Changing Ecotypes of Dengue Virus 2 Serotype in Nigeria and the Emergence of Cosmopolitan and Asian I Lineages, 1966–2019

Dengue virus (DENV) is a leading mosquito-borne virus with a wide geographical spread and a major public health concern. DENV serotype 1 (DENV-1) and serotype 2 (DENV-2) were first reported in Africa in 1964 in Ibadan, Nigeria. Although the burden of dengue is unknown in many African countries, DENV-2 is responsible for major epidemics. In this study, we investigated the activities of DENV-2 to determine the circulating strains and to appraise the changing dynamics in the epidemiology of the virus in Nigeria. Nineteen DENV-2 sequences from 1966–2019 in Nigeria were retrieved from the GenBank of the National Center of Biotechnology Information (NCBI). A DENV genotyping tool was used to identify the specific genotypes. The evolutionary history procedure was performed on 54 DENV-2 sequences using MEGA 7. There is a deviation from Sylvatic DENV-2 to other genotypes in Nigeria. In 2019, the Asian I genotype of DENV-2 was predominant in southern Edo State, located in the tropical rainforest region, with the first report of the DENV-2 Cosmopolitan strain. We confirmed the circulation of other non-assigned genotypes of DENV-2 in Nigeria. Collectively, this shows that DENV-2 dynamics have changed from Sylvatic transmission reported in the 1960s with the identification of the Cosmopolitan strain and Asian lineages. Sustained surveillance, including vectorial studies, is required to fully establish the trend and determine the role of these vectors

Evaluation of the antimicrobial susceptibility testing process in clinical microbiology laboratories at Niamey, Niger

Background: Risk assessment is the means of identifying and evaluating potential errors or problems that may occur in testing process. The aim of this study was to perform risk assessment of antimicrobial susceptibility testing (AST) process in clinical microbiology laboratories of Niamey, Niger Republic.Methodology: We conducted a descriptive cross-sectional study from October 1 to December 31, 2019, to evaluate AST performance in seven clinical microbiology laboratories at Niamey, the capital city of Niger republic. The evaluation focused on the determination of the criticality index (CI) of each critical point (frequency of occurrence of anomalies, severity of the process anomaly, and detectability of the anomaly during the process) in the AST process and the performance of the AST through an observation sheet using two reference strains; Escherichia coli ATCC 25922 and Staphylococcus aureus ATCC 29213.Results: The criticality index (CI) was greater than 6 for most of the critical points related to material, medium, equipment, method and labour for the AST process in all the laboratories. A range of 18-100% errors on the inhibition zone diameters of the reference strains were observed. Major and/or minor categorization (Sensitive S, Intermediate I and Resistance R) discrepancies were found at all the laboratories for either one or both reference strains. The antibiotics most affected by the S/I/R discrepancies were trimethoprim (100%), vancomycin (100%), amoxicillin (80%) and amoxicillin + clavulanic acid (70%).Conclusion: This study showed a deficiency in the control of critical control points that impacts the performance of the AST reported by the laboratories in Niger. Corrective actions are needed to improve the performance of AST in clinical microbiology laboratories in Niger.   French title: Evaluation du processus de réalisation de l’antibiogramme dans les laboratoires d’analyses de biologie médicale de la ville de Niamey, Niger Contexte: L'évaluation des risques est le moyen d'identifier et d'évaluer les erreurs ou les problèmes potentiels qui peuvent survenir dans le processus de test. L'objectif de cette étude était de réaliser une évaluation des risques du processus d'antibiogramme (ABG) dans les laboratoires de microbiologie clinique de Niamey, en République du Niger.Méthodologie: Nous avons mené une étude transversale descriptive du 1er octobre au 31 décembre 2019 pour évaluer la performance des ABG dans sept laboratoires de microbiologie clinique à Niamey, capitale de la république du Niger. L'évaluation a porté sur la détermination de l'indice de criticité (IC) de chaque point critique (fréquence d'apparition des anomalies, gravité de l'anomalie du processus et détectabilité de l'anomalie au cours du processus) dans le processus et la performance des AGB à travers une fiche d'observation en utilisant deux souches de référence; Escherichia coli ATCC 25922 et Staphylococcus aureus ATCC 29213.Résultats: L'indice de criticité était supérieur à 6 pour la plupart des points critiques liés au matériel, au milieu, à l'équipement, à la méthode et à la main-d'oeuvre pour le processus AST dans tous les laboratoires. Une fourchette d'erreurs de 18 à 100% sur les diamètres des zones d'inhibition des souches de référence a été observée. Des écarts de catégorisation majeurs et/ou mineurs (Sensible: S, Intermédiaire: I et Résistance: R) ont été constatés dans tous les laboratoires pour l'une ou les deux souches de référence. Les  antibiotiques les plus touchés par les écarts S/I/R étaient la triméthoprime (100%), la vancomycine (100%), l'amoxicilline (80%) et l'amoxicilline + acide clavulanique (70%).Conclusion: Cette étude a montré une déficience dans le contrôle des points de contrôle critiques qui a un impact sur la performance de l'antibiogramme rapportée par les laboratoires au Niger. Des actions correctives sont nécessaires pour améliorer la performance des ABG dans les laboratoires de microbiologie clinique au Niger

