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.

Serological Evidence Supporting the Occurrence of Ehrlichia chaffeensis or a Closely Related Species in Brazilian Dogs.

Ehrlichia chaffeensis is a tick-borne bacterium that causes human monocytotropic ehrlichiosis, an emerging life-threatening disease in humans transmitted by Amblyomma americanum. Although most studies have reported bacterial isolations and clinical cases in the United States, their occurrence is not restricted to North America. Some studies in the Southern Cone of South America have molecularly detected a close phylogenetic relative of E. chaffeensis in ticks and wild mammals. Even so, many gaps must be filled to confirm the presence of this agent in the region. To add new data on this issue, we report the first detection of specific anti-E. chaffeensis antibodies in dogs collected from all regions of Brazil. By means of IFA and ELISA with crude and specific antigens of E. chaffeensis, sera from 1134 dogs were analyzed. Serological analyses using ELISA showed nine (0.7%) seropositive dogs, with seven of them exhibiting IFA titers ranging from 160 to 5120. All regions of Brazil had at least one seropositive dog. Our results support the evidence for the occurrence of E. chaffeensis in South America. As dogs have a close relationship with humans, they can be used as an environmental sentinel for these infections because they can act as a bridge to human parasitism or infection with ehrlichial agents.

Epidemiological aspects of the occurrence of anti-Sarcocystis spp. andanti-Toxoplasma gondii antibodies in dogs in the Northern region of Brazil / Aspectos epidemiológicos da ocorrência de anticorpos anti-Sarcocystisspp. e anti-Toxoplasma gondii em cães da região norte do Brasil

We conducted aseroepidemiological study on the occurrence of anti-Sarcocystisspp. and anti-Toxoplasma gondii antibodies in dogs from family farming properties in the municipality of Ji-Paraná, Rondônia.Blood samples were collected from apparently healthy dogs between September 2012 and November 2013. In total, 181 blood serum samples were analyzed using an indirect immunofluorescence assay, among which 57 (31.49%) and 20 (11.04%) were positive for anti-T. gondii and anti-Sarcocystis spp., respectively. Statistical analyses showed that the type of food fed to the dogs was associated with the occurrence of anti-Sarcocystisspp. antibodies. In contrast, age and access to bovine carcasses were the risk factors for anti-T. gondii.The high occurrence of seropositive dogs for Sarcocystis spp. and T. gondii evidences the wide distribution of these agents in the studied area, possibly due to human and animal exposure to these protozoan species. In addition, anti-T. gondii antibodies were directly proportional to dog age. The increase in the number of positive animals with age was statistically significant. Furthermore, high antibody titers (up to 800) against Sarcocystis spp. in dogs suggest the possibility of recent exposure, in addition to environmental contamination by oocysts/sporocysts eliminated by the feces of these animals.

Conduzimos um estudo soroepidemiológico sobre a ocorrência de anticorpos anti- Sarcocystis spp. e anti-Toxoplasma gondiiem cães de propriedades de agricultura familiar no município de Ji-Paraná, Rondônia. Amostras de sangue foram coletadas de cães aparentemente saudáveis, entre setembro de 2012 e novembro de 2013. Ao todo, foram analisados 181 soros sanguíneos por meio do ensaio de imunofluorescência indireta, sendo positivas 57 (31,49%) e 20 (11,04%) amostras para anticorpos anti-T. gondii e anti-Sarcocystis spp., respectivamente. As análises estatísticas demonstraram que o tipo de alimentação fornecida aos cães esteve associado à ocorrência de anticorpos anti-Sarcocystis spp. Em contraste a idade e o acesso à carcaça bovina foram fatores de risco para a presença de anticorpos anti-T. gondii. A alta ocorrência de cães soropositivos para Sarcocystis spp. e T. gondii evidencia a ampla distribuição desses agentes na área estudada, possivelmente devido à exposição humana e animal a essas espécies de protozoários. Além disso, o resultado dos anticorpos anti-T. gondii relacionados a idade do cão mostraram diferença estatística, com aumento significativo no número de animais positivos com a idade. Além disso, altos títulos de anticorpos (até 800) contra Sarcocystis spp. em cães sugerem a possibilidade de exposição recente, além da contaminação ambiental por oocistos/esporocistos eliminados pelas fezes desses animais.

