Evaluation of ten (10) SARS-CoV-2 rapid serological tests in comparison with WANTAI SARS-CoV-2 ab ELISA in Burkina Faso, West Africa.

BACKGROUND: The aim of this study was to evaluate the performance of ten (10) SARS-CoV-2 serological rapid diagnostic tests in comparison with the WANTAI SARS-CoV-2 Ab ELISA test in a laboratory setting. MATERIALS AND METHODS: Ten (10) SARS-CoV-2 serological rapid diagnostic tests (RDTs) for SARS-CoV-2 IgG/IgM were evaluated with two (2) groups of plasma tested positive for one and negative for the other with the WANTAI SARS-CoV-2 Ab ELISA. The diagnostic performance of the SARS-CoV-2 serological RDTs and their agreement with the reference test were calculated with their 95% confidence intervals. RESULTS: The sensitivity of serological RDTs ranged from 27.39 to 61.67% and the specificity from 93.33 to 100% compared to WANTAI SARS-CoV-2 Ab ELISA test. Of all the tests, two tests (STANDARD Q COVID-19 IgM/IgG Combo SD BIOSENSOR and COVID-19 IgG/IgM Rapid Test (Zhejiang Orient Gene Biotech Co., Ltd)) had a sensitivity greater than 50%. In addition, all ten tests had specificity greater than or equal to 93.33% each. The concordance between RDTs and WANTAI SARS-CoV-2 Ab ELISA test ranged from 0.25 to 0.61. CONCLUSION: The SARS-CoV-2 serological RDTs evaluated show low and variable sensitivities compared to the WANTAI SARS-CoV-2 Ab ELISA test, with however a good specificity. These finding may have implications for the interpretation and comparison of COVID-19 seroprevalence studies depending on the type of test used.

Buruli Ulcer, a Prototype for Ecosystem-Related Infection, Caused by Mycobacterium ulcerans.

Buruli ulcer is a noncontagious disabling cutaneous and subcutaneous mycobacteriosis reported by 33 countries in Africa, Asia, Oceania, and South America. The causative agent, Mycobacterium ulcerans, derives from Mycobacterium marinum by genomic reduction and acquisition of a plasmid-borne, nonribosomal cytotoxin mycolactone, the major virulence factor. M. ulcerans-specific sequences have been readily detected in aquatic environments in food chains involving small mammals. Skin contamination combined with any type of puncture, including insect bites, is the most plausible route of transmission, and skin temperature of <30°C significantly correlates with the topography of lesions. After 30 years of emergence and increasing prevalence between 1970 and 2010, mainly in Africa, factors related to ongoing decreasing prevalence in the same countries remain unexplained. Rapid diagnosis, including laboratory confirmation at the point of care, is mandatory in order to reduce delays in effective treatment. Parenteral and potentially toxic streptomycin-rifampin is to be replaced by oral clarithromycin or fluoroquinolone combined with rifampin. In the absence of proven effective primary prevention, avoiding skin contamination by means of clothing can be implemented in areas of endemicity. Buruli ulcer is a prototype of ecosystem pathology, illustrating the impact of human activities on the environment as a source for emerging tropical infectious diseases.

[Characteristics of parent-adolescent communication about sexuality and HIV in Bobo-Dioulasso, Burkina Faso]. / Caractéristiques de la communication parents-adolescentes sur la sexualité et le VIH à Bobo-Dioulasso, Burkina Faso.

Adolescent females are a key target audience in the fight against sexually transmitted infections and HIV in sub-Saharan Africa. One issue is that families in Africa play a very limited role in sex education. The objective of this study was to examine parent-child communication from a qualitative perspective by exploring the characteristics and quality of parent-child communication. A cross-sectional study was conducted between April and September 2009 in Bobo-Dioulasso (Burkina Faso). The study included 40 parent-child pairs (50% of in-school children and 50% of out-of-school children). Individual interviews and focus groups were conducted. The data were analyzed using Stata version 9.1 (quantitative data) and QSR Nvivo 2.0 (qualitative data). The study found that 74% (14/19) of out-of-school children communicated with their parents, compared to just 45% of in-school children (p = 0.07). Mother-child communication was found to be the most common type of parent-child communication, with 59% (13/22) of families who communicated about sexuality and HIV preferring mother-child communication. Further research is needed to identify the factors determining better communication among out-of-school children.