Best-bet integrated strategies for containing drug-resistant trypanosomes in cattle

Background African animal trypanosomosis is a major constraint to the rearing of productive livestock in the sub-humid Sudan-Sahel zone of West Africa where cotton is grown. Trypanosomosis is mainly controlled using trypanocidal drugs, but the effective use of drugs is threatened by the development of widespread resistance. This study tested integrated best-bet strategies for containment and/ or reversal of trypanocide resistance in villages in south-east Mali where resistance has been reported. Methods Four sentinel villages each from an intervention area (along the road from Mali to Burkina Faso) and a control area (along the road from Mali to Côte d’Ivoire) were selected for the study. Tsetse control was based on deltamethrin-treated stationary attractive devices and targeted cattle spraying between March 2008 and November 2009. Trypanosome-positive cattle were selectively treated with 3.5 mg/kg diminazene aceturate. Strategic helminth control using 10 mg/kg albendazole was also undertaken. During the intervention, tsetse densities along drainage lines, trypanosome infections and faecal egg counts in risk cattle (3 to 12 months of age) were monitored. Results Catch reductions of 66.5 % in Glossina palpalis gambiensis and 90 % in G. tachinoides were observed in the intervention area. Trypanosome prevalence was significantly (p < 0.05) lower in the intervention area (2.3 %; 1.3-3.6 %) compared to the control area (17.3 %; 14.8-20.1 %). Albendazole treatment resulted in a faecal egg count reduction of 55.6 % and reduced trypanosome infection risk (2.9 times lower than in the placebo group) although not significantly (p > 0.05). Further studies are required before confirming the existence of albendazole resistant strongyles in the study area. Conclusion Integration of best-bet strategies in areas of multiple drug- resistance is expected to reduce trypanosome infection risk thus contributing to containment of trypanocidal drug resistance. Integrated best-bet strategies could therefore be considered a viable trypanosomosis control option especially in areas where multiple drug-resistance has been reported

La bronchiolite aiguë du nourrisson: à propos de 112 cas hospitalisés au département pédiatrie du CHU Gabriel Touré

Introduction : Les infections respiratoires aiguës (IRA) constituent l’une des principales causes de morbidité et de mortalité infantile au Mali. L’objectif de ce travail était de décrire les aspects épidémiologiques, cliniques et thérapeutiques de la bronchiolite aiguë du nourrisson au service de pédiatrie du CHU Gabriel Touré. Méthodologie: Il s’agit d’une étude rétrospective portant sur 112 nourrissons âgés de 1 à 24 mois hospitalisés dans le service de pédiatrie générale pour bronchiolite aiguë. L’étude s’étalait sur une période d’un an (du 1er janvier au 31 décembre 2012). Résultats: Les nourrissons âgés de 1 à 6 mois étaient majoritaires (69%). L’âge moyen des patients était de 6 mois avec des extrêmes de 1 et 24 mois. Le sexe masculin était prédominant (63%) avec un sex-ratio de 1,73. Le principal motif de consultation était la difficulté respiratoire (87%). Le pic d’hospitalisation était au mois de novembre (33%). Les principaux signes cliniques en plus des sibilants étaient la toux, la rhinorrhée et la détresse respiratoire (97%). La fièvre était présente dans 38% des cas et la cyanose chez 4% des patients. La saturation en oxygène était inférieure à 94% chez 50% des patients. La kinésithérapie respiratoire a été faite chez un tiers des malades. Tous les malades avaient reçu une corticothérapie et une nébulisation avec du salbutamol et sérum physiologique. Une antibiothérapie a été faite chez 85% des patients. La durée moyenne d’hospitalisation était de 6 jours avec des extrêmes de 1 et 30 jours. Le taux de guérison était de 98 %. Conclusion : La bronchiolite est une pathologie bénigne et fréquente chez le nourrisson de 1 à 6 mois qui évolue favorablement dans la majorité des cas

Targeted Next Generation Sequencing for malaria research in Africa:Current status and outlook

