance in pathogens co-habiting the gastro-intestinal tract of animals. This study was to determine the Extended Spectrum Beta-Lactamase (ESBL) and Metallo-Beta-Lactamase (MBL) production in Pseudomonas species from cattle fecal samples. Cattle dungs were collected from the University of Ibadan Cattle Ranch and the Pseudomonas species isolated using Pseudomonas Base Agar with Pseudomonas CN Selective Supplement were identified using standard tests. Phenotypic detection of ESBL and MBL was by double disk synergy test and Ethylene Di-amine Tetra Acetic Acid Combined Disk Test respectively. Antibiotics susceptibility tests was done using the disc diffusion technique against ten antibiotics. A total of 144 Pseudomonas species were isolated and identified as P. aeruginosa (71.5%), P. fluorescens (19.4%) and P. stutzeri (9.1%) and 19 (37.1%) produced ESBL including P. aeruginosa (15), P. fluorescens (2) and P. stutzeri (2) while, one (6.7%) ESBL P. aeruginosa produced MBL. All the ESBL producers were resistant to cefotaxime and trimethoprim; resistance of P. aeruginosa to ciprofloxacin was 93.3% and to ceftazidime was 80.0%, while it was 13.3% (colistin) and 6.7% (imipenem). The ESBL producing P. fluorescens were resistant to ceftazidime, ciprofloxacin and trimethoprim, likewise, the ESBL producing P. stutzeri showed resistance to gentamicin, ciprofloxacin and trimethoprim. The production of ESBL and MBL observed among the Pseudomonas species in this study with high level of resistance to some antibiotics portend public health risk, hence a need for caution in the use of antibiotics in animal husbandry. DOI: http://dx.doi.org/10.5281/zenodo.363080

Antibiotics Susceptibility Profile of Listeria Species Isolated from Poultry Wastes and Fishpond Water from Private and Institutional Farms in Ibadan, Nigeria

Introduction: Untreated waste being discharged into the environment due to proliferation of poultry and fish farms can constitute a public health threat to human. Listeria, an emerging pathogen is commonly associated with food. This study aimed at determining the antibiotic resistant pattern of Listeria species isolated from poultry droppings and fish pond water in Ibadan. Materials and Methods: Poultry waste and fishpond water samples were collected between April and July, 2016. Listeria Selective Agar was used to isolate Listeria species and identified using conventional methods. Antimicrobial susceptibility testing was done using the Kirby-Bauer disk diffusion method against ampicillin, cloxacillin, amoxicillin, streptomycin, ceftriazone, chloramphenicol, ciprofloxacin, ofloxacin, sulfamethoxazole-trimethoprim and tetracycline. Results: A total of forty samples were collected and 105 Listeria spp. isolated; 62.9% from poultry waste and 37.1% from fishpond water. From the institutional fish pond and poultry waste, 18.1% and 51.4% isolates were obtained while from private fish pond and poultry waste, 19.1% and 11.4% isolates respectively were obtained. The isolates were Listeria monocytogenes (27.6%), L. innocua (8.6%), L. ivanovii (16.2%) and other Listeria spp. (47.6%). All the isolates were resistant to ampicillin and ciprofloxacin; all L. monocytogenes, L. innocua and L. ivanovii were resistant to ceftriaxone, but 89.3% of the other Listeria spp. showed resistance. Furthermore, all (100%) the isolates obtained from private poultry farm were resistant to chloramphenicol while 53.3% were resistant to trimethoprim/sulfamethoxazole. Also, 12.2% of the isolates were found to be resistant to a combination of six antibiotics including: Ampicillin, amoxicillin, ceftriaxone, streptomycin and trimethoprim/sulfamethoxazole. Conclusion: This study revealed that poultry waste and fish pond water from the selected farms could be a potential source for the transmission of multi-drug resistant bacteria to humans. Proper treatment of poultry waste and fish pond water should be ensured before discharge into the environment

Direct detection of Mycobacterium tuberculosis with nitrate reductase assay and microscopic observation drug susceptibility

