Molecular detection and characterization of Bartonella spp. in small mammals in the Amazonia and Cerrado biomes, midwestern Brazil.

Although Bartonella spp. have been worldwide described in rodents and bats, few studies have reported these agents in marsupials. The present work aimed to investigate the occurrence and genetic diversity of Bartonella in small mammals (rodents, marsupials, and bats) and associated ectoparasites in two ecoregions (Amazonia and Cerrado biomes) in midwestern Brazil. For this purpose, DNA samples from 378 specimens of small mammals (128 rodents, 111 marsupials, and 139 bats) and 41 fleas (Siphonaptera) were screened for the Bartonella genus employing a quantitative real-time PCR assay (qPCR) based on the nuoG (nicotinamide adenine dinucleotide dehydrogenase gamma subunit) gene. Then, positive samples in qPCR were submitted to conventional PCR (cPCR) assays targeting the gltA, ftsZ, and rpoB genes. One (0.78 %) rodent, 23 (16.54 %) bats, and 3 (7.31 %) fleas showed positive results in the qPCR for Bartonella sp. After cPCR amplification and sequencing, 13 partial Bartonella DNA sequences of the following genes were obtained only from bats´ blood samples: 9 gltA (citrate synthase), 3 ftsZ (cell division protein), and 1 rpoB (RNA polymerase beta subunit). The maximum likelihood inference based on the gltA gene positioned the obtained sequences in three different clades, closely related to Bartonella genotypes previously detected in other bat species and bat flies sampled in Brazil and other countries from Latin America. Similarly, the ftsZ sequences clustered in two different clades with sequences described in bats from Brazil, other countries from Latin America, and Georgia (eastern Europe). Finally, the Bartonella rpoB from a specimen of Lophostoma silvicolum clustered with a Bartonella sp. sequence obtained from a Noctilio albiventris (KP715475) from French Guiana. The present study provided valuable insights into the diversity of Bartonella genotypes infecting bats from two ecoregions (Amazonia and Cerrado) in midwestern Brazil and emphasized that further studies should be conducted regarding the description and evaluation of different lineages of Bartonella in wild small mammals and their ectoparasites in different Brazilian biomes.

Diversity of rickettsiae in ticks (Acari: Ixodidae) collected from wild vertebrates in part of the Amazon, Cerrado, and Pantanal biomes in Brazil.

Ticks parasitizing 102 wild animals in the states of Mato Grosso and Goiás, Brazil were collected between 2015 and 2018. A total of 2338 ticks (865 males, 541 females, 823 nymphs, and 109 larvae) belonging to four genera (Amblyomma, Dermacentor, Haemaphysalis, and Rhipicephalus) and at least 21 species were identified. DNA extraction and a molecular survey for rickettsial agents were performed on 650 ticks. The results revealed parasitism by the following species: Rickettsia amblyommatis in Amblyomma cajennense s.s., A. cajennense s.l., Amblyomma coelebs, Amblyomma humerale, Amblyomma longirostre, Amblyomma nodosum, Amblyomma scalpturatum, Amblyomma sculptum, and Amblyomma romitii; Rickettsia parkeri in Amblyomma nodosum, Amblyomma ovale, Amblyomma scalpturatum, and Amblyomma triste; Rickettsia rhipicephali in Haemaphysalis juxtakochi; Rickettsia sp. in A. cajennense s.s., A. nodosum, and A. sculptum, and lastly, 'Candidatus Rickettsia andeanae' in Amblyomma parvum and Rhipicephalus microplus. This study expands the body of knowledge about tick parasitism among wild animals, including new data concerning tick-host associations, and provides information about the epidemiology of tick-borne pathogens in the Center-West region of Brazil.

Phylogenetic relationships, distribution, and conservation of Roosmalens' dwarf porcupine, Coendouroosmalenorum Voss & da Silva, 2001 (Rodentia, Erethizontidae).

