Knowledge, attitudes and practices towards yaws in endemic areas of Ghana, Cameroon and Côte d'Ivoire.

Yaws, caused by Treponema pallidum ssp. pertenue, remains a significant public health concern in tropical regions of West Africa and the South Pacific, primarily affecting children in remote areas with limited access to hygiene and sanitation. In this study, conducted in three endemic countries of West Africa where yaws remains a significant public health concern (Ghana, Cameroon, and Côte d'Ivoire), we aimed to assess the knowledge, attitudes, and practices related to yaws among community members, community health workers (CHWs), and traditional healers. The study revealed variations in the perception of causes of yaws among community members: the majority or participants in Ghana attributed yaws to germs (60.2%); in Cameroon the most reported form of transmission was contact with or drinking infected water sources (44.6%); and in Côte d'Ivoire both of these answers were also the most prevalent (60.3% germs and 93.% water sources). A substantial proportion of participants in Côte d'Ivoire also associated yaws with witchcraft and divine punishment (44.8%). Only a small proportion of individuals in Ghana and Côte d'Ivoire correctly identified contact with an infected person as a form of transmission (11.9% and 20.7%, respectively) and less than half in Cameroon (42.6%), although more than 98% of all participants reported avoidance behaviours towards yaws infected people due to fear of getting infected. Most participants expressed a preference for seeking care at hospitals (49.2%, 60.6%, 86.2%) or health care professionals including doctors and nurses (58.5%, 41,5% and 17.2%) if they were diagnosed with yaws, although a quarter of participants in Côte d'Ivoire also sought support from traditional healers. The CHWs interviewed were generally well-trained on yaws causes and treatment options, although they often reported low availability of treatment and diagnostic tests for yaws. Our findings underscore the need for community education, awareness campaigns, ongoing CHW training, and improved access to yaws treatment and diagnostic resources. The data also suggest that collaboration with traditional healers, who usually hold a highly esteemed position in the society, such as giving training on yaws causes and transmission or exchanging knowledge on treatment options, could be beneficial in certain regions, particularly in Côte d'Ivoire.

Using 16s rRNA sequencing to characterize the microbiome of tropical cutaneous ulcer disease: insights into the microbial landscape and implications for diagnosis and treatment.

Cutaneous ulcers are common in yaws-endemic areas. Although often attributed to 'Treponema pallidum subsp. pertenue' and Haemophilus ducreyi, quantitative PCR has highlighted a significant proportion of these ulcers are negative for both pathogens and are considered idiopathic. This is a retrospective analysis utilising existing 16S rRNA sequencing data from two independent yaws studies that took place in Ghana and the Solomon Islands. We characterized bacterial diversity in 38 samples to identify potential causative agents for idiopathic cutaneous ulcers. We identified a diverse bacterial profile, including Arcanobacterium haemolyticum, Campylobacter concisus, Corynebacterium diphtheriae, Staphylococcus spp. and Streptococcus pyogenes, consistent with findings from previous cutaneous ulcer microbiome studies. No single bacterial species was universally present across all samples. The most prevalent bacterium, Campylobacter ureolyticus, appeared in 42% of samples, suggesting a multifactorial aetiology for cutaneous ulcers in yaws-endemic areas. This study emphasizes the need for a nuanced understanding of potential causative agents. The findings prompt further exploration into the intricate microbial interactions contributing to idiopathic yaw-like ulcers, guiding future research toward comprehensive diagnostic and therapeutic strategies.

Management of diseases in a ruminant livestock production system: a participatory appraisal of the performance of veterinary services delivery, and utilization in Ghana.

INTRODUCTION: Sustainable livestock production remains crucial for attainment of food security globally and for safeguarding the livelihoods of many households in low- and -middle income countries. However, the high prevalence of infectious livestock diseases, coupled with inadequate provision and adoption of effective control measures, leads to reduced livestock productivity, increased animal mortalities, and emergence of antimicrobial resistant pathogens. This study sought to assess the management strategies employed by farmers for priority diseases affecting their animals and the utilization and performance of veterinary services. METHODS: We conducted the study in three districts, namely, Mion, Pru East, and Kwahu Afram Plains South Districts, which represent the main livestock production belts in Ghana. We used questionnaires in surveys, to collect pertinent data from 350 ruminant livestock farmers and 13 professional veterinary officers (VOs) in the study districts. Additionally, we conducted seven focus group discussions (FGDs) with 65 livestock farmers in the study districts. The survey data was analyzed, and we describe the distribution of the priority livestock diseases, the disease management strategies employed, and the performance of veterinary services in Ghana. We also analyzed the raw FGD transcript texts deductively based on the study objectives. To validate findings across the different datasets, we used triangulation. RESULTS: Almost all the farmers (98%) reared small ruminants, with about 25% also rearing cattle. The main priority livestock diseases identified includes pestes-des-petits-ruminants and mange infection in sheep and goats, as well as contagious bovine pleuropneumonia and foot-and-mouth-disease in cattle. We found that majority (82%) of the farmers relied on treatment, while only 20% opted for vaccination services. Additionally, the veterinary system in Ghana did not adequately regulate the antimicrobial medications employed by farmers to manage diseases. Thus, in most of the cases, the medicines applied by farmers were not useful for the target diseases. Although our findings show the farmers perceived VOs to perform highly compared to informal providers on most of the attributes evaluated including medicine availability and quality, treatment effectiveness, advisory services, service affordability, and competence, only 33% utilized VOs services. The majority of the farmers (51%) used the services of informal providers, who were better in proximity and popularity with farmers. CONCLUSIONS: The livestock sector in Ghana faces a substantial challenge due primarily to vaccine-preventable diseases. Even though VOs demonstrated superior performance on key veterinary service performance indicators, their services are underutilized by livestock farmers. Additionally, the absence of regulatory oversight by the veterinary system over antimicrobials utilized in animal production contributes to their misapplication by livestock farmers, posing a considerable risk to both public health and food security. It is thus imperative to introduce new initiatives that enhance the uptake of animal vaccines and better antimicrobial stewardship to ensure sustainable livestock production.

