Detection of Anaplasma and Ehrlichia bacteria in humans, wildlife, and ticks in the Amazon rainforest.

Tick-borne bacteria of the genera Ehrlichia and Anaplasma cause several emerging human infectious diseases worldwide. In this study, we conduct an extensive survey for Ehrlichia and Anaplasma infections in the rainforests of the Amazon biome of French Guiana. Through molecular genetics and metagenomics reconstruction, we observe a high indigenous biodiversity of infections circulating among humans, wildlife, and ticks inhabiting these ecosystems. Molecular typing identifies these infections as highly endemic, with a majority of new strains and putative species specific to French Guiana. They are detected in unusual rainforest wild animals, suggesting they have distinctive sylvatic transmission cycles. They also present potential health hazards, as revealed by the detection of Candidatus Anaplasma sparouinense in human red blood cells and that of a new close relative of the human pathogen Ehrlichia ewingii, Candidatus Ehrlichia cajennense, in the tick species that most frequently bite humans in South America. The genome assembly of three new putative species obtained from human, sloth, and tick metagenomes further reveals the presence of major homologs of Ehrlichia and Anaplasma virulence factors. These observations converge to classify health hazards associated with Ehrlichia and Anaplasma infections in the Amazon biome as distinct from those in the Northern Hemisphere.

Capybara and Brush Cutter Involvement in Q Fever Outbreak in Remote Area of Amazon Rain Forest, French Guiana, 2014.

We investigated a Q fever outbreak that occurred in an isolated area of the Amazon Rain Forest in French Guiana in 2014. Capybara fecal samples were positive for Coxiella burnetii DNA. Being near brush cutters in use was associated with disease development. Capybaras are a putative reservoir for C. burnetii.

DNA barcoding of the rodent genus Oligoryzomys (Cricetidae: Sigmodontinae): mitogenomic-anchored database and identification of nuclear mitochondrial translocations (Numts).

DNA barcoding has become a standard method for species identification in taxonomically complex groups. An important step of the barcoding process is the construction of a library of voucher-based material that was properly identified by independent methods, free of inaccurate identification, and paralogs. We provide here a cytochrome oxidase I (mt-Co1) DNA barcode database for species of the genus Oligoryzomys, based on type material and karyotyped specimens, and anchored on the mitochondrial genome of one species of Oligoryzomys, O. stramineus. To evaluate the taxonomic determination of new COI sequences, we assessed species intra/interspecific genetic distances (barcode gap), performed the General Mixed Yule Coalescent method (GMYC) for lineages' delimitation, and identified diagnostic nucleotides for each species of Oligoryzomys. Phylogenetic analyses of Oligoryzomys were performed on 2 datasets including 14 of the 23 recognized species of this genus: a mt-Co1 only matrix, and a concatenated matrix including mt-Co1, cytochrome b (mt-Cytb), and intron 7 of the nuclear fibrinogen beta chain gene (i7Fgb). We recovered nuclear-mitochondrial translocated (Numts) pseudogenes on our samples and identified several published sequences that are cases of Numts. We analyzed the rate of non-synonymous and synonymous substitution, which were higher in Numts in comparison to mtDNA sequences. GMYC delimitations and DNA barcode gap results highlight the need for further work that integrate molecular, karyotypic, and morphological analyses, as well as additional sampling, to tackle persistent problems in the taxonomy of Oligoryzomys.

Streblid bat flies (Diptera) and other ectoparasites on bats (Mammalia: Chiroptera) from French Guiana / Moscas estréblidas (Diptera) e outros ectoparasitos de morcegos (Mammalia: Chiroptera) na Guiana Francesa

Abstract: Recent field surveys of bats (Chiroptera) in various localities of French Guiana have been accompanied by the collection and preservation of ectoparasites, mainly bat flies (Diptera: Streblidae and Nycteribiidae). Most specimens of ectoparasites was collected haphazardly during the course of bats inventories, but systematic surveys on the whole chiropteran community were realized at five opportunities. Concerning Streblidae, 813 individuals have been examined, which represent 46 species and/or subspecies belonging to 15 genera and 6 taxa for confirmation and/or future description. For Nycteribiidae, 44 individuals of 3 identified species and 2 for confirmation and/or future description. Other ectoparasites have been found (Hemiptera and acarids), which are also listed with details on their bat-host, place and date of collect. For six species of bats in which at least 10 animals were carrying ectoparasites, a brief description of the frequencies of their ectoparasites provides some preliminary characteristics of their infracommunities.

