Samuel Kekeli Agordzo

Background: Antimicrobial resistance (AMR) remains a global health threat, and the use of antibiotics in animal husbandry, particularly poultry production, is well documented. There is, therefore, a need to investigate the prevalence of priority bacterial pathogens as reservoirs of AMR genes in poultry. We aimed to ascertain the prevalence and resistant profiles of Extended-spectrum Beta-lactamase (ESBL)-producing and Carbapenem-resistant E. coli in retail poultry in Ghana. Methods: One hundred and sixty-five (165) poultry caecal content samples were collected from three vendors. After overnight culture and purity plating on MacConkey agar, strain identification was performed using the VITEK 2 Compact System. Antibiotic susceptibility testing and confirmation of ESBL-producing and carbapenem-resistant strains were conducted with the combination disc test. The laboratory data and antibiogram generation were managed using WHONET 2024 software. Prevalence and antibiotic resistance patterns were analyzed and presented using both Microsoft Excel 365 and the AMR package in R Studio. Results: Of the 165 poultry caecal content samples examined, an overall E. coli prevalence of 88.5% (n=146) was obtained. The prevalence of ESBL-producing and carbapenem-resistant E. coli was 90.4% (n = 132/146) and 1.4% (n = 2/146), respectively. All isolates demonstrated 100% resistance to amoxicillin/clavulanic acid and temocillin, and 99.0% resistance to ampicillin and cefpodoxime. Susceptibility to ceftazidime/avibactam, meropenem, and tigecycline was 100%. Monthly, between 80% and 100% of the isolates were resistant to amoxicillin-clavulanic acid, ampicillin, aztreonam, cefpodoxime, cefepime, and temocillin throughout the year. Conclusions: The prevalence of multidrug-resistant ESBL-producing E. coli in the caecal contents of retail poultry is high. Also, the resistance patterns of these isolates to all antibiotics except ceftazidime/avibactam and tigecycline throughout the year are of great concern. These observations raise questions about the use of antibiotics in poultry farming in Ghana. While veterinary officers are encouraged to intensify education and monitoring, poultry retailers are advised to adhere to good hygiene practices.

Background and Aim. RNA extraction is a commonly used technique in molecular biology. In recent years, commercially available RNA extraction kits have largely replaced conventional approaches. However, these commercial kits are expensive and are not readily available in many resource-constrained institutions and laboratories. This study therefore compared the performance of the conventional acid guanidinium thiocyanate-phenol-chloroform (AGPC) extraction method to QIAamp® Viral RNA Mini Kit (QIAGEN, Cat. No. 52906) and OxGEn RNA Kit (OxGEn Molecular Solutions, GE-009) to build an in-house RNA extraction technique from blood and oral swab samples. Method. In a comparative experimental cross-sectional study, RNA was extracted from oral swabs and blood samples from 25 healthy individuals at the Department of Molecular Medicine, KNUST. RNA was extracted by the manual AGPC extraction method and commercial RNA extraction kits. The quantity (ng/μl) and purities (260/280 nm) of the extracted RNA were measured spectrophotometrically using the IMPLEN NanoPhotometer® N60. The presence of RNA in the extracts was confirmed using 2% agarose gel electrophoresis. Statistical analyses were conducted using R language. Results. The yield of RNA extracted from blood and oral swab samples using modified AGPC was significantly higher compared to the commercial methods ( < 0.0001). However, the purity of RNA extracted by the manual AGPC method from blood was significantly lower than the commercial methods ( < 0.0001). Moreover, the purity from oral swabs using the manual AGPC method was significantly lower compared to QIAamp ( < 0.0001) and the OxGEn kits method ( < 0.001). Conclusion. The modified manual AGPC method has a very high yield of RNA extracts using blood samples, which could serve as an alternate cost-effective method for RNA extraction in resource-limited laboratories; however, its purity may not be suitable for downstream processes. Moreover, the manual AGPC method may not be suitable for extracting RNA from oral swab samples. Future investigation is needed to improve the purity of the manual AGPC RNA extraction method and also confirmation of the obtained results by PCR amplification and RNA purity verification by sequencing.

This cross-sectional study seeks to investigate the bacterial contamination of poultry products (eggs) produced from selected farms and sold within the major markets in the New Juaben Municipality in the Eastern region of Ghana. A total of 100 eggs were sampled randomly from October to December, 2018 from five poultry farms and five retail markets in the municipality for microbial evaluation. The collected swabs of eggshells and its contents collected at various times of the day were cultured on suitable media and standard microbiological test were performed to identify the isolated organisms. The entire eggshell samples were positive for bacteria with only 13 (13%) having growth in the egg content. The genera of bacteria recorded are Staphylococcus, Escherichia, Klebsiella, Proteus and Citrobacter. The mean total viable count of bacteria from both shell and content sampled from poultry farms and retail markets were 13.2×10⁷cfu/ml and 8.0 ×10⁷cfu/ml (p>0.05) respectively. These were higher than the acceptable value of 10×10⁵ gm/cfu. The study also showed that the level of microbial contamination of the eggs was highest in the afternoon as compared to the morning and evening. The bacteria isolates and the sheer numbers from both the eggshells and the egg content confirm the poor hygienic conditions under which both the poultry farms and markets were kept. Even though serious human pathogens were not prevalent in this study, it is recommended that strict hygienic protocols be adhered to on the poultry farms and in the markets.

