Antimicrobial, Multi-Drug and Colistin Resistance in Enterobacteriaceae in Healthy Pigs in the Greater Accra Region of Ghana, 2022: A Cross-Sectional Study.

There is little published information on antimicrobial resistance (AMR) in animals in Ghana. We determined the prevalence and factors associated with AMR, multi-drug resistance (MDR-resistance to ≥3 antimicrobial classes) and colistin resistance in Enterobacteriaceae in healthy pigs in Accra, Ghana. Rectal swabs obtained from the pigs on 20 farms from January to March 2022, were examined for Escherichia coli, Enterobacter spp. and Klebsiella pneumoniae. AMR was determined using standard microbiological techniques and the mcr-1 gene detected through molecular analysis. Enterobacteriaceae were isolated from 197 of 200 pigs: these comprised 195 E. coli isolates, 38 Enterobacter spp. and 3 K. pneumoniae, either singly or combined. Over 60% of E. coli were resistant to tetracycline, with 27% and 34% being resistant to amoxicillin/clavulanic acid and ampicillin, respectively; 23% of E. coli and 5% of Enterobacter spp. exhibited MDR phenotypes. Phenotypic colistin resistance was found in 8% of E. coli and Enterobacter spp., with the mcr-1 gene detected in half. Our study findings should be incorporated into on-going AMR, MDR and colistin resistance surveillance programs in Ghana. We further advocate for tailored-specific education for pig farmers on animal antimicrobial use and for strengthened regulatory policy on antimicrobial usage and monitoring in the animal production industry.

Antimicrobial Resistance of Escherichia coli from Broilers, Pigs, and Cattle in the Greater Kumasi Metropolis, Ghana.

Globally, resistance to antimicrobial drugs in food animals is on the rise. Escherichia coli of livestock, though commensal in nature, serves as reservoir for antimicrobial resistance genes with the potential of disseminating them. This study sought to examine the antimicrobial resistance profiles of Escherichia coli in broilers, pigs, and cattle in the Kumasi Metropolis and undertake molecular characterisation of the resistances. Faecal E. coli isolates (n = 48) were obtained from 10 broiler farms, (n = 43) from 15 pig farms, and (n = 42) from cattle from the Kumasi Abattoir using standard bacteriological techniques. The Kirby-Bauer disc diffusion method was employed in testing the sensitivities of 133 E. coli isolates to 15 antimicrobials. All 48 isolates from broilers presented no resistance to amoxicillin/clavulanic acid and ceftiofur. A 100% resistance to meropenem was observed in pig and cattle isolates. Multidrug resistance (MDR) across animal groups was 95.8% (n = 46), 95.3% (n = 41), and 64.3% (n = 27) for broilers, pigs, and cattle, respectively. Twenty-eight isolates presenting phenotypic resistance to aminopenicillins and cephalosporins were screened for the presence of extended-spectrum beta-lactamase (ESBL) genes by PCR. One isolate from poultry and another from cattle tested positive for the blaCTX-M ESBL gene. There were no positives for the blaTEM and blaSHV ESBL genes. Commensal E. coli of food animal origin represents an important reservoir of antimicrobial resistance that transfers resistance to pathogenic and nonpathogenic microbes affecting humans and animals. There is an urgent need to institute routine surveillance for the establishment of the mechanisms and molecular orientation of resistance in these organisms.

Aflatoxins in randomly selected groundnuts (Arachis hypogaea) and its products from some local markets across Ghana: Human risk assessment and monitoring.

A random assessment and human risk analysis were conducted on 80 groundnut pastes and raw groundnuts from some local markets across the different agroecological zones of Ghana. Total aflatoxins (AFtotal) and aflatoxins (AFB1, AFB2, AFG1, and AFG2) were analyzed using the High-Performance Liquid Chromatography (HPLC) method. Out of 80 samples investigated, 49 (61.25 %) tested positive for AFB1 and ranged from 0.38 ± 0.04-230.21 ± 22.14 µg/kg. The same proportion was recorded for total aflatoxins (AFtotal) and ranged between 0.38 ± 0.02-270.51 ± 23.14 µg/kg. Limits of AFB1 and total aflatoxins (AFtotal) for the Ghana Standards Authority (GSA) (5 and 10 µg/kg) and the European Food Safety Authority (EFSA) (2 and 4 µg/kg), were used as checks. A total of 33 (41.25 %) samples were above the limits for both. Risk assessments recorded for Estimated Daily Intake (EDI), Margin of Exposure (MOE), potency, cancer risk, and population risks ranged 0.087-0.380 µg/Kg.bw/day, 1052.630-4597.700, 0-0.00396 ng Aflatoxins kg-1bwday-1 and, 1.5 × 10-3 - 7.9 × 10-4 respectively for total aflatoxins. While for aflatoxins B1 (AFB1), ranges of values of 0.068-0.300 µg/Kg.bw/day, 1333.33-5882.35, 0-0.00396 ng aflatoxins kg/bw/day and, 1.19 × 10-3 - 6.34 × 10-4 corresponded for Estimated Daily Intake (EDI), Margin of Exposure (MOE), potency, cancer risk, and population risk respectively. There were risks of adverse health effects involved in the consumption of groundnuts for all age groups investigated since MOE values were all below 10,000.