Bioverm® in the Control of Nematodes in Beef Cattle Raised in the Central-West Region of Brazil.

Cooperia, Haemonchus and Oesophagostomum are the genera of gastrointestinal parasitic nematodes most prevalent in cattle and constitute a serious problem in cattle breeding due to the impact they have on meat and milk production and the high costs of control measures. The objective of the present work was to evaluate the efficiency of Bioverm® (Duddingtonia flagrans) in the control of gastrointestinal parasitism of young cattle raised in the field, in the Central-West region of Brazil. The experiment was conducted on a farm located in the municipality of Jangada, MT, where 18 cattle, Nelore and Aberdeen Angus breeds, aged six to ten months, were randomly divided into two groups (treated group and control group) and distributed in paddocks of Brachiaria decumbens, naturally infested by larvae of gastrointestinal nematodes. The animals in the treated group received 1g of Bioverm® for each 10 kg of body weight, administered daily with commercial feed, throughout a period of six months. In the control group, each animal received 1 g of rice bran for each 10 kg of body weight, without Bioverm®, added to the feed. Stool and pasture samples were collected every two weeks. The treated group showed a significant reduction (p < 0.05) in values of eggs per gram of feces (EPG) and a significant gain of body weight (p < 0.05) when compared to the control group. The fungal formulation Bioverm® was effective in pasture decontamination and consequently in reducing the occurrence of reinfection by nematodes. The animals treated with Bioverm® showed a lower parasitic load and greater weight gain.

Serological evidence of Ehrlichia minasensis infection in Brazilian dogs.

Ehrlichia spp. are important tick-borne pathogens of animals in Brazil, and Ehrlichia canis is the most prevalent species infecting dogs. Moreover, Ehrlichia minasensis has also recently been identified as a novel ehrlichial agent that infects cattle in Brazil. The objective of this study was to determine whether dogs could be infected by E. minasensis. To investigate this possibility, sera (n = 429) collected from dogs in the Pantanal region were retrospectively analyzed for the presence of antibodies against E. canis and E. minasensis. Canine sera were screened by two isolates of E. canis in indirect immunofluorescence assay (IFA) and the majority (n = 298; 69.4%) had antibodies with endpoint titers ranging from 80 to 327,680. In order to further confirm E. canis-specific antibodies, IFA positive sera were analyzed by ELISA using E. canis-specific peptides (i.e. TRP19 and TRP36 US/BR/CR), which detected E. canis antibodies in 80.2% (239/298) of the dog sera. Fifty-nine (13.7%) samples had detectable antibodies to E. canis by IFA but were negative by E. canis peptide ELISA. These sera were then tested by E. minasensis IFA (Cuiaba strain) as antigen and 67.8% (40/59) were positive (titers ranging from 80 to 20,480). Eleven sera had antibody titers against E. minasensis at least two-fold higher than observed for E. canis and suggests that these dogs were previously infected with E. minasensis. The results of the present study suggest that multiple ehrlichial agents infect dogs in Brazil, which highlights the need to consider different Ehrlichia spp. in Brazilian dogs, particularly in areas where dogs are frequently exposed to multiple tick species. This investigation is the first to provide serologic evidence of E. minasensis infection in dogs from Brazil.

Occurrence of autogeny in a population of Ornithodoros fonsecai (Acari: Argasidae).

Ornithodoros fonsecai is an argasid tick that is endemic to Brazil and has been described in the municipality of Bonito, state of Mato Grosso do Sul. Some specimens of this species were found in a cave in the municipality of Nobres, state of Mato Grosso. The specific identification of this population was confirmed by means of morphology and molecular biology. The mitochondrial 16S rDNA partial sequence of this species from Nobres has been deposited in GenBank (MK158949). The objective of this study was to elucidate the biology of O. fonsecai from Nobres, and to report autogeny in this tick population. Along three laboratory generations was observed molting of first nymphal instar to the second instar without feeding, a typical behavior of species included in the subgenus Alectorobius. The first generation (F1) presented five nymphal instars (N1 to N5), and most of adults emerged through molting of N5. The last nymphal instar of second generation (F2) was N4, but most of adults emerged from N3. In the third generation (F3) the last nymphal instar was N5, with most of the adults emerging from N4. In F2, some females (n = 20) originated from N3 began laying eggs without a blood meal. It was observed that those N3 fed twice before they molted to autogenic females. However, autogenic behavior occurred in relation to third generation females (F3) with specimens originating from N4 (n = 12) that were fed only once as nymphs. This behavior has already been reported as obligatory for the genera Otobius and Antricola, while it is facultative for one species of genus Argas and for four species of genus Ornithodoros. However, the present report provides the first record of facultative autogeny for a species of Ornithodoros in Brazil.

