Three new species of Eimeria Schneider 1875 in the montane grass mouse, Akodon montensis (Rodentia: Cricetidae: Sigmodontinae), and redescription of Eimeria zygodontomyis Lainson and Shaw 1990 from southeastern Brazil.

We describe three new coccidian species of the genus Eimeria Schneider 1875 (Apicomplexa: Eimeriidae) and redescribe and report Eimeria zygodontomyis Lainson and Shaw, 1990 in the montane grass mouse, Akodon montensis Thomas, 1913 from the Serra dos Órgãos National Park in southeastern Brazil. Sporulated oocysts of Eimeria zygodontomyis are ellipsoidal to cylindrical with a 0.6 (0.5-0.8) µm thick very delicate bi-layered wall; length × width (n = 49) 18.3 × 12.5 (16-20 × 11-13) µm; length/width ratio of 1.4 (1.2-1.6); 1 polar granule occasionally present; micropyle, residuum both absent. Sporocysts are ellipsoidal; length × width 8.5 × 5.2 (8-11 × 5-6) µm; length/width ratio of 1.5 (1.3-1.7) µm; Stieda body is prominent; sub-Stieda body is absent; sporocyst residuum is compact. Sporulated oocysts of Eimeria montensis n. sp. are spheroidal to subspheroidal with a 1.2 (1.0-1.4) µm thick bi-layered wall; outer layer lightly pitted; length × width (n = 30) 16.3 × 12.5 (15-17 × 13-15) µm; length/width ratio of 1.3 (1.0-1.4); 1 polar granule present; micropyle, residuum both absent. Sporocysts are ellipsoidal; length × width 7.2 × 5.1 (6-8 × 4-6) µm; length/width ratio of 1.4 (1.2-1.6); Stieda body is present, sub-Stieda body is absent; sporocyst residuum consists of small, scattered granules. Sporulated oocysts of Eimeria uricanensis n. sp. are ovoidal to pyriform with a 1.4 ( 1.3-1.6) µm thick bi-layered wall; outer layer lightly pitted; length × width (n = 40) 26.6 × 18.6 (23-30 × 17-20) µm; length/width ratio of 1.4 (1.3-1.6); 1 polar granule present; micropyle, residuum both absent. Sporocysts are ellipsoidal, length × width 13.3 × 8.0 (10-16 × 7-9) µm; length/width ratio of 1.7 (1.5-1.9); Stieda body, sub-Stieda body both absent; sporocyst residuum consists of a cluster of granules, forming a spheroid mass. Sporulated oocysts of Eimeria parnasiensis n. sp. are subspheroidal to ellipsoidal with a 1.8 ( 1.3-2.4) µm thick bi-layered wall; outer layer lightly pitted; length × width (n = 54) 28.2 × 21.9 (26-32 × 19-28) µm; length/width ratio of 1.3 (1.2-1.4); 1 polar granule present; micropyle is absent; oocyst residuum is present and consists of a cluster of granules of varying thickness. Sporocysts are ovoidal, tapering towards the Stieda body; length × width 12.2 × 7.6 (10-13 × 6-9) µm; length/width ratio of 1.6 (1.4-1.9); Stieda body is present; sub-Stieda body is absent; sporocyst residuum is present and consists of an aggregate of thin granules.

Co-circulation of Araraquara and Juquitiba Hantavirus in Brazilian Cerrado.

Hantavirus cardiopulmonary syndrome is an emerging serious disease in the Americas, transmitted from wild rodents to humans through inhalation of aerosol containing virus. Herein, we characterized two distinct hantaviruses circulating in rodent species form Central Plateau, Midwestern region of Brazil in the Cerrado (savanna-like) biome, an area characterized by small trees and grasses adapted to climates with long dry periods. In this study, we identified the co-circulation of the Araraquara virus and a possible new lineage of the Juquitiba virus (JUQV) in Oligoryzomys nigripes. The implications of co-circulation are still unknown, but it can be the key for increasing viral diversity or emergence of new species through spillover or host switching events leading to co-infection and consequently recombination or reassortment between different virus species. Phylogenetic analyses based on the complete S segment indicated that, alongside with Oligoryzomys mattogrossae rodents, O. nigripes species could also have a whole as JUQV reservoir in the Cerrado biome. Although these rodents' species are common in the Cerrado biome, they are not abundant demonstrating how complex and different hantavirus enzootic cycles can be in this particular biome.