Exposure of domestic swine to influenza A viruses in Ghana suggests unidirectional, reverse zoonotic transmission at the human-animal interface.

Influenza A viruses (IAVs) have both zoonotic and anthroponotic potential and are of public and veterinary importance. Swine are intermediate hosts and 'mixing vessels' for generating reassortants, progenies of which may harbour pandemic propensity. Swine handlers are at the highest risk of becoming infected with IAVs from swine but there is little information on the ecology of IAVs at the human-animal interface in Africa. We analysed and characterized nasal and throat swabs from swine and farmers respectively, for IAVs using RT-qPCR, from swine farms in the Ashanti region, Ghana. Sera were also analysed for IAVs antibodies and serotyped using ELISA and HI assays. IAV was detected in 1.4% (n = 17/1,200) and 2.0% (n = 2/99) of swine and farmers samples, respectively. Viral subtypes H3N2 and H1N1pdm09 were found in human samples. All virus-positive swine samples were subtyped as H1N1pdm09 phylogenetically clustering closely with H1N1pdm09 that circulated among humans during the study period. Phenotypic markers that confer sensitivity to Oseltamivir were found. Serological prevalence of IAVs in swine and farmers by ELISA was 3.2% (n = 38/1,200) and 18.2% (n = 18/99), respectively. Human H1N1pdm09 and H3N2 antibodies were found in both swine and farmers sera. Indigenous swine influenza A viruses and/or antibodies were not detected in swine or farmers samples. Majority (98%, n = 147/150) of farmers reported of not wearing surgical mask and few (4%, n = 6) reported to wear gloves when working. Most (n = 74, 87.7%) farmers reported of working on the farm when experiencing influenza-like illness. Poor husbandry and biosafety practices of farmers could facilitate virus transmission across the human-swine interface. Farmers should be educated on the importance of good farm practices to mitigate influenza transmission at the human-animal interface.

Understanding attitude, practices and knowledge of zoonotic infectious disease risks among poultry farmers in Ghana.

Zoonotic infectious diseases (ZIDs) are increasing globally, and livestock farmers in low- and middle-income countries are at particularly high risk. An evaluation of farmer's behaviour on farms can be used to identify the risk factors and to develop tailored control strategies. This study documents the knowledge of zoonotic poultry diseases (ZPD) among 152 poultry farm workers (respondents) from 76 farms in the Ashanti region of Ghana and assessed their on-farm attitude and practices that increase their risk to exposure of ZPD. The median age of respondents was 29 years, 91.4% (n = 139) had a formal education, and 80.9% (n = 123) had worked on the farm for more than 1 year. The majority of farms (n = 69, 90.8%) had multiple flocks and 27.6% (n = 21) kept other animals, of which 57.1% (n = 12) were pigs. The majority of respondents had good knowledge about poultry diseases but not about ZPD. A higher level of education and longer work experience improved respondents' knowledge of poultry and ZPD. Although respondents identified the wearing of personal protective equipment (PPE) as a major ZPD preventive measure, the majority did not put that knowledge into practice. Most farms (71.1%, n = 54) had no footbath and 55.3% (n = 42) deposited farm-waste on the farm. While 97.4% (n = 148) of respondents washed their hands after working, only 48.7% (n = 74) wore protective footwear, 2.7% (n = 4) wore overalls, 2% (n = 3) wore nose masks and none (n = 0) wore gloves. The husbandry practices and attitude of farmers expose them to pathogens on the farm and increase their risk of becoming infected with ZPD in the sub-region. The results from this study could be used to promote human health among farm workers in Ghana.

Health risk assessment and levels of toxic metals in fishes (Oreochromis noliticus and Clarias anguillaris) from Ankobrah and Pra basins: Impact of illegal mining activities on food safety.