Soropositividade para Leptospira sp. e sorogrupos predominantes em cães do Pantanal Brasileiro / Seropositivity for Leptospira sp. and predominant serogroups in dogs from the Brazilian Pantanal

Em face à grande importância que a leptospirose possui no contexto sanitário mundial, tanto no aspecto humano como animal, este estudo teve por objetivo realizar a pesquisa de anticorpos anti-Leptospira sp. pela técnica de Soroaglutinação Microscópica (SAM) em 429 amostras de soros de cães provenientes de quatro municípios (Poconé/MT, Santo Antônio de Leverger/MT, Barão de Melgaço/MT e Corumbá/MS) localizados na região do Pantanal Brasileiro, bem como foram verificadas possíveis associações entre os resultados dos exames sorológicos e respostas aos questionários epidemiológicos aplicados aos proprietários. Do total de cães avaliados pela SAM (título ≥100), verificou-se que 34 (7,93%; IC 95%: 5,63%-11,00%) cães tinham anticorpos anti-Leptospira sp. Os títulos encontrados variaram entre 100 e 1600 e todos os municípios analisados tinham cães sororreagentes ao agente pesquisado. O sorogrupo reator mais frequente foi o Icterohaemorrhagiae, seguido pelo Australis. Por outro lado, foram observadas menores proporções de cães reagentes aos sorogrupos Tarassovi, Hebdomadis, Autumnalis e Grippotyphosa. As variáveis associadas com a ocorrência de leptospirose foram habitat rural (P<0,01) e área alagável (P=0,01). Estes resultados demonstram que os cães da região pantaneira tiveram contato com agentes do gênero Leptospira, o que representa uma informação relevante para a saúde pública local devido à importância zoonótica da doença.

Given the great importance that leptospirosis has the global health context, both in human and animal aspect, this study aimed to search for antibodies anti-Leptospira sp. by the technique of microscopic agglutination test (MAT) in 429 samples of sera from dogs from four municipalities (Poconé/MT, Santo Antônio de Leverger/MT, Barão de Melgaço/MT and Corumbá/MS) located in the Brazilian Pantanal region, in order for determine associations between the results of the serological tests and answers to epidemiological questionnaires applied to owners. Of the total dogs evaluated by MAT (titer ≥100), it was verified that 34 (7.93%, 95% CI: 5.63% -11.00%) dogs had antibodies against Leptospira sp. The titers found ranged from 100 to 1600 and all municipalities analyzed had seroreactive dogs for the investigated agent. The most frequent serogroup reactor was Icterohaemorrhagiae, followed by Australis. On the other hand, smaller proportions of reactive dogs were observed for serogroups Tarassovi, Hebdomadis, Autumnalis and Grippotyphosa. The variables associated with the occurrence of leptospirosis were rural habitat (P<0.01) and flooded area (P=0.01). These results demonstrate that dogs from the Pantanal region had contact with agents of the genus Leptospira, which represents information relevant to local public health due to the zoonotic importance of the disease.

Soropositividade para Leptospira sp. e sorogrupos predominantes em cães do Pantanal Brasileiro / Seropositivity for Leptospira sp. and predominant serogroups in dogs from the Brazilian Pantanal

Em face à grande importância que a leptospirose possui no contexto sanitário mundial, tanto no aspecto humano como animal, este estudo teve por objetivo realizar a pesquisa de anticorpos anti-Leptospira sp. pela técnica de Soroaglutinação Microscópica (SAM) em 429 amostras de soros de cães provenientes de quatro municípios (Poconé/MT, Santo Antônio de Leverger/MT, Barão de Melgaço/MT e Corumbá/MS) localizados na região do Pantanal Brasileiro, bem como foram verificadas possíveis associações entre os resultados dos exames sorológicos e respostas aos questionários epidemiológicos aplicados aos proprietários. Do total de cães avaliados pela SAM (título 100), verificou-se que 34 (7,93%; IC 95%: 5,63%-11,00%) cães tinham anticorpos antiLeptospira sp. Os títulos encontrados variaram entre 100 e 1600 e todos os municípios analisados tinham cães sororreagentes ao agente pesquisado. O sorogrupo reator mais frequente foi o Icterohaemorrhagiae, seguido pelo Australis. Por outro lado, foram observadas menores proporções de cães reagentes aos sorogrupos Tarassovi, Hebdomadis, Autumnalis e Grippotyphosa. As variáveis associadas com a ocorrência de leptospirose foram habitat rural (P 0,01) e área alagável (P=0,01).