Background: We aimed at investigating the impact of malaria on the haematological parameters of residents from different demographic settlements in the Ashanti Region of Ghana. Malaria parasites trigger changes in certain haematological parameters, which may result in a number of clinical manifestations. Differences in demographic settlements, such as rural, peri-urban and urban settlements may also influence these changes, but this has not been extensively studied in Ghana. Methods: We conducted a hospital-based, cross-sectional study from January to December 2018 in three different settlements. A total of 598 participants were recruited. Blood smears were examined to detect and quantify malaria parasitaemia, while haematological parameters were measured using a haematology analyser. Results: Participants from the rural settlement had the highest malaria prevalence (21.3%) compared to urban (11.8%) and peri-urban areas (13.3%); however, the peri-urban area had the highest median parasite density (568; IQR=190.0-1312.0). Age was significantly associated with the odds of malaria positivity (OR: 0.97; CI:0.96 — 0.99; p=4.96*10-4). When haematological parameters of the malaria-infected study participants were compared to the parameters of uninfected participants, red blood cell count (p=0.017), haemoglobin (p=0.0165), haematocrit (p=0.0015), mean corpuscular volume (p=0.0014), plateletcrit (p<0.0001) and platelet count (p<0.0001) were all significantly lower in the malaria infected group. In addition to age, haemoglobin and plateletcrit levels were also inversely correlated with the odds of testing positive for malaria, suggesting that children who were anaemic and/or thrombocytopaenic were likely to be infected. After fitting the data to a logistic regression model comprising the three variables, the model correctly categorised 78% of uninfected study participants, but only 50% of the malaria-positive participants. Conclusions: Study participants who were positive for malaria were younger and had low haemoglobin and plateletcrit levels compared to uninfected individuals. Further studies are needed to more precisely elucidate the relationship between malaria infection,demographic and haematological parameters.

Background: Toxoplasma gondii is an obligate, intracellular, apicomplexan parasite that causes toxoplasmosis. Although the global prevalence of toxoplasmosis has been estimated to be approximately 30%, there is limited seroprevalence data in Ghana, with a dearth of information on the impact of T. gondii on haematological parameters in exposed persons. Methods: Questionnaires were administered to 300 consenting individuals to obtain demographic information and assessment of their risk of exposure to T. gondii. Using anti- T. gondii IgG/IgM combo test kits, seropositivity to parasite-specific IgG and/or IgM was determined. A haematological analyser was used to measure haematological parameters. Results: There was an overall seroprevalence of 50.3% (n=151), with 49.7% (n=149) of the study participants seropositive for IgG and 1% (n=3) testing positive for IgM. Furthermore, the observed seroprevalence among pregnant women was 56.4% (n=62). With regard to settlement type, a seroprevalence of 55.6% was observed in the rural community, 50.6% in the peri-urban community and 47.1% in the urban community. The study identified cat ownership, contact with cat litter, contact with raw meat [RR (95% CI: 1.76 (1.23-2.53), 1.66 (1.03-2.67), 1.25(1.00-1.57)] and age (p<0.001) as risk factors for infection. Analyses of haematological data revealed significant reduction in the white blood cell, lymphocytes and mean corpuscular volume levels in seropositive males (p=0.0223, 0.0275, and 0.0271) respectively. Only the mean corpuscular volume of seropositive females reduced significantly as compared to the seronegative counterparts (p=0.0035). Conclusions: About half of the study population, including women of reproductive age carried antibodies against T. gondii, raising concerns about the risk of congenital toxoplasmosis and anaemia. We, therefore, recommend that screening for Toxoplasma gondii be included in the routine screening of pregnant women seeking antenatal care and further investigation should be conducted on the haematological implications of infection in humans.

