Multidrug-Resistant Escherichia coli from Raw Cow Milk in Namwala District, Zambia: Public Health Implications.

Escherichia coli (E. coli), a major foodborne disease-causing pathogen found in raw cow milk, has even far more reaching public health ramifications as it encodes for antimicrobial resistance (AMR). This study aimed to identify multidrug-resistant (MDR) E. coli from raw cow's milk and evaluate their antimicrobial-resistant profiles. In total, 418 pooled raw cow milk samples were collected from milk collection centers and analysed using standard culture methods to isolate E. coli. Antimicrobial Susceptibility Testing (AST) was conducted using the Kirby Bauer disk diffusion method and PCR was used to identify cefotaxime (CTX) resistant genes. Overall isolation of E. coli was 51.2% (214/418) with MDR observed in 21% (45/214) of isolates across different antibiotic combinations. Resistance was observed towards ampicillin (107/214, 50%), tetracycline (86/214, 40.1%), trimethoprim/sulfamethoxazole (61/214, 28.5%), and amoxicillin/clavulanic acid (CTX) (50/214, 23.4%). Notably, 15% (32/214) resistance to CTX was observed, while 12.6% (27/214) exhibited resistance to imipenem. The blaCTX-M and blaTEM genes were detected in CTX-resistant isolates. The findings of MDR E. coli that harbour blaCTX-M and blaTEM genes in raw cow's milk indicate serious public health risks for consumers.

Determinants of Knowledge, Attitude, and Practices of Veterinary Drug Dispensers toward Antimicrobial Use and Resistance in Main Cities of Malawi: A Concern on Antibiotic Stewardship.

Antimicrobial resistance (AMR) is an emerging challenge to global public health. The use of antibiotics in the veterinary field is one of the contributing factors to AMR mostly due to poor knowledge, attitudes, and practices (KAP) of dispensers. Veterinary drug dispensers are expected to guide clients on indications, contraindications, and withdrawal periods of veterinary drugs. This study assessed veterinary drug dispensers' KAP toward AMR and associated potential contributing factors. A cross-sectional study, using a structured questionnaire, was conducted in three main cities of Malawi, namely Mzuzu, Lilongwe, and Blantyre. A total of 68 agrovet shops were selected using a simple random sampling technique. The KAP level was presented descriptively. Bivariate and multivariable analyses were run to investigate the relationships between the independent and outcome variable. Overall, the KAP score for knowledge, attitude, and practices was 46.7%, 49.2%, and 41.6%, respectively. The significant determinants of the knowledge were the practice of asking for a written prescription (OR: 16.291, 95% CI: 11.6-24.2) (p = 0.024), female (OR: 0.609, 95% CI: 0.3-0.9) (p = 0.001), and old age (≥35) (OR: 0.227, 95% CI: 0.1-0.5) (p = 0.04). Poor knowledge, negative attitude, and poor practices were observed among most of the participants. Sensitization and training on AMR and antimicrobial stewardship are recommended to address the KAP score gaps and the observed determinants among veterinary drug dispensers.

Seroprevalence and Associated Risk Factors of Rift Valley Fever in Livestock from Three Ecological Zones of Malawi.

The epidemiology of Rift Valley fever (RVF) is poorly understood in Malawi. Here, a cross-sectional study was conducted (March-June 2020) to investigate the seroprevalence and potential risk factors of RVF virus (RVFV) in cattle, goats, and sheep in three ecological zones of Malawi. A total of 1523 serum samples were tested for anti-RVFV IgG and IgM antibodies by ELISA. Additionally, a questionnaire survey was used to assess potential RVF risk factors. The overall seroprevalence was 17.14% (261/1523; 95% CI = 15.33-19.11) for individual livestock and 33.24% (120/361; 95% CI = 28.18-38.11) for the livestock herd. Seroprevalence was significantly high in sheep (25.68%, 95% CI = 19.31-33.26) compared with cattle (21.35%, 95% CI = 18.74-24.22) and goats (7.72%, 95% CI = 5.72-10.34), (p = 0.047). At the individual livestock level, the risk was elevated in female livestock (OR: 1.74, 95% CI = 1.08-12.82) (p = 0.016), while at the herd level, areas receiving approximately 1001-1500 mm of rainfall (OR: 2.47, 95% CI = 1.14-5.37) (p = 0.022), areas of rainfall amount greater than approximately 1600 mm (OR: 2.239, 95% CI = 1.07-8.82) (p = 0.023), and mixed species herds (OR: 10.410, 95% CI = 3.04-35.59) (p = 0.001), were significant risk factors. The detection of IgM antibodies confirmed active circulation of RVFV in Malawi. Therefore, monitoring of RVF in animals, humans, and vectors using a "One Health" approach, along with community sensitization among the high-risk populations, could help mitigate the threat posed by this zoonotic disease in Malawi.