Arsenic (As), mercury (Hg), Cadmium (Cd) and lead (Pb) are toxic heavy metals that naturally occur in the ecosystem. Their levels are on the rise due to anthropogenic activities posing threat to aquatic wildlife and humans. In Ghana, pollution of some water bodies has led to unsafe consumption of riverine fishes as well as a shortage of treated potable water principally because the cost of treating polluted water has become expensive across the country. This study aimed to assess the As, Hg, Pb and Cd concentrations in water and fishes from rivers Pra and Ankobrah where activities of artisanal gold mining were carried out resulting in gross pollution of the water bodies. An experimental study was performed to ascertain the levels of As, Hg, Pb and Cd in fish species of Nile tilapia (Oreochromis noliticus) and mudfish (Clarias anguillaris) and aquatic media (water) from the Pra and River Ankobrah basins using the Atomic Absorption Spectrophotometer (AAS) (Varian AA240FS). Both river water samples recorded ranges of 0- 0.0040, 0.0060- 0.0387, 0 - 0.0020, 0.006-0.0093 mg/l for Cadmium, Lead, Arsenic and Mercury respectively. For Cadmium and Arsenic, their levels were comparable (p > 0.05). However, detected values for Lead and Mercury were no comparable (p < 0.05). Toxic metals concentrations in the rivers decreased in the order of Hg > Pb > Cd > As. For the fish samples, values ranged 0-0.08, 0.04-0.42, 0-0.04, and 0.40- 0.60 mg/kg for Cadmium, Lead, Arsenic and Mercury respectively. Generally, appreciably high values were obtained for Mercury. Toxic metals concentrations in the rivers decreased in the order of Hg > Pb > Cd > As. Human health risk assessment from heavy metal exposure through fish consumption from the Rivers for both children and adults showed no significant non-carcinogenic adverse health risk to humans since all calculated values for Hazard Quotient (HQ) were <1. Nonetheless, Target Hazard Quotient (THQ) values calculated for children and adult exposure to Cadmium and Mercury were>1 which implied a likely cause of adverse effects during a person's lifetime.

Potential health risk assessment of toxic metals contamination in clay eaten as pica (geophagia) among pregnant women of Ho in the Volta Region of Ghana.

INTRODUCTION: Geophagia although pleasurable and somewhat a necessity among pregnant women, also comes along with its own attendant problems such as exposure to potentially hazardous substances like bacteria, fungi, helminthes and ova, radioactive materials, and toxic elemental minerals in the soil depending on the geographical location. METHODOLOGY: This study evaluated the potential health risk involved during the exposure of pregnant women to toxic elemental minerals via the consumption of clay as pica (geophagia). Elemental mineral analysis was carried out using Buck Scientific 210VGP Flame Atomic Absorption Spectrophotometer (Buck Scientific, Inc. East Norwalk, USA). Risk assessment methods were also used to ascertain the various risks factors and the overall risk level. RESULTS: Concentrations of the macro elements investigated were 1.38 ± 1.5, 2.40 ± 1.5, 7.74 ± 1.5, 4.01 ± 1.0, 13.24 ± 2.2 and 13.76 ± 2.1 mg/Kg for iron (Fe), copper (Cu), zinc (Zn), potassium (K), magnesium (Mg) and sodium (Na) respectively. While that for the micro elements were 1.63 ± 0.03 µg/Kg, 4.72 ± 0.8, 0.53 ± 0.02 and 1.85 ± 0.3 mg/kg respectively for arsenic (As), manganese (Mn), lead (Pb) and nickel (Ni). Estimated Daily Intake (EDI), Hazard Quotient (HQ), Target Hazard Quotient (THQ) and Total Target Hazard Quotient (TTHQ) values ranged 0.611-5.44 (mg/kg Bw/day), 6.26 × 10- 4 - 106.5, 0.067-10.34 and 15 respectively. CONCLUSION: There is the likelihood of posing adverse health problems when clay samples obtained from Anfoega which is sited in the Volta region of Ghana is consumed due to the fact that the HQ's of these elemental minerals were > 1 which points to high content of Manganese (Mn) and Nickel (Ni). It is also likely to cause adverse health problems in an individual's life time since THQ for Arsenic, Lead and Nickel were above 1. Ultimately, the cumulative effect of these toxicants were exceedingly great (≤ 15) which implied a high level of unsafety associated with this clay. Per the results from this study, it is not safe for pregnant women to consume clay as pica since these toxic elements may cause detrimental effects on the foetus of the unborn child.