Farmers' valuation and willingness to pay for vaccines to protect livestock resources against priority infectious diseases in Ghana.

INTRODUCTION: Livestock vaccination coverage rates remain low in many lower and middle income countries despite effective vaccines being commonly available. Consequently, many preventable infectious livestock diseases remain highly prevalent, causing significant animal mortalities and threatening farmers' livelihood and food security. This study sought to assess farmers' maximum willingness to pay (WTP) for contagious bovine pleuropneumonia (CBPP), and peste-des-petits-ruminants (PPR) vaccination of cattle, and sheep and goats, respectively. METHODS: Overall, 350 ruminant livestock farmers were randomly selected from three districts located in the northern, middle and southern farming belts of Ghana. We implemented a double-bounded dichotomous contingent valuation experiment, where farmers indicated their WTP for vaccinating each livestock specie(s) owned at randomly assigned price points. WTP responses were analyzed using maximum likelihood estimation, and factors influencing WTP were assessed using censored regression analysis accounting for village-level clustering. RESULTS: Mean WTP for CBPP vaccination was USD 1.43 or Ghanaian Cedi (GHC) 8.63 (95% CI: GHC 7.08-GHC 10.19) per cattle. Mean WTP for PPR vaccination was USD 1.17 or GHC 7.02 (95% CI: GHC 5.99-GHC 8.05) per sheep, and USD 1.1 or GHC 6.66 (95% CI: GHC 5.89-GHC 7.44) per goat. WTP was positively associated with resilience, limited knowledge about vaccines (assessed prior to WTP experiment), farmland size, and male gender, after adjusting for other covariates. To attain 70% vaccination coverage in Ghana, vaccination costs should be no larger than GHC 5.30 (USD 0.88) for CBPP per cattle and GHC 3.89 (USD 0.65) and GHC 3.67 (USD 0.61), respectively, for PPR vaccines per sheep and goat. CONCLUSIONS: Ruminant livestock farmers in Ghana value vaccination highly, and are, on average, willing to pay vaccination costs that exceed the prevailing market prices (GHC 6 for CBPP and GHC 5 for PPR vaccination) to protect their livestock resources. To achieve 70% coverage, only minor subsidies would likely be required. These results suggest that effective disease control in these settings should be possible with appropriate distribution strategies.

Relationship between animal health and livestock farmers' wellbeing in Ghana: beyond zoonoses.

INTRODUCTION: Livestock production is a key livelihood source for many people in developing countries. Poor control of livestock diseases hamper livestock productivity, threatening farmers' wellbeing and food security. This study estimates the effect of livestock mortalities attributable to disease on the wellbeing of livestock farmers. METHODS: Overall, 350 ruminant livestock farmers were randomly selected from three districts located in the north, middle and southern belts of Ghana. Mixed-effect linear regression models were used to estimate the relationship between animal health and farmer wellbeing. Farmer wellbeing was assessed using the WHOQOL-BREF tool, as the mean quality-of-life in four domains (physical, psychological, social, and environmental). Animal health was assessed as annual livestock mortalities to diseases adjusted for herd size, and standardized in tropical livestock units to account for different ruminant livestock species. We adjusted for the potential confounding effect of farmers' age, sex, educational attainment, farmland size, socio-economic status, perception of disease risk to herd, satisfaction with health, previous experience of disease outbreaks in herds, and social support availability by including these as fixed effects, and community as random effects, in a pre-specified model. RESULTS: Our results showed that farmers had a median score of 65.5 out of 100 (IQR: 56.6 to 73.2) on the wellbeing scale. The farmers' reported on average (median) 10% (IQR: 0 to 23) annual herd mortalities to diseases. There was a significantly negative relationship between increasing level of animal disease-induced mortality in herds and farmers' wellbeing. Specifically, our model predicted an expected difference in farmers' wellbeing score of 7.9 (95%CI 1.50 to 14.39) between a farmer without any herd mortalities to diseases compared to a (hypothetical) farmer with 100% of herd mortalities caused by diseases in a farming year. Thus, there is a reduction of approximately 0.8 wellbeing points of farmers, for the average of 10% disease-induced herd mortalities experienced. CONCLUSIONS: Disease-induced livestock mortalities have a significant negative effect on farmers' wellbeing, particularly in the physical and psychological domains. This suggests that veterinary service policies addressing disease risks in livestock, could contribute to improving the wellbeing of livestock dependent populations, and public food security.

