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Opossums are synanthropic marsupials able to interchange among wild, peri­urban and urban environments, playing an epidemiologically important role as hosts for emerging pathogens and ectoparasites of relevance in public health. The present study aimed to detect and molecularly characterize vector-borne agents in a population of common opossums (Didelphis marsupialis) from the Island of São Luís do Maranhão, northeastern Brazil. Of the 45 animals analyzed, one (2.22%) was positive in the nested PCR assay based on the 18S rRNA gene of piroplasmids. The obtained sequence was phylogenetically positioned in a clade containing sequences of Babesia sp. previously detected in Didelphis aurita, Didelphis albiventris and associated ticks from Brazil. Eight (17.77%) samples were positive in PCR for Ehrlichia spp. based on the dsb gene; four samples were sequenced and positioned into a new clade, sister to E. minasensis and Ehrlichia sp. clade detected in Superorder Xenarthra mammals. No samples tested positive in the screening PCR assays based on the 16S rRNA gene of Anaplasma spp. Two samples were positive in the qPCR for Bartonella spp. based on the nuoG gene. Seven animals (15.56%) were positive in the nPCR based on the 16S rRNA gene of hemoplasmas. Of these, three were positive in a PCR based on the 23S rRNA gene. The phylogenies based on both 16S rRNA and 23S rRNA genes corroborated to each other and positioned the sequences in the same clade of hemoplasmas previously detected in D. aurita and D. albiventris sampled in Brazil. Finally, three (6.66%) animals were positive in the PCR for Hepatozoon spp.; the obtained 18S rRNA sequence was positioned into the H. felis clade.The present study showed, for the first time, the circulation of piroplasmids, Hepatozoon spp., Ehrlichia spp., hemoplasmas and Bartonella spp. in D. marsupialis sampled in northeastern Brazil, with description of putative novel genotypes of Ehrlichia and Hepatozoon and copositivity by different vector-borne agents. The present work consolidates the "South American Marsupialia" piroplasmid clade, adding one more genotype of Babesia sp. to this clade.

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Ticks are vectors for several pathogens, including bacteria belonging to the Rickettsia genus, such as Rickettsia rickettsii and Rickettsia parkeri, the causative agents of spotted fever. The aim of the present study was to investigate the tick species richness and rickettsial agents associated with wild birds captured in the Humaita Forest Reserve, Acre, in the Western Amazon region. Wild birds were captured with ornithological nets for visual inspection with the purpose of collecting ticks, which were identified through morphological analyses and molecular tests for several genes (12S rDNA, 16S rDNA, gltA, ompA, and sca4). A total of 607 wild birds were captured, 12% of which were parasitized by 268 ticks of the Amblyomma genus, with new host-parasite associations reported for Amblyomma calcaratum, Amblyomma geayi, Amblyomma longirostre, Amblyomma naponense, Amblyomma nodosum, and Amblyomma varium. Of the total ticks collected, 113 were tested for the presence of rickettsial DNA fragments, with 19 testing positive for R. parkeri in A. geayi, Rickettsia tamurae-like in Amblyomma sp., and Rickettsia amblyommatis in A. geayi, A. longirostre, and Amblyomma sp. We detected R. tamurae-like in Amblyomma larvae for the first time in the Western Brazilian Amazon biome, and registered spotted fever group rickettsiae, although the relevance of the detected species in a public health context should be further explored in South America, as well as new host-parasite interactions in this underexplored region.