A new species of Eimeria Schneider, 1875 from the Serra dos Órgãos National Park, Rio de Janeiro, Brazil, with notes on its endogenous development in the montane grass mouse, Akodon montensis Thomas, 1913 (Rodentia: Sigmodontinae).

A total of 53 specimens of the montane grass mouse, Akodon montensis Thomas, 1913 were collected in the Serra dos Órgãos National Park (SONP) in November 2014 and July 2015. The fecal material was analyzed, and a prevalence of 7.5% was recorded for a new coccidian species of the genus Eimeria Schneider, 1875, with part of its endogenous development recorded in the small intestine. The oocysts of a new coccidian species of genus Eimeria are ellipsoidal to subspherical. The wall is bi-layered, c. 1.5 µm (1.3-1.6 µm) thick, outer layer rough. Oocyst (n = 126) mean length is 25.3 µm (21.0-28.0 µm), with a width of 20.2 µm (17.0-22.0 µm) and mean length/width (L:W) ratio of 1.3 (1.2-1.4). Polar granule is present, with the oocyst residuum as a large spherical to subspherical globule. Sporocyst shape (n = 126) is ellipsoidal, with a mean length of 11.8 µm (9.3-14.4 µm), width of 7.9 µm (6.7-9.3 µm), and mean L:W ratio of 1.5 (1.4-1.7). Sporocysts with nipple-like Stieda body and sub-Stieda body are absent. A sporocyst residuum formed by several globules, usually along the sporocyst wall. This is the first record of Eimeria in the montane grass mouse from Brazil.

Association of Bartonella Species with Wild and Synanthropic Rodents in Different Brazilian Biomes.

Bartonella spp. comprise an ecologically successful group of microorganisms that infect erythrocytes and have adapted to different hosts, which include a wide range of mammals, besides humans. Rodents are reservoirs of about two-thirds of Bartonella spp. described to date; and some of them have been implicated as causative agents of human diseases. In our study, we performed molecular and phylogenetic analyses of Bartonella spp. infecting wild rodents from five different Brazilian biomes. In order to characterize the genetic diversity of Bartonella spp., we performed a robust analysis based on three target genes, followed by sequencing, Bayesian inference, and maximum likelihood analysis. Bartonella spp. were detected in 25.6% (117/457) of rodent spleen samples analyzed, and this occurrence varied among different biomes. The diversity analysis of gltA sequences showed the presence of 15 different haplotypes. Analysis of the phylogenetic relationship of gltA sequences performed by Bayesian inference and maximum likelihood showed that the Bartonella species detected in rodents from Brazil was closely related to the phylogenetic group A detected in other cricetid rodents from North America, probably constituting only one species. Last, the Bartonella species genogroup identified in the present study formed a monophyletic group that included Bartonella samples from seven different rodent species distributed in three distinct biomes. In conclusion, our study showed that the occurrence of Bartonella bacteria in rodents is much more frequent and widespread than previously recognized. IMPORTANCE: In the present study, we reported the occurrence of Bartonella spp. in some sites in Brazil. The identification and understanding of the distribution of this important group of bacteria may allow the Brazilian authorities to recognize potential regions with the risk of transmission of these pathogens among wild and domestic animals and humans. In addition, our study accessed important gaps in the biology of this group of bacteria in Brazil, such as its low host specificity, high genetic diversity, and relationship with other Bartonella spp. detected in rodents trapped in America. Considering the diversity of newly discovered Bartonella species and the great ecological plasticity of these bacteria, new studies with the aim of revealing the biological aspects unknown until now are needed and must be performed around the world. In this context, the impact of Bartonella spp. associated with rodents in human health should be assessed in future studies.

Trypanosoma cruzi TcI and TcII transmission among wild carnivores, small mammals and dogs in a conservation unit and surrounding areas, Brazil.

