Knowledge, attitudes, practices and risk perception of cattle farmers in Namibia to antibiotic use and resistance.

The misuse of antibiotics in livestock contributes to the emergence of antibiotic resistant bacteria. The development of antibiotic resistance in bacteria of animal origin is of public health concern since drug resistant pathogens in animals can spread to humans. Several countries have therefore introduced regulations to restrict the use of antibiotics in livestock. Namibia was the first African country to ban the use of antibiotics as growth promoters and restrict the use of antibiotics to the treatment of bacterial diseases with a prescription from a veterinarian. However, there are limited studies on antibiotic stewardship among farmers in the country. The aim of this study was to determine the antibiotic awareness and usage of cattle farmers in Oshikoto region Namibia. A questionnaire on antibiotic knowledge, attitudes and practices (KAP) was administered to 274 randomly selected cattle farmers in Namibia. To calculate the KAP scores, one mark was awarded for each correct answer and zero was awarded for incorrect or unsure responses. Respondents having ≥ 50% correct answers were considered as having good KAP scores. The KAP score ranged from 8 to 73%. Forty-three per cent of the respondents had good KAP scores. Good KAP was associated with age, farm type and education status. Semi-commercial/commercial farmers had significantly better KAP than post and homestead farmers. The findings of this study indicate that there is a need to improve antibiotic usage and resistance education programmes in order to promote rational antibiotic usage.

Non-prescription sale and dispensing of antibiotics for prophylaxis in broiler chickens in Lusaka District, Zambia: findings and implications on one health.

Background: The irrational use of antibiotics in humans and livestock has contributed to the emergence of antimicrobial resistance (AMR). This study investigated the commonly sold and dispensed antibiotics for prophylaxis in broiler chickens in pharmacy and agro-veterinary shop personnel in the Lusaka District of Zambia. Methods: This cross-sectional study was conducted from August 2023 to October 2023 among 200 veterinary medicine dispensers in the Lusaka District of Zambia using a simulated farmer or mystery shopper approach. Data analysis was performed using IBM Statistical Package for Social Sciences version 23.0. Results: Out of the 200 medicine outlets investigated, 23 (11.5%) were agro-veterinary shops, while 177 (88.5%) were community pharmacies. A total of 165 community pharmacies and agro-veterinary shops provided veterinary services in the Lusaka District and sold medicines without prescription giving a 100% non-prescription sale. Of the 178 medicines dispensed for prophylaxis, 88.5% were antibiotics, while 13.5% were vitamins. The most dispensed antibiotic drug for prophylaxis in broiler chickens was oxytetracycline (30.34%), amoxicillin (17.98%) and gentamicin/doxycycline (10.67%). Conclusions: This study revealed a high sale and dispensing of antibiotics for prophylaxis in broiler chickens of which oxytetracycline, amoxicillin and gentamicin/doxycycline were commonly dispensed. There is a need for increased regulatory enforcement of selling antibiotics without a prescription as this may predispose poultry to the development of AMR and possible transmission of superbugs to humans and the environment. Educational interventions should be provided to community pharmacy and agro-veterinary personnel on adhering to antimicrobial stewardship practices when dispensing poultry antibiotics.

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.

Quantitative risk assessment for human Taenia solium taeniasis/cysticercosis exposure through consumption of pork in Mpwapwa District of Dodoma Region, Tanzania.

Background and Aim: Pig farming is a livelihood activity undertaken by many rural communities in Tanzania. However, pigs in rural communities become infected with Taenia solium, a zoonotic parasite leading to porcine cysticercosis (PCC). Thus, routine meat inspection is fundamental in ensuring that the meat consumed is PCC-free. However, routine meat inspection is constrained by low sensitivity as a diagnostic test. Regardless of its low sensitivity, at the local level, no substitute tool would potentially lead to meat being risk-free for human infection. This study aimed at quantifying the risk of humans getting exposed to T. solium taeniasis through the consumption of pork approved safe for human consumption by employing a quantitative risk assessment (QRA) model. Materials and Methods: A cross-sectional study employing a quantitative risk assessment technique was conducted to quantify the risk of human infection in Mpwapwa District through exposure to T. solium infection through consumption of officially inspected pork. The input parameters in this study were simulated in @risk software to obtain the risk of exposure and the risk factors for exposure to T. solium taeniasis. Results: The risk of one getting exposed to T. solium taeniasis/cysticercosis (TSTC) through consumption of pork approved for human consumption was found to be 0.018 (95% confidence interval [CI] = 0.00-0.0250). Likewise, the probability that a cyst is localized in a pork portion was found to be the most influencing input risk factor of getting exposed to TSTC. Furthermore, the probability of developing T. solium taeniasis was estimated to be 0.73605 (95% CI = 0-0.950) when infected undercooked pork portion is consumed and 0.99652 (95% CI = 0.98161-0.99908) from consuming raw pork portion. Likewise, about 47 (95% CI = 42-52) people who consumed undercooked pork and 26 (95% CI = 22-30) who consumed raw pork would get infected in Mpwapwa District per year. Conclusion: The results from this study are anticipated to create public awareness of the problem and promote the use of one-health approach in the control and prevention of the consumption of infected pork.

