Implementation of a National Reference Laboratory for Buruli Ulcer Disease in Togo

Background: In a previous study PCR analysis of clinical samples from suspected cases of Buruli ulcer disease (BUD) from Togo and external quality assurance (EQA) for local microscopy were conducted at an external reference laboratory in Germany. The relatively poor performance of local microscopy as well as effort and time associated with shipment of PCR samples necessitated the implementation of stringent EQA measures and availability of local laboratory capacity. This study describes the approach to implementation of a national BUD reference laboratory in Togo. Methodology: Large scale outreach activities accompanied by regular training programs for health care professionals were conducted in the regions "Maritime'' and "Central,'' standard operating procedures defined all processes in participating laboratories (regional, national and external reference laboratories) as well as the interaction between laboratories and partners in the field. Microscopy was conducted at regional level and slides were subjected to EQA at national and external reference laboratories. For PCR analysis, sample pairs were collected and subjected to a dry-reagent-based IS2404-PCR (DRB-PCR) at national level and standard IS2404 PCR followed by IS2404 qPCR analysis of negative samples at the external reference laboratory. Principal Findings: The inter-laboratory concordance rates for microscopy ranged from 89% to 94%; overall, microscopy confirmed 50% of all suspected BUD cases. The inter-laboratory concordance rate for PCR was 96% with an overall PCR case confirmation rate of 78%. Compared to a previous study, the rate of BUD patients with non-ulcerative lesions increased from 37% to 50%, the mean duration of disease before clinical diagnosis decreased significantly from 182.6 to 82.1 days among patients with ulcerative lesions, and the percentage of category III lesions decreased from 30.3% to 19.2%. Conclusions: High inter-laboratory concordance rates as well as case confirmation rates of 50% (microscopy), 71% (PCR at national level), and 78% (including qPCR confirmation at external reference laboratory) suggest high standards of BUD diagnostics. The increase of non-ulcerative lesions, as well as the decrease in diagnostic delay and category III lesions, prove the effect of comprehensive EQA and training measures involving also procedures outside the laboratory

Augmentation de la résistance aux antibiotiques des Entérobactéries isolées à l’Institut National d’Hygiène de Lomé de 2010 à 2017: Increase in antibiotic resistance of Enterobacteriaceae isolated at the National Institute of Hygiene of Lomé from 2010 to 2017

Introduction: La résistance des Entérobactéries aux antibiotiques est un problème d’importance croissante en pratique médicale. L’objectif de cette étude était de déterminer le profil de résistance aux antibiotiques des Entérobactéries isolées à l’institut national d’hygiène (INH) de Lomé et d’analyser son évolution dans le temps. Méthodes: Il s’agissait d’une analyse rétrospective, sur une période de huit ans (2010-2017), portant sur l’ensemble des souches d’Entérobactéries isolées des prélèvements pathologiques analysés au laboratoire de bactériologie de l’INH. Résultats: Au total, 5910 Entérobactéries ont été isolées majoritairement des urines (59,59%), avec une prédominance d’Escherichia coli (63,93%) suivie de Klebsiella spp (22,86%). Entre 2010 et 2017, le taux de résistance des souches d’Escherichia coli a augmenté significativement de 18,69% à 39,26% (p< 0,0001) à la Ceftazidime ; de 1,68% à 40,22% à la Ceftriaxone (p< 0,0001) et de 42,37% à 63,23% (p< 0,0001) à la Ciprofloxacine. La résistance des souches de Klebsiella spp à la Ceftazidime a augmenté significativement de 25,26% à 42,54% (p< 0,0001) et celle à la Ceftriaxone de 2,17% à 41,79% (p< 0,0001) respectivement de 2010 à 2017. Conclusion: L’augmentation de la résistance des Entérobactéries aux antibiotiques et surtout l’évolution des résistances aux Céphalosporines de 3e Génération et aux Fluoroquinolones est un phénomène réel. Ceci exposera à des difficultés de prise en charge thérapeutique et nécessite la mise en place des dispositions idoines. Background: Antibiotic resistance in Enterobacteriaceae is a growing problem in medical practice. The objective of this study was to determine the antibiotic resistance profile of Enterobacteriaceae isolated at the National Institute of Hygiene (INH) of Lomé and to analyse its evolution over time. Method: This was a retrospective analysis, over a period of eight years (2010-2017), of all strains of Enterobacteriaceae isolated from pathological samples analysed in the bacteriology laboratory of the INH. Results: A total of 5910 Enterobacteriaceae were isolated mainly from urine (59.59%), with a predominance of Escherichia coli (63.93%) followed by Klebsiella spp (22.86%). Between 2010 and 2017, the resistance rate of Escherichia coli strains increased significantly from 18.69% to 39.26% (p<0.0001) to Ceftazidime; from 1.68% to 40.22% to Ceftriaxone (p<0.0001) and from 42.37% to 63.23% (p<0.0001) to Ciprofloxacin. Resistance of Klebsiella spp strains to Ceftazidime increased significantly from 25.26% to 42.54% (p< 0.0001) and to Ceftriaxone from 2.17% to 41.79% (p< 0.0001) respectively from 2010 to 2017. Conclusion: The increase in antibiotic resistance in Enterobacteriaceae and especially the evolution of resistance to 3rd generation cephalosporins and fluoroquinolones is a real phenomenon. This will lead to difficulties in therapeutic management and requires the implementation of appropriate measures