Background: We aimed at investigating the impact of malaria on the haematological parameters of residents from different demographic settlements in the Ashanti Region of Ghana. Malaria parasites trigger changes in certain haematological parameters, which may result in a number of clinical manifestations. Differences in demographic settlements, such as rural, peri-urban and urban settlements may also influence these changes, but this has not been extensively studied in Ghana. Methods: We conducted a hospital-based, cross-sectional study from January to December 2018 in three different settlements. A total of 598 participants were recruited. Blood smears were examined to detect and quantify malaria parasitaemia, while haematological parameters were measured using a haematology analyser. Results: Participants from the rural settlement had the highest malaria prevalence (21.3%) compared to the urban (11.8%) and peri-urban areas (13.3%); however, the peri-urban area had the highest median parasite density (568; IQR=190.0-1312.0). Age was significantly associated with the odds of malaria positivity (OR: 0.97; CI:0.96 — 0.99). When haematological parameters of the malaria-infected study participants were compared to the parameters of uninfected participants, red blood cell count (p=0.017), haemoglobin (p=0.0165), haematocrit (p=0.0015), mean corpuscular volume (p=0.0014), plateletcrit (p<0.0001) and platelet count (p<0.0001) were all significantly lower in the malaria infected group. In addition to age, haemoglobin and plateletcrit levels were also inversely correlated with the odds of testing positive for malaria, suggesting that children who were anaemic and/or thrombocytopaenic were likely to be infected. After fitting the data to a logistic regression model comprising the three variables, the model correctly categorised 78% of uninfected study participants, but only 50% of the malaria-positive participants. Conclusions: Study participants who were positive for malaria were younger and had low haemoglobin and plateletcrit levels compared to uninfected individuals. Further studies are needed to more precisely elucidate the relationship between malaria infection,demographic and haematological parameters.

Background: Toxoplasma gondii is an obligate, intracellular, apicomplexan parasite that causes toxoplasmosis. Although the global prevalence of toxoplasmosis has been estimated to be approximately 30%, there is limited seroprevalence data in Ghana, with a dearth of information on the impact of T. gondii on haematological parameters in exposed persons. Methods: Questionnaires were administered to 300 consenting individuals to obtain demographic information and assessment of their risk of exposure to T. gondii. Using anti-T. gondii IgG/IgM combo test kits, seropositivity to parasite-specific IgG and/or IgM was determined. A haematological analyser was used to measure haematological parameters. Results: The participants included 58 males and 242 females, and ranged in age from 6 months to 84 years, with a median age of 27 years. There was an overall seroprevalence of 50.3% (n=151), with 49.7% (n=149) of the study participants seropositive for IgG and 1% (n=3) testing positive for IgM. Furthermore, the observed seroprevalence among pregnant women was 56.4% (n=62). With regards to the different communities in which the hospitals were located, a seroprevalence of 55.6% was observed in the rural community, 50.6% in the peri-urban community and 47.1% in the urban community. The study identified cat ownership, contact with cat litter [RR (95% CI: 1.76 (1.23-2.53), 1.66 (1.03-2.67), 1.25(1.00-1.57)] and age (p<0.001) as risk factors for infection. Analyses of haematological data also revealed significant differences between the red blood cell counts (p=0.038) and mean corpuscular volumes (p=0.0007) of seropositive and seronegative study participants. Conclusions: About half of the study population, including a significant number of women of reproductive age carried antibodies against T. gondii, raising questions about the risk of congenital toxoplasmosis, as well as possible links to anaemia. We, therefore, recommend that screening for Toxoplasma gondii be included in the routine screening of pregnant women seeking antenatal care.

Background: This study aimed at investigating haematological changes in malaria patients across different demographic settlements. Malaria parasites trigger changes in certain haematological parameters, which may result in a number of clinical manifestations. Differences in demographic settlements, such as rural, peri-urban and urban settlements, may also influence these changes, but this has rarely been studied. Methods: We conducted a hospital-based, cross-sectional study from January to December 2018 in three different settlements. A total of 598 participants were recruited. Giemsa-stained blood smears were examined to detect and quantify malaria parasitaemia, while haematological parameters were measured using a haematology analyser. Results: The rural settlement had the highest malaria prevalence compared to the other study communities (p=0.009). The difference in parasite densities across the three communities was also significant (p=0.0149). When the malaria-infected population was compared to the uninfected, there were differences in red blood cell count (p=0.0170), haemoglobin levels (p=0.0165), mean corpuscular volume (p=0.0139) and platelet counts (p<0.0001). The difference in median white blood cell (p-value <0.0001), neutrophil (p-value <0.0001) and lymphocyte (p-value <0.0269) count were significantly higher in infected patients from the peri-urban area compared to malaria patients from the rural and urban areas. There were also significant differences in platelet (p=0.0002), plateletcrit (p=0.0041), mean platelet volume (p=0.0009) and platelet large cell ratio (p=0.0046) levels between patients from the urban, peri-urban and rural areas. Conclusions: Patients infected with malaria generally had low red blood cell, haemoglobin and platelets in comparison to uninfected patients. There were also significant differences in several haematological parameters between malaria-infected patients from the three demographic settlements. Atypical results from routine haematological assays, especially findings of anaemia and thrombocytopenia, may be indicative of malaria and, in cases where the infection is asymptomatic, may improve diagnosis by prompting a more thorough search for the parasite in the peripheral circulation.