Human Health Risk Assessment from Lead Exposure through Consumption of Raw Cow Milk from Free-Range Cattle Reared in the Vicinity of a Lead-Zinc Mine in Kabwe.

Lead (Pb) contamination in the environment affects both humans and animals. Chronic exposure to Pb via dietary intake of animal products such as milk from contaminated areas poses a health risk to consumers; therefore, the present study investigated Pb contamination in cow milk and its health risk impact on humans through consumption of milk from cattle reared in the proximity of a Pb−Zn mine in Kabwe, Zambia. Fresh milk samples were collected from cows from Kang'omba (KN), Kafulamse (KF), Mpima (MP), Mukobeko (MK), and Munga (MN) farming areas. Pb determination was performed using Graphite Flame Absorption Atomic Spectrophotometry (GFAAS). Cow milk Pb levels showed different concentration patterns according to season, distance, and location of the farms from the Pb−Zn mine. The overall mean Pb levels were ranged 0.60−2.22 µg/kg and 0.50−4.24 µg/kg in the wet and dry seasons, respectively. The mean Pb concentration, chronic daily intake (CDIs), target hazard quotients (THQs), and incremental lifetime cancer risk (ILCR) results obtained were all within the permissible limits of 20 µg/kg, 3 and 12.5 µg/kg-BW/day, <1 and 10−4 to10−6, respectively. In conclusion, although Pb was detected in milk from cows reared in Kabwe, the health risk effects of Pb exposure associated with the consumption of milk in both adults and children were negligible.

Metallothionein from Wild Populations of the African Catfish Clarias gariepinus: From Sequence, Protein Expression and Metal Binding Properties to Transcriptional Biomarker of Metal Pollution.

Anthropogenic pollution with heavy metals is an on-going concern throughout the world, and methods to monitor release and impact of heavy metals are of high importance. With a view to probe its suitability as molecular biomarker of metal pollution, this study has determined a coding sequence for metallothionein of the African sharptooth catfish Clarias gariepinus. The gene product was recombinantly expressed in Escherichia coli in presence of Zn(II), Cd(II), or Cu, and characterised by Electrospray Ionisation Mass Spectrometry and elemental analysis. C. gariepinus MT displays typical features of fish MTs, including 20 conserved cysteines, and seven bound divalent cations (Zn(II) or Cd(II)) when saturated. Livers from wild C. gariepinus fish collected in all three seasons from four different sites on the Kafue River of Zambia were analysed for their metal contents and for MT expression levels by quantitative PCR. Significant correlations were found between Zn and Cu levels and MT expression in livers, with MT expression clearly highest at the most polluted site, Chililabombwe, which is situated in the Copperbelt region. Based on our findings, hepatic expression of MT from C. gariepinus may be further developed as a major molecular biomarker of heavy metal pollution resulting from mining activities in this region.

Sediment Metal Contamination in the Kafue River of Zambia and Ecological Risk Assessment.

Zambia's Kafue River receives wastes from various sources, resulting in metal pollution. This study determined the degree of contamination of 13 metals (Al, Cr, Mn, Fe, Co, Ni, Cu, Zn, As, Se, Cd, Hg and Pb) in Kafue River sediment and the associated ecological risks at six sites in three different seasons. The level of contamination for most metals showed significant site and seasonal differences. The contamination factor and pollution load index indicated that concentrations of most metals particularly copper (Cu), cobalt (Co), manganese (Mn) and arsenic (As) were very high at sites within the Copperbelt mining area. The geoaccumulation index showed an absence of anthropogenic enrichment with Cd and Hg at all the study sites and extreme anthropogenic enrichment with Cu at sites in the Copperbelt mining area. Potential ecological risk showed that Cu and As were likely to cause adverse biological effects to aquatic organisms in the Copperbelt mining region of the Kafue River.

Seroprevalence of Canine Parvovirus in Dogs in Lusaka District, Zambia.

Canine parvovirus (CPV) enteritis is a highly contagious enteric disease of young dogs. Limited studies have been done in Zambia to investigate the prevalence of CPV in dogs. Blood was collected from dogs from three veterinary clinics (clinic samples, n = 174) and one township of Lusaka (field samples, n = 56). Each dog's age, sex, breed, and vaccination status were recorded. A haemagglutination assay using pig erythrocytes and modified live parvovirus vaccine as the antigen was used. Antibodies to CPV were detected in 100% of dogs (unvaccinated or vaccinated). The titres ranged from 160 to 10240 with a median of 1280. Vaccinated dogs had significantly higher antibody titres compared to unvaccinated (p < 0.001). There was a significant difference in titres of clinic samples compared to field samples (p < 0.0001) but not within breed (p = 0.098) or sex (p = 0.572). Multiple regression analysis showed that only age and vaccination status were significant predictors of antibody titres. The presence of antibody in all dogs suggests that the CPV infection is ubiquitous and the disease is endemic, hence the need for research to determine the protection conferred by vaccination and natural exposure to the virus under local conditions.