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Visceral Leishmaniasis is a serious public health problem and dogs are considered to be the main source of infection in urban areas. In Brazil, this disease is present in all regions, but with high concentration of cases in the Northeast, and the state of Maranhão is considered to be an endemic region. The aim of this study was to conduct an epidemiological, spatial, molecular and serological survey on Leishmania infantum among domestic dogs in the municipality of Belágua, Maranhão. Blood samples were collected from dogs and questionnaires were applied to their owners to obtain epidemiological data and risk factors relating to this zoonosis in the region. The coordinates of the dogs' homes were obtained to produce a disease risk map. Serological diagnoses were made using the indirect immunofluorescence reaction (IFAT) and the dual-path platform chromatographic immunoassay test (DPP®) (Bio-Manguinhos/FIOCRUZ, Brazil). A molecular investigation was undertaken using the polymerase chain reaction (PCR). Georeferencing was performed using the global positioning system (GPS) and cases of canine visceral leishmaniasis in the municipality were spatially represented and analyzed using QGIS version 3.16.6 (QGIS Development Team, 2021). A total of 205 blood samples were collected, of which 122 (59.51%) were seroreactive for L. infantum through IFAT, while the DPP test showed 84 reactive samples (40.97%). IFAT and DPP detected 16 positive animals simultaneously. One sample that was seroreactive through IFAT was also positive through PCR. In the clinical evaluation, it was observed that among the seropositive dogs, 112 (91.80%) were symptomatic and 10 (8.20%) were asymptomatic. In the spatial analysis, the Kernel density estimator enabled determination of the place at greatest risk of occurrence of the disease. The areas with the highest concentrations of cases were in districts with large quantities of precarious housing and lack of basic sanitation. This was the first report on the occurrence of L. infantum among dogs in the municipality of Belágua. The results show that canine visceral leishmaniasis is well dispersed in this municipality, thus putting at risk the human population.

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Glanders is an infectious disease of equids caused by Burkholderia mallei, a facultative intracellular non-mobile Gram-negative bacterium that can be transmitted to other animals and humans. This study described glanders cases reported in the state of Maranhão, located within an Amazon-Cerrado transition region in northeastern Brazil, from 2007 to 2017. A database in an electronic spreadsheet (Microsoft Excel 2010) was developed containing information on the number of positive animals according to age, sex, purpose, year and month, municipality and mesoregion of origin of the animal. The descriptive analyzis was performed on data provided by the State Agency for Agricultural and Livestock Protection of the state of Maranhão (AGED), and by the official private laboratory. As the database did not have information data about negative animals, possible risk factors could not be evaluated. Among the total of 62,555 equids were evaluated by means of the complement fixation test (CFT), 59,036 were horses, 2,981 mules and 538 donkeys. Thirty-five samples (0.06%) reacted in the CFT. Five additional samples were by the western blot technique and three of them were positive. All the reactive horses (Equuscaballus) were from rural areas. Results presented here indicate that glanders may be endemic in the state of Maranhão and is a public health concern.

O mormo é uma doença infecciosa de equídeos causada por Burkholderia mallei, uma bactéria Gram-negativa intracelular, não móvel e facultativa que pode ser transmitida a outros animais e humanos. O objetivo deste estudo foi descrever os casos de mormo relatados no estado do Maranhão, em uma região de transição Amazônia-Cerrado do Nordeste do Brasil, no período de 2007 a 2017. Um banco de dados em uma planilha eletrônica (Microsoft Excel 2010) foi desenvolvido contendo informações sobre o número de animais positivos de acordo com a idade, sexo, finalidade, ano, mês, municípios e mesorregiões de origem do animal. A análise descritiva foi realizada sobre os dados fornecidos pela Agência Estadual de Defesa Agropecuária do Estado do Maranhão (AGED), e pelo laboratório privado oficial. Como o banco de dados não possui informações sobre animais negativos, possíveis fatores de risco não puderam ser avaliados. Entre o total de 62.555 equídeos foram avaliados com o teste de fixação do complemento (CFT), 59.036 eram cavalos, 2.981 mulas e 538 jumentos. Trinta e cinco (0,06%) amostras reagiram ao CFT. Cinco amostras adicionais foram testadas pela técnica de western blot e três foram positivas. Todos os animais positivos foram cavalos (Equus caballus) da área rural. Os resultados indicam que o mormo pode ser endêmico no estado do Maranhão e uma preocupação de saúde pública.

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Chiropterans play an important role in the maintenance of the environmental balance, since they are pollinators, seed dispersers and predators. They contribute to transmission and spreading of microorganisms such as helminths, fungi, protozoa, bacteria and virus. The aim of the present study was to investigate natural filariid infection among bats in the Legal Amazon region, Brazil, by means of parasitological and molecular analyses. Blood samples were collected from 82 bats for blood smears and for DNA extraction via the polymerase chain reaction (PCR) assay. Microfilariae were observed in blood smears from Carollia perspicillata (2), Artibeus lituratus (1), Artibeus fimbriatus (2), Dermanura gnoma (2) and Glossophaga soricina (1). Five positive samples were detected through the PCR assay and four of these were also positive in blood smears. From genome sequencing and comparative analysis with sequences deposited in GenBank, one sample showed 99.31% similarity to the species Litomosoides brasiliensis. The present study expands the geographical distribution of L. brasiliensis, to include the state of Maranhão as an area of occurrence of this species and includes D. gnoma and A. fimbriatus as hosts in Brazil.