Effectiveness of Routine BCG Vaccination on Buruli Ulcer Disease: A Case-Control Study in the Democratic Republic of Congo, Ghana and Togo

Background: The only available vaccine that could be potentially beneficial against mycobacterial diseases contains live attenuated bovine tuberculosis bacillus (Mycobacterium bovis) also called Bacillus Calmette-Guerin (BCG). Even though the BCG vaccine is still widely used, results on its effectiveness in preventing mycobacterial diseases are partially contradictory, especially regarding Buruli Ulcer Disease (BUD). The aim of this case-control study is to evaluate the possible protective effect of BCG vaccination on BUD. Methodology: The present study was performed in three different countries and sites where BUD is endemic: in the Democratic Republic of the Congo, Ghana, and Togo from 2010 through 2013. The large study population was comprised of 401 cases with laboratory confirmed BUD and 826 controls, mostly family members or neighbors. Principal Findings: After stratification by the three countries, two sexes and four age groups, no significant correlation was found between the presence of BCG scar and BUD status of individuals. Multivariate analysis has shown that the independent variables country (p = 0.31),sex (p = 0.24),age (p = 0.96),and presence of a BCG scar (p = 0.07) did not significantly influence the development of BUD category I or category II/III. Furthermore, the status of BCG vaccination was also not significantly related to duration of BUD or time to healing of lesions. Conclusions: In our study, we did not observe significant evidence of a protective effect of routine BCG vaccination on the risk of developing either BUD or severe forms of BUD. Since accurate data on BCG strains used in these three countries were not available, no final conclusion can be drawn on the effectiveness of BCG strain in protecting against BUD. As has been suggested for tuberculosis and leprosy, well-designed prospective studies on different existing BCG vaccine strains are needed also for BUD

Effectiveness of Routine BCG Vaccination on Buruli Ulcer Disease: A Case-Control Study in the Democratic Republic of Congo, Ghana and Togo

Background: The only available vaccine that could be potentially beneficial against mycobacterial diseases contains live attenuated bovine tuberculosis bacillus (Mycobacterium bovis) also called Bacillus Calmette-Guerin (BCG). Even though the BCG vaccine is still widely used, results on its effectiveness in preventing mycobacterial diseases are partially contradictory, especially regarding Buruli Ulcer Disease (BUD). The aim of this case-control study is to evaluate the possible protective effect of BCG vaccination on BUD. Methodology: The present study was performed in three different countries and sites where BUD is endemic: in the Democratic Republic of the Congo, Ghana, and Togo from 2010 through 2013. The large study population was comprised of 401 cases with laboratory confirmed BUD and 826 controls, mostly family members or neighbors. Principal Findings: After stratification by the three countries, two sexes and four age groups, no significant correlation was found between the presence of BCG scar and BUD status of individuals. Multivariate analysis has shown that the independent variables country (p = 0.31),sex (p = 0.24),age (p = 0.96),and presence of a BCG scar (p = 0.07) did not significantly influence the development of BUD category I or category II/III. Furthermore, the status of BCG vaccination was also not significantly related to duration of BUD or time to healing of lesions. Conclusions: In our study, we did not observe significant evidence of a protective effect of routine BCG vaccination on the risk of developing either BUD or severe forms of BUD. Since accurate data on BCG strains used in these three countries were not available, no final conclusion can be drawn on the effectiveness of BCG strain in protecting against BUD. As has been suggested for tuberculosis and leprosy, well-designed prospective studies on different existing BCG vaccine strains are needed also for BUD

Comparative Characterization of Vibrio cholerae O1 from Five Sub-Saharan African Countries Using Various Phenotypic and Genotypic Techniques.