The New World porcupines of the genus Coendou comprise 16 species of arboreal nocturnal rodents. Some of these species are poorly known and have not been included in phylogenetic analyses. Based on recently collected specimens with associated tissue from the Brazilian Amazonia, we investigate the distribution and phylogenetic relationships of Roosmalens' dwarf porcupine, Coendouroosmalenorum, using an integrative approach using mitochondrial gene sequences and morphological data from new specimens and localities. Our results recovered C.roosmalenorum in the subgenus Caaporamys. However, analyses of our molecular and combined datasets produced different topologies. The new record shows the presence of C.roosmalenorum 480 km to the southeast of the Rio Madeira and 95 km away from Rio Juruena in Mato Grosso state, indicating a wider distribution in southern Amazonia than suspected. All known records of C.roosmalenorum are in the Madeira biogeographical province, to which it might be endemic.

Diversity of rickettsiae in ticks (Acari: Ixodidae) collected from wild vertebrates in part of the Amazon, Cerrado, and Pantanal biomes in Brazil / Diversidade de riquétsias em carrapatos (Acari: Ixodidae) coletados de vertebrados silvestres em parte dos biomas Amazônia, Cerrado e Pantanal no Brasil

Ticks parasitizing 102 wild animals in the states of Mato Grosso and Goiás, Brazil were collected between 2015 and 2018. A total of 2338 ticks (865 males, 541 females, 823 nymphs, and 109 larvae) belonging to four genera (Amblyomma, Dermacentor, Haemaphysalis, and Rhipicephalus) and at least 21 species were identified. DNA extraction and a molecular survey for rickettsial agents were performed on 650 ticks. The results revealed parasitism by the following species: Rickettsia amblyommatis in Amblyomma cajennense s.s., A. cajennense s.l., Amblyomma coelebs, Amblyomma humerale, Amblyomma longirostre, Amblyomma nodosum, Amblyomma scalpturatum, Amblyomma sculptum, and Amblyomma romitii; Rickettsia parkeri in Amblyomma nodosum, Amblyomma ovale, Amblyomma scalpturatum, and Amblyomma triste; Rickettsia rhipicephali in Haemaphysalis juxtakochi; Rickettsia sp. in A. cajennense s.s., A. nodosum, and A. sculptum, and lastly, 'Candidatus Rickettsia andeanae' in Amblyomma parvum and Rhipicephalus microplus. This study expands the body of knowledge about tick parasitism among wild animals, including new data concerning tick-host associations, and provides information about the epidemiology of tick-borne pathogens in the Center-West region of Brazil.(AU)

Foram coletados carrapatos parasitando 102 animais silvestres nos estados de Mato Grosso e Goiás, Brasil, entre 2015 e 2018. Um total de 2338 carrapatos (865 machos, 541 fêmeas, 823 ninfas e 109 larvas) pertencentes a quatro gêneros (Amblyomma, Dermacentor, Haemaphysalis e Rhipicephalus) e pelo menos 21 espécies foram identificadas. A extração de DNA e pesquisa molecular para agentes riquétsiais foram realizadas em 650 carrapatos. Os resultados revelaram parasitismo pelas seguintes espécies: Rickettsia amblyommatis em Amblyomma cajennense s.s., A. cajennense s.l., Amblyomma coelebs, Amblyomma humerale, Amblyomma longirostre, Amblyomma nodosum, Amblyomma scalpturatum, Amblyomma sculptum e Amblyomma romitii; Rickettsia parkeri em Amblyomma nodosum, Amblyomma ovale, Amblyomma scalpturatum e Amblyomma triste; Rickettsia rhipicephali em Haemaphysalis juxtakochi; Rickettsia sp. em A. cajennense s.s., A. nodosum e A. sculptum e, por último, 'Candidatus Rickettsia andeanae' em Amblyomma parvum e Rhipicephalus microplus. Este estudo amplia o conhecimento sobre o parasitismo de carrapatos em animais silvestres, incluindo-se novos dados sobre associações carrapato-hospedeiro. Fornece, ainda, informações sobre a epidemiologia de patógenos transmitidos por carrapatos na região Centro-Oeste do Brasil.(AU)

Distance sampling surveys reveal 17 million vertebrates directly killed by the 2020's wildfires in the Pantanal, Brazil.