Targeted Next Generation Sequencing (TNGS) is an efficient and economical Next Generation Sequencing (NGS) platform and the preferred choice when specific genomic regions are of interest. So far, only institutions located in middle and high-income countries have developed and implemented the technology, however, the efficiency and cost savings, as opposed to more traditional sequencing methodologies (e.g. Sanger sequencing) make the approach potentially well suited for resource-constrained regions as well. In April 2018, scientists from the Plasmodium Diversity Network Africa (PDNA) and collaborators met during the 7th Pan African Multilateral Initiative of Malaria (MIM) conference held in Dakar, Senegal to explore the feasibility of applying TNGS to genetic studies and malaria surveillance in Africa. The group of scientists reviewed the current experience with TNGS platforms in sub-Saharan Africa (SSA) and identified potential roles the technology might play to accelerate malaria research, scientific discoveries and improved public health in SSA. Research funding, infrastructure and human resources were highlighted as challenges that will have to be mitigated to enable African scientists to drive the implementation of TNGS in SSA. Current roles of important stakeholders and strategies to strengthen existing networks to effectively harness this powerful technology for malaria research of public health importance were discussed

In vitro susceptibility to pyrimethamine of DHFR I164L single mutant Plasmodium falciparum

<p>Abstract</p> <p>Background</p> <p>Recently, <it>Plasmodium falciparum </it>parasites bearing <it>Pfdhfr </it>I164L single mutation were found in Madagascar. These new mutants may challenge the use of antifolates for the intermittent preventive treatment of malaria during pregnancy (IPTp). Assays with transgenic bacteria suggested that I164L parasites have a wild-type phenotype for pyrimethamine but it had to be confirmed by testing the parasites themselves.</p> <p>Methods</p> <p>Thirty <it>Plasmodium falciparum </it>clinical isolates were collected in 2008 in the south-east of Madagascar. A part of <it>Pfdhfr </it>gene encompassing codons 6 to 206 was amplified by PCR and the determination of the presence of single nucleotide polymorphisms was performed by DNA sequencing. The multiplicity of infection was estimated by using an allelic family-specific nested PCR. Isolates that appeared monoclonal were submitted to culture adaptation. Determination of IC_50sto pyrimethamine was performed on adapted isolates.</p> <p>Results</p> <p>Four different <it>Pfdhfr </it>alleles were found: the 164L single mutant-type (N = 13), the wild-type (N = 7), the triple mutant-type 51I/59R/108N (N = 9) and the double mutant-type 108N/164L (N = 1). Eleven out 30 (36.7%) of <it>P. falciparum </it>isolates were considered as monoclonal infection. Among them, five isolates were successfully adapted in culture and tested for pyrimethamine <it>in vitro </it>susceptibility. The wild-type allele was the most susceptible with a 50% inhibitory concentration (IC₅₀) < 10 nM. The geometric mean of IC₅₀of the three I164L mutant isolates was 6-fold higher than the wild-type with 61.3 nM (SD = 3.2 nM, CI95%: 53.9-69.7 nM). These values remained largely below the IC₅₀of the triple mutant parasite (13,804 nM).</p> <p>Conclusion</p> <p>The IC₅₀s of the I164L mutant isolates were significantly higher than those of the wild-type (6-fold higher) and close from those usually reported for simple mutants S108N (roughly10-fold higher than wild type). Given the observed values, the determination of IC₅₀s directly on parasites did not confirm what has been found on transgenic bacteria. The prevalence increase of the <it>Pfdhfr </it>I164L single mutant parasite since 2006 could be explained by the selective advantage of this allele under sulphadoxine-pyrimethamine pressure. The emergence of highly resistant alleles should be considered in the future, in particular because an unexpected double mutant-type allele S108N/I164L has been already detected.</p

Fc gamma receptor IIa-H131R polymorphism and malaria susceptibility in sympatric ethnic groups, Fulani and Dogon of Mali.