Microbiological quality and antimicrobial resistance of Bacteria species recovered from ready-to-eat food, water samples, and palm swabs of food vendors in Accra, Ghana.

This study sought to investigate microbial quality and antimicrobial resistance of bacteria species from Ready-to-Eat (RTE) food, water, and vendor palm swab samples. Between 2019 and 2020, RTE food, water and vendor palm swab samples were collected from food vending sites in Accra, Ghana. Samples were cultured and confirmed using the Matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF). Antimicrobial susceptibility testing (AST) was conducted using disk diffusion method. Beta-lactamase and Diarrheagenic Escherichia coli (DEC) genes were determined using Polymerase Chain Reaction (PCR). Total plate count (TPC) and Total coliform count (TCC) were performed on food and water samples. In total, 179 RTE food, 72 water and 10 vendor palm swab samples were collected. Enterobacter spp. (16.8 %), Citrobacter spp. (10.1 %), Enterococcus faecalis (7.8 %), Pseudomonas spp. (6.7 %) and Klebsiella pneumoniae (4.0 %) occurred in food. Isolates from water and palm were Klebsiella pneumoniae (20.8 %), Aeromonas spp. (16.7 %) and Enterobacter cloacae (11.1 %). Resistance to Amoxicillin-clavulanate, Tetracycline, Azithromycin, Sulfamethoxazole-trimethoprim, and Nitrofurantoin were common among Enterobacterales. High mean TPC and TCC showed in some RTE food and different water types used in vending depicting their unsafe condition for consumption and usage. The blaSHV and blaTEM genes were present in some Enterobacterales from food and water. The lt gene was identified in two food samples. AMR organisms associated with nosocomial infections in the samples investigated, calls for continuous surveillance in the food industry in Ghana. Also, the unsafe outcome of RTE food and water depicts the need for the enforcement of Ghana's food safety laws.

Access to vaccination services for priority ruminant livestock diseases in Ghana: Barriers and determinants of service utilization by farmers.

INTRODUCTION: Livestock diseases are a major constraint to agricultural productivity, frequently causing significant livelihood losses for farmers, and negatively affecting public food safety and security. Vaccines provide an effective and profitable means for controlling most infectious livestock diseases, but remain underutilized. This study sought to assess the barriers and determinants of vaccination utilization for priority livestock diseases in Ghana. METHODS: We conducted a mixed-method study involving a quantitative survey with ruminant livestock farmers (N = 350) and seven focus group discussions (FGD) involving 65 ruminant livestock farmers. The survey data were analyzed, and distribution of barriers to vaccination access described. We evaluated the determinants of vaccination utilization (any use of vaccination against contagious-bovine-pleuropneumonia (CBPP) and peste-des-petits-ruminants (PPR) in 2021) using logistic regression analyses at the 0.05 significance level. FGD transcripts were analyzed deductively. We used triangulation to achieve convergence across the different datasets and analyses. RESULTS: The farmers kept an average (median) of 5 tropical livestock units (TLUs) of ruminant livestock (IQR=2.6-12.0) that were on average 8 kilometers (IQR=1.9-12.4) away from veterinary officers (VOs). Only 16% (56/350) of herds were vaccinated against the diseases. Most farmers (274/350) had limited knowledge on vaccines against CBPP and PPR infections, 63% (222/350) perceived low risk of these diseases to their herds. About half of farmers reported experiencing outbreaks of either disease in the study year (2021). Farmers scored on average 80.5 out of 98 (IQR=74-85) on the RS-14 resilience scale. After adjusting for farmers' livestock rearing experience, herd size, sex, wealth status, distance to VOs, previous disease outbreaks, and perceived risk of the diseases, vaccination utilization was negatively associated with limited knowledge (aOR=0.19, 95%CI=0.08-0.43), and positively associated with personal exposure to outbreaks in the study year (aOR=5.26, 95%CI=2.01-13.7) and increasing resilience (aOR=1.13, 95%CI=1.07-1.19). FGDs revealed farmer misconceptions about vaccines, costs of vaccines, and timely access to vaccines from VOs as additional barriers. CONCLUSIONS: Acceptability, affordability, accessibility, and availability of vaccine services represent the main barriers to vaccines utilization by ruminant livestock farmers in Ghana. Given that limited knowledge regarding the value of vaccination and shortfalls in veterinary service supply are of central importance for both the demand and supply side, more collaboration between the different stakeholders in a transdisciplinary manner to effectively address the low vaccination utilization problem is needed.