Confirming Autochthonous Buruli Ulcer Cases in Burkina Faso, West Africa.

Environmental Mycobacterium ulcerans causes a disabling skin disease called Buruli ulcer. Recent studies completed the knowledge of the evolving geographic extension and epidemiology of Buruli ulcer in West Africa, where Côte d'Ivoire is reporting the highest number of cases. We report seven polymerase chain reaction-documented patients in Burkina Faso, a neighboring country of Côte d'Ivoire, where previously Buruli ulcer cases were confirmed primarily using clinical arguments.

Epidemiology and microscopic diagnosis of tuberculosis in pigs and small ruminants slaughtered at Bobo-Dioulasso abattoir, Burkina Faso.

Bovine tuberculosis (bTB) is a zoonotic, infectious, chronic and contagious disease, caused by Mycobacterium bovis that mainly affects cattle. This pathology has a negative impact on animals and animal products trade. Unfortunately, in Burkina Faso where agriculture and livestock sectors represent around 80% of the socio-economic activities, the real situation of the disease is not well known especially in small ruminants and swine. Thus, our study focused on both the epidemiology and the microbiological diagnosis of tuberculosis (TB) in small ruminants and pigs slaughtered at Bobo-Dioulasso abattoir. A prospective study was conducted between August 2017 and December 2017. Epidemiological data collection was performed during routine meat inspection; moreover, samples were taken and transported to the Bacteriology laboratory of Centre Muraz for microbiological analyses. This diagnosis consisted in search of Acid Fast Bacilli (AFB) using the hot Ziehl-Neelsen staining. Out of a total of 14 648 small ruminants and 2430 pigs slaughtered during the study period, 156 and 17 had lesions suggestive of bTB with prevalence of 1.07% and 0.7%, respectively. Females and those between 2 and 4 years old were mainly infected. The most affected organs were: lungs, liver, spleen and lymph nodes. Finally, microscopy revealed 43.35% (75/173) of positive cases for AFB. These results confirm the presence of bTB in small ruminants and pigs in Burkina Faso. Efforts must still be made in the fight against this zoonosis in order to limit its economic and public health impacts.

[Vancomycin-based Lowenstein-Jensen selective medium for reducing contamination of mycobacterial cultures by spore-forming bacteria]. / Milieu sélectif de Lowenstein-Jensen à base de vancomycine pour la réduction des contaminations de cultures de mycobactéries par les bactéries sporulantes.

INTRODUCTION: despite the development of new methods, culture on solid medium is the gold standard for the diagnosis of tuberculosis. However, this method is associated with increased rates of contamination of cultures by spore-forming bacteria. These bacteria are generally sensitive to vancomycin and to a combinsation of vancomycin, colistin, nystatin, and trimethoprim (VCNT). The purpose of this study was to assess the effectiveness of VCNT-based selective Lowenstein-Jensen (LJ) medium in reducing contamination of cultures by spore-forming bacteria. METHODS: sputum samples, collected from the 120 TB and non-TB patients included in the study between October 2016 and May 2017, were decontaminated with the modified Petroff method. Decontamination pellets were inoculated onto conventional LJ media and selective VCNT-based LJ medium containing 10µg/ml vancomycin. Fifteen strains of spore-forming bacteria were inoculated onto the same media in order to assess their sensitivity to VCNT. RESULTS: the contamination of cultures on VCNT-based LJ medium containing 10µg/ml of vancomycin and LJ medium were 11.66% (14/120) and 39.16% (47/120) with p <0.0001, respectively. Sensitivity of spore-forming bacteria to VCNT decreased with the increasing of culture incubation time. CONCLUSION: VCNT-based selective LJ medium containing 10µg/ml vancomycin led to a significant reduction in the rate of culture contamination. This environment could contribute to improve the quality of mycobacterial cultures and thus bacteriological diagnosis of tuberculosis.