Anthropogenic factors have significantly influenced the frequency, duration, and intensity of meteorological drought in many regions of the globe, and the increased frequency of wildfires is among the most visible consequences of human-induced climate change. Despite the fire role in determining biodiversity outcomes in different ecosystems, wildfires can cause negative impacts on wildlife. We conducted ground surveys along line transects to estimate the first-order impact of the 2020 wildfires on vertebrates in the Pantanal wetland, Brazil. We adopted the distance sampling technique to estimate the densities and the number of dead vertebrates in the 39,030 square kilometers affected by fire. Our estimates indicate that at least 16.952 million vertebrates were killed immediately by the fires in the Pantanal, demonstrating the impact of such an event in wet savanna ecosystems. The Pantanal case also reminds us that the cumulative impact of widespread burning would be catastrophic, as fire recurrence may lead to the impoverishment of ecosystems and the disruption of their functioning. To overcome this unsustainable scenario, it is necessary to establish proper biomass fuel management to avoid cumulative impacts caused by fire over biodiversity and ecosystem services.

Molecular survey of Cryptosporidium spp. in calves from the state of Mato Grosso, Brazil / Levantamento molecular de Cryptosporidium spp. em bezerros do estado de Mato Grosso, Brasil

Cryptosporidium spp. is a protozoan that infects a wide range of vertebrate hosts; it has been reported to be the cause of severe illness or death in livestock worldwide, which leads to decreased performance and production losses, especially in young animals. This study investigated the presence of Cryptosporidiumin calves from beef farms in the state of Mato Grosso, midwestern Brazil. For this purpose, fecal samples from 237 animals aged ≤ 45 days, raised in 20 rural properties were subjected to DNA extraction and nested polymerase chain reaction (nPCR) targeting 18S ribosomal RNA (18S rRNA) gene followed by sequencing. Additionally, positive samples, previously identified as Cryptosporidium parvum by sequencing and phylogenetic analyses based on 18S rRNA gene, were subsequently analyzed focusing the amplification and sequencing using nPCR of a fragment of the 60 kDa glycoprotein (gp60) gene. Of the 237 fecal samples analyzed by PCR (18S rRNA), 50 (21.1%) fecal samples were positive for Cryptosporidium spp., while 14 (70%) of the 20 properties had at least one positive animal. The following Cryptosporidium species were detected: C. bovis, C. parvum, and C. ryanae. Thereafter, two potentially zoonotic subtypes (IIaA15G2R1 and IIaA16G3R1) of C. parvum were identified based on gp60 gene sequences. This study resulted in the detection of subtype IIaA16G3R1 for the first time in South America and showed a wide distribution of the protozoan in beef farms in the studied area of the State.(AU)