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The species of the genus Leishmania are protozoa that are widely distributed from Asia to the Americas, affecting humans and wild and domestic animals. Little is known about infection by Leishmania in bats in the state of Maranhão, Brazil. The objective of this study was to investigate the presence of Leishmania in bats in Maranhão. Blood samples were collected from bat species for parasitological diagnosis. Samples of spleen and liver were collected for molecular analysis. All the blood cultures were negative. In two blood smears, organisms similar to amastigotes of Leishmania sp. were detected. Of the 116 samples, two spleen samples were positive and showed similarity to Leishmania infantum. Therefore, further studies are needed to elucidate whether bats take part in the epidemiological chain of leishmaniasis.

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With increased national and international transportation, humans as well as domestic and wild animals are able to get from one end to the other within hours, accidently carrying transboundary vectors and pathogens. We report here, the introduction of Amblyomma coelebs in Rio Grande do Sul, the southernmost state of Brazil, via a man travelling to the Carajás National Forest in the state of Pará (2661 km away). Upon return to his residence in Porto Alegre, RS, he realized that he was parasitized by one tick. The tick was identified as a nymph of Amblyomma sp., placed in a Biological Oxygen Demand incubator, and after ecdysis was identified as a male of Amblyomma coelebs. The tick's DNA was screened for Rickettsia spp., where Rickettsia amblyommatis DNA was detected. The parasitized man was seropositive to spotted fever group rickettsiae, by immunofluorescence assay, in a single serum sample collected 45 days after the tick-bite. Amblyomma coelebs is found in some Brazilian states, but Parana is the southernmost state in Brazil where it occurs. This species has already been reported to carry some species of Rickettsia, including species of the spotted fever group. It may risk public health if established in Rio Grande do Sul, with the introduction of tick and pathogenic rickettsias.

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BACKGROUND: Small mammals (rodents and marsupials) have been poorly explored for the occurrence of apicomplexan (genus Hepatozoon and genera of the order Piroplasmorida) and Anaplasmataceae agents in Brazil. Thus, this study investigated the occurrence of Hepatozoon spp., Piroplasmorida, and Anaplasmataceae agents in small mammals in seven forest fragments in Brazil. METHODS: During 2015-2018, small mammals were captured in six forest fragments in the State of São Paulo (Cerrado and Atlantic Forest biomes) and one fragment in the State of Mato Grosso do Sul (Pantanal biome). Mammal blood, liver, spleen, and lung samples were tested molecularly for the presence of DNA of Hepatozoon, Piroplasmorida, and Anaplasmataceae agents. RESULTS: A total of 524 mammals were captured, comprising seven species of marsupials, 14 rodents, two carnivores, and one Cingulata. Four novel haplotypes (1, 2, 3, 4) of Hepatozoon spp. were detected in small mammals from different biomes. In São Paulo state, haplotype 1 was detected in rodents from Cerrado and a transition area of Cerrado and Atlantic Forest biomes, whereas haplotype 2 was detected in rodents from the Atlantic Forest biome. On the other hand, haplotypes 3 and 4 were restricted to rodents and marsupials, respectively, from the Pantanal biome of Mato Grosso do Sul. No host species shared more than one haplotype. Despite these distinct geographical and host associations, our phylogenetic analyses indicated that the four Hepatozoon haplotypes belonged to the same clade that contained nearly all haplotypes previously reported on rodents and marsupials, in addition to several reptile-associated haplotypes from different parts of the world. No mammal samples yielded detectable DNA of Piroplasmorida agents. On the other hand, the Anaplasmataceae-targeted polymerase chain reaction (PCR) assay amplified a sequence 100% identical to the Wolbachia pipientis endosymbiont of the rodent filarid Litomosoides galizai. CONCLUSIONS: We report a variety of Hepatozoon haplotypes associated with small mammals in three Brazilian biomes: Cerrado, Atlantic Forest, and Pantanal. Through phylogenetic analyses, the Hepatozoon agents grouped in the rodent-marsupial-reptile large clade of Hepatozoon spp. from the world. The detection of a W. pipientis associated with the rodent filarid L. galizai indicates that the rodent was infected by filarial nematodes.

