Molecular prevalence, genetic characterization and patterns of Toxoplasma gondii infection in domestic small mammals from Cotonou, Benin.

Toxoplasmosis, one of the most prevalent parasitic infections in humans and animals, is caused by the intracellular protozoan parasite Toxoplasma gondii. Small mammals play a key role as intermediate reservoir hosts in the maintenance of the T. gondii life cycle. In this study, we estimated the molecular prevalence and provide genetic diversity data for T. gondii in 632 small mammals sampled in four areas of Cotonou city, Benin. Both the brain and heart of each individual were screened through T. gondii-targeting qPCR, and positive samples were then genotyped using a set of 15 T. gondii-specific microsatellites. Prevalence data were statistically analyzed in order to assess the relative impact of individual host characteristics, spatial distribution, composition of small mammal community, and urban landscape features. An overall T. gondii molecular prevalence of 15.2% was found and seven genotypes, all belonging to the Africa 1 lineage, could be retrieved from the invasive black rat Rattus rattus and the native African giant shrew Crocidura olivieri. Statistical analyses did not suggest any significant influence of the environmental parameters used in this study. Rather, depending on the local context, T. gondii prevalence appeared to be associated either with black rat, shrew, or mouse abundance or with the trapping period. Overall, our results highlight the intricate relationships between biotic and abiotic factors involved in T. gondii epidemiology and suggest that R. rattus and C. olivieri are two competent reservoirs for the Africa 1 lineage, a widespread lineage in tropical Africa and the predominant lineage in Benin.

Title: Prévalence moléculaire, caractérisation génétique et schémas d'infection par Toxoplasma gondii chez les petits mammifères domestiques de Cotonou, Bénin. Abstract: La toxoplasmose, l'une des infections parasitaires les plus répandues chez l'homme et les animaux, est causée par le parasite protozoaire intracellulaire Toxoplasma gondii. Les petits mammifères jouent un rôle clé en tant qu'hôtes réservoirs intermédiaires dans le maintien du cycle de vie de T. gondii. Dans cette étude, nous estimons sa prévalence moléculaire et fournissons des données sur sa diversité génétique chez 632 petits mammifères échantillonnés dans quatre localités de la ville de Cotonou. Le cerveau et le cœur de chaque individu ont été analysés par qPCR ciblant T. gondii, et les échantillons positifs ont ensuite été génotypés à l'aide d'un ensemble de 15 microsatellites spécifiques à T. gondii. Les données de prévalence ont été analysées statistiquement afin d'évaluer l'impact relatif des caractéristiques individuelles de l'hôte, de la distribution spatiale, de la composition de la communauté des petits mammifères ainsi que des caractéristiques du paysage urbain. Une prévalence moléculaire globale de T. gondii de 15,2 % a été estimée et sept génotypes, tous appartenant à la lignée Africa 1, ont pu être extraits du rat noir Rattus rattus, espèce envahissante, et de la musaraigne Crocidura olivieri, espèce indigène. Les analyses statistiques n'ont pas suggéré d'influence significative des paramètres environnementaux utilisés dans cette étude. Au contraire, selon le contexte local, la prévalence de T. gondii semble être associée à l'abondance de rats noirs, de musaraignes ou de souris ainsi qu'à la période de piégeage. Dans l'ensemble, nos résultats mettent en évidence les relations complexes entre les facteurs biotiques et abiotiques impliqués dans l'épidémiologie de T. gondii et suggèrent que R. rattus et C. olivieri sont deux réservoirs compétents pour la lignée Africa 1, une lignée répandue en Afrique tropicale et prédominante au Bénin.

Fine-scale prevalence and genetic diversity of urban small mammal-borne pathogenic Leptospira in Africa: A spatiotemporal survey within Cotonou, Benin.

Leptospirosis is a zoonotic disease that is caused by spirochete bacteria of the genus Leptospira. Around the world, one million people each year are infected, leading to 60,000 deaths. Infection occurs through contact with environmental pathogens excreted by mammals (notably rodents). Data on Leptospira and leptospirosis in Africa are rather scarce, especially in urban habitats though these appear to be favourable environments for the pathogen circulation and human contamination. Using qPCR, DNA sequencing as well as MST/VNTR approaches, we examined Leptospira occurrence and genetic diversity in 779 commensal small mammals that were sampled over 2 years in the city centre of Cotonou, Benin, from three neighbourhoods with contrasting socio-environmental conditions. Overall prevalence reached 9.1%. However, very marked variations in both space and time were observed, with local peaks of high prevalence but no clear seasonal pattern. In most sites that could be regularly sampled, Leptospira-positive rodents were found at least once, thus confirming the widespread circulation of the pathogen within small mammal communities of Cotonou. Interestingly, an unusual diversity of small mammal-borne Leptospira species and genotypes was retrieved, with up to four species and three different genovars within the same neighbourhood, and even instances of two species and two genovars identified simultaneously within the same household. To our knowledge, such a high genetic diversity has never been described at such a fine scale, a fortiori in Africa and, more generally, within an urban environment. Altogether, our results underline that much remains unknown about leptospirosis as well as the associated infectious risk in African cities where the disease may be massively over-looked.

Genetic Characterization of Seoul Virus in the Seaport of Cotonou, Benin.

Seoul virus is a zoonotic pathogen carried by the brown rat Rattus norvegicus. Information on its circulation in Africa is limited. In this study, the virus was detected in 37.5% of brown rats captured in the Autonomous Port of Cotonou, Benin. Phylogenetic analyses place this virus in Seoul virus lineage 7.

Pathogenic Leptospira and water quality in African cities: A case study of Cotonou, Benin.

Leptospirosis is a waterborne zoonosis (60,000 infections and 1 million deaths annually). Knowledge about the disease in the urban context is surprisingly rare, especially in Africa. Here, we provide the first study of leptospires in waters within an African city. A simple centrifugation-based method was developed to screen waterborne leptospires from remote or poorly areas. Major ions, trace elements, stable isotopes and pathogenic Leptospira were then seasonally investigated in 193 water samples from three neighborhoods of Cotonou (Benin) with different socio-environmental and hydrographic characteristics. Firstly, no leptospire was detected in tap waters. Secondly, although surface contamination cannot be excluded, one groundwater well was found leptospire positive. Thirdly, pathogenic Leptospira mainly contaminated surface waters of temporary and permanent ponds (9.5% and 27.3% of total prevalence, respectively). Isotopic signatures suggest that leptospires occurred in pond waters formed at the beginning of the rainy season following low to moderate rainfall events. Nevertheless, Leptospira-containing waters possess physico-chemical characteristics that are similar to the spectrum of waters sampled throughout the three sites, thus suggesting that Cotonou waters are widely compatible with Leptospira survival. The frequent contact with water exposes Cotonou inhabitants to the risk of leptospirosis which deserves more attention from public health authorities.

Two new African siblings of Pulveroboletusravenelii (Boletaceae).

This paper sorts out the taxonomy of species affiliated with Pulveroboletusravenelii in the Guineo-soudanian and Zambezian woodlands of Africa. Morphological and genetic characters of African Pulveroboletus collections were studied and compared to those of North American and Asian species. A phylogenetic analysis showed that the African specimens form a subclade, sister to the Asian and American taxa. Although clamp connections have previously never been reported from Pulveroboletus, all specimens of the African subclade show very small clamp connections. Two new African species, Pulveroboletusafricanus sp. nov. and P.sokponianus sp. nov., are described and illustrated. Comments concerning morphology and identification, as well as distribution and ecology, are given for both species.