Cryptosporidium spp. é um protozoário que infecta uma grande variedade de hospedeiros vertebrados, sendo descrito como causa de doenças graves ou morte na pecuária em todo o mundo, o que leva à diminuição do desempenho e à perda de produção, especialmente em animais jovens. Este estudo investigou a presença de Cryptosporidium em bezerros de fazendas de bovinocultura de corte no estado de Mato Grosso, Centro-Oeste do Brasil. Para esse fim, amostras fecais de 237 animais com idade ≤ 45dias, criados em 20 propriedades rurais foram submetidas à extração de DNA e uma "nested" da reação em cadeia pela polimerase (nPCR) direcionada ao gene 18S RNA ribossomal (18S rRNA), seguido do sequenciamento. Adicionalmente, amostras positivas, previamente identificadas como Cryptosporidium parvum pelo sequenciamento e análises filogenéticas baseadas no gene 18S rRNA, foram posteriormente analisadas usando a nPCR buscando a amplificação e sequenciamento de um fragmento do gene de uma glicoproteína de 60 kDa (gp60). Das 237 amostras fecais analisadas pela PCR (18S rRNA), 50 (21,1%)amostras fecais foram positivas para Cryptosporidium spp., enquanto 14 (70%) das 20 propriedades tinham ao menos um animal positivo. As seguintes espécies de Cryptosporidium foram detectadas: C. bovis, C. parvum e C. ryanae. Posteriormente, dois subtipos potencialmente zoonóticos (IIaA15G2R1e IIaA16G3R1) foram identificados. Este estudo resultou na detecção do subtipo IIaA16G3R1 pela primeira vez na América do Sul e mostrou uma ampla distribuição do protozoário em rebanhos de corte na área estudada do Estado.(AU)

Molecular survey of Cryptosporidium spp. in calves from the state of Mato Grosso, Brazil / Levantamento molecular de Cryptosporidium spp. em bezerros do estado de Mato Grosso, Brasil

Cryptosporidium spp. is a protozoan that infects a wide range of vertebrate hosts; it has been reported to be the cause of severe illness or death in livestock worldwide, which leads to decreased performance and production losses, especially in young animals. This study investigated the presence of Cryptosporidiumin calves from beef farms in the state of Mato Grosso, midwestern Brazil. For this purpose, fecal samples from 237 animals aged ≤ 45 days, raised in 20 rural properties were subjected to DNA extraction and nested polymerase chain reaction (nPCR) targeting 18S ribosomal RNA (18S rRNA) gene followed by sequencing. Additionally, positive samples, previously identified as Cryptosporidium parvum by sequencing and phylogenetic analyses based on 18S rRNA gene, were subsequently analyzed focusing the amplification and sequencing using nPCR of a fragment of the 60 kDa glycoprotein (gp60) gene. Of the 237 fecal samples analyzed by PCR (18S rRNA), 50 (21.1%) fecal samples were positive for Cryptosporidium spp., while 14 (70%) of the 20 properties had at least one positive animal. The following Cryptosporidium species were detected: C. bovis, C. parvum, and C. ryanae. Thereafter, two potentially zoonotic subtypes (IIaA15G2R1 and IIaA16G3R1) of C. parvum were identified based on gp60 gene sequences. This study resulted in the detection of subtype IIaA16G3R1 for the first time in South America and showed a wide distribution of the protozoan in beef farms in the studied area of the State.

Cryptosporidium spp. é um protozoário que infecta uma grande variedade de hospedeiros vertebrados, sendo descrito como causa de doenças graves ou morte na pecuária em todo o mundo, o que leva à diminuição do desempenho e à perda de produção, especialmente em animais jovens. Este estudo investigou a presença de Cryptosporidium em bezerros de fazendas de bovinocultura de corte no estado de Mato Grosso, Centro-Oeste do Brasil. Para esse fim, amostras fecais de 237 animais com idade ≤ 45dias, criados em 20 propriedades rurais foram submetidas à extração de DNA e uma "nested" da reação em cadeia pela polimerase (nPCR) direcionada ao gene 18S RNA ribossomal (18S rRNA), seguido do sequenciamento. Adicionalmente, amostras positivas, previamente identificadas como Cryptosporidium parvum pelo sequenciamento e análises filogenéticas baseadas no gene 18S rRNA, foram posteriormente analisadas usando a nPCR buscando a amplificação e sequenciamento de um fragmento do gene de uma glicoproteína de 60 kDa (gp60). Das 237 amostras fecais analisadas pela PCR (18S rRNA), 50 (21,1%)amostras fecais foram positivas para Cryptosporidium spp., enquanto 14 (70%) das 20 propriedades tinham ao menos um animal positivo. As seguintes espécies de Cryptosporidium foram detectadas: C. bovis, C. parvum e C. ryanae. Posteriormente, dois subtipos potencialmente zoonóticos (IIaA15G2R1e IIaA16G3R1) foram identificados. Este estudo resultou na detecção do subtipo IIaA16G3R1 pela primeira vez na América do Sul e mostrou uma ampla distribuição do protozoário em rebanhos de corte na área estudada do Estado.