The global increase in tuberculosis drug resistant which is a threat to its control, require low cost method of diagnosis and detection. Available conventional and molecular methods consume time, and are expensive for countries with high disease burden. Nitrate Reductase Assay (NRA) and Microscopic Observation Drug Susceptibility (MODS) performance to directly detect tuberculosis resistance to four drugs was evaluated. The NRA (liquid and solid) and MODS performance of smear-positive sputum samples were evaluated; Sensitivities and specificities were compared with Proportion Method (PM). Sensitivity and specificity of liquid NRA (LNRA) were 90% and 98% (rifampicin), 81.8% and 100% (isoniazid), 88.9% and 98.1% (streptomycin), and 57.1% and 94.4% (ethambuthol). Also, the sensitivity and specificity for solid NRA (SNRA) were 69.2% and 98.3% (rifampicin); 100% and 100% (isoniazid); 88.9% and 95.2% (streptomycin); 70% and 80.6% (ethambuthol). Moreover, For MODS, rifampicin and isoniazid sensitivity and specificity was 100%, it was 100% and 98.1% for streptomycin, and 71.4% and 98.2% for ethambuthol. At day 14, the results available for LNRA, SNRA and MODS were 93%, 68.5% and 100% respectively. The agreement between LNRA and PM was 97% (RIF, INH and SM) and 90% (EMB). For SNRA, it was 93% (RIF), 100% (INH), 94% (SM) and 89% (EMB). While for MODS, it was 100% (RIF and INH), 98% (SM) and 95% (EMB). Direct NRA and MODS are sensitive, reliable and fast for antituberculosis drug susceptibility; they have potential to effectively and reliably detect drug resistant tuberculosis in the low resource countries. DOI: http://dx.doi.org/10.5281/zenodo.429588

Patterns of drug resistance in Mycobacterium tuberculosis from tuberculosis patients in Ibadan Nigeria

The success of the global tuberculosis (TB) control program has been threatened with drug resistant strains emergence; especially the Multidrug Resistant Tuberculosis (MDR-TB). Despite that Nigeria is one of the countries with high tuberculosis burden, little is known on the magnitude of MDR-TB in the country. This study was to determine drug resistant patterns of Mycobacterium tuberculosis isolated from patients that attended Directly Observed Treatment Short-course (DOTS) centres in Ibadan, Nigeria. Sputum samples collected from confirmed TB patients were processed using the N-acetyl L-cysteine-sodium hydroxide decontamination method. Direct drug susceptibility test was carried out against rifampicin, isoniazid, ethambuthol and streptomycin. Out of the 319 samples collected, 149 (46.7%) were culture positive and susceptibility test was completed for 101 (67.8%) isolates, out of which any resistance and mono-resistance to rifampicin was 23.8% and 8.9% respectively. In all 11.9% MDR-TB was observed comprising 30.8% (acquired), and 8.3% (primary) while, 3.96% showed resistance to all tested drugs. The patterns of MDR-TB in this study indicates that active case findings as well as expansion of drug susceptibility testing is required to effectively control TB, drug resistant strains and to forestall the transmission and spread of the drug-resistant TB in the society. DOI: http://dx.doi.org/10.5281/zenodo.428850

Antibiotic resistance and virulence genes in Campylobacter species from pig and cattle samples in Ibadan, Nigeria

Campylobacter species are the leading cause of gastroenteritis worldwide with incidence cases higher than those caused by Salmonella. This study was designed to determine the antibiotic resistance patterns and virulence genes detection in selected Multi-Drug Resistant (MDR) strains of Campylobacter species isolated from pigs and cattle dungs. Stool samples were collected from pigs and cattle at the teaching and research farm of the University of Ibadan, Nigeria. Isolation and identification of Campylobacter species were made using modified charcoal cefoperazone deoxycholate agar and standard biochemical tests. Antibiotic susceptibility test was carried out using the disk diffusion technique. The hipO and cadF virulence genes were detected using a multiplex-polymerase chain reaction. The occurrence of Campylobacter species in pigs and cattle was 90.0% and 95.0%, respectively. In pigs, C. coli had the highest occurrence, while it was C. jejuni in cattle. The antibiotic resistance patterns showed that 1.3% and 1.2% of the isolates from pigs and cattle, respectively were resistant to all the antibiotics tested. Among the selected MDR strains, cadF genes were detected in 76.5% (pigs) and 75.0% (cattle). However, hipO genes were detected in 11.8% (pigs) and 50.0% (cattle) isolates. The cadF gene was detected in all the Campylobacter species, while hipO gene was detected only in C. jejuni. In conclusion, the pigs and cattle faecal wastes harbored virulent and multidrug-resistant Campylobacter species. Hence, the indiscriminate discharge of untreated animal faecal wastes into the environment and water bodies should be discouraged. DOI: http://dx.doi.org/10.5281/zenodo.1090957

Extended Spectrum Beta Lactamase (ESBL) producing enteric bacteria from hospital wastewater, Ibadan, Nigeria