Given the great importance that leptospirosis has the global health context, both in human and animal aspect, this study aimed tosearch for antibodies anti-Leptospira sp. by the technique of microscopic agglutination test (MAT) in 429 samples of sera from dogsfrom four municipalities (Poconé/MT, Santo Antônio de Leverger/MT, Barão de Melgaço/MT and Corumbá/MS) located in the BrazilianPantanal region, in order for determine associations between the results of the serological tests and answers to epidemiologicalquestionnaires applied to owners. Of the total dogs evaluated by MAT (titer ≥100), it was verified that 34 (7.93%, 95% CI: 5.63%-11.00%) dogs had antibodies against Leptospira sp. The titers found ranged from 100 to 1600 and all municipalities analyzed hadseroreactive dogs for the investigated agent. The most frequent serogroup reactor was Icterohaemorrhagiae, followed by Australis.On the other hand, smaller proportions of reactive dogs were observed for serogroups Tarassovi, Hebdomadis, Autumnalis andGrippotyphosa. The variables associated with the occurrence of leptospirosis were rural habitat (P<0.01) and flooded area (P=0.01).These results demonstrate that dogs from the Pantanal region had contact with agents of the genus Leptospira, which representsinformation relevant to local public health due to the zoonotic importance of the disease.

Prevalence of Neospora caninum and Toxoplasma gondii antibodies in dogs in the municipality of Poconé, state of Mato Grosso, Brazil / Prevalência de anticorpos contra Neospora caninum e Toxoplasma gondii no município de Poconé, estado de Mato Grosso, Brasil

This study evaluated the presence of Neospora caninum and Toxoplasma gondii antibodies in 320 domestic dogs living in urban and rural areas of the municipality of Poconé, located in the Pantanal wetlands of the state of Mato Grosso, Brazil. Anti- N. caninum (cut-off point 50) and anti- T. gondii (cut-off point 16) antibodies were detected by means of immunofluorescence assays (IFA), using NC-Bahia and RH strains as antigens, respectively. Anti- N. caninum antibodies were detected in 69 (21.56%; 95% CI: 17.27%-26.56%) dogs, 31 (44.93%) of which lived in urban areas and 38 (55.07%) in rural areas, and endpoint titers ranged from 50 to 3200. Anti- T. gondii antibodies were found in 132 (41.25%; 95% CI: 35.84%-46.87%) dogs, 58 (43.94%) from urban areas and 74 (56.06%) from rural areas, and endpoint titers ranged from 16 to 8192. A total of 33 dogs (10.3%) (12 urban and 21 rural animals) reacted to both agents (P <0.05). This study showed a higher prevalence of anti- T. gondii compared to anti- N. caninum antibodies in the dogs living in the Pantanal region, suggesting that both protozoans circulate in the studied region. This information is relevant, in view of its implications for animal and public health.

Este estudo avaliou a presença de anticorpos contra N. caninum e T. gondii em 320 cães domésticos das áreas urbana e rural do município de Poconé, no estado do Mato Grosso, região pantaneira do Pantanal. A Reação de Imunofluorescência Indireta (RIFI) foi utilizada para detectar anticorpos anti- N. caninum (ponto de corte igual a 50) e anticorpos anti- T. gondii (ponto de corte igual a 16) utilizando as cepas NC-Bahia e RH como antígenos, respectivamente. Anticorpos anti- N. caninum foram detectados em 69 (21,56%; IC 95%: 17,27%-26,56%) cães, dos quais 31 (44,93%) eram da área urbana e 38 (55,07%) eram da área rural e os títulos variaram entre 50 a 3200. Em relação aos anticorpos contra T. gondii 132 (41,25%; IC 95%: 35,84%-46,87%) cães foram sororeagentes sendo que 58 (43,94%) eram da área urbana e 74 (56,06%) da área rural e os títulos variaram entre 16 a 8192. Um total de 33 (10,31%) (12 cães urbanos e 21 cães rurais) cães reagiram para ambos os agentes testados pela RIFI (P <0,05). Este estudo mostrou uma maior ocorrência de anticorpos contra T. gondii em relação a N. caninum nos cães avaliados, mostrando que na região do Pantanal há circulação desses importantes protozoários. Essa informação é relevante dada sua importância para a saúde animal e pública

