Assessing the mechanisms of multi-drug resistant non-typhoidal Salmonella (NTS) serovars isolated from layer chicken farms in Nigeria.

BACKGROUND: In Nigeria, there have been reports of widespread multiple antimicrobial resistance (AMR) amongst Salmonella isolated from poultry. To mitigate the impact of mortality associated with Salmonella on their farms, farmers resort to the use of antimicrobials without sound diagnostic advice. We conducted this study to describe the AMR patterns, mechanisms and genetic similarities within some Salmonella serovars isolated from different layer farms. METHOD: We determine the AMR profiles of two hundred Salmonella isolates, selected based on frequency, serovar, and geographical and sample type distribution. We also assessed the mechanisms of multi-drug resistance for specific genetic determinants by using PCR protocols and gene sequence analysis. Pulsed-field gel electrophoresis (PFGE) was conducted on seven selected serovars to determine their genetic relatedness. RESULTS: Of 200 isolates, 97 (48.5%) revealed various AMR profiles, with the multiple antibiotic resistance (MAR) index ranging from 0.07-0.5. Resistance to ciprofloxacin was common in all the multi-drug resistant isolates, while all the isolates were susceptible to cefotaxime, ceftazidime, and meropenem. Genotypic characterization showed the presence of resistance genes as well as mutations in the nucleotide genes with subsequent amino acid substitutions. Fifteen isolates (43%) of S. Kentucky were indistinguishable, but were isolated from four different states in Nigeria (Ogun, n = 9; Kaduna, n = 6; Plateau, n = 3, and: Bauchi, n = 2). PFGE revealed 40 pulsotype patterns (Kentucky, n = 12; Larochelle, n = 9; Virchow, n = 5; Saintpaul, n = 4; Poona, n = 3; Isangi, n = 2, and; Nigeria, n = 2). CONCLUSION: This study recorded strictly related but diversely distributed Salmonella serovars with high AMR rates in poultry. We recommend strict regulation on antimicrobial use and regular monitoring of AMR trends among bacteria isolated from animals and humans to inform public policy.

Determination of antimicrobial use in commercial poultry farms in Plateau and Oyo States, Nigeria.

BACKGROUND: Indiscriminate use of antimicrobials for the prevention and treatment of bacterial infection in animals is a common practice in Nigeria as in other developing countries. These antimicrobials are purchased over the counter without restrictions and often administered in form of medicated feedstuffs. In Nigeria, like most developing countries, antimicrobial prescription data are not routinely collected or reported at the farm level, instead import data are used in reporting antimicrobial consumption. Farmers can be useful sources of data on the use of antimicrobial agents by class, animal species, production type and age. The objective of the study was to determine the knowledge, attitude and practices of poultry farmers on antimicrobial resistance and to generate data on antimicrobial use (AMU) in poultry farms in Plateau and Oyo states in accordance with the guidelines of the World Organization for Animal Health (WOAH). METHODS: A questionnaire used by the Food and Agriculture Organization (FAO) of the United Nations in Ghana was adopted and modified to collect data on the knowledge, attitude and practices of farmers on AMR and AMU and to collect AMU data from selected poultry farms. A focus group discussion (FGD) was conducted in Plateau state with poultry farmers and representatives from the state veterinary services, using a checklist. The aim of the FGD was to have an idea on antimicrobial use among poultry farmers and to generate additional questions that might be added to the questionnaire. Stratified random sampling technique was used to select 50 farms from Plateau and Oyo states, using the list of registered poultry farms in the two states as sampling frame. RESULTS: Ninety eight percent (98%) of farmers gave antibiotics as prophylactic treatment to day old chicks. There were 47 different products used in the two states within the study period. We observed that five classes of antibiotics (Tetracyclines, Penicillins, Aminoglycosides, Polypeptides and Fluoroquinolone) were used in the two states. A total of 351 kg of active ingredients from seven different classes, namely: tetracyclines, penicillins, aminoglycosides, polypeptide, fluoroquinolones, amphenicol and macrolides were recorded from the two states. Some products contained cocktail of antibiotics, having up to six different classes with very high concentration of active ingredients which are not in the list of registered antimicrobials reported to WOAH. CONCLUSION: The concept used for this survey proved that the approach can be applied for AMU surveillance in the animal health sector. It also provided insight on farmers' knowledge and practices with regards to the use of antimicrobials which is missing in the national import data. The need for "stronger" antibiotics was identified as one of the drivers of antibiotic resistance.

Perceptions and practices of farmers of indigenous poultry towards Salmonella infections in North-Central Nigeria.

Background: Salmonella infections continue to be of global concern to poultry health, productivity, and public health. About 44% of the poultry in Nigeria are indigenous and kept in close interaction with farmers who are mostly rural dwellers and have limited access to veterinary and extension services. Aim: The perceptions and practices of farmers of indigenous poultry toward Salmonella infections were assessed to obtain and document baseline data that can be used to create awareness among farmers about these infections and their attendant public health implications. Methods: A cross-sectional approach using a multistage sampling method was used in this survey. A total of 419 farmers keeping indigenous poultry were interviewed using a pre-tested electronic questionnaire in three randomly selected states within North-Central Nigeria. Data were analyzed using descriptive and regression analysis. Results: Out of the 419 respondents, 138 (32.9%), 141 (33.7%), and 140 (33.4%) were from Benue, Kwara, and Plateau States, respectively. Of the 419, 55.4% were females, 40.8% were above 40 years, and 35.8% have over 10 years of farming experience. The majority of the poultry are not housed (58.5%) and farmers predominantly rear chickens (51.8%). Also, 49.9% of the birds were 1-6 months with 41.5% of the flock sizes being 11-20. Respondents had a poor level of perception toward Salmonella infection as the majority did not know that Salmonella affects poultry (89.3%) and that Salmonella infections are zoonotic (94.5%). Significant (p = 0.000) associations existed between categorized perception score and age, educational status, family size, and farming experience of farmers. There were significant (p = 0.000) associations of categorized practice scores with gender, age, education status, family size, and farming experience of farmers. Conclusion: This study has revealed the poor perception of farmers on Salmonella infections and has highlighted their practices. There is a need to raise awareness about these infections to improve indigenous poultry health and productivity as well as public health.