Knowledge, Attitudes, and Practices of Community Pharmacy Professionals on Poultry Antibiotic Dispensing, Use, and Bacterial Antimicrobial Resistance in Zambia: Implications on Antibiotic Stewardship and WHO AWaRe Classification of Antibiotics.

Globally, the inappropriate dispensing and use of antibiotics in animals has contributed to the development of bacterial antimicrobial resistance (AMR). In Zambia, there is insufficient information among community pharmacy professionals on antibiotic use (ABU) and AMR in food-producing animals. This study assessed community pharmacy professionals' knowledge, attitudes, and practices regarding poultry antibiotic dispensing, use, and bacterial AMR in the Lusaka district of Zambia. A cross-sectional study was conducted among 178 community pharmacy professionals between February and April 2022 using a semi-structured questionnaire. Data were analyzed using Stata version 17. Of the total participants (n = 178), 51.1% (n = 91) were pharmacists. The most dispensed antibiotic was oxytetracycline, a Watch antibiotic, mainly without prescriptions. Good knowledge of ABU and AMR was associated with work experience for more than one year (p = 0.016), while good practices were associated with male gender (p = 0.039) and work experience of more than one year (p = 0.011). The study found moderate knowledge, positive attitudes, and moderate practices of pharmacy professionals on poultry ABU and AMR. There was high dispensing of poultry antibiotics without prescriptions, which calls for strict implementation of antimicrobial stewardship and surveillance programs in poultry production in Zambia to reduce AMR.

Evidence of Community-Wide Spread of Multi-Drug Resistant Escherichia coli in Young Children in Lusaka and Ndola Districts, Zambia.

Increased antimicrobial resistance (AMR) has been reported for pathogenic and commensal Escherichia coli (E. coli), hampering the treatment, and increasing the burden of infectious diarrhoeal diseases in children in developing countries. This study focused on exploring the occurrence, patterns, and possible drivers of AMR E. coli isolated from children under-five years in Zambia. A hospital-based cross-sectional study was conducted in the Lusaka and Ndola districts. Rectal swabs were collected from 565 and 455 diarrhoeic and healthy children, respectively, from which 1020 E. coli were cultured and subjected to antibiotic susceptibility testing. Nearly all E. coli (96.9%) were resistant to at least one antimicrobial agent tested. Further, 700 isolates were Multi-Drug Resistant, 136 were possibly Extensively-Drug Resistant and nine were Pan-Drug-Resistant. Forty percent of the isolates were imipenem-resistant, mostly from healthy children. A questionnaire survey documented a complex pattern of associations between and within the subgroups of the levels of MDR and socio-demographic characteristics, antibiotic stewardship, and guardians' knowledge of AMR. This study has revealed the severity of AMR in children and the need for a community-specific-risk-based approach to implementing measures to curb the problem.

Assessment of Knowledge, Attitudes, and Practices towards Rift Valley Fever among Livestock Farmers in Selected Districts of Malawi.

Rift Valley fever (RVF) is a mosquito-borne viral zoonosis whose cases go unreported in endemic areas without active surveillance. Information on the knowledge, attitude, and practice of RVF among livestock farmers remains speculative in Malawi. A cross-section survey using a semi-structured questionnaire (n = 400) was conducted in eight districts of Malawi to capture information on knowledge, attitude, and management practices (KAP) regarding RVF. The average KAP score was calculated from total scores for knowledge, attitude, and practices and then assessed. The association between the level of knowledge and factors of knowledge, factors of attitude, and factors of practices was determined using Pearson chi-square. Multivariate analysis was used to determine the predictors of knowledge. Participants had an overall poor knowledge (score = 17.94%), negative attitude (score = 9.40%), and poor management practices (score = 41.23%) towards RVF. Only 8.25% (33/400) of participants had sufficient knowledge of RVF. The study found that the cause of abortion (OR: 3.86 (95% CI: 1.14-13.05)) (p = 0.030) and knowledge on transmission of RVFV (OR: 5.65 (95% CI: 1.76-18.12)) (p = 0.004) were predictors of insufficient knowledge of RVF. The current study reported that participants had insufficient knowledge and a negative attitude despite displaying limited management practices towards RVF. Therefore, this study recommends community sensitization to RVF and advocates for the importance of reporting suspected cases to relevant authorities for proper management.