Genetic diversity and molecular survey of Trypanosoma (Megatrypanum ) theileri in cattle in Brazil's western Amazon region / Diversidade genética e levantamento molecular de Trypanosoma (Megatrypanum) theileri em bovinos na região da Amazônia Ocidental do Brasil

Abstract Trypanosoma (Megatrypanum) theileri is a flagellated protozoan that infects ruminants and it displays high genetic diversity. In this study, we investigated the prevalence rates of this protozoan based on hemoculture and molecular diagnosis. The isolates of T. theileri thus obtained were characterized by molecular markers SSU rDNA and gGAPDH and molecular diagnosis based on Cathepsin L-like gene (PCR-TthCATL). The PCR-TthCATL and hemoculture indicated an overall prevalence rate of 8.13%, and the CATL derived sequence named IB was identified for the first time in cattle in the western Amazon region, as well as IF in Brazil. We also describe a possible new PCR-TthCATL derived sequence in cattle, designated IL.

Resumo Trypanosoma (Megatrypanum) theileri é um protozoário flagelado que infecta ruminantes e apresenta alta diversidade genética. Neste estudo, investigamos as taxas de prevalência deste protozoário com base na hemocultura e no diagnóstico molecular. Os isolados de T . theileri obtidos foram caracterizados pelos marcadores moleculares SSU rDNA e gGAPDH e o diagnóstico molecular foi baseado no gene do tipo Catepsina L (PCR-TthCATL). O PCR-TthCATL e a hemocultura indicaram uma taxa de prevalência total de 8,13% e a sequência derivada do gene Catepsina L denominada IB de T. theileri foi identificada pela primeira vez em bovinos da Amazônia Ocidental, bem como a IF no Brasil. Também descrevemos uma possível nova sequência derivada da PCR-TthCATL em bovinos, designada IL.

Genetic diversity and molecular survey of Trypanosoma (Megatrypanum ) theileri in cattle in Brazil's western Amazon region.

Trypanosoma (Megatrypanum) theileri is a flagellated protozoan that infects ruminants and it displays high genetic diversity. In this study, we investigated the prevalence rates of this protozoan based on hemoculture and molecular diagnosis. The isolates of T. theileri thus obtained were characterized by molecular markers SSU rDNA and gGAPDH and molecular diagnosis based on Cathepsin L-like gene (PCR-TthCATL). The PCR-TthCATL and hemoculture indicated an overall prevalence rate of 8.13%, and the CATL derived sequence named IB was identified for the first time in cattle in the western Amazon region, as well as IF in Brazil. We also describe a possible new PCR-TthCATL derived sequence in cattle, designated IL.

Seropositivity for Rickettsia spp. and Ehrlichia spp. in the human population of Mato Grosso, Central-Western Brazil.

INTRODUCTION: The epidemiology of Rickettsia and Ehrlichia species infection is underestimated in Mato Grosso State. METHODS:: Serum samples obtained during a Dengue outbreak in 2011-2012 were tested via indirect immunofluorescence and/or ELISA. RESULTS:: Samples from 19/506 (3.8%) patients presented antibodies for at least one of three Rickettsia species; 2/506 (0.4%) samples reacted against Ehrlichia canis. Most afflicted patients are residents of cities from the south-central region of the state, where these diseases have been reported in animals. CONCLUSIONS:: These results show serological evidence of human exposure to Rickettsia and Ehrlichia species in Mato Grosso State.