Rural area of the Brazilian Pantanal wetlands associated with the occurrence of anti-Leishmania spp. antibodies in dogs / Área rural do Pantanal brasileiro associada à ocorrência de anticorpos anti-Leishmania spp. em cães

An evaluation was made of the presence of anti-Leishmania infantum chagasi antibodies in domestic dogs from the urban and rural areas of Brazil's Pantanal wetland region using serological techniques. A total of 429 dogs were sampled in three areas of the Pantanal biome, including the municipalities of Poconé, Santo Antônio de Leverger, and Barão de Melgaço, in the state of Mato Grosso, and in the municipality of Corumbá, in the state of Mato Grosso do Sul. The immunofluorescence assay (IFA) was used to detect antibodies (cut-off point 40) using Leishmania infantum chagasi antigen. Because of the possibility of cross-reactivity between species of the genus Leishmania, samples that were positive in the IFA against L. infantum chagasi were also tested by IFA in the same conditions, using L. amazonensis and L. braziliensis antigens. IFA-positive samples to L. infantum chagasi were also evaluated using Enzyme-Linked Immunosorbent Assay (ELISA). The results showed the presence of antibodies against L. infantum chagasi in 23 (5.36%; 95% CI: 3.50%-8.05%) dogs and at least one seroreactive dog was found in each of the municipalities evaluated in this study. Antibody titers ranged from 40 to 5,120, and all IFA positive samples were positive in the ELISA. Among the 23 positive dogs, nine were also were seroreactive for L. amazonensis and L. braziliensis antigens. The occurrence of anti- L. infantum chagasi antibodies in dogs was higher in rural areas (7.06%) than in urban areas (2.50%) (P < 0.05). Based on this study, we concluded that dogs from rural areas of the Pantanal wetlands were in contact with Leishmania species, which is relevant information given their importance to public health.(AU)

Neste trabalho foi realizada uma avaliação sobre a presença de anticorpos anti-Leishmania infantum chagasi em cães domésticos das áreas urbanas e rurais da região do Pantanal brasileiro usando técnicas sorológicas. Um total de 429 cães foram amostrados em três áreas do bioma do Pantanal, incluindo os municípios de Poconé, Santo Antônio de Leverger e Barão de Melgaço, em Mato Grosso, e o município de Corumbá, em Mato Grosso do Sul. A reação de imunofluorescência indireta (RIFI) foi utilizada para detectar anticorpos (ponto de corte de 40) de Leishmania infantum chagasi como antígeno. Devido à possibilidade de reação cruzada entre as espécies do gênero Leishmania, as amostras positivas na RIFI para L. infantum chagasi foram também avaliadas na RIFI utilizando L. amazonensis e L. braziliensis como antígenos. As amostras positivas na RIFI para L. infantum chagasi foram avaliadas utilizando o ensaio de imunoadsorção ligado à enzima (ELISA). Os resultados mostraram a presença de anticorpos contra L. infantum chagasi em 23 (5,36%; IC 95%: 3,50% -8,05%) cães e pelo menos um cão soro-reativo foi encontrado em todos os municípios avaliados neste estudo. Os títulos de anticorpos variaram de 40 a 5.120 e todas as amostras positivas na RIFI foram positivas no ELISA. Entre os 23 cães positivos, nove também reagiram para L. amazonensis e L. braziliensis. A ocorrência de anticorpos anti-L. infantum chagasi em cães foi maior nas áreas rurais (7,06%) do que nas áreas urbanas (2,50%) (P < 0,05). Com base neste estudo, concluímos que cães de áreas rurais do Pantanal tiveram contato com espécies de Leishmania, o que é uma informação relevante, dada a sua importância para a saúde pública.(AU)