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Chiropterans play an important role in the maintenance of the environmental balance, since they are pollinators, seed dispersers and predators. They contribute to transmission and spreading of microorganisms such as helminths, fungi, protozoa, bacteria and virus. The aim of the present study was to investigate natural filariid infection among bats in the Legal Amazon region, Brazil, by means of parasitological and molecular analyses. Blood samples were collected from 82 bats for blood smears and for DNA extraction via the polymerase chain reaction (PCR) assay. Microfilariae were observed in blood smears from Carollia perspicillata (2), Artibeus lituratus (1), Artibeus fimbriatus (2), Dermanura gnoma (2) and Glossophaga soricina (1). Five positive samples were detected through the PCR assay and four of these were also positive in blood smears. From genome sequencing and comparative analysis with sequences deposited in GenBank, one sample showed 99.31% similarity to the species Litomosoides brasiliensis. The present study expands the geographical distribution of L. brasiliensis, to include the state of Maranhão as an area of occurrence of this species and includes D. gnoma and A. fimbriatus as hosts in Brazil.(AU)

Os quirópteros desempenham um papel importante na manutenção do equilíbrio ambiental, pois são polinizadores, dispersores de sementes e predadores. Contribuem para a transmissão e disseminação de microrganismos, como helmintos, fungos, protozoários, bactérias e vírus. O objetivo do presente estudo foi investigar a infecção natural por filariídeos em morcegos na região da Amazônia Legal, Brasil, por meio de análises parasitológicas e moleculares. Amostras de sangue foram coletadas de 82 morcegos para confecção de esfregaços e para extração de DNA para a Reação em Cadeia da Polimerase (PCR). Microfilárias foram observadas em esfregaços sanguíneos de Carollia perspicillata (2), Artibeus lituratus (1), Artibeus fimbriatus (2), Dermanura gnoma (2) e Glossophaga soricina (1). Cinco amostras positivas foram detectadas por meio do ensaio de PCR, e quatro destas também foram positivas em esfregaços de sangue. A partir do sequenciamento do genoma e análise comparativa com sequências depositadas no GenBank, uma amostra apresentou 99,31% de similaridade com a espécie Litomosoides brasiliensis. O presente estudo expande a distribuição geográfica de L. brasiliensis, abrangendo o estado do Maranhão como área de ocorrência desta espécie, e inclui D. gnoma e A. fimbriatus como hospedeiros no Brasil.(AU)

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Trypanosoma cruzi, the etiological agent for Chagas disease, is widely distributed in the Americas. Its hosts are humans and wild and domestic mammals, and its vectors are triatomine insects. Studies have indicated that domestic dogs are sentinel animals in the epidemiology of Chagas disease in endemic regions, including states in the Legal Amazon region of Brazil. In São Luís, the capital of Maranhão, a non-endemic state, the existence of a domestic cycle involving domestic rats has been proven, along with a wild cycle maintained by didelphids. However, no studies on T. cruzi infection in domestic animals in this locality have been conducted. The aim of this study was to investigate occurrence of T. cruzi in dogs living in the Itaqui Bacanga district of São Luís, Maranhão, by means of serological and molecular tests. Blood samples were obtained from 330 dogs and structured epidemiological questionnaires were applied to their keepers. These samples were used in the indirect immunofluorescent antibody test (IFAT), enzyme-linked immunosorbent assay (ELISA) and polymerase chain reaction (PCR). Fisher's exact test was used for statistical calculations with the aim of identifying risk factors. Out of the 330 animals, 105 (31.8%) were reactive in IFAT, 46 (13.0%) in ELISA and 20 (6.0%) in both serological tests. The results were not significant (p > 0.05) when submitted to statistical analysis for the studied variables. From PCR, 58 samples (17.5%) were found to be positive and, of these, one (0.3%) showed similarity to T. cruzi after sequencing. These data demonstrate that dogs were exposed to and infected by T. cruzi. Thus, they can be considered sentinel animals for Chagas disease in the locality studied, which signals that there is a need for epidemiological surveillance actions.

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