Patterns and drivers of antibiotic use in small-scale broiler production systems in Lilongwe District, Malawi.

Reducing the inappropriate use of antibiotics in food animals is a global priority to address antimicrobial resistance (AMR). We investigated practices and factors associated with antibiotic use in small-scale commercial broiler farms in Lilongwe district, Malawi. We used structured questionnaires to collect data on recent antibiotic use practices among 128 broiler farmers, who kept between 50 and 1 000 birds, from December 2022 to March 2023. Logistic regression analysis was used to identify risk factors associated with antibiotic use. Over half (53.1 %, n=68) of the farms reported using antibiotics at least once in the previous production cycle. Overall, 11 different types of antibiotics were used either for treatment and/or preventive purposes, with oxytetracycline (88.2 %), erythromycin (29.4 %), and enrofloxacin (26.5 %) reported as the frequently used. One-third of all antibiotic formulations contained multiple active antibiotic ingredients, with 12 % containing four antibiotics. Covariates associated with an increased likelihood of antibiotic use include disease incidence (OR=13.8, 95 % CI 5.27-42.50, p<0.001) and entry of wild birds into poultry houses (OR=3.56, 95 % CI =1.44-9.61, p=0.008). Our study highlights inappropriate usage of antibiotics, largely associated with reduced biosecurity and disease incidence. These findings underscore the need to strengthen veterinary services, reinforce regulations on antibiotic access and use, and farmer education programs promoting proper husbandry, biosecurity, and responsible antibiotic use.

Direct detection of Mycobacterium bovis by a dry loop-mediated isothermal amplification assay in cattle samples collected during routine abattoir examination in Malawi.

The lack of quick, accurate, and low-cost detection methods has hindered the active control strategies for bovine tuberculosis (bTB) in resource-limited countries with a high burden of disease. We developed a dry loop-mediated isothermal amplification (LAMP) assay for rapid and specific detection of Mycobacterium bovis, the principal causative agent of bTB, and evaluated the efficacy of the assay using suspected bTB samples collected during routine meat inspection at major regional abattoirs in Malawi. Template genomic DNA was extracted directly from the granulomatous bTB-like lesion (crude extracted DNA), as well as growth from the incubated mycobacterial growth indicator tubes (MGIT). Field results were visualized by the naked eye within 40 min following a color change of the amplified products. The sensitivity and specificity of the dry LAMP assay while using 152 DNA samples extracted from MGIT with confirmed M. bovis results were 98% and 88%, respectively. When 43 randomly selected crude DNA samples from lesions were used, the sensitivity and specificity of the dry LAMP assay were 100% and 75%, respectively. Our LAMP assay offers the potential to meet the demands for a low-cost and rapid field detection tool for bTB in resource-limited countries in which bTB is endemic.

Burden, Antibiotic Resistance, and Clonality of Shigella spp. Implicated in Community-Acquired Acute Diarrhoea in Lilongwe, Malawi.

Although numerous studies have investigated diarrhoea aetiology in many sub-Saharan African countries, recent data on Shigella species' involvement in community-acquired acute diarrhoea (CA-AD) in Malawi are scarce. This study investigated the incidence, antibiotic susceptibility profile, genotypic characteristics, and clonal relationships of Shigella flexneri among 243 patients presenting with acute diarrhoea at a District Hospital in Lilongwe, Malawi. Shigella spp. were isolated and identified using standard microbiological and serological methods and confirmed by identifying the ipaH gene using real-time polymerase chain reaction. The isolates' antibiotic susceptibility to 20 antibiotics was determined using the VITEK 2 system according to EUCAST guidelines. Genes conferring resistance to sulfamethoxazole (sul1, sul2 and sul3), trimethoprim (dfrA1, dfrA12 and dfrA17) and ampicillin (oxa-1 and oxa-2), and virulence genes (ipaBCD, sat, ial, virA, sen, set1A and set1B) were detected by real-time PCR. Clonal relatedness was assessed using ERIC-PCR. Thirty-four Shigella flexneri isolates were isolated (an overall incidence of 14.0%). All the isolates were fully resistant to sulfamethoxazole/trimethoprim (100%) and ampicillin (100%) but susceptible to the other antibiotics tested. The sul1 (79%), sul2 (79%), sul3 (47%), dfrA12 (71%) and dfrA17 (56%) sulfonamide and trimethoprim resistance genes were identified; Oxa-1, oxa-2 and dfrA1 were not detected. The virulence genes ipaBCD (85%), sat (85%), ial (82%), virA (76%), sen (71%), stx (71%), set1A (26%) and set1B (18%) were detected. ERIC-PCR profiling revealed that the Shigella isolates were genetically distinct and clonally unrelated, indicating the potential involvement of genetically distinct S. flexneri in CA-AD in Malawi. The high percentage resistance to ampicillin and sulfamethoxazole/trimethoprim and the presence of several virulence determinants in these isolates emphasises a need for continuous molecular surveillance studies to inform preventive measures and management of Shigella-associated diarrhoeal infections in Malawi.

Microbiological hazard identification and exposure assessment of food prepared and served in rural households of Lungwena, Malawi.

The presence of food-borne pathogens, Escherichia coli 0157:H7, Staphylococcus aureus, Salmonella species, Campylobacter jejuni and non-pathogenic E. coli, in 132 home cooked food samples consisting of maize flour porridge (MFP), (n=41), fish (n=37), vegetables (n=28), beans (n=13) and "Others" (n=13), collected from 6 villages in Lungwena, Malawi was investigated. It was found that 35% of the food samples were contaminated with one or more pathogens; with 48%, 8%, 61% and 23% of the food samples being found to harbour E. coli, pathogenic E. coli 0157: H7, S aureus and Salmonella species, respectively. C. jejuni was not detected in any food sample. Using a 95% level of significance, pathogen concentration among food categories demonstrated a statistical difference (p=0.001). Distribution of pathogens among villages was also found to be significant (p=0.03). MFP was the most contaminated food. Practices that promote the spread of the pathogens in the rural household kitchens were investigated. Food was thought to be contaminated as a result of poor food handling, preparation and storage practices.