Isolation, discrimination, and molecular detection of Listeria species from slaughtered cattle in Namwala District, Zambia.

BACKGROUND: The food industry is increasingly becoming more scrutinized, given the frequency and intensity with which zoonotic diseases are being reported. Pathogen tracking has become more applicable with regards food safety. It is in this regard that the present study was formulated to track Listeria species. in freshly slaughtered cattle carcasses by utilizing standard and molecular biological techniques. METHODS: A cross-sectional study design was conducted from March to December 2020 with 200 samples being equally collected in the rainy and dry seasons. A total of 180 and 20 swabs were aseptically collected from carcasses and the environment respectively. Samples were first subjected to pre-enrichment in half-strength Fraser broth followed by enrichment in full strength Fraser broth and subsequent plating on Listeria agar. Listeria growth characteristics were identified up to species level based on their morphological and biochemical characteristics. Further, molecular detection and phylogenetic analysis was conducted. Quantitative proportionate survey data were analyzed using Stata Version 15 software to estimate crude prevalence taking into account complex design at abattoir level. Factors associated with contamination were characterized using logistic regression. Sequences were analyzed using, Genetyyx version 12 and phylogenetic Mega. RESULTS: Of the 200 samples, 19 were positive for Listeria species identified as L.innocua 14/19 (73.7%) and L. monocytogenes 5/19 (26.3%). All isolates were from freshly slaughtered carcasses, and none from environment. Siginificant differences in contamination levels were observed based on season: rainy season yielded 14 (73.6%) whilst the dry season 5 (26.3%). The L. monocytogenes strains showed a high degree of homogeneity on phylogenetic analysis and clustered based on abattoir. Seasonality was identified as a major determinant influencing contamination based on the final logistic regression model. CONCLUSION: This study found evidence of L. monocytogenes contamination on traditionally raised beef carcasses across various abattoirs surveyed. The failure to find Listeria contamination on the abattoir environment may to a greater extent intimate cattle carccases as primary sources of contamination. However, a more comprerehnsive study incorporating different geographical regions is needed to conclusively ascertain these present findings.

Antibiotic Resistance Patterns of Listeria Species Isolated from Broiler Abattoirs in Lusaka, Zambia.

L. monocytogenes is a public health threat linked to fast foods such as broiler chickens. This study aimed to verify the occurrence of Listeria species in chickens from abattoirs and evaluate their antimicrobial resistance. In total, 150 broiler carcass swabs distributed as cloacal (n = 60), exterior surface (n = 60), and environmental (n = 30) were collected. Listeria species were characterized using biochemical tests and PCR. We conducted antibiotic resistance tests using the disc diffusion and Etest (Biomerieux, Durham, NC, USA) methods. Overall isolation of Listeria species was 15% (23/150) 95% CI (10.16-22.33), 2% (3/150) 95% CI (0.52-6.19) and 13% (20/150) 95% CI (8.53-20.08) came from environmental swabs and carcass swabs, respectively. Proportions of positive Listeria isolates were L. monocytogenes 74% (17/23), L. welshimeri 22% (5/23), and L. innocua 4% (1/23). Listeria species from the exterior carcass swabs was 61% (14/23), cloacal swabs 26% (6/23), and environmental swabs 3% (3/23). L. monocytogenes had the greatest resistance percentage to the following antibiotics: clindamycin (61%, 10/23), tetracycline 30% (7/23), and erythromycin 13%, (3/23). Isolation of L. monocytogenes in relatively high numbers, including the antimicrobial profiles, suggests a potential risk of the pathogen remaining viable in the food continuum and a public health risk to would-be consumers.

Awareness of Antimicrobial Resistance and Associated Factors among Layer Poultry Farmers in Zambia: Implications for Surveillance and Antimicrobial Stewardship Programs.

Antimicrobial resistance (AMR) is a global public health problem affecting animal and human medicine. Poultry production is among the primary sources of income for many Zambians. However, the increased demand for poultry products has led to a subsequent increase in antimicrobial use. This study assessed the awareness of AMR and associated factors among layer poultry farmers in Zambia. A cross-sectional study was conducted among 77 participants from September 2020 to April 2021. Data was analysed using Stata version 16.1. The overall awareness of AMR among the farmers was 47% (n = 36). The usage of antibiotics in layer poultry production was high at 86% (n = 66). Most antibiotics were accessed from agrovets (31%, n = 24) and pharmacies (21%, n = 16) without prescriptions. Commercial farmers were more likely to be aware of AMR compared to medium-scale farmers (OR = 14.07, 95% CI: 2.09-94.70), as were farmers who used prescriptions to access antibiotics compared to those who did not (OR = 99.66, 95% CI: 7.14-1391.65), and farmers who did not treat market-ready birds with antibiotics compared to those who did (OR = 41.92, 95% CI: 1.26-1396.36). The awareness of AMR among some layer farmers was low. Therefore, policies that promote the rational use of antibiotics need to be implemented together with heightened surveillance activities aimed at curbing AMR.

