Seroprevalence and renal carriage of pathogenic Leptospira in livestock in Cotonou, Benin.

BACKGROUND: Leptospirosis is a zoonotic disease. It is particularly prevalent in tropical countries and has major consequences for human and animal health. In Benin, the disease's epidemiology remains poorly understood, especially in livestock, for which data are lacking. OBJECTIVES: To characterise Leptospira seroprevalence and locally circulating serogroups in livestock from Cotonou and to estimate the prevalence of Leptospira renal carriage in cattle. METHODS: We conducted a cross-sectional study in February 2020 during which livestock were sampled at an abattoir and in an impoverished city district. We analysed blood samples from 279 livestock animals (i.e. cattle, sheep, goats and pigs) using the microscopic agglutination test. Additionally, samples of renal tissue from 100 cattle underwent 16s rRNA (rrs) real-time PCR analysis. RESULTS: For the 131 cattle, 85 sheep, and 50 goats tested, seroprevalence was 18% (95% confidence interval [CI] [12%, 26%]), 9% (95% CI [4%, 17%] and 2% (95% CI [0%, 9%]), respectively, and most of the seropositive animals were associated with 1:100 titres. All 13 pigs were seronegative. Leptospira DNA was found in the renal tissue of 10% (95% CI [5%, 18%]) of the cattle tested (n = 100). Leptospira borgpetersenii was the main species present (n = 7), but Leptospira interrogans (n = 2) and Leptospira kirschneri (n = 1) were also detected. Various serogroups (Canicola, Grippotyphosa, Sejroe, Icterohaemorrhagiae, Pomona, Pyrogenes, Australis and Autumnalis) were detected using microscopic agglutination test without a clear predominance of any of them. CONCLUSIONS: These results suggest that abattoir workers and people living in close contact with livestock in poor urban areas are exposed to the risk of Leptospira infection.

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.

A hotspot of Toxoplasma gondii Africa 1 lineage in Benin: How new genotypes from West Africa contribute to understand the parasite genetic diversity worldwide.

Through international trades, Europe, Africa and South America share a long history of exchanges, potentially of pathogens. We used the worldwide parasite Toxoplasma gondii to test the hypothesis of a historical influence on pathogen genetic diversity in Benin, a West African country with a longstanding sea trade history. In Africa, T. gondii spatial structure is still non-uniformly studied and very few articles have reported strain genetic diversity in fauna and clinical forms of human toxoplasmosis so far, even in African diaspora. Sera from 758 domestic animals (mainly poultry) in two coastal areas (Cotonou and Ouidah) and two inland areas (Parakou and Natitingou) were tested for T. gondii antibodies using a Modified Agglutination Test (MAT). The hearts and brains of 69 seropositive animals were collected for parasite isolation in a mouse bioassay. Forty-five strains were obtained and 39 genotypes could be described via 15-microsatellite genotyping, with a predominance of the autochthonous African lineage Africa 1 (36/39). The remaining genotypes were Africa 4 variant TUB2 (1/39) and two identical isolates (clone) of Type III (2/39). No difference in terms of genotype distribution between inland and coastal sampling sites was found. In particular, contrarily to what has been described in Senegal, no type II (mostly present in Europe) was isolated in poultry from coastal cities. This result seems to refute a possible role of European maritime trade in Benin despite it was one of the most important hubs during the slave trade period. However, the presence of the Africa 1 genotype in Brazil, predominant in Benin, and genetic analyses suggest that the triangular trade was a route for the intercontinental dissemination of genetic strains from Africa to South America. This supports the possibility of contamination in humans and animals with potentially imported virulent strains.

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.

Bartonella spp. in Small Mammals, Benin.

This study aimed to investigate the prevalence and genetic diversity of Bartonella organisms in small mammals in Cotonou, Benin. We captured 163 rodents and 12 insectivores and successfully detected Bartonella DNA from 13 of the 175 small mammal individuals. Bartonella spp., identical or closely related to Bartonella elizabethae, Bartonella tribocorum, and Bartonella rochalimae, was detected. A potential new Bartonella species, proposed as Candidatus Bartonella mastomydis, was found in three Mastomys individuals and genetically characterized by targeting two housekeeping genes (rpoB and gltA) and the intergenic species region. However, 20.8% of gray rats were found to be infected with Bartonella spp., and none of the black rats analyzed was positive. This work may be important from a public health point of view due to the zoonotic nature of the Bartonella species detected and warrants further investigation on the unknown zoonotic potential of this newly proposed Bartonella species.

Detection of Rickettsia felis, Rickettsia typhi, Bartonella Species and Yersinia pestis in Fleas (Siphonaptera) from Africa.

UNLABELLED: Little is known about the presence/absence and prevalence of Rickettsia spp, Bartonella spp. and Yersinia pestis in domestic and urban flea populations in tropical and subtropical African countries. METHODOLOGY/PRINCIPAL FINDINGS: Fleas collected in Benin, the United Republic of Tanzania and the Democratic Republic of the Congo were investigated for the presence and identity of Rickettsia spp., Bartonella spp. and Yersinia pestis using two qPCR systems or qPCR and standard PCR. In Xenopsylla cheopis fleas collected from Cotonou (Benin), Rickettsia typhi was detected in 1% (2/199), and an uncultured Bartonella sp. was detected in 34.7% (69/199). In the Lushoto district (United Republic of Tanzania), R. typhi DNA was detected in 10% (2/20) of Xenopsylla brasiliensis, and Rickettsia felis was detected in 65% (13/20) of Ctenocephalides felis strongylus, 71.4% (5/7) of Ctenocephalides canis and 25% (5/20) of Ctenophthalmus calceatus calceatus. In the Democratic Republic of the Congo, R. felis was detected in 56.5% (13/23) of Ct. f. felis from Kinshasa, in 26.3% (10/38) of Ct. f. felis and 9% (1/11) of Leptopsylla aethiopica aethiopica from Ituri district and in 19.2% (5/26) of Ct. f. strongylus and 4.7% (1/21) of Echidnophaga gallinacea. Bartonella sp. was also detected in 36.3% (4/11) of L. a. aethiopica. Finally, in Ituri, Y. pestis DNA was detected in 3.8% (1/26) of Ct. f. strongylus and 10% (3/30) of Pulex irritans from the villages of Wanyale and Zaa. CONCLUSION: Most flea-borne infections are neglected diseases which should be monitored systematically in domestic rural and urban human populations to assess their epidemiological and clinical relevance. Finally, the presence of Y. pestis DNA in fleas captured in households was unexpected and raises a series of questions regarding the role of free fleas in the transmission of plague in rural Africa, especially in remote areas where the flea density in houses is high.