Culturomics Discloses Anti-Tubercular Enterococci Exclusive of Pulmonary Tuberculosis: A Preliminary Report.

Mycobacterium tuberculosis causes pulmonary tuberculosis, a deadly infection of which the clinical expression and prognosis are not fully understood at the individual level, apart from genetic susceptibility traits. We investigated whether individual gut microbiota may correlate with pulmonary tuberculosis status. Culturomics investigations of gut microbiota in two pulmonary tuberculosis patients and two controls in Burkina Faso found 60 different bacterial species in patients and 97 in controls, including 45 in common. Further analysis of the results at the individual level indicated seven bacteria, including Enterococcus mundtii and Enterococcus casseliflavus, which were exclusively cultured in controls. Blind quantitative PCR-based exploration of faeces samples in two cohorts in Burkina Faso and in France confirmed a nonsignificant association of E. mundtii and E. casseliflavus with controls. Further in vitro explorations found four E. mundtii and E. casseliflavus strains inhibiting the growth of M. tuberculosis strains representative of four different lineages as well as Mycobacterium africanum, Mycobacterium canettii, and Mycobacterium bovis, in an inoculum-dependent manner. Heat-killed E. mundtii or E. casseliflavus were ineffective. These unprecedented observations of direct interactions between gut E. mundtii and E. casseliflavus with M. tuberculosis complex mycobacteria suggest that gut microbiota may modulate the expression of pulmonary tuberculosis.

Identification of spore-forming bacteria isolated from contaminated Lowenstein Jensen media and effectiveness of Vancomycin to reduce Mycobacterial culture contamination in Burkina-Faso.

The type of commensal microorganisms can influence the efficiency of sputum decontamination for TB diagnosis. A basic characterization of contaminants from LJ contaminated media showed that Gram positive Spore Forming Bacteria (SFB) were the major contaminants. This study aims to identify the species of this contaminants and to evaluate the effectiveness of VCNT at 10 µg of vancomycin to reduce mycobacterial culture contamination mainly linked to SFB. Fifty-three SFB isolated between February 2016 and May 2017 were used. The effectiveness of LJ with VCNT at 10 µg of Vancomycin were evaluated with sputum collected in the same period. SFB had been stored at -20 °C and identified after subculture onto 5% sheep blood Columbia agar and incubated at 37 °C during 24 h. Bacteria cells and isolated colonies were described. API 50CH/B was performed and MALDI-TOF MS was used for external quality control. Thirty- five (66%) isolates representing 4 genera (Bacillus, Paenibacillus, Brevisbacillus and Lysinibacillus) including 10 species were identified. The most important species were Bacillus cereus (30%) and Bacillus licheniformis (21%). Eighteen (34%) isolates were non-reactive Bacillus. The overall contamination rate on LJ with VCNT at 10 µg of vancomycin was statistically lower than which without VCNT (18.7% versus 43.8%) (p = 0.01). The most important SFB identified were B. cereus and B. licheniformis. Almost all identified strains were similar to those currently isolated in fermented traditional food suggesting in part food related contaminants. VCNT containing 10 µg of vancomycin is a good alternative method to reduce mycobacterial culture contamination.

A protocol for culturing environmental strains of the Buruli ulcer agent, Mycobacterium ulcerans.

