Litomosoides brasiliensis (Nematoda: Onchocercidae) infecting chiropterans in the Legal Amazon region, Brazil.

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.

Parasitoses intestinais e fatores de risco associados em crianças em um município do Nordeste Brasileiro / Intestinal parasites and risk factors associated f in children in a Brazilian Northeast municipality

Introdução: os parasitos gastrintestinais são causas de morbidade em crianças, podendo determinar quadros de anemia e diarréria. Podem estar associados as condições socioeconômicas, de saneamento básico e práticas de higiene. Objetivo: o objetivo desta investigação foi identificar as parasitoses intestinais e os fatores de risco associados em crianças menores de 12 anos cadastradas em duas Unidades Básicas de Saúde do município de Caxias, estado do Maranhão. Metodologia: neste estudo transversal 422 exames coproparasitológicos pela técnica de Hoffman, Pons e Janner foram realizados. Para comparar as proporções utilizou-se o teste Qui-quadrado ou teste exato de Fisher. Empregou-se análises univariadas e multivariadas por regressão logística para avaliar a associação entre a infecção por enteroparasitos e os fatores de risco. Resultados: a frequência de parasitos intestinais foi 49,05%, sendo predominante o Ascaris lumbricoides (68,18%) e a Entamoeba sp. (65,38%). Conforme a análise estatística, ingerir água sem tratamento domiciliar e não lavar as mãos antes das refeições podem ser considerados como fatores de risco para as enteropatias parasitárias. Conclusão: ressalta-se a necessidade de implementar políticas públicas que visem reduzir as altas taxas destes parasitos, e que contribuam para melhorar a qualidade de vida dessa população.

Introduction: intestinal parasites are cause of morbidity in children and can determine anemia and diarrhea. It can be associated to social economical conditions, basic sanitation and hygiene. Objective: the objective of this research was to identify tne intestinal parasitosis and the risk factors associated in children less than 12 years old registered in two Basic Health Units in Caxias municipality, state of Maranhão. Metodology: in this cross-sectional study 422 fecal exams were performed by the technique of Hoffman, Pons and Janner in 422 children. To compare the proportions, the chi-square test or Fisher's exact test was used. Univariate and multivariate analyzes were used by logistic regression to evaluate the association between enteroparasite infection and risk factors. Results: the frequency of intestinal parasites was 49.05%, being predominant Ascaris lumbricoides (68.8%) and Entamoeba sp. (65.38%). According to statistical analysis, drinking water without home treatment and not washing hands before meals, can be considered as risk factors for parasitic enteropathies. Conclusion: It's necessary to implement public policies aimed at reducing the high rates of intestinal parasites, which contributes to improve the quality of life of this population.

Diagnosis and epidemiology of Leishmania infantum in domestic cats in an endemic area of the Amazon region, Brazil.

Visceral leishmaniasis is a zoonotic disease caused by Leishmania infantum for which dogs are the main reservoir. In South America, presence of this disease is expanding along with increasing dispersion of its principal vector, the sand-fly Lutzomyia longipalpis. Feline leishmaniasis is an emerging disease in domestic cats, but epidemiological studies in endemic areas of the Amazon region of Brazil are scarce and the role of cats as reservoirs of L. infantum has been debated. The aim of this study was to investigate L. infantum infection in cats living in the Amazon biome region, using serological and molecular methods. A total of 105 cats were subjected to clinical examination and blood samples were taken for immunofluorescent-antibody (IFAT) serological evaluation, to determine anti-Leishmania antibody titers. Conventional PCR and Sanger's sequencing targeting L. infantum chitinase and Leishmania species ribosomal internal transcribed spacer (ITS-1) encoding genes were performed on conjunctival swabs from these cats. Seropositivity was detected in 32 animals (30.48%), thus confirming that contact between these cats and the parasite was occurring. PCR followed by amplicon sequencing showed that three samples (2.86%) were positive for a chitinase gene and six (5.71%) were positive for the ITS-1 gene. Parasite-positive diagnoses presented a statistically significant association with free access to the streets (p = 0.0111), cohabitation with dogs affected previously by VL (p = 0.0006) and absence of backyard cleaning and garbage collection (p = 0.00003). These results emphasize that cats should be included in epidemiological surveys of leishmaniasis, especially in endemic areas, if not as the reservoir host (unproven), at least as a "sentinel host" that is useful for revealing situations of endemic circulation of L. infantum. Moreover, in these areas, feline leishmaniasis needs to be considered in the differential diagnosis among domestic cats presenting alopecia, rarefied hair, lacerations and ulcerative dermatitis.

Exposure of Baixadeiro horses to Rickettsia spp. and to ticks infected by Rickettsia amblyommatis in the Baixada Maranhense micro-region, Maranhão, Brazil / Exposição dos cavalos Baixadeiro a Rickettsia spp. e carrapatos infectados pela Rickettsia amblyommatis na Baixada Maranhense, microrregião do Estado do Maranhão, Brasil

ABSTRACT: The aim of this study was to investigate exposure of Baixadeiro horses to Rickettsia spp. and to ticks infected by Rickettsia in the Baixada Maranhense (lowlands) micro-region, state of Maranhão. A total of 258 horses were tested for Rickettsia rickettsii, Rickettsia amblyommatis and Rickettsia bellii using the immunofluorescence assay (IFA). Overall, 58.91% (152/258) of the horses were seroreactive for at least one Rickettsia species, and 85.27% of the horses were infested with one or more species of tick, which were identified as Dermacentor nitens (93.63%), Amblyomma cajennense sensu stricto (4.55%) and Rhipicephalus (Boophilus) microplus (1.82%). These ticks were subjected to DNA extraction and were tested using the polymerase chain reaction (PCR), targeting two rickettsia genes: citrate synthase gene (gltA) and 190kDa outer membrane protein gene (ompA). Three specimens of A. cajennense s.s. were positive. BLAST analyses on the nucleotide sequences obtained from the PCR products showed that these were 99-100% identical to the corresponding sequences of R. amblyommatis. Thus, results indicate that R. amblyommatis and/or a strain very close to this is circulating in ticks in this micro-region.

RESUMO: O objetivo deste estudo foi verificar a exposição e a infecção de cavalos baixadeiro e carrapatos por Rickettsia spp. na micro-região da Baixada Maranhense do Estado Maranhão. Um total de 258 cavalos foi testado pelo ensaio de Imunofluorescência Indireta (IFI) para Rickettsia rickettsii, Rickettsia amblyommatis e Rickettsia bellii. Deste total, 58,91% (152/258) foram sororreativos para pelo menos uma espécie de Rickettsia sp., e 85,27% cavalos estavam infestados por uma ou mais espécies de carrapatos identificados como Dermacentor nitens (93,63