Rural area of the Brazilian Pantanal wetlands associated with the occurrence of anti-Leishmania spp. antibodies in dogs / Área rural do Pantanal brasileiro associada à ocorrência de anticorpos anti-Leishmania spp. em cães

An evaluation was made of the presence of anti-Leishmania infantum chagasi antibodies in domestic dogs from the urban and rural areas of Brazil's Pantanal wetland region using serological techniques. A total of 429 dogs were sampled in three areas of the Pantanal biome, including the municipalities of Poconé, Santo Antônio de Leverger, and Barão de Melgaço, in the state of Mato Grosso, and in the municipality of Corumbá, in the state of Mato Grosso do Sul. The immunofluorescence assay (IFA) was used to detect antibodies (cut-off point 40) using Leishmania infantum chagasi antigen. Because of the possibility of cross-reactivity between species of the genus Leishmania, samples that were positive in the IFA against L. infantum chagasi were also tested by IFA in the same conditions, using L. amazonensis and L. braziliensis antigens. IFA-positive samples to L. infantum chagasi were also evaluated using Enzyme-Linked Immunosorbent Assay (ELISA). The results showed the presence of antibodies against L. infantum chagasi in 23 (5.36%; 95% CI: 3.50%-8.05%) dogs and at least one seroreactive dog was found in each of the municipalities evaluated in this study. Antibody titers ranged from 40 to 5,120, and all IFA positive samples were positive in the ELISA. Among the 23 positive dogs, nine were also were seroreactive for L. amazonensis and L. braziliensis antigens. The occurrence of anti- L. infantum chagasi antibodies in dogs was higher in rural areas (7.06%) than in urban areas (2.50%) (P < 0.05). Based on this study, we concluded that dogs from rural areas of the Pantanal wetlands were in contact with Leishmania species, which is relevant information given their importance to public health.(AU)

Neste trabalho foi realizada uma avaliação sobre a presença de anticorpos anti-Leishmania infantum chagasi em cães domésticos das áreas urbanas e rurais da região do Pantanal brasileiro usando técnicas sorológicas. Um total de 429 cães foram amostrados em três áreas do bioma do Pantanal, incluindo os municípios de Poconé, Santo Antônio de Leverger e Barão de Melgaço, em Mato Grosso, e o município de Corumbá, em Mato Grosso do Sul. A reação de imunofluorescência indireta (RIFI) foi utilizada para detectar anticorpos (ponto de corte de 40) de Leishmania infantum chagasi como antígeno. Devido à possibilidade de reação cruzada entre as espécies do gênero Leishmania, as amostras positivas na RIFI para L. infantum chagasi foram também avaliadas na RIFI utilizando L. amazonensis e L. braziliensis como antígenos. As amostras positivas na RIFI para L. infantum chagasi foram avaliadas utilizando o ensaio de imunoadsorção ligado à enzima (ELISA). Os resultados mostraram a presença de anticorpos contra L. infantum chagasi em 23 (5,36%; IC 95%: 3,50% -8,05%) cães e pelo menos um cão soro-reativo foi encontrado em todos os municípios avaliados neste estudo. Os títulos de anticorpos variaram de 40 a 5.120 e todas as amostras positivas na RIFI foram positivas no ELISA. Entre os 23 cães positivos, nove também reagiram para L. amazonensis e L. braziliensis. A ocorrência de anticorpos anti-L. infantum chagasi em cães foi maior nas áreas rurais (7,06%) do que nas áreas urbanas (2,50%) (P < 0,05). Com base neste estudo, concluímos que cães de áreas rurais do Pantanal tiveram contato com espécies de Leishmania, o que é uma informação relevante, dada a sua importância para a saúde pública.(AU)