Serological and Molecular Surveillance of Infectious Bronchitis Virus Infection in Free-Range Chickens and Guinea Fowls in the Ga-East District of Ghana.

Infectious bronchitis is an economically important disease with worldwide distribution. Information is available on the presence of infectious bronchitis virus in commercial chicken in parts of Ghana but there is no information on free-range poultry and guinea fowls in the country. Possible IBV infections among free-range chickens and guinea fowls in Abokobi and Frafraha communities in the Ga-East district of the Greater Accra Region of Ghana were investigated using serology and PCR. Blood, tracheal, and cloacal swabs were obtained from 219 free-range chickens and guinea fowls with no respiratory symptoms and no history of IBV vaccination. Sera were evaluated for IBV antibodies by ELISA using commercial IBV test kit from IDEXX, Inc., USA. Swab samples were evaluated for S1 glycoprotein gene by one-step RT PCR. All the swab samples tested negative for IBV. 16% of all tested sera were positive for IBV. IBV seroprevalence in guinea fowls was 0%. 21.2% of sera from local chickens were positive for IBV. The seroprevalence of IBV among local chickens from Frafraha was 30% and that of Abokobi was 7.7%. This study shows exposure of local chickens in the study communities to IBV.

Geographically weighted regression of land cover determinants of Plasmodium falciparum transmission in the Ashanti Region of Ghana.

BACKGROUND: Malaria is a mosquito-borne parasitic disease that causes severe mortality and morbidity, particularly in Sub-Saharan Africa. As the vectors predominantly bite between dusk and dawn, risk of infection is determined by the abundance of P. falciparum infected mosquitoes in the surroundings of the households. Remote sensing is commonly employed to detect associations between land use/land cover (LULC) and mosquito-borne diseases. Due to challenges in LULC identification and the fact that LULC merely functions as a proxy for mosquito abundance, assuming spatially homogenous relationships may lead to overgeneralized conclusions. METHODS: Data on incidence of P. falciparum parasitaemia were recorded by active and passive follow-up over two years. Nine LULC types were identified through remote sensing and ground-truthing. Spatial associations of LULC and P. falciparum parasitaemia rate were described in a semi-parametric geographically weighted Poisson regression model. RESULTS: Complete data were available for 878 individuals, with an annual P. falciparum rate of 3.2 infections per person-year at risk. The influences of built-up areas (median incidence rate ratio (IRR): 0.94, IQR: 0.46), forest (median IRR: 0.9, IQR: 0.51), swampy areas (median IRR: 1.15, IQR: 0.88), as well as banana (median IRR: 1.02, IQR: 0.25), cacao (median IRR: 1.33, IQR: 0.97) and orange plantations (median IRR: 1.11, IQR: 0.68) on P. falciparum rate show strong spatial variations within the study area. Incorporating spatial variability of LULC variables increased model performance compared to the spatially homogenous model. CONCLUSIONS: The observed spatial variability of LULC influence in parasitaemia would have been masked by traditional Poisson regression analysis assuming a spatially constant influence of all variables. We conclude that the spatially varying effects of LULC on P. falciparum parasitaemia may in fact be associated with co-factors not captured by remote sensing, and suggest that future studies assess small-scale spatial variation of vegetation to circumvent generalised assumptions on ecological associations that may in fact be artificial.

Differing effects of HbS and HbC traits on uncomplicated falciparum malaria, anemia, and child growth.

The high prevalence of hemoglobin S (HbS) in Africa and hemoglobin C (HbC) in parts of West Africa is caused by the strong protection against severe falciparum malaria during childhood. Much less is known about the effect of HbS and especially HbC on Plasmodium falciparum infection, uncomplicated malaria, and anemia. A total of 1070 children from the Ashanti Region, Ghana, were enrolled at the age of 3 months and visited monthly until 2 years of age. The effects of the beta-globin genotype on the age-dependent incidence of malaria, levels of parasitemia, and hemoglobin as well as physical development were analyzed by population-averaged models. Infants with HbAS were protected from uncomplicated malaria (P < .005) and anemia (P < .001), had lower age-adjusted parasite densities (P < .001), and higher age-adjusted hemoglobin levels compared with children with the HbAA genotype (P = .004). In contrast, HbAC carriers had lower hemoglobin levels (P < .033) and were not protected against malaria or anemia. Notably, infants with HbAS were also significantly protected against stunting compared with carriers of HbAA or HbAC. This indicates differing mechanisms of protection against malaria of HbAS and HbAC and might help to understand why HbC is restricted to distinct areas of West Africa.