Resumo: Pesquisas de campo recentes de morcegos (Chiroptera) em várias localidades da Guiana Francesa foram acompanhadas pela coleta e preservação de ectoparasitas, principalmente moscas ectoparasitas (Diptera: Streblidae e Nycteribiidae). A maioria dos espécimes de ectoparasitos foi coletada aleatoriamente durante o curso dos inventários de morcegos, mas pesquisas sistemáticas em toda a comunidade de quirópteros foram realizadas em cinco oportunidades. Sobre Streblidae, 813 indivíduos foram examinados, dos quais representam 46 espécies e/ou subespécies pertencentes a 15 gêneros e 6 táxons para posterior confirmação específica e/ou descrição. Para Nycterbiidae, 44 indivíduos de 3 espécies e 2 para posterior confirmação específica e/ou descrição. Outros ectoparasitos foram coletados (hemípteros polictenídeos e ácaros), os quais também foram listados com detalhes sobre seus hospedeiros, localidade e data de coleta. Para seis espécies de morcegos com mais de 10 morcegos infestados a descrição da composição de cada infracomunidade encontrada e sua frequência são apresentados.

Reproduction and growth in a Neotropical arboreal mouse: Oecomys rutilus (Sigmodontinae: Cricetidae) in French Guiana / Reprodução e crescimento de um rato arbóreo neotropical: Oecomys rutilus (Sigmodontinae: Cricetidae) na Guiana Francesa

Abstract: Postnatal growth and development of the small Neotropical mouse Oecomys rutilus (Sigmodontinae: Cricetidae) were investigated from birth to day 143, in the laboratory. Morphometric measurements at age of 3 days, of both sexes combined, revealed body weight to be 3.4 ± 0.3 g, mean tail length as 27.4 ± 1.1 mm, and mean hind foot length as 9.3 ± 0.7 mm. Body weight was found to increase steadily until at least 69 days, whereas the instantaneous growth rates of other measurements declined earlier: the daily growth of hind foot length declined to a minimum at age of 24 days, and the growth of tail and of ear declined by the age of 33 days. Average litter size for 12 captive births was 2.5, ranging from 2 to 3. The preserved eye crystalline lens was weighted in 23 captive-born animals of known age, allowing a rough estimate of the age of reproduction in wild-caught animals. Based on the inferred relation between eye-lens weight and age, the youngest reproductive (pregnant) wild-caught females had an estimated age of 90 and 95 days.

Resumo: O crescimento pós-natal e o desenvolvimento do pequeno roedor neotropical Oecomys rutilus (Sigmodontinae: Cricetidae) foram investigados desde o nascimento até o dia 143, em laboratório. Mensurações morfométricas na idade de 3 dias, de ambos os sexos combinados, revelaram a massa corporal de 3,4 ± 0,3 g, comprimento médio da cauda de 27,4 ± 1,1 mm, e comprimento médio do pé de 9,3 ± 0,7 mm. A massa do corpo aumentou rapidamente até o dia 69, enquanto as taxas de crescimento instantâneo de outras medidas diminuíram mais cedo: o crescimento diário do pé diminuiu para o mínimo na idade de 24 dias, e os crescimentos da cauda e da orelha diminuíram na idade de 33 dias. O tamanho de prole médio para 12 nascimentos em cativeiro foi de 2,5, variando de 2 a 3. As lentes preservadas do cristalino do olho foram pesadas em 23 animais nascidos em cativeiro de idades conhecidas, permitindo fazer uma estimativa da idade de reprodução em animais capturados na natureza. Baseado na relação inferida entre peso da lente do cristalino e idade, as mais jovens fêmeas (prenhes) capturadas na natureza tiveram uma estimativa de 90 a 95 dias de idade.