Investigation of potential risk factors associated with Salmonella presence in commercial laying hen farms in Nigeria.

BACKGROUND: In 2012/13, the Federal government of Nigeria approved the surveillance of salmonellae in commercial poultry farms with the aim of generating baseline data for the development of a control programme. That baseline provided an opportunity to investigate potential risk factors in commercial layer farms. METHODS: Five hundred and twenty-three farms were evaluated for the presence of Salmonella. Each farmer was interviewed through a structured questionnaire. Univariate analysis identified 26 variables potentially associated with Salmonella presence on the farms, with different levels of significance. To simultaneously evaluate the effect of these variables, a multivariable logistic regression model was implemented. RESULTS: Farmers that are member of the Poultry Association of Nigeria (PAN) emerged as a risk factor, as well as being farmer as exclusive occupation. The risk of Salmonella increased in farms with a combined housing system as compared to farms with a single housing system. Moreover, in terms of farm management the use of plastic egg crates had a higher risk of Salmonella presence compared to paper crates. Higher risk of contamination was also identified in farms having regular contact with animal health workers, since this could be indicative of animal health problems requiring continuous assistance. Farms with no previous outbreak of salmonellosis were associated with an increase risk, since most of the farms in this category were newly established. With regard to feeding, the use of coccidiostats and commercial feeds were associated with the spread of Salmonella. Unexpectedly, the presence of other farms at distances further than 1 km away was associated with an increase in Salmonella presence as compared to farms which were in closer proximity. Close proximity among farms could favour the application of stricter biosecurity measures. CONCLUSIONS: The results of the survey can be considered a baseline for implementing effective measures aimed at reducing the Salmonella prevalence in Nigerian laying hen farms.

Salmonella serovars and their distribution in Nigerian commercial chicken layer farms.

Commercial poultry farms (n° 523), located in all the six regions of Nigeria were sampled with a view to generate baseline information about the distribution of Salmonella serovars in this country. Five different matrices (litter, dust, faeces, feed and water) were collected from each visited farm. Salmonella was isolated from at least one of the five matrices in 228 farms, with a farm prevalence of 43.6% (CI95[39.7-48.3%]). Altogether, 370 of 2615 samples collected (14.1%, CI95[12.8; 15.5%]) contained Salmonella. Considering the number of positive farms and the number of positive samples, it was evident that for the majority of the sampled farms, few samples were positive for Salmonella. With regard to the matrices, there was no difference in Salmonella prevalence among the five matrices considered. Of the 370 isolates serotyped, eighty-two different serotypes were identified and Salmonella Kentucky was identified as having the highest isolation rate in all the matrices sampled (16.2%), followed by S. Poona and S. Elisabethville. S. Kentucky was distributed across the country, whereas the other less frequent serovars had a more circumscribed diffusion. This is one of few comprehensive studies on the occurrence and distribution of Salmonella in commercial chicken layer farms from all the six regions of Nigeria. The relatively high prevalence rate documented in this study may be attributed to the generally poor infrastructure and low biosecurity measures in controlling stray animals, rodents and humans. Data collected could be valuable for instituting effective intervention strategies for Salmonella control in Nigeria and also in other developing countries with a similar poultry industry structure, with the final aim of reducing Salmonella spread in animals and ultimately in humans.

Motile Salmonella serotypes causing high mortality in poultry farms in three South-Western States of Nigeria.

This study was carried out to identify the Salmonella serotypes causing high mortality in chickens in Lagos, Ogun and Oyo states, Nigeria. Chickens presented for postmortem examination during disease outbreaks that were characterised by high mortality (40 per cent to 80 per cent) in poultry farms in the study area were examined from January to December, 2013. Samples of the lungs, heart, liver, spleen, kidneys, proventriculus, intestine and caecum were collected from suspected cases of salmonellosis, for bacterial culture and identification. Salmonella isolates were confirmed using PCR and serotyped using the Kauffman-White scheme. Twenty-six day-old pullets were raised to two weeks and inoculated orally with 0.2 mL of 1×108 colony forming units of Salmonella Zega identified in the present study to determine their pathogenicity, while another 26 served as control. The Salmonella serotypes were S Zega (n=13; 35.14 per cent), Salmonella Kentucky (n=9; 24.32 per cent), Salmonella Herston (n=6; 16.22 per cent), Salmonella Nima (n=4; 10.81 per cent), Salmonella Telelkebir (n=3; 8.11 per cent), Salmonella Colindale (n=1; 2.70 per cent) and Salmonella Tshiongwe (n=1; 2.70 per cent). Clinical signs in both natural and experimental infections were acute (70 per cent) and chronic (30 per cent), and included weakness, anorexia, yellowish diarrhoea, pasted vents, somnolescence and mortality, while gross lesions showed marked pulmonary congestion and oedema, necrotic foci in the myocardium; the liver, spleen and kidneys were markedly enlarged and had subcapsular multifocal necrosis. There were catarrhal proventriculitis and enteritis, and haemorrhagic typhlitis. While most of the serotypes identified in the present study have been isolated from poultry sources from commercial farms in Nigeria, to the best of the authors' knowledge, they have not been previously reported to cause high mortality in chickens in the study area.