Effectiveness and profitability of preventive veterinary interventions in controlling infectious diseases of ruminant livestock in sub-Saharan Africa: a scoping review.

Agriculture in general, and livestock production in particular, serve as a livelihood source for many people in sub-Saharan Africa (SSA). In many settings, lack of control of infectious diseases hampers livestock productivity, undermining the livelihood of rural populations. This scoping review sought to identify veterinary interventions previously evaluated as well as their relative effectiveness in controlling infectious livestock diseases. To be included, papers had to be written in English, German or French, and had to describe the effectiveness and/or profitability of preventive veterinary intervention(s) against anthrax, blackleg, bovine tuberculosis, brucellosis, contagious bovine pleuropneumonia, contagious caprine pleuropneumonia, foot-and-mouth disease, goat pox, lumpy skin disease, pasteurellosis, peste des petits ruminants, and/or sheep pox in any SSA country. Of the 2748 publications initially screened, 84 met our inclusion criteria and were analyzed. Most of the studies (n = 73, 87%) evaluated the effectiveness and/or profitability of vaccination, applied exclusively, applied jointly with, or compared to strategies like deworming, antimicrobial treatment, surveillance, feed supplementation, culling and dipping in reducing morbidity and/or mortality to livestock diseases. The effectiveness and/or profitability of antimicrobial treatment (n = 5), test and slaughter (n = 5), and use of lay animal health workers (n = 1) applied exclusively, were evaluated in the other studies. Vaccination was largely found to be both effective and with positive return on investment. Ineffective vaccination was mainly due to loss of vaccine potency under unfavorable field conditions like adverse weather events, cold chain failure, and mismatch of circulating pathogen strain and the vaccines in use.In summary, vaccination is the most effective and profitable means of controlling infectious livestock diseases in SSA. However, to achieve effective control of these diseases, its implementation must integrate pathogen surveillance, and optimal vaccine delivery tools, to overcome the reported field challenges.

Magnitude and associated factors of latent tuberculosis infection due to Mycobacterium tuberculosis complex among high-risk groups in urban Bobo-Dioulasso, Burkina Faso.

Objectives: To determine the prevalence and risk factors for latent tuberculosis infection (LTBI) among three high-risk groups - household contacts of TB index cases, healthcare workers and slaughterhouse workers - in Bobo-Dioulasso, Burkina Faso. Methods: Participants were recruited to this cross-sectional study from March to July 2020 after giving informed consent. Sociodemographic, clinical and biological data were collected using a structured questionnaire. The QuantiFERON-TB Gold Plus test (QFT-Plus) and the tuberculin skin test (TST) were used for detection of LTBI. Bivariate and multivariate logistic regression analyses were performed to identify risk factors for LTBI. Results: The prevalence of LTBI among 101 participants (age range 15-68 years) was 67.33% [95% confidence interval (CI) 57.27-76.33] and 84.16% (95% CI 75.55-90.66) based on QFT-Plus and TST results, respectively. Compared with healthcare workers and household contacts of TB index cases, the prevalence of LTBI among slaughterhouse workers was significantly higher for both QTF-Plus (96.8%; P<0.001) and TST (100%; P=0.003). Working in a slaughterhouse [adjusted odds ratio (AOR) 1.095, 95% CI 1.00-2.036], smoking (AOR 4.214, 95% CI 1.051-16.899), ≥15 years of exposure (AOR 5.617, 95% CI 1.202-32.198), having an animal at home (AOR 2.735, 95% CI 1.102-6.789) and protozoal infection (AOR 2.591, 95% CI 1.034-6.491) were significantly associated with LTBI on the QFT-Plus assay. Conclusion: The prevalence of LTBI was high in all three groups, particularly slaughterhouse workers. The risk factors identified could form the basis of targeted intervention.

Microbial Safety of Beef Along Beef Value Chains in the Ashaiman Municipality of Ghana.