Contaminations and fastidiousness of M. ulcerans may have both hamper isolation of strains from environmental sources. We aimed to optimize decontamination and culture of environmental samples to circumvent both limitations. Three strains of M. ulcerans cultured onto Middlebrook 7H10 at 30 °C for 20 days yielded a significantly higher number of colonies in micro-aerophilic atmosphere compared to ambient atmosphere, 5% CO2 and anaerobic atmosphere. In a second step, we observed that M. ulcerans genome uniquely encoded chitinase, fucosidase and A-D-GlcNAc-diphosphoryl polyprenol A-3-L-rhamnosyl transferase giving M. ulcerans the potential to metabolize chitine, fucose and N-acetyl galactosamine (NAG), respectively. A significant growth-promoting effect of 0.2 mg/mL chitin (p < 0.05), 0.01 mg/mL N-acetyl galactosamine (p < 0.05), 0.01 mg/mL fucose (p < 0.05) was observed with M. ulcerans indicating that NAG alone or combined with fucose and chitin could complement Middlebrook 7H10. Finally, the protocol combining 1% chlorhexidine decontamination with micro-aerophilic incubation on Middlebrook 7H10 medium containing chitin (0.2%), NAG (0.01%) and fucose (0.01%) medium and auto-fluorescence detection of colonies allowed for the isolation of one mycolactone-encoding strain from Thryonomys swinderianus (aulacode) feces specimens collected near the Kossou Dam, Côte d'Ivoire. We propose that incubation of chlorhexidine-decontaminated environmental specimens on Middlebrook 7H10-enriched medium under micro-aerophilic atmosphere at 30 °C may be used for the tentative isolation of M. ulcerans strains from potential environmental sources.

High-Throughput Carbon Substrate Profiling of Mycobacterium ulcerans Suggests Potential Environmental Reservoirs.

BACKGROUND: Mycobacterium ulcerans is a close derivative of Mycobacterium marinum and the agent of Buruli ulcer in some tropical countries. Epidemiological and environmental studies pointed towards stagnant water ecosystems as potential sources of M. ulcerans, yet the ultimate reservoirs remain elusive. We hypothesized that carbon substrate determination may help elucidating the spectrum of potential reservoirs. METHODOLOGY/PRINCIPAL FINDINGS: In a first step, high-throughput phenotype microarray Biolog was used to profile carbon substrates in one M. marinum and five M. ulcerans strains. A total of 131/190 (69%) carbon substrates were metabolized by at least one M. ulcerans strain, including 28/190 (15%) carbon substrates metabolized by all five M. ulcerans strains of which 21 substrates were also metabolized by M. marinum. In a second step, 131 carbon substrates were investigated, through a bibliographical search, for their known environmental sources including plants, fruits and vegetables, bacteria, algae, fungi, nematodes, mollusks, mammals, insects and the inanimate environment. This analysis yielded significant association of M. ulcerans with bacteria (p = 0.000), fungi (p = 0.001), algae (p = 0.003) and mollusks (p = 0.007). In a third step, the Medline database was cross-searched for bacteria, fungi, mollusks and algae as potential sources of carbon substrates metabolized by all tested M. ulcerans; it indicated that 57% of M. ulcerans substrates were associated with bacteria, 18% with alga, 11% with mollusks and 7% with fungi. CONCLUSIONS: This first report of high-throughput carbon substrate utilization by M. ulcerans would help designing media to isolate and grow this pathogen. Furthermore, the presented data suggest that potential M. ulcerans environmental reservoirs might be related to micro-habitats where bacteria, fungi, algae and mollusks are abundant. This should be followed by targeted investigations in Buruli ulcer endemic regions.

Impact of pre-analytical factors on mycobacterium cultures contaminations rates in Burkina Faso, West Africa.

INTRODUCTION: For a high quality level diagnosis, mycobacterium culture must comply with the pre-analytical and analytical conditions recommended by the WHO and the country National Tuberculosis Program (NTP). In this study, we determined whether temperature and duration of sputum storage were associated with culture contamination in Burkina Faso. METHODS: Sputa were collected in 5 districts labs in Burkina Faso. Temperature and duration of sputum storage were recorded. After the collection, sputa were decontaminated using Petroff modified method, and the pellet was inoculated on LJ media and LJ media supply with 2% sodium pyruvate. Risk of culture contamination associated with temperature and duration of sputum storage was measured by Chi2 test and logistic regression. RESULTS: Out of 404 specimens, 61% (246/404) were stored between 2 and 8°C, and 15% (61/404) were processed within three days. The global contamination rate was 24%, with only 8% for samples respecting WHO recommendations, up to 35% for others. Storage at room temperature was associated with a significantly higher risk of contamination compared to storage at 2-8°C (OR 2.24, p = 0.001, IC 95%). CONCLUSION: The recommendations about the temperature and the duration of sputum storage before cultures are not completely respected. This leads to high contamination rate of mycobacterium culture. It will be necessary to take logistics measures in peripherals health services or to develop more selective medium for mycobacterium culture in low income countries.