It has been previously shown that there are some interethnic differences in susceptibility to malaria between two sympatric ethnic groups of Mali, the Fulani and the Dogon. The lower susceptibility to Plasmodium falciparum malaria seen in the Fulani has not been fully explained by genetic polymorphisms previously known to be associated with malaria resistance, including haemoglobin S (HbS), haemoglobin C (HbC), alpha-thalassaemia and glucose-6-phosphate dehydrogenase (G6PD) deficiency. Given the observed differences in the distribution of FcγRIIa allotypes among different ethnic groups and with malaria susceptibility that have been reported, we analysed the rs1801274-R131H polymorphism in the FcγRIIa gene in a study of Dogon and Fulani in Mali (n = 939). We confirm that the Fulani have less parasite densities, less parasite prevalence, more spleen enlargement and higher levels of total IgG antibodies (anti-CSP, anti-AMA1, anti-MSP1 and anti-MSP2) and more total IgE (P < 0.05) compared with the Dogon ethnic group. Furthermore, the Fulani exhibit higher frequencies of the blood group O (56.5%) compared with the Dogon (43.5%) (P < 0.001). With regard to the FcγRIIa polymorphism and allele frequency, the Fulani group have a higher frequency of the H allele (Fulani 0.474, Dogon 0.341, P < 0.0001), which was associated with greater total IgE production (P = 0.004). Our findings show that the FcγRIIa polymorphism might have an implication in the relative protection seen in the Fulani tribe, with confirmatory studies required in other malaria endemic settings

In silico characterisation of putative Plasmodium falciparum vaccine candidates in African malaria populations.

Genetic diversity of surface exposed and stage specific Plasmodium falciparum immunogenic proteins pose a major roadblock to developing an effective malaria vaccine with broad and long-lasting immunity. We conducted a prospective genetic analysis of candidate antigens (msp1, ama1, rh5, eba175, glurp, celtos, csp, lsa3, Pfsea, trap, conserved chrom3, hyp9, hyp10, phistb, surfin8.2, and surfin14.1) for malaria vaccine development on 2375 P. falciparum sequences from 16 African countries. We described signatures of balancing selection inferred from positive values of Tajima's D for all antigens across all populations except for glurp. This could be as a result of immune selection on these antigens as positive Tajima's D values mapped to regions with putative immune epitopes. A less diverse phistb antigen was characterised with a transmembrane domain, glycophosphatidyl anchors between the N and C- terminals, and surface epitopes that could be targets of immune recognition. This study demonstrates the value of population genetic and immunoinformatic analysis for identifying and characterising new putative vaccine candidates towards improving strain transcending immunity, and vaccine efficacy across all endemic populations

Identification of pyrimethamine- and chloroquine-resistant Plasmodium falciparum in Africa between 1984 and 1998: genotyping of archive blood samples

<p>Abstract</p> <p>Background</p> <p>Understanding the geographical distribution of drug resistance of <it>Plasmodium falciparum </it>is important for the effective treatment of malaria. Drug resistance has previously been inferred mainly from records of clinical resistance. However, clinical resistance is not always consistent with the parasite's genetic resistance. Thus, molecular identification of the parasite's drug resistance is required. In Africa, clinical resistance to pyrimethamine (Pyr) and chloroquine (CQ) was evident before 1980 but few studies investigating the genetic resistance to these drugs were conducted before the late 1990s. In this study, genotyping of genes involved in resistance to Pyr and CQ was performed using archive blood samples from Africa between 1984 and 1998.</p> <p>Methods</p> <p>Parasite DNA was extracted from <it>P. falciparum</it>-infected blood smears collected from travellers returning to Japan from Africa between 1984 and 1998. Genotypes of the dihydrofolate reductase gene (<it>dhfr</it>) and CQ-resistance transporter gene (<it>pfcrt) </it>were determined by polymerase chain reaction amplification and sequencing.</p> <p>Results</p> <p>Genotyping of <it>dhfr </it>and <it>pfcrt </it>was successful in 59 and 80 samples, respectively. One wild-type and seven mutant <it>dhfr </it>genotypes were identified. Three <it>dhfr </it>genotypes lacking the S108N mutation (NRSI, ICSI, IRSI; amino acids at positions 51, 59, 108, and 164 with mutations underlined) were highly prevalent before 1994 but reduced after 1995, accompanied by an increase in genotypes with the S108N mutation. The <it>dhfr </it>IRNI genotype was first identified in Nigeria in 1991 in the present samples, and its frequency gradually increased. However, two double mutants (ICNI and NRNI), the latter of which was exclusively found in West Africa, were more frequent than the IRNI genotype. Only two <it>pfcrt </it>genotypes were found, the wild-type and a Southeast Asian type (CVIET; amino acids at positions 72-76 with mutations underlined). The CVIET genotype was already present as early as 1984 in Tanzania and Nigeria, and appeared throughout Africa between 1984 and 1998.</p> <p>Conclusions</p> <p>This study is the first to report the molecular identification of Pyr- and CQ-resistant genotypes of <it>P. falciparum </it>in Africa before 1990. Genotyping of <it>dhfr </it>and <it>pfcrt </it>using archive samples has revealed new aspects of the evolutionary history of Pyr- and CQ-resistant parasites in Africa.</p