Food from animal sources continues to be a significant food safety hazard. This study determined the microbial quality and safety of beef along beef value chains with case studies in the Ashaiman Municipality of Ghana. Raw beef samples were collected from four slaughter slabs in the Ashaiman Municipality and analyzed using standard microbiological methods to determine the quality and prevalence of specific pathogens, including Salmonella species, Listeria monocytogenes (L. monocytogenes), and Brucella species, as well as Toxoplasma gondii (T. gondii), Cyclospora cayetanensis (C. cayetanensis), and Cryptosporidium parvum (C. parvum). Data regarding food safety knowledge and practices were collected and observed from stakeholders (cattle farmers, butchers, and beef retailers). Salmonella typhimurium was isolated from 7.5% (6/80) of the total raw beef samples. However, L. monocytogenes, Brucella spp., T. gondii, C. cayetanensis, and C. parvum were not isolated in this study. The mean level of microbial contamination of beef from the slaughter slabs/abattoir [5.2 Log10 colony-forming unit (CFU)/g] was not significantly different (p > 0.05) from the mean level observed at retail points (5.4 Log10 CFU/g). However, the mean coliform count of 4.3 Log10 CFU/g recorded at retail shops exceeded the permissible limits of 104 CFU/g (4 Log10 CFU/g) required by the Ghana Standards Authority for safety of meat and carcasses. Knowledge on food safety was at average level for butchers and retailers. Unhygienic practices and poor sanitary conditions at the abattoirs and retail shops observed could be the main contributing factors to microbial contamination of raw beef. Continuous education for meat handlers on issues of food safety and monitoring of slaughter activities will reduce the rate and level of contamination of beef.

Genetic diversity and drug resistance profiles of Mycobacterium tuberculosis complex isolates from patients with extrapulmonary tuberculosis in Ghana and their associated host immune responses.

Objectives: This study sought to determine the genetic diversity and drug resistance profiles of Mycobacterium tuberculosis complex (MTBC) isolates from extrapulmonary tuberculosis (EPTB) patients in Ghana, and their associated immune responses. Methods: Spoligotyping was performed on 102 MTBC isolates from EPTB patients. Lineages/sub-lineages were assigned by comparing spoligotyping patterns primarily with the SITVIT2 database and subsequently with the TB-Lineage online tool for unknown isolates in SITVIT2. Drug susceptibility testing was performed using MGIT (BD BACTEC 960), Lowenstein-Jensen media (indirect proportion method), and GenoType MTBDRplus/MTBDRsl assays. Differential cytokine levels in the serum of 20 EPTB patients infected with MTBC lineage 4 were determined using the Luminex multiplex immunoassay. Results: Around 95% (97/102) of isolates were Mycobacterium tuberculosis, predominantly lineage 4 (95%; 92/97). Of the lineage 4 isolates, the majority were sub-lineage Cameroon (37%, 34/92). Prevalence was significantly higher in the 15-34 years age group among EPTB patients infected with lineage 4 strains (p = 0.024). Fifteen isolates were resistant to at least one anti-TB drug tested. Decreased levels of IL-1ß, IL-17A, and IFN-α were observed in individuals infected with Cameroon sub-lineages compared with other lineage 4 sub-lineages. Conclusions: Our study confirms Cameroon (SIT61) as the most common spoligotype causing human EPTB in Ghana, and that it is associated with decreased serum IL-1ß, IL-17A, and IFN-α.

Antimicrobial resistance and genomic analysis of staphylococci isolated from livestock and farm attendants in Northern Ghana.

BACKGROUND: The emergence of antimicrobial resistant bacteria in food producing animals is of growing concern to food safety and health. Staphylococci are common inhabitants of skin and mucous membranes in humans and animals. Infections involving antibiotic resistant staphylococci are associated with increased morbidity and mortality, with notable economic consequences. Livestock farms may enable cross-species transfer of antibiotic resistant staphylococci. The aim of the study was to investigate antimicrobial resistance patterns of staphylococci isolated from livestock and farm attendants in Northern Ghana using phenotypic and genotypic methods. Antimicrobial susceptibility testing was performed on staphylococci recovered from livestock and farm attendants and isolates resistant to cefoxitin were investigated using whole genome sequencing. RESULTS: One hundred and fifty-two staphylococci comprising S. sciuri (80%; n = 121), S. simulans (5%; n = 8), S. epidermidis (4%; n = 6), S. chromogens (3%; n = 4), S. aureus (2%; n = 3), S. haemolyticus (1%; n = 2), S. xylosus (1%; n = 2), S. cohnii (1%; n = 2), S. condimenti (1%; n = 2), S. hominis (1%; n = 1) and S. arlettae (1%; n = 1) were identified. The isolates showed resistance to penicillin (89%; n = 135), clindamycin (67%; n = 102), cefoxitin (19%; n = 29), tetracycline (15%; n = 22) and erythromycin (11%; n = 16) but showed high susceptibility to gentamicin (96%; n = 146), sulphamethoxazole/trimethoprim (98%; n = 149) and rifampicin (99%; n = 151). All staphylococci were susceptible to linezolid and amikacin. Carriage of multiple resistance genes was common among the staphylococcal isolates. Genome sequencing of methicillin (cefoxitin) resistant staphylococci (MRS) isolates revealed majority of S. sciuri (93%, n = 27) carrying mecA1 (which encodes for beta-lactam resistance) and the sal(A) gene, responsible for resistance to lincosamide and streptogramin. Most of the MRS isolates were recovered from livestock. CONCLUSION: The study provides insights into the genomic content of MRS from farm attendants and livestock in Ghana and highlights the importance of using whole-genome sequencing to investigate such opportunistic pathogens. The finding of multi-drug resistant staphylococci such as S. sciuri carrying multiple resistant genes is of public health concern as they could pose a challenge for treatment of life-threatening infections that they may cause.