A Study of Naturally Acquired Canine Babesiosis Caused by Single and Mixed Babesia Species in Zambia: Clinicopathological Findings and Case Management.

A retrospective and prospective analysis of clinical records of dogs diagnosed with Babesia infections was carried out for the years 2000 to 2013 from practices in Lusaka, Zambia. Records of 363 dogs with confirmed Babesia infections were analysed using demographic factors including sex, breed, age, and clinical signs in relation to haematological findings and Babesia species. The clinical and laboratory findings observed are described as well as Babesia species identification. The study included 18 breeds and the highest proportion were mongrels (32.2%), males representing 64.5% of the population. The most common presenting problems were anorexia (65.3%) and lethargy/weakness (65.3%). The most common clinical signs were fever (87.3%), pallor (52.3%), lymphadenopathy (47.4%), and presence of ticks (44.9%). Anaemia (96.4%) and nucleated erythrocytes (42.2%) were the most common laboratory findings. A mixed infection of Babesia rossi and Babesia gibsoni was present in 59.7% of dogs, whilst 8% and 32.2% had B. rossi and B. gibsoni as a single infection, respectively. Case management mainly involved therapy with tetracyclines and imidocarb and was usually accompanied by clinical improvement. This study highlights, for the first time, the presence of B. gibsoni in natural dog populations in Zambia, where previously only B. rossi was reported.

The nucleotide sequence of metallothioneins (MT) in liver of the Kafue lechwe (Kobus leche kafuensis) and their potential as biomarkers of heavy metal pollution of the Kafue River.

The study determined heavy metal concentrations and MT1 nucleotide sequence [phylogeny] in liver of the Kafue lechwe. Applicability of MT1 as a biomarker of pollution was assessed. cDNA-encoding sequences for lechwe MT1 were amplified by RT-PCR to characterize the sequence of MT1 which was subjected to BLAST searching at NCBI. Phylogenetic relationships were based on pairwise matrix of sequence divergences calculated by Clustal W. Phylogenetic tree was constructed by NJ method using PHILLIP program. Metals were extracted by acid digestion and concentrations of Cr, Co, Cu, Zn, Cd, Pb, and Ni were determined using an AAS. MT1 mRNA expression levels were measured by quantitative comparative real-time RT-PCR. Lechwe MT1 has a length of 183bp, which encode for MT1 proteins of 61AA, which include 20 cysteines. Nucleotide sequence of lechwe MT1 showed identity with sheep MT (97%) and cattle MT1E (97%). Phylogenetic tree revealed that lechwe MT1 was clustered with sheep MT and cattle MT1E. Cu and Ni concentrations and MT1 mRNA expression levels of lechwe from Blue Lagoon were significantly higher than those from Lochinvar (p<0.05). Concentrations of Cd and Cu, Co and Cu, Co and Pb, Ni and Cu, and Ni and Cr were positively correlated. Spearman's rank correlations also showed positive correlations between Cu and Co concentrations and MT mRNA expression. PCA further suggested that MT mRNA expression was related to Zn and Cd concentrations. Hepatic MT1 mRNA expression in lechwe can be used as biomarker of heavy metal pollution.

Metal and metalloid levels and bio-accumulation characteristics in soil, sediment, land plants and hippopotami (Hippopotamus amphibius L) from the South Luangwa National Park, Zambia.

Hippopotami (Hippopotamus amphibius L) are large semi-aquatic mammals that can be exposed to metals and metalloid from both terrestrial and aquatic environments. Therefore, knowledge of metal and metalloid accumulation characteristics in hippopotami living in the national park is important from ecotoxicological point of view. Levels of toxic metals (Cd, Pb and Hg) and metalloid (As) in hippopotami liver from the South Luangwa National Park in Zambia were far lower compared to the established values of toxic levels in cattle. No temporal variations of metal levels in hippopotami were observed, probably because of good management condition and the lack of anthropogenic activities around the national park. However, hippopotami liver accumulated significantly higher concentrations of Hg compared to soil, sediment and their food (plants), most likely due to a process of biomagnification throughout a trophic chain. Moreover, hippopotami liver and land plants showed significantly higher Cd levels than those of soil. These results strongly suggest that hippopotami liver accumulate higher levels of these metals if surrounding environment is contaminated. Levels of Cr and Ni in hippopotami liver were higher compared to other toxic metals. Since this is the first report to show the Cr and Ni levels and bio-accumulation characteristics of Hg and Cd in hippopotami, we concluded that continuous monitoring and evaluation of toxic effects of these metals on hippopotami should be conducted.