Mycobacterium bovis in Burkina Faso: epidemiologic and genetic links between human and cattle isolates.

BACKGROUND: In sub-Saharan Africa, bovine tuberculosis (bTB) is a potential hazard for animals and humans health. The goal of this study was to improve our understanding of bTB epidemiology in Burkina Faso and especially Mycobacterium bovis transmission within and between the bovine and human populations. METHODOLOGY/PRINCIPAL FINDINGS: Twenty six M. bovis strains were isolated from 101 cattle carcasses with suspected bTB lesions during routine meat inspections at the Bobo Dioulasso and Ouagadougou slaughterhouses. In addition, 7 M. bovis strains were isolated from 576 patients with pulmonary tuberculosis. Spoligotyping, RDAf1 deletion and MIRU-VNTR typing were used for strains genotyping. The isolation of M. bovis strains was confirmed by spoligotyping and 12 spoligotype signatures were detected. Together, the spoligotyping and MIRU-VNTR data allowed grouping the 33 M. bovis isolates in seven clusters including isolates exclusively from cattle (5) or humans (1) or from both (1). Moreover, these data (genetic analyses and phenetic tree) showed that the M. bovis isolates belonged to the African 1 (Af1) clonal complex (81.8%) and the putative African 5 (Af5) clonal complex (18.2%), in agreement with the results of RDAf1 deletion typing. CONCLUSIONS/SIGNIFICANCE: This is the first detailed molecular characterization of M. bovis strains from humans and cattle in Burkina Faso. The distribution of the two Af1 and putative Af5 clonal complexes is comparable to what has been reported in neighbouring countries. Furthermore, the strain genetic profiles suggest that M. bovis circulates across the borders and that the Burkina Faso strains originate from different countries, but have a country-specific evolution. The genetic characterization suggests that, currently, M. bovis transmission occurs mainly between cattle, occasionally between cattle and humans and potentially between humans. This study emphasizes the bTB risk in cattle but also in humans and the difficulty to set up proper disease control strategies in Burkina Faso.

[Advantages and drawbacks of expectoration decontamination methods for tuberculosis and anti-tuberculosis drug resistance diagnosis]. / Avantages et limites des méthodes de décontamination des expectorations pour le diagnostic de la tuberculose et des résistances aux antituberculeux.

In the actual context of increasing tuberculosis and anti-tuberculosis drug resistance, the laboratory diagnosis of Mycobacterial infections remain the primordial objective of control and surveillance of human tuberculosis. The diagnosis and following of tuberculosis in resource limited settings are done by microscopy Ziehl-Neelsen method which is poor sensitive (20-53%) and have poor specificity because it's can't distinguish tuberculosis mycobacterium and atypical tuberculoid mycobacterium. Mycobacterium culture on solid media is the gold standard method for tuberculosis and anti-tuberculosis drug resistance diagnosis. Here, the challenge is that expectorations using for culture contain mycobacterium and others contaminating bacteria responsible of culture contamination. Many different methods of homogenization and decontamination of sputum specimens for culturing exist and each laboratory had to do a choice of the better method to optimize isolating of mycobacterium. This review is a summary of homogenization and decontamination methods described in literature and used by certain laboratories for diagnosis of TB by culture. However, it's essential for each laboratory to conduct evaluation of the different methods and do the choice of the appropriate one by taking into account factors such as the feasibility and cost effectively. Nine methods of decontaminations are described in this review taking account of their advantages, drawbacks and their feasibility in resource limited settings.