Non-tuberculous mycobacteria, not Mycobacterium bovis, are a significant cause of TB-like lesions observed in slaughtered cattle in Ghana.

Objectives: The aim was to isolate and identify the species of mycobacteria causing tuberculous-like (TB-like) lesions in cattle in Ghana. Methods: Between 2019 and 2020, 68 bovine tissue samples with TB-like lesions, identified during post slaughter examination, were obtained from four major abattoirs close to border towns in Ghana. The samples were cultured on Lowenstein-Jensen medium. Isolated bacteria were characterized by Ziehl-Neelsen staining and observation for acid-fast bacilli (AFB) under a microscope. DNA was extracted from AFB-positive isolates, and mycobacterial speciation was performed by line probe assay using GenoType Mycobacterium CM and also with mycobacterial 16S rRNA gene amplification and sequencing. Results: No Mycobacterium bovis was identified; however 53 bacterial isolates were obtained, of which 41 were non-tuberculous mycobacteria (NTM) strains and 12 were gram-positive bacteria. The predominant NTM species was M. fortuitum (43.9%, 18/41), with the rest being M. novocastrense, M. terrae, M. flavescens, M. holsaticum, M. cosmeticum, M. virginiense, M. intracellulare, M. mageritense, M. minnesotensis, M. duvalii, M. lehmannii, and M. koreense. Conclusions: In cattle, NTM contribute significantly to lesions observed during slaughter examination and may be an important cause of zoonotic tuberculosis. A One Health surveillance of NTM in Ghana would provide insights into their clinical significance.

Safety of Retailed Poultry: Analysis of Antibiotic Resistance in Escherichia coli From Raw Chicken and Poultry Fecal Matter From Selected Farms and Retail Outlets in Accra, Ghana.

Purpose: To assess the safety of retailed poultry using the prevalence of antibiotic resistance in Escherichia coli (E. coli), a dominant intestinal microflora. Methods: Two medium-scale farms and 8 well-known retail outlets within the La-Nkwantanang Madina municipality in Accra were purposively selected for sampling from January to March 2020. We randomly sampled raw chicken (n = 25) and poultry fecal matter (n = 50). All samples were immediately transported on ice to the laboratory for analysis within 12 hours after collection. Conventional culture techniques, biochemical tests, and matrix-assisted laser desorption/ionization-time-of-flight mass spectrometry (MALDI-TOF-MS) were used for isolation and identification. The antimicrobial susceptibility of isolated E. coli strains (n = 36) was tested using the Kirby Bauer disk diffusion method. Results: Antimicrobial resistance in E. coli ranged from 10.7 % (cefotaxime) to 82.1% (tetracycline) in fecal matter and 0% (gentamicin & cefotaxime) to 62.5% (tetracycline) in chicken. The prevalence of antimicrobial resistant E. coli in fecal samples was higher than in chicken for almost all antibiotics tested, except for cefoxitin, cefuroxime, and ceftazidime. Multidrug resistance was 57.1% in E. coli from fecal samples compared to 62.5% in chicken. Conclusion: The high level of resistance to E. coli in fecal matter is of public health concern because cross-contamination often occurs during slaughter and processing. This calls for close surveillance and strict adherence to Hazard Analysis and Critical Control Point (HACCP) principles in the chicken production chain to prevent the transmission of antimicrobial-resistant E. coli strains through the food chain.

Diarrhoeagenic E. coli occurrence and antimicrobial resistance of Extended Spectrum Beta-Lactamases isolated from diarrhoea patients attending health facilities in Accra, Ghana.