This study was conducted to investigate the occurrence of antibiotic resistance, including β-lactamase and extended spectrum β-lactamase production among enteric bacteria isolated from hospital wastewater from selected hospital within Ibadan. Physico-chemical analysis of hospital wastewater samples was done, enteric bacteria were isolated and identified using convectional biochemical tests while the selection of potential ESBL-producing bacteria was carried out using disc diffusion method and ESBL detection using double synergy test. The turbidity of the wastewater samples ranged between 4.45-6.5 NTU and total suspended solids ranged between 3.4- 45.5 mg/L. While electrical conductivity was between114.25-214 µs/m, the biological oxygen demand was between 25.8-31.25 mg/L and chemical oxygen demand ranged between 41.25-45.38 mg/L. Of the 200 bacteria isolated 35(17.5%) produced ESBL; 14(40%) from the tertiary hospital and 21(60%) from private hospital out of which 85.7%, 80% and 65.7% showed resistance to sulphamethxazole/Trimetoprim, streptomycin and tetracycline respectively, while resistance to meropenem (8.6%) was low. Among the ESBL-producing isolates, K. pneumonia had the highest (15(42.8%) rate of occurrence. This study revealed a need for hospital wastewater to be properly treated before discharged into water bodies and the environment to forestall the indiscriminate discharge of wastewater harbouring ESBL-producing bacteria

Direct detection of Mycobacterium tuberculosis with nitrate reductase assay and microscopic observation drug susceptibility

The global increase in tuberculosis drug resistant which is a threat to its control, require low cost method of diagnosis and detection. Available conventional and molecular methods consume time, and are expensive for countries with high disease burden. Nitrate Reductase Assay (NRA) and Microscopic Observation Drug Susceptibility (MODS) performance to directly detect tuberculosis resistance to four drugs was evaluated. The NRA (liquid and solid) and MODS performance of smear-positive sputum samples were evaluated; Sensitivities and specificities were compared with Proportion Method (PM). Sensitivity and specificity of liquid NRA (LNRA) were 90% and 98% (rifampicin), 81.8% and 100% (isoniazid), 88.9% and 98.1% (streptomycin), and 57.1% and 94.4% (ethambuthol). Also, the sensitivity and specificity for solid NRA (SNRA) were 69.2% and 98.3% (rifampicin); 100% and 100% (isoniazid); 88.9% and 95.2% (streptomycin); 70% and 80.6% (ethambuthol). Moreover, For MODS, rifampicin and isoniazid sensitivity and specificity was 100%, it was 100% and 98.1% for streptomycin, and 71.4% and 98.2% for ethambuthol. At day 14, the results available for LNRA, SNRA and MODS were 93%, 68.5% and 100% respectively. The agreement between LNRA and PM was 97% (RIF, INH and SM) and 90% (EMB). For SNRA, it was 93% (RIF), 100% (INH), 94% (SM) and 89% (EMB). While for MODS, it was 100% (RIF and INH), 98% (SM) and 95% (EMB). Direct NRA and MODS are sensitive, reliable and fast for antituberculosis drug susceptibility; they have potential to effectively and reliably detect drug resistant tuberculosis in the low resource countries. DOI: http://dx.doi.org/10.5281/zenodo.429588

The problem of resistance in Mycobacterium tuberculosis may be underestimated in Africa.

The true burden of tuberculosis (TB) and particularly multidrug-resistant (MDR) TB in Sub-Saharan Africa has remained underestimated

Nontuberculous Mycobacteria Isolated from Tuberculosis Suspects in Ibadan, Nigeria

In Nigeria, one of the highest tuberculosis (TB) burdened nations, sputum smear microscopy is routinely employed for TB diagnosis at Directly Observed Treatment Short-Course (DOTS) Centers. This diagnostic algorithm does not differentiate Mycobacterium tuberculosis complex (MTC) from nontuberculous mycobacteria (NTM). Between December 2008 and January 2009, consecutive patients diagnosed with TB were screened for inclusion at 10 DOTS centers in Ibadan, Nigeria. To verify Mycobacterium species in patients diagnosed, we cultured and identified mycobacterial isolates using PCR, line probe assay, and spoligotyping techniques. From 48 patients screened, 23 met the inclusion criteria for the study. All the 23 study patients had a positive culture. Overall, we identified 11/23 patients (48%) with MTC only, 9/23 (39%) with NTM only, and 3/23 (13%) with evidence of both MTC and NTM. Strains of MTC identified were Latin American Mediterranean (LAM) genotype (n=12), M. africanum (n=1), and the genotype family T (n=1). Four M. avium-intracellulare-M. scrofulaceum complexes, one M. chelonae complex, one M. abscessus, and one M. intracellulare were identified. Our findings underscore the need to incorporate molecular techniques for more precise diagnosis of TB at DOTS centers to improve clinical outcomes and safe guard public health, particularly in TB endemic countries