We used standardized methodologies to characterize Vibrio cholerae O1 isolates from Guinea, Democratic Republic of the Congo (DRC), Togo, Côte d'Ivoire and Mozambique. We investigated 257 human isolates collected in 2010 to 2013. DRC isolates serotyped O1 Inaba, while isolates from other countries serotyped O1 Ogawa. All isolates were biotype El Tor and positive for cholera toxin. All isolates showed multidrug resistance but lacked ciprofloxacin resistance. Antimicrobial susceptibility profiles of isolates varied between countries. In particular, the susceptibility profile of isolates from Mozambique (East-Africa) included resistance to ceftriaxone and was distinctly different to the susceptibility profiles of isolates from countries located in West- and Central-Africa. Molecular subtyping of isolates using pulsed-field gel electrophoresis (PFGE) analysis showed a complex relationship among isolates. Some PFGE patterns were unique to particular countries and clustered by country; while other PFGE patterns were shared by isolates from multiple countries, indicating that the same genetic lineage is present in multiple countries. Our data add to a better understanding of cholera epidemiology in Africa

Wissenschaftliche Begleitung des Vorhabens 'Eindampfanlage zur Behandlung von Deponiesickerwaessern beim Zweckverband Sondermuellplaetze Mittelfranken (ZVSMM) Abschlussbericht

Available from TIB Hannover: RN 5905(2268) / FIZ - Fachinformationszzentrum Karlsruhe / TIB - Technische InformationsbibliothekSIGLEDEGerman

Dynamics of cholera epidemics from Benin to Mauritania.

The countries of West Africa are largely portrayed as cholera endemic, although the dynamics of outbreaks in this region of Africa remain largely unclear.To understand the dynamics of cholera in a major portion of West Africa, we analyzed cholera epidemics from 2009 to 2015 from Benin to Mauritania. We conducted a series of field visits as well as multilocus variable tandem repeat analysis and whole-genome sequencing analysis of V. cholerae isolates throughout the study region. During this period, Ghana accounted for 52% of the reported cases in the entire study region (coastal countries from Benin to Mauritania). From 2009 to 2015, we found that one major wave of cholera outbreaks spread from Accra in 2011 northwestward to Sierra Leone and Guinea in 2012. Molecular epidemiology analysis confirmed that the 2011 Ghanaian isolates were related to those that seeded the 2012 epidemics in Guinea and Sierra Leone. Interestingly, we found that many countries deemed "cholera endemic" actually suffered very few outbreaks, with multi-year lulls.This study provides the first cohesive vision of the dynamics of cholera epidemics in a major portion of West Africa. This epidemiological overview shows that from 2009 to 2015, at least 54% of reported cases concerned populations living in the three urban areas of Accra, Freetown, and Conakry. These findings may serve as a guide to better target cholera prevention and control efforts in the identified cholera hotspots in West Africa

Summary of phenotypic data for <i>V</i>. <i>cholerae</i> O1 isolates.

<p>* Isolates were determined to be resistant to antimicrobial agents at the following MIC breakpoints: ampicillin (Amp), ≥16 μg/ml; ceftriaxone (Cro), ≥2 μg/ml; ≥16 μg/ml; cotrimoxazole (Cot), ≥4 μg/ml; chloramphenicol (Chl), ≥16 μg/ml; nalidixic acid (Nal), ≥32 μg/ml; ciprofloxacin (Cip), ≥2 μg/ml; tetracycline (Tet), ≥8 μg/ml; erythromycin (Ery), ≥4 μg/ml; nitrofurantoin (Nit), ≥64 μg/ml.</p><p>Summary of phenotypic data for <i>V</i>. <i>cholerae</i> O1 isolates.</p

Snapshot of dendrogram of PFGE (<i>Not</i>I digestion) patterns for <i>V</i>. <i>cholerae</i> O1 isolates.

<p>Snapshot of dendrogram of PFGE (<i>Not</i>I digestion) patterns for <i>V</i>. <i>cholerae</i> O1 isolates.</p

Map of Africa showing the countries described in the current study.

<p>Map of Africa showing the countries described in the current study.</p