INTRODUCTION: Diarrhoea accounts for high morbidity and mortality in children and adults worldwide. Extended Spectrum Beta-Lactamase-Producing Enterobacteriaceae (ESBL-PE) and Diarrhoeagenic Escherichia coli (DEC) contribute to prolonged hospitalization because of their resistance and virulence properties aiding in the spread of diarrhoeal disease and delayed treatment. AIM: To determine DEC and the antimicrobial resistance of ESBL-PE isolated among diarrhoea patients attending two health facilities in Ghana. METHODS: Stool samples were collected from 122 diarrhoeal patients who attended Maamobi General Hospital and Kaneshie Polyclinic between January 2019 and March 2020. Identification of bacteria was performed by using the Matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS). Using disk diffusion, antimicrobial susceptibility testing (AST) was conducted and interpreted according to the 2018 CLSI guidelines. Detection of ESBL and DEC genes was performed using Polymerase chain reaction (PCR). RESULTS: A total of 80.3% (98/122) Enterobacteriaceae was recovered from the patients in the study with an overall ESBL occurrence of 20.4% (20/98), predominantly among E. coli showed 13.2% (10/76), Klebsiella pneumoniae,35.7%(5/14) and Proteus mirabilis, 57.1%(4/7). Among the ESBL genes detected, blaTEM (n = 14) was common, followed by blaCTX-M (n = 13) and blaSHV (n = 4). Thirty-four E. coli isolates possessed the heat labile (Lt) gene of Enterotoxigenic E. coli (ETEC). CONCLUSION: Our findings confirm the existence of DEC and the antimicrobial resistance patterns of ESBL-PE among stool isolates, limiting the options of commonly used drugs for diarrhoeal treatment in Ghana. Routine laboratory testing in health care facilities and strengthened surveillance systems among hospital networks are encouraged for a better understanding of their epidemiology and clinical implications.

LAMP4yaws: Treponema pallidum, Haemophilus ducreyi loop mediated isothermal amplification - protocol for a cross-sectional, observational, diagnostic accuracy study.

INTRODUCTION: Yaws, caused by the bacterium Treponema pallidum subsp. pertenue, is a neglected tropical disease targeted for eradication by 2030. Improved diagnostics will be essential to meet this goal. Diagnosis of yaws has relied heavily on clinical and serological tools. However, the presence of coendemic cutaneous skin ulcer diseases, such as lesions caused by Haemophilus ducreyi (HD), means these techniques do not provide a reliable diagnosis. Thus, new diagnostic tools are needed. Molecular tools such as PCR are ideal, but often expensive as they require trained technicians and laboratory facilities, which are often not available to national yaws programmes. METHODS AND ANALYSIS: The LAMP4yaws project is a cross-sectional, observational, diagnostic accuracy study of a combined Treponema pallidum (TP) and HD loop mediated isothermal amplification (TPHD-LAMP) test performed under real world conditions in three endemic countries in West Africa. Individuals with serologically confirmed yaws will be recruited in Cameroon, Côte d'Ivoire and Ghana. Each participant will provide paired swabs, one of which will be sent to the respective national reference laboratory for yaws quantitative PCR and the other will be tested for both TP and HD using the TPHD-LAMP test at local district laboratories. Sensitivity and specificity of the TPHD-LAMP test will be calculated against the reference standard qPCR. We will also assess the acceptability, feasibility and cost-effectiveness of the test. We anticipate that results from this study will support the adoption of the TPHD-LAMP test for use in global yaws eradication efforts. ETHICS AND DISSEMINATION: We have received ethical approval from all relevant institutional and national ethical committees. All participants, or their parents or guardians, must provide written informed consent prior to study enrolment. Study results will be published in an open access journal and disseminated with partners and the World Health Organization. TRIAL REGISTRATION NUMBER: NCT04753788.

Drug susceptibility profiles and factors associated with non-tuberculous mycobacteria species circulating among patients diagnosed with pulmonary tuberculosis in Tanzania.

BACKGROUND: While most Non-tuberculous mycobacteria (NTM) are saprophytic, several species have been associated with human diseases, from localized infection to disseminated diseases. Pulmonary NTM infections lead to TB-like disease called NTM pulmonary disease (NTM-PD). Due to variation in treatment options among NTM species, it is necessary to identify the species and determine drug susceptibility profiles to inform the choice of appropriate regimen for the disease. DESIGN: A total of 188 culture-positive isolates from patients diagnosed with TB were screened for NTM at the Central Tuberculosis Reference Laboratory. All NTM were further speciated using GenoType® Mycobacterium-Common Mycobacterium and Additional species (GenoType® CM/AS) kit. Mycobacteria avium complex (MAC) and Mycobacteria abscessus complex (MABC) which could not be identified with the test to species were subjected to GenoType® Mycobacteria NTM-DR for further speciation. Using the same test, identified MAC and MABC were genotyped to determine the drug susceptibility profile for each isolate to macrolide and aminoglycosides. RESULTS: Of all isolates identified as mycobacteria, 24 (13%) were NTM. Fifteen isolates could be identified to species level of which prevalent species was M. avium sub. intracellulare 4 (27%). A total of 10 isolates were MAC (n = 6) and MABC (n = 4) were subjected to GenoType® Mycobacteria NTM-DR for determination of macrolide and aminoglycoside susceptibility. Three of the four MABC had a mutation at the T28 position of the erm (41). All MAC were susceptible to both drugs. CONCLUSION: In this study, MAC was the most frequently isolated NTM species followed by MABC. While all MAC and MABC identified, were susceptible to aminoglycosides, three MABC were resistant to the macrolides due to mutation at position 28 of the erm (41) gene. For this, it is important for clinicians need to rule out NTM, understand species and their drug susceptibility for optimal case management.