A global perspective of antibiotic-resistant Listeria monocytogenes prevalence in assorted ready to eat foods: A systematic review.

BACKGROUND AND AIM: Listeria monocytogenes in ready-to-eat (RTE) foods remains consistently under-reported globally. Nevertheless, several independent studies conducted to investigate have elucidated the prevalence and antibiotic resistance profiles of L. monocytogenes in RTE-associated foods and their antibiotic resistance profiles. Given the rapid increase in consumption of RTE foods of both animal and plant origin, it is imperative to know the prevalence deductive data focusing on how much of L. monocytogenes is present in RTE foods, which is critical for food safety managers and retailers to assess the possible risk posed to end-users. In addition, valuable insight and another angle to the depth of the problem, we conducted a systematic review and meta-analysis to synthesize available data regarding the prevalence of L. monocytogenes in RTE foods and antibiotic resistance profiles. MATERIALS AND METHODS: We conducted a meta-analysis study of L. monocytogenes and antibiotic resistance to clinically relevant antibiotics to determine the extent of L. monocytogenes contamination in RTE foods and antibiotic resistance profiles. The primary search terms, also known as keywords used, were restricted to peer-reviewed and review articles, and databases, including Google Scholars, Science-Direct, and Scopus, were searched. The inclusion of articles meeting eligibility criteria published between 2010 and 2020 after title, abstract, and full article screening. Data analysis was performed at multiple stages using quantitative meta-analysis reviews. RESULTS: L. monocytogenes pooled proportion/prevalence was highest in chicken products determined at (22%) followed by various but uncategorized RTE foods at 21%. Regarding antibiotic resistance, profiling's highest pooled prevalence resistance was observed in penicillin at 80% resistance, followed by cephalosporin at 47%. CONCLUSION: Within its limitations, this study has attempted to provide insight into the pooled proportion/prevalence of L. monocytogenes in RTE foods and the antibiotic resistance profile at the global level. Determining the proportion/prevalence of L. monocytogenes in RTE foods across the globe and antibiotic resistance profile is essential for providing quality food and reducing public health problems due to unsuccessful treatment of foodborne illness. This study provides insight into the pooled prevalence of L. monocytogenes in RTE foods and the antibiotic resistance profile. The results of this study partly endeavored to help appropriate authorities strengthen their preventive measures on specific RTE foods that are most likely to be contaminated with L. monocytogenes and antibiotic resistance profiles.

Evaluation of Bacterial Contamination in Dressed Chickens in Lusaka Abattoirs.

Objective: The aim of this study was to evaluate bacterial contamination and the risk factors associated with contamination of poultry during processing. Despite the rapid growth of the poultry industry, the presence of high levels of pathogenic bacteria contaminants, such as Escherichia coli and Salmonella, pose serious public health concerns in dressed chickens. These infections negatively affect the product's shelf life. Methods: A cross sectional design was used to study two main poultry abattoirs in Lusaka. The processing line was used to collect biological samples along with the acquisition of risk-associated data using a structured questionnaire. Data collected both from biological sources and the risk analysis were entered into Excel and analysed in STATA version 14 for windows. Results: Escherichia coli and Salmonella contamination was detected in 70 and 2.5% of the selected dressed chickens (n = 80), respectively. The number of total coliforms and Escherichia coli were observed to be significantly higher in samples from washed carcasses than pre-washed carcasses (65 and 35%, respectively). In addition, this study revealed that among the anthropogenic and exposure risk factors, bacterial contamination levels resulted mainly from a lack of hygienic practices. This included hand washing and an increased frequency of slaughters per day (>15,000). Conclusion: This study indicates that the water used for dressing chickens is probably the major cause of high levels of cross-contamination. The results also highlight the issues that need to be addressed to improve environmental and carcass hygiene in a poultry abattoir. Significance: Critical findings in this study are that contamination sources may be variable and hygienic practices may play a major role. In this particular study, the reuse of contaminated water was a case in point. Accordingly, there is need for both the water source and the water being used for processing to be tested.