Seropositivity for Rickettsia spp. and Ehrlichia spp. in the human population of Mato Grosso, Central-Western Brazil

Abstract INTRODUCTION The epidemiology of Rickettsia and Ehrlichia species infection is underestimated in Mato Grosso State. METHODS: Serum samples obtained during a Dengue outbreak in 2011-2012 were tested via indirect immunofluorescence and/or ELISA. RESULTS: Samples from 19/506 (3.8%) patients presented antibodies for at least one of three Rickettsia species; 2/506 (0.4%) samples reacted against Ehrlichia canis. Most afflicted patients are residents of cities from the south-central region of the state, where these diseases have been reported in animals. CONCLUSIONS: These results show serological evidence of human exposure to Rickettsia and Ehrlichia species in Mato Grosso State.

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)

Ehrlichia canis DNA in domestic cats parasitized by Rhipicephalus sanguineus sensu lato (s. l. ) ticks in Brazil ­ case report / Felinos domésticos parasitados por carrapato Rhipicephalus sanguineus sensu lato (s. l. ) infectados por Ehrlichia canis no Brasil ­ relato de caso

Ectoparasites can transmit pathogens, including bacteria such as Ehrlichia sp., which trigger infectious diseases in domestic animals. Little is known about the epidemiology of feline ehrlichiosis, although several studies have focused on elucidating the pathogenesis and transmission of this disease. This paper presents the first mutual infection by Ehrlichia sp. between a domestic cat and a Rhipicephalus sanguineus sensu lato (s.l.) tick removed from the animal. The cat and tick were tested by Polymerase Chain Reaction (PCR) to detect the dsb gene, and the analyzed sequences revealed samples 100% identical to E. canis. Based on this report, we discussed the importance of cats as E. canis reservoirs s and their position in the cycle of transmission between dogs and cats in Brazil.(AU)

Os ectoparasitos são capazes de transmitir patógenos incluindo algumas bactérias, como a Ehrlichia sp., causando doenças em animais domésticos. Pouco se conhece sobre a epidemiologia da erliquiose felina, embora alguns estudos já tenham sido realizados para elucidar a sua patogenia e transmissão. Este trabalho relata a primeira infecção mútua por Ehrlichia sp. entre um felino doméstico e o carrapato Rhipicephalus sanguineus sensu lato (s.l.) removido deste animal. Ambos foram testados pela Reação em Cadeia pela Polimerase (PCR) para detectar o gene dsb, e as sequências analisadas confirmaram amostras 100% idênticas à E. canis. Baseado neste relato é discutida a importância dos gatos como vetores de E. canis e sua posição no ciclo de transmissão de carrapatos entre cães e gatos no Brasil.(AU)

Molecular assessment of the transplacental transmission of Toxoplasma gondii, Neospora caninum, Brucella canis and Ehrlichia canis in dogs.

Given the fact that numerous microbial species can be detected in pregnant female dogs, the objective of this study was to assess the transplacental transmission of Brucella canis, Ehrlichia canis, Neospora caninum and Toxoplasma gondii in stillborn puppies. This study involved 41 stillborn puppies, 78.6% of which were positive for T. gondii, 52.4% for N. caninum and 59.5% for B. canis. E. canis was not detected in any of the analyzed puppies. Pregnancy is an important physiological condition for the transmission of infectious agents to puppies and transplacental transmission may be epidemiologically relevant in the spread of these opportunistic agents.

Serological evidence of exposure to tick-borne agents in opossums (Didelphis spp. ) in the state of São Paulo, Brazil / Evidência sorológica de exposição a agentes transmitidos por carrapatos em gambás (Didelphis spp. ) no estado de São Paulo, Brasil

Abstract This work involved a serological investigation of tick-borne pathogens in opossums in eight municipalities of the state of São Paulo, Brazil. Serum samples from 109 opossums (91 Didelphis aurita and 18 Didelphis albiventris) were tested to detect antibodies to Rickettsia rickettsii (Taiaçu strain, 1:64 cut-off) and Ehrlichia canis (São Paulo strain, 1:40 cut-off), by indirect immunofluorescence assay (IFA); and against Borrelia burgdorferi (strain G39/40) by enzyme-linked immunosorbent assay (ELISA). The presence of antibodies to anti-R. rickettsii, anti-E. canis and anti-B. burgdorferi was detected in 32 (29.35%), 16 (14.67%) and 30 (27.52%) opossums, respectively. Opossum endpoint titers ranged from 64 to 1,024 for R. rickettsii, from 40 to 160 for E. canis, and from 400 to >51,200 for B. burgdorferi. These serological results suggest that opossums have been exposed to Rickettsia spp., Ehrlichia spp., and B. burgdorferi-related agents in the state of São Paulo. Our study underscores the need for further research about these agents in this study area, in view of the occurrence of Spotted Fever and Baggio-Yoshinari Syndrome disease in humans in the state of São Paulo, Brazil.