Utility of anti-Mycobacterium tuberculosis antibody (ab905) for detection of mycobacterial antigens in formalin-fixed paraffin-embedded tissues from clinically and histologically suggestive extrapulmonary tuberculosis cases.

Background: The detection of acid-fast bacilli in extrapulmonary tissue samples is challenging due to its paucibacillary nature. The present study assessed the utility of immunohistochemistry (IHC) using anti-Mycobacterium tuberculosis antibody (ab905) for detecting the presence of mycobacterial antigens in archived formalin-fixed paraffin-embedded (FFPE) tissues. Methods: FFPE tissues [surgical biopsies (n = 32) and post-mortem tissues (n = 8)] from clinically and histologically suggestive extrapulmonary tuberculosis (EPTB) cases at the Korle Bu Teaching Hospital, Accra, Ghana from 2015 to 2020 were stained with IHC (anti-Mycobacterium tuberculosis antibody) and Ziehl-Neelsen (ZN) stain. The staining outcomes of IHC and ZN were compared, and their sensitivity and specificity determined against histopathology as reference standard. Results: Lymph nodes were about 40% (16/40) of the samples analyzed. IHC stained positive in 43.8% (7/16) biopsies and 87.5% (4/5) post-mortem samples ranging from 43.8% (7/16) in lymph nodes to 80% (4/5) in gastrointestinal organs. The overall sensitivity for IHC was 52.50% (95% CI: 36.13%-68.49%) and 0% (95% CI: 0.00%-8.81%) for ZN. Specificity was 72.50% (95% CI: 56.11%-85.40%) and 75% (95% CI: 58.80-87.31%) for IHC and ZN respectively. Conclusions: IHC using anti-Mycobacterium tuberculosis antibody (ab905) can detect mycobacterial antigens in diverse range of paucibacillary extrapulmonary tissue sections. It is potentially a useful tool for the diagnosis of EPTB in FFPE tissues in a routine pathology laboratory.

Coping with Adversity: Resilience Dynamics of Livestock Farmers in Two Agroecological Zones of Ghana.

Despite the increasing occurrence of adverse events including droughts and conflicts, livestock farmers in Ghana continue to raise animals to support their livelihoods and the national economy. We assessed the resilience of cattle farmers (CF) to adverse events they faced using a cross-sectional survey of 287 CF in two agroecological zones in Ghana. Resilience to adversities was assessed using the Resilience Scale (RS-14). Resilience scores and categories were computed and factors that explained variations in resilience categories assessed. The farmers kept, on average, 31 cattle per household, with a majority (91%) also growing crops. Key adverse events confronting them in both districts were animal disease outbreaks, pasture shortages, and theft, with 85% (240/287) losing, on average, seven cattle (15% of the herd size) over a one-year period. The mean resilience score was 71 (SD = 8) out of 98; 52% were highly resilient. Resilience was higher in the southern district (72 versus 70), albeit not statistically significant (p = 0.06). The resilience significantly improved with age, each unit increase in cattle in the herd, and having experience raising livestock (p < 0.001). The CF have relatively high resilience to adverse events affecting their productivity. The findings provide relevant information for implementing mitigation measures to improve production by reducing animal mortalities through high-quality veterinary services.

Multiplex Recombinase Polymerase Amplification Assay for Simultaneous Detection of Treponema pallidum and Haemophilus ducreyi in Yaws-Like Lesions.

Yaws is a skin debilitating disease caused by Treponema pallidum subspecies pertenue with most cases reported in children. World Health Organization (WHO) aims at total eradication of this disease through mass treatment of suspected cases followed by an intensive follow-up program. However, effective diagnosis is pivotal in the successful implementation of this control program. Recombinase polymerase amplification (RPA), an isothermal nucleic acid amplification technique offers a wider range of differentiation of pathogens including those isolated from chronic skin ulcers with similar characteristics such as Haemophilus ducreyi (H. ducreyi). We have developed a RPA assay for the simultaneous detection of Treponema pallidum (T. pallidum) and H. ducreyi (TPHD-RPA). The assay demonstrated no cross-reaction with other pathogens and enable detection of T. pallidum and H. ducreyi within 15 min at 42 °C. The RPA assay was validated with 49 clinical samples from individuals confirmed to have yaws by serological tests. Comparing the developed assay with commercial multiplex real-time PCR, the assay demonstrated 94% and 95% sensitivity for T. pallidum and H. ducreyi, respectively and 100% specificity. This simple novel TPHD-RPA assay enables the rapid detection of both T. pallidum and H. ducreyi in yaws-like lesions. This test could support the yaws eradication efforts by ensuring reliable diagnosis, to enable monitoring of program success and planning of follow-up interventions at the community level.