The role of Pleistocene climate change in the genetic variability, distribution and demography of Proechimys cuvieri and P. guyannensis (Rodentia: Echimyidae) in northeastern Amazonia.

The spiny rats, genus Proechimys, have the highest species richness within the Echimyidae family, as well as species with high genetic variability. The genus distribution includes tropical South America and Central America south to Honduras. In this study, we evaluate the phylogeographic histories of Proechimys guyannensis and P. cuvieri using cytochrome b, in a densely sampled area in northeastern Amazon where both species are found in sympatry in different environments. For each species, Bayesian and Maximum Likelihood phylogenetic analysis were congruent and recovered similar clades in the studied area. Bayesian phylogenetic analysis using a relaxed molecular clock showed that these clusters of haplotypes diversified during Pleistocene for both species. Apparently, the large rivers of the region did not act as barriers, as some clades include specimens collected from opposite banks of Oiapoque, Araguari and Jari rivers. Bayesian skyline plot analysis showed recent demographic expansion in both species. The Pleistocene climatic changes in concert with the geologic changes in the Amazon fan probably acted as drivers in the diversification that we detected in these two spiny rats. Proechimys cuvieri and P. guyannensis show genetic structure in the eastern part of the Guiana region. Greater genetic distances observed in P. guyannensis, associated with highly structured groups, suggest that more detailed studies of systematics and ecology should be directed to this species.

In Cold Blood: Compositional Bias and Positive Selection Drive the High Evolutionary Rate of Vampire Bats Mitochondrial Genomes.

Mitochondrial genomes of animals have long been considered to evolve under the action of purifying selection. Nevertheless, there is increasing evidence that they can also undergo episodes of positive selection in response to shifts in physiological or environmental demands. Vampire bats experienced such a shift, as they are the only mammals feeding exclusively on blood and possessing anatomical adaptations to deal with the associated physiological requirements (e.g., ingestion of high amounts of liquid water and iron). We sequenced eight new chiropteran mitogenomes including two species of vampire bats, five representatives of other lineages of phyllostomids and one close outgroup. Conducting detailed comparative mitogenomic analyses, we found evidence for accelerated evolutionary rates at the nucleotide and amino acid levels in vampires. Moreover, the mitogenomes of vampire bats are characterized by an increased cytosine (C) content mirrored by a decrease in thymine (T) compared with other chiropterans. Proteins encoded by the vampire bat mitogenomes also exhibit a significant increase in threonine (Thr) and slight reductions in frequency of the hydrophobic residues isoleucine (Ile), valine (Val), methionine (Met), and phenylalanine (Phe). We show that these peculiar substitution patterns can be explained by the co-occurrence of both neutral (mutational bias) and adaptive (positive selection) processes. We propose that vampire bat mitogenomes may have been impacted by selection on mitochondrial proteins to accommodate the metabolism and nutritional qualities of blood meals.

iDNA screening: Disease vectors as vertebrate samplers.

In the current context of global change and human-induced biodiversity decline, there is an urgent need for developing sampling approaches able to accurately describe the state of biodiversity. Traditional surveys of vertebrate fauna involve time-consuming and skill-demanding field methods. Recently, the use of DNA derived from invertebrate parasites (leeches and blowflies) was suggested as a new tool for vertebrate diversity assessment. Bloodmeal analyses of arthropod disease vectors have long been performed to describe their feeding behaviour, for epidemiological purposes. On the other hand, this existing expertise has not yet been applied to investigate vertebrate fauna per se. Here, we evaluate the usefulness of hematophagous dipterans as vertebrate samplers. Blood-fed sand flies and mosquitoes were collected in Amazonian forest sites and analysed using high-throughput sequencing of short mitochondrial markers. Bloodmeal identifications highlighted contrasting ecological features and feeding behaviour among dipteran species, which allowed unveiling arboreal and terrestrial mammals of various body size, as well as birds, lizards and amphibians. Additionally, lower vertebrate diversity was found in sites undergoing higher levels of human-induced perturbation. These results suggest that, in addition to providing precious information on disease vector host use, dipteran bloodmeal analyses may represent a useful tool in the study of vertebrate communities. Although further effort is required to validate the approach and consider its application to large-scale studies, this first work opens up promising perspectives for biodiversity monitoring and eco-epidemiology.

Virus hébergés par les rongeurs en Guyane française.

Among mammals, rodents play a key role in the emergence of viral diseases. In French Guiana, with 36 rodent species recorded in various ecosystems (pristine forests, savannas, anthropized environments), some natural habitats today encounter anthropogenic perturbations that induce changes in community structure and population dynamics. These modifications are sometimes associated with the circulation and emergence of viral pathogens. For 10 years, investigations on the circulation of two rodent-borne viruses, Hantavirus and Mammarenavirus, are underway in rodent populations as well as in humans for hantavirus. These investigations identified viruses from both genera in their potential reservoirs and allow describing the most favourable habitats for the reservoirs of hantavirus where the risk of viral emergence may be higher. We suggest to investigate how anthropic perturbations in rodent communities can drive the emergence of viruses that are currently confined to a small scale and search for evidence of infection in the human population.

Rodent-borne viruses in French Guiana.

Among mammals, rodents play a key role in the emergence of viral diseases. In French Guiana, with 36 rodent species recorded in various ecosystems (pristine forests, savannas, anthropized environments), some natural habitats today encounter anthropogenic perturbations that induce changes in community structure and population dynamics. These modifications are sometimes associated with the circulation and emergence of viral pathogens. For 10 years, investigations on the circulation of two rodent-borne viruses, Hantavirus and Mammarenavirus, are underway in rodent populations as well as in humans for hantavirus. These investigations identified viruses from both genera in their potential reservoirs and allow describing the most favourable habitats for the reservoirs of hantavirus where the risk of viral emergence may be higher. We suggest to investigate how anthropic perturbations in rodent communities can drive the emergence of viruses that are currently confined to a small scale and search for evidence of infection in the human population.

Guerrerostrongylus marginalis n. sp. (Trichostrongyloidea: Heligmonellidae) from the Guianan arboreal mouse (Oecomys auyantepui) from French Guiana.

Based on the number and arrangement of cuticular ridges and configuration of the dorsal ray, nematode specimens collected from the small intestine of eight Guianan arboreal mice, Oecomys auyantepui (Rodentia: Sigmodontinae), in French Guiana are herein described and characterized. Guerrerostrongylus marginalis n. sp. (Heligmosomoidea: Heligmonellidae) shows a synlophe consisting of more than 40 ridges and a unique bursal arrangement with ray 8 (externo-dorsal) extending to the edge of the bursal margin, and appearing more prominent than the dorsal ray. This bursal arrangement is common in members of Hassalstrongylus Durette-Desset, 1971, but uncommon in the other four species in Guerrerostrongylus Sutton & Durette-Desset, 1991. The placement of the new species in Guerrerostrongylus is based on the number and nature of cuticular ridges and the ray arrangement and symmetry of the caudal bursa. Diagnostic characteristics of Guerrerostrongylus marginalis n. sp. include the length of ray 8 relative to bursal margin, the relative size of the spicules and vestibule, and the number of eggs in the uterus. We propose an amendment to the generic diagnosis of Guerrerostrongylus to modify the characters of the long rays 6 (postero-lateral), rays 8 (externo-dorsal), and dorsal ray as diagnostic, since at least ray 6 appears to be short in two different species in the genus, namely G. ulysi Digiani, Notarnicola & Navone, 2012 and G. marginalis n. sp.

Identification of lymphocytic choriomeningitis mammarenavirus in house mouse (Mus musculus, Rodentia) in French Guiana.

Thirty-seven house mice (Mus musculus, Rodentia) caught in different localities in French Guiana were screened to investigate the presence of lymphocytic choriomeningitis mammarenavirus (LCMV). Two animals trapped in an urban area were found positive, hosting a new strain of LCMV, that we tentatively named LCMV "Comou". The complete sequence was determined using a metagenomic approach. Phylogenetic analyses revealed that this strain is related to genetic lineage I composed of strains inducing severe disease in humans. These results emphasize the need for active surveillance in humans as well as in house mouse populations, which is a rather common rodent in French Guianese cities and settlements.

First record of Myotis albescens (Chiroptera, Vespertilionidae) in French Guiana.

BACKGROUND: Myotis albescens occurs from Mexico southward to Uruguay and Argentina. The species is known for all South American countries except French Guiana and Chile. NEW INFORMATION: Based on one specimen recently collected in French Guiana we fill part of the gap in the distribution of the species in South America. Myotis albescens occurs in the Guiana Shield with other four congeners, from which it can be distinguished by external and skull traits. As an aid to future identifications, we provide a key to this assemblage.

Lack of dissemination of acquired resistance to ß-lactams in small wild mammals around an isolated village in the Amazonian forest.

In this study, we quantitatively evaluated the spread of resistance to ß-lactams and of integrons in small rodents and marsupials living at various distances from a point of antibiotic's use. Rectal swabs from 114 animals were collected in Trois-Sauts, an isolated village in French Guiana, and along a 3 km transect heading through the non-anthropized primary forest. Prevalence of ticarcillin-resistant enterobacteria was 36% (41/114). Klebsiella spp., naturally resistant to ticarcillin, were found in 31.1% (23/73) of animals from the village and in an equal ratio of 31.7% (13/41) of animals trapped along the transect. By contrast Escherichia coli with acquired resistance to ticarcillin were found in 13.7% (10/73) of animals from the village and in only 2.4% (1/41) of those from the transect (600 m from the village). There was a huge diversity of E. coli and Klebsiella pneumoniae strains with very unique and infrequent sequence types. The overall prevalence of class 1 integrons carriage was 19.3% (22/114) homogenously distributed between animals from the village and the transect, which suggests a co-selection by a non-antibiotic environmental factor. Our results indicate that the anthropogenic acquired antibiotic resistance did not disseminate in the wild far from the point of selective pressure.

Retroviral envelope gene captures and syncytin exaptation for placentation in marsupials.

Syncytins are genes of retroviral origin captured by eutherian mammals, with a role in placentation. Here we show that some marsupials-which are the closest living relatives to eutherian mammals, although they diverged from the latter ∼190 Mya-also possess a syncytin gene. The gene identified in the South American marsupial opossum and dubbed syncytin-Opo1 has all of the characteristic features of a bona fide syncytin gene: It is fusogenic in an ex vivo cell-cell fusion assay; it is specifically expressed in the short-lived placenta at the level of the syncytial feto-maternal interface; and it is conserved in a functional state in a series of Monodelphis species. We further identify a nonfusogenic retroviral envelope gene that has been conserved for >80 My of evolution among all marsupials (including the opossum and the Australian tammar wallaby), with evidence for purifying selection and conservation of a canonical immunosuppressive domain, but with only limited expression in the placenta. This unusual captured gene, together with a third class of envelope genes from recently endogenized retroviruses-displaying strong expression in the uterine glands where retroviral particles can be detected-plausibly correspond to the different evolutionary statuses of a captured retroviral envelope gene, with only syncytin-Opo1 being the present-day bona fide syncytin active in the opossum and related species. This study would accordingly recapitulate the natural history of syncytin exaptation and evolution in a single species, and definitely extends the presence of such genes to all major placental mammalian clades.

Patawa Virus, a New Arenavirus Hosted by Forest Rodents in French Guiana.

Molecular screening of rodents from French Guiana has detected a new arenavirus, named "Patawa," in two Oecomys species (Muridae, Sigmodontinae). Further investigations are needed to better understand the circulation of this virus in rodent and human populations and its public health impact.

Evolution of spatially coexpressed families of type-2 vomeronasal receptors in rodents.

The vomeronasal organ (VNO) is an olfactory structure for the detection of pheromones. VNO neurons express three groups of unrelated G-protein-coupled receptors. Type-2 vomeronasal receptors (V2Rs) are specifically localized in the basal neurons of the VNO and are believed to sense protein pheromones eliciting specific reproductive behaviors. In murine species, V2Rs are organized into four families. Family-ABD V2Rs are expressed monogenically and coexpress with family-C V2Rs of either subfamily C1 (V2RC1) or subfamily C2 (V2RC2), according to a coordinate temporal diagram. Neurons expressing the phylogenetically ancient V2RC1 coexpress family-BD V2Rs or a specific group of subfamily-A V2Rs (V2RA8-10), whereas a second neuronal subset (V2RC2-positive) coexpresses a recently expanded group of five subfamily-A V2Rs (V2RA1-5) along with vomeronasal-specific Major Histocompatibility Complex molecules (H2-Mv). Through database mining and Sanger sequencing, we have analyzed the onset, diversification, and expansion of the V2R-families throughout the phylogeny of Rodentia. Our results suggest that the separation of V2RC1 and V2RC2 occurred in a Cricetidae ancestor in coincidence with the evolution of the H2-Mv genes; this phylogenetic event did not correspond with the origin of the coexpressing V2RA1-5 genes, which dates back to an ancestral myomorphan lineage. Interestingly, the evolution of receptors within the V2RA1-5 group may be implicated in the origin and diversification of some of the V2R putative cognate ligands, the exocrine secreting peptides. The establishment of V2RC2, which probably reflects the complex expansion and diversification of family-A V2Rs, generated receptors that have probably acquired a more subtle functional specificity.

The captured retroviral envelope syncytin-A and syncytin-B genes are conserved in the Spalacidae together with hemotrichorial placentation.

Syncytins are fusogenic envelope (env) genes of retroviral origin that have been captured for a function in placentation. Multiple independent events of syncytin gene capture were found to have occurred in primates, rodents, lagomorphs, carnivores, and ruminants. In the mouse, two syncytin-A and -B genes are present, which trigger the formation of the two-layered placental syncytiotrophoblast at the maternal-fetal interface, a structure classified as hemotrichorial. Here, we identified syncytin-A and -B orthologous genes in the genome of all Muroidea species analyzed, thus dating their capture back to about at least 40 million years ago, with evidence that they evolved under strong purifying selection. We further show, in the divergent Spalacidae lineage (blind mole rats [Spalax]), that both syncytins have conserved placenta-specific expression, as revealed by RT-PCR analysis of a panel of Spalax galili tissues, and display fusogenic activity, using ex vivo cell-cell fusion assays. Refined analysis of the placental architecture and ultrastructure revealed that the Spalax placenta displays a hemotrichorial organization of the interhemal membranes, as similarly observed for other Muroidea species, yet with only one trophoblastic cell layer being clearly syncytialized. In situ hybridization experiments further localized syncytin transcripts at the level of these differentiated interhemal membranes. These findings argue for a role of syncytin gene capture in the establishment of the original hemotrichorial placenta of Muroidea, and more generally in the diversity of placental structures among mammals.

Retroviral envelope syncytin capture in an ancestrally diverged mammalian clade for placentation in the primitive Afrotherian tenrecs.

Syncytins are fusogenic envelope (env) genes of retroviral origin that have been captured for a function in placentation. Syncytins have been identified in Euarchontoglires (primates, rodents, Leporidae) and Laurasiatheria (Carnivora, ruminants) placental mammals. Here, we searched for similar genes in species that retained characteristic features of primitive mammals, namely the Malagasy and mainland African Tenrecidae. They belong to the superorder Afrotheria, an early lineage that diverged from Euarchotonglires and Laurasiatheria 100 Mya, during the Cretaceous terrestrial revolution. An in silico search for env genes with full coding capacity within a Tenrecidae genome identified several candidates, with one displaying placenta-specific expression as revealed by RT-PCR analysis of a large panel of Setifer setosus tissues. Cloning of this endogenous retroviral env gene demonstrated fusogenicity in an ex vivo cell-cell fusion assay on a panel of mammalian cells. Refined analysis of placental architecture and ultrastructure combined with in situ hybridization demonstrated specific expression of the gene in multinucleate cellular masses and layers at the materno-fetal interface, consistent with a role in syncytium formation. This gene, which we named "syncytin-Ten1," is conserved among Tenrecidae, with evidence of purifying selection and conservation of fusogenic activity. To our knowledge, it is the first syncytin identified to date within the ancestrally diverged Afrotheria superorder.