Resumo O presente estudo investigou evidência sorológica de agentes transmitidos por carrapatos em gambás em oito municípios do Estado de São Paulo, Brasil. Amostras de soro de 109 gambás (91 Didelphis aurita e 18 Didelphis albiventris) foram testadas para detecção de anticorpos contra Rickettsia rickettsii (cepa Taiaçu, ponto de corte 1:64) Ehrlichia canis (cepa São Paulo, ponto de corte 1:40), pela reação de imunofluorescência indireta (RIFI); e contra Borrelia burgdorferi (cepa G39/40) pelo teste imunoenzimático (ELISA). A presença de anticorpos anti-R. rickettsii, anti-E. canis e anti-B. burgdorferi foi detectada em 32 (29,35%), 16 (14,67%) e 30 (27,52%) gambás, respectivamente. Os títulos finais variaram de 64 a 1.024 para R. rickettsii, de 40 a 160 para E. canis, e de 400 a >51.200 para B. burgdorferi. Esses resultados sugerem que os gambás foram expostos a agentes relacionados à Rickettsia spp., Ehrlichia spp., e B. burgdorferi no Estado de São Paulo. Neste estudo salienta a necessidade de novas pesquisas sobre esses agentes nessas áreas de trabalho, devido à ocorrência da Febre Maculosa e da Síndrome Baggio-Yoshinari em humanos no Estado de São Paulo, Brasil.

Serological evidence of exposure to tick-borne agents in opossums (Didelphis spp.) in the state of São Paulo, Brazil.

This work involved a serological investigation of tick-borne pathogens in opossums in eight municipalities of the state of São Paulo, Brazil. Serum samples from 109 opossums (91 Didelphis aurita and 18 Didelphis albiventris) were tested to detect antibodies to Rickettsia rickettsii (Taiaçu strain, 1:64 cut-off) and Ehrlichia canis (São Paulo strain, 1:40 cut-off), by indirect immunofluorescence assay (IFA); and against Borrelia burgdorferi (strain G39/40) by enzyme-linked immunosorbent assay (ELISA). The presence of antibodies to anti-R. rickettsii, anti-E. canis and anti-B. burgdorferi was detected in 32 (29.35%), 16 (14.67%) and 30 (27.52%) opossums, respectively. Opossum endpoint titers ranged from 64 to 1,024 for R. rickettsii, from 40 to 160 for E. canis, and from 400 to >51,200 for B. burgdorferi. These serological results suggest that opossums have been exposed to Rickettsia spp., Ehrlichia spp., and B. burgdorferi-related agents in the state of São Paulo. Our study underscores the need for further research about these agents in this study area, in view of the occurrence of Spotted Fever and Baggio-Yoshinari Syndrome disease in humans in the state of São Paulo, Brazil.

Molecular detection of tick-borne protozoan parasites in a population of domestic cats in midwestern Brazil.

Some tick-borne pathogens that infect domestic cats have been considered emergent in veterinary medicine. Occurrences of Hepatozoon spp., Babesia spp. and Cytauxzoon spp. have been described in several regions of Brazil. This paper offers a comprehensive analysis of the 18S rRNA gene of a Hepatozoon sp. strain detected in domestic cats in the metropolitan area of Cuiabá, in Midwestern Brazil. Based on a molecular analysis, we detected the presence of Hepatozoon species circulating among cats in this region. The aforementioned strain is closely related to other isolates of H. felis detected in wild felids. Moreover, a phylogenetic analysis indicates that this genotype is grouped into a clade of 18S rRNA sequences previously described for the genus Hepatozoon in wild felids around the world. Hepatozoon felis strains detected in cats from Spain and Israel showed, respectively, 98% and 97% identity to our sequence and are clustered on a separate branch of the phylogenetic tree. This finding suggests a high diversity of Hepatozoon genotypes occurring in cats in Europe and South America. None of the analyzed cats were positive for Babesia spp. or Cytauxzoon spp. by PCR analysis.