Aiming to better understand the ecological aspects of Trypanosoma cruzi transmission cycles, wild carnivores, small mammals and dogs were examined for T. cruzi infection in the Serra da Canastra National Park region, Brazil. Isolates were genotyped using mini-exon gene and PCR-RFLP (1f8 and H3) genomic targets. Trypanosoma cruzi transmission was well established in the area and occurred in both wild and peridomestic environments. Dog seroprevalence was 29·4% (63/214) and TcI and TcII genotypes, besides mixed infections were observed. Only TcI was detected in wild mammals. Marsupials displayed lower relative abundance, but a high prevalence of positive haemocultures (4/22), whereas rodents displayed positive haemocultures (9/113) mainly in the abundant Akodon montensis and Cerradomys subflavus species. The felid Leopardus pardalis was the only carnivore to display positive haemoculture and was captured in the same region where the small mammal prevalence of T. cruzi infection was high. Two canid species, Chrysocyon brachyurus and Cerdocyon thous, were serologically positive for T. cruzi infection (4/8 and 8/39, respectively), probably related to their capacity to exploit different ecological niches. Herein, dog infection not only signals T. cruzi transmission but also the genotypes present. Distinct transmission strategies of the T. cruzi genotypes are discussed.

Integrating databases for spatial analysis of parasite-host associations and the novel Brazilian dataset.

Incomplete information on parasites, their associated hosts, and their precise geographical location hampers the ability to predict disease emergence in Brazil, a continental-sized country characterised by significant regional disparities. Here, we demonstrate how the NCBI Nucleotide and GBIF databases can be used as complementary databases to study spatially georeferenced parasite-host associations. We also provide a comprehensive dataset of parasites associated with mammal species that occur in Brazil, the Brazilian Mammal Parasite Occurrence Data (BMPO). This dataset integrates wild mammal species' morphological and life-history traits, zoonotic parasite status, and zoonotic microparasite transmission modes. Through meta-networks, comprising interconnected host species linked by shared zoonotic microparasites, we elucidate patterns of zoonotic microparasite dissemination. This approach contributes to wild animal and zoonoses surveillance, identifying and targeting host species accountable for disproportionate levels of parasite sharing within distinct biomes. Moreover, our novel dataset contributes to the refinement of models concerning disease emergence and parasite distribution among host species.

A multi-attribute approach to evaluating the impact of biostimulants on crop performance.

An ever-growing collection of commercial biostimulants is becoming available in a wide variety of forms and compositions to improve crop performance. Given the intricate nature of deciphering the underlying mechanisms of commercial products, which typically comprise various biological components, it is crucial for research in this area to have robust tools to demonstrate their effectiveness in field trials. Here, we took a multi-attribute approach to evaluating the impact of biostimulants on crop performance. First, we assessed the impact of a biostimulant on the soil and rhizosphere microbiomes associated to crops in eight reference farms, including corn (3 farms), soybean (2), cotton (2) and sugarcane (1), in different biomes and production contexts in Brazil and Paraguay. Second, we modeled a set of integrated indicators to measure crop responses to biostimulant application, including five analytical themes as follows: i) crop development and production (9 indicators), ii) soil chemistry (9), iii) soil physics (5), iv) soil biology (6) and v) plant health (10). Amplicon 16S rRNA and ITS sequencing revealed that the use of the biostimulant consistently changes the structure of bacterial and fungal communities associated with the production system for all evaluated crops. In the rhizosphere samples, the most responsive bacterial taxa to biostimulant application were Prevotella in cotton; Prauserella and Methylovirgula in corn; and Methylocapsa in sugar cane. The most responsive fungal taxa to biostimulant use were Arachnomyces in soybean and cotton; and Rhizophlyctis in corn. The proposed integrated indicators yielded highly favorable positive impact indices (averaging at 0.80), indicating that biostimulant-treated fields correlate with better plant development and crop performance. Prominent indices were observed for indicators in four themes: soil biology (average index 0.84), crop production (0.81), soil physics (compaction reduction 0.81), and chemical fertility (0.75). The multi-attribute approach employed in this study offers an effective strategy for assessing the efficacy of biostimulant products across a wide range of crops and production systems.

Molecular epidemiology of Laguna Negra virus, Mato Grosso State, Brazil.

We associated Laguna Negra virus with hantavirus pulmonary syndrome in Mato Grosso State, Brazil, and a previously unidentified potential host, the Calomys callidus rodent. Genetic testing revealed homologous sequencing in specimens from 20 humans and 8 mice. Further epidemiologic studies may lead to control of HPS in Mato Grosso State.

Detection of the first incidence of Akodon paranaensis naturally infected with the Jabora virus strain (Hantavirus) in Brazil.

We characterised hantaviruses circulating in different Akodon rodent species collected in midwestern Santa Catarina (SC), southern Brazil, where the Jabora hantavirus (JABV) strain was first identified in Akodon montensis. Genetic and phylogenetic analyses based on a partial S segment indicated that, in SC, Akodon paranaensis and A. montensis carried the same type of hantavirus. Additionally, we conducted the first genomic characterisation of the complete S segment from the Brazilian JABV strain. This is the first report of A. paranaensis infected with the JABV.

Trypanosoma Species in Small Nonflying Mammals in an Area With a Single Previous Chagas Disease Case.

Trypanosomatids are hemoflagellate parasites that even though they have been increasingly studied, many aspects of their biology and taxonomy remain unknown. The aim of this study was to investigate the Trypanosoma sp. transmission cycle in nonflying small mammals in an area where a case of acute Chagas disease occurred in Mangaratiba municipality, Rio de Janeiro state. Three expeditions were conducted in the area: the first in 2012, soon after the human case, and two others in 2015. Sylvatic mammals were captured and submitted to blood collection for trypanosomatid parasitological and serological exams. Dogs from the surrounding areas where the sylvatic mammals were captured were also tested for T. cruzi infection. DNA samples were extracted from blood clots and positive hemocultures, submitted to polymerase chain reaction targeting SSU rDNA and gGAPDH genes, sequenced and phylogenetic analysed. Twenty-one wild mammals were captured in 2012, mainly rodents, and 17 mammals, mainly marsupials, were captured in the two expeditions conducted in 2015. Only four rodents demonstrated borderline serological T. cruzi test (IFAT), two in 2012 and two in 2015. Trypanosoma janseni was the main Trypanosoma species identified, and isolates were obtained solely from Didelphis aurita. In addition to biological differences, molecular differences are suggestive of genetic diversity in this flagellate species. Trypanosoma sp. DID was identified in blood clots from D. aurita in single and mixed infections with T. janseni. Concerning dogs, 12 presented mostly borderline serological titers for T. cruzi and no positive hemoculture. In blood clots from 11 dogs, T. cruzi DNA was detected and characterized as TcI (n = 9) or TcII (n = 2). Infections by Trypanosoma rangeli lineage E (n = 2) and, for the first time, Trypanosoma caninum, Trypanosoma dionisii, and Crithidia mellificae (n = 1 each) were also detected in dogs. We concluded that despite the low mammalian species richness and degraded environment, a high Trypanosoma species richness species was being transmitted with the predominance of T. janseni and not T. cruzi, as would be expected in a locality of an acute case of Chagas disease.

Socioecological vulnerability and the risk of zoonotic disease emergence in Brazil.

In developing countries, outbreaks of zoonotic diseases (ZDs) result from intertwined ecological, socioeconomic, and demographic processes that shape conditions for (i) increased contact between vulnerable human population and wildlife in areas undergoing environmental degradation and (ii) the rapid geographic spread of infections across socially vulnerable regions. In Brazil, recent increases in environmental and social vulnerabilities, amplified by economic and political crises, are potential triggers for outbreaks. We discuss Brazilian features that favor outbreaks and show a novel quantitative method for zoonotic risk assessment. Using data on nine ZDs from 2001 to 2019, we found that the most significant causal variables were vegetation cover and city remoteness. Furthermore, 8 of 27 states presented low-level risk of ZD outbreaks. Given the ZD-bushmeat connection, we identified central hunted mammals that should be surveilled to prevent spillover events. The current challenge is to coordinate intersectoral collaboration for effective One Health management in megadiverse countries with high social vulnerability and growing environmental degradation like Brazil.

Trypanosomatid Richness Among Rats, Opossums, and Dogs in the Caatinga Biome, Northeast Brazil, a Former Endemic Area of Chagas Disease.

Parasites are important components of the immense n-dimensional trophic network that connects all living beings because they, among others, forge biodiversity and deeply influence ecological evolution and host behavior. In this sense, the influence of Trypanosomatidae remains unknown. The aim of this study was to determine trypanosomatid infection and richness in rats, opossums, and dogs in the semiarid Caatinga biome. We submitted DNA samples from trypanosomatids obtained through axenic cultures of the blood of these mammals to mini exon multiplex-PCR, Sanger, and next-generation sequencing targeting the 18S rDNA gene. Phylogenetic analyses were performed to identify genetic diversity in the Trypanosomatidae family. Shannon, Simpson, equability, and beta-diversity indices were calculated per location and per mammalian host. Dogs were surveyed for trypanosomatid infection through hemocultures and serological assays. The examined mammal species of this area of the Caatinga biome exhibited an enormous trypanosomatid species/genotypes richness. Ten denoised Operational Taxonomic Units (ZOTUs), including three species (Trypanosoma cruzi, Trypanosoma rangeli and Crithidia mellificae) and one Trypanosoma sp. five genotypes/lineages (T. cruzi DTU TcI, TcII, and TcIV; T. rangeli A and B) and four DTU TcI haplotypes (ZOTU1, ZOTU2, ZOTU5, and ZOTU10 merged), as well as 13 Amplicon Sequence Variants (ASVs), including five species (T. cruzi, T. rangeli, C. mellificae, Trypanosoma dionisii, and Trypanosoma lainsoni), five genotypes/lineages (same as the ZOTUs) and six DTU TcI haplotypes (ASV, ASV1, ASV2, ASV3, ASV5 and ASV13), were identified in single and mixed infections. We observed that trypanosomatids present a broad host spectrum given that species related to a single host are found in other mammals from different taxa. Concomitant infections between trypanosomatids and new host-parasite relationships have been reported, and this immense diversity in mammals raised questions, such as how this can influence the course of the infection in these animals and its transmissibility. Dogs demonstrated a high infection rate by T. cruzi as observed by positive serological results (92% in 2005 and 76% in 2007). The absence of positive parasitological tests confirmed their poor infectivity potential but their importance as sentinel hosts of T. cruzi transmission.

Pygmy rice rat as potential host of Castelo dos Sonhos Hantavirus.

To study the dynamics of wild rodent populations and identify potential hosts for hantavirus, we conducted an eco-epidemiologic study in Campo Novo do Parecis, Mato Grosso State, Brazil. We detected and genetically characterized Castelo dos Sonhos virus found in a species of pygmy rice rat (Oligoryzomys utiaritensis).

Colonization process of the Brazilian common vesper mouse, Calomys expulsus (Cricetidae, Sigmodontinae): a biogeographic hypothesis.

Riverine barriers have been associated to genetic diversification and speciation of several taxa. The Rio São Francisco is one of the largest rivers in South America, representing the third largest river basin in Brazil and operating as a geographic barrier to gene flow of different taxa. To evaluate the influence of the Rio São Francisco in the speciation of small rodents, we investigated the genetic structure of Calomys expulsus with phylogenetic and network analyses of cytochrome b DNA. Our results suggested that C. expulsus can be divided into 3 subpopulations, 2 on the left and another one on the right bank of this river. The time of divergence of these subpopulations, using a Bayesian framework, suggested colonization from the south to the north/northeast. Spatial analysis using a clustering method and the Monmonier's algorithm suggested that the Rio São Francisco is a biogeographic barrier to gene flow and indicated that this river may play a role in the incipient speciation process of these subpopulations.

Composition and structure of the helminth community of rodents in matrix habitat areas of the Atlantic forest of southeastern Brazil.

The predominant landscape of the Atlantic Forest of the Brazilian state of Rio de Janeiro is made up of forest fragments surrounded by a matrix of modified habitat, which may influence the occurrence and distribution of host species and their parasites in comparison with the original continuous forest. The present study describes the structure, composition, and diversity of the helminth community found in rodents in two areas of an open matrix of different status of conservation. The abundance, intensity, and prevalence were calculated for each helminth species in rodent species. The influence of biotic and abiotic factors on the abundance and prevalence of the helminth species was also investigated. Community structure was analyzed based on the beta diversity and a bipartite network. Nine helminth species were recovered from Akodon cursor, Necromys lasiurus and Mus musculus, with the greatest helminth species richness being recorded in A. cursor (S = 8), followed by N. lasiurus (S = 6), and M. musculus (S = 3). Only three of the helminths recorded in A. cursor had been recorded previously in this rodent in the Atlantic Forest, where 12 different helminths have been recorded, so that the other five are new occurrences for this rodent. All the helminth species of N. lasiurus had been reported previously in this rodent in the Cerrado and Caatinga regions. Mus musculus was infected with the same helminths as the local fauna. Host species and locality were the most important factors influencing helminth abundance and prevalence. Beta-diversity was high for infracommunities indicating more substitutions of helminth species than losses among individuals. Three helminths species were shared by the three host species. The reduced beta-diversity observed in the component communities was consistent with the overlap observed in the helminth fauna of the host species.

Orthohantavirus Survey in Indigenous Lands in a Savannah-Like Biome, Brazil.

In Brazil, the first confirmed cases of hantavirus cardiopulmonary syndrome in Indigenous populations occurred in 2001. The purpose of this study was to determine the seroprevalence of orthohantavirus infections in the Utiariti Indigenous land located in the southeastern region of the Brazilian Amazon. In December 2014 and 2015, a survey was conducted using an enzyme-linked immunosorbent assay in nine villages belonging to the Haliti-Paresí Indigenous communities. A total of 301 participants were enrolled in the study. Of the two study cohorts, the one from 2014 showed a prevalence of 12.4%, whereas the one from 2015 had a serum prevalence of 13.4%. Analysis of the paired samples of 110 Indigenous people who participated in both stages of the study enabled identification of four individuals who had seroconverted during the study period. Identifying the circulation of orthohantaviruses in the Utiariti Indigenous land highlights a serious public health problem in viral expansion and highlights the need to implement preventive measures appropriate to the sociocultural reality of these communities.

Hantaviruses and hantavirus pulmonary syndrome, Maranhao, Brazil.

To confirm circulation of Anajatuba virus in Maranhao, Brazil, we conducted a serologic survey (immunoglobulin G ELISA) and phylogenetic studies (nucleocapsid gene sequences) of hantaviruses from wild rodents and persons with hantavirus pulmonary syndrome. This virus is transmitted by Oligoryzomys fornesi rodents and is responsible for hantavirus pulmonary syndrome in this region.

Investigation of Bartonella spp. in brazilian mammals with emphasis on rodents and bats from the Atlantic Forest.

The Bartonella species are zoonotic agents that infect mammals and are transmitted by arthropod vectors. Approximately 18 distinct genotypes cause diseases in humans, and may be spread by both domestic and wild animals. In Brazil, Bartonella genotypes have been identified in several species of wild mammals, and in the present study, we analyzed samples from non-human primates (marmosets), marsupials, rodents, and bats, and compared them with the genotypes described in mammals from Brazil, to examine the distribution of Bartonella genotypes in two impacted areas of Rio de Janeiro state, in southeastern Brazil. We used polymerase chain reaction (PCR) methods to detect the Bartonella DNA using partial sequences of the gltA, ftsZ, and groEL genes. We generated Bayesian inference and maximum likelihood trees to characterize the positive PCR samples and infer the phylogenetic relationships of the genotypes. A total of 276 animals were captured, including 110 bats, 91 rodents, 38 marsupials, and 37 marmosets. The DNA of Bartonella was amplified from tissue samples collected from 12 (4.34%) of the animals, including eight rodents - Akodon cursor (5/44) and Nectomys squamipes (3/27) - and four bats, Artibeus lituratus (3/58) and Carollia perspicillata (1/15). We identified Bartonella genotypes closely related to those described in previous studies, as well as new genotypes in both the rodent and the bat samples. Considering the high diversity of the Bartonella genotypes and hosts identified in the present study, further research is needed to better understand the relationships between the different Bartonella genotypes and their vectors and host species. The presence of Bartonella in the wild rodents and bats from the study area indicates that the local human populations may be at risk of infection by Bartonella due to the spillover of these strains from the wild environment to domestic and peri-domestic environments.

Small Mammals as Carriers/Hosts of Leptospira spp. in the Western Amazon Forest.

Leptospira is a bacteria that causes leptospirosis and is transmitted through water, soil, or mud that is contaminated by the urine of infected animals. Although it is mainly associated with the urban environment, Leptospires also circulate in rural and wild environments. This study aimed to investigate the role of small mammals in leptospirosis epidemiology in the western Amazon, Brazil. In total, 103 animals from 23 species belonging to the orders Didelphimorphia and Rodentia were captured. Blood, kidney, and urine samples were collected and Microscopic Agglutination Test (MAT), lipL32 PCR, secY sequencing, and culturing were conducted. MAT was reactive on 1/15 sera, and no bacterial isolate was obtained. PCR yielded 44.7% positive samples from 16 species. Twenty samples were genetically characterized and identified as L. interrogans (n = 12), L. noguchii (n = 4), and L. santarosai (n = 4). No statistical association was found between the prevalence of infection by Leptospira spp. in small mammals within carrier/hosts species, orders, study area, and forest strata. Our results indicate a high prevalence of pathogenic Leptospira spp. in several rodent and marsupial species and report the first evidence of Leptospira spp. carrier/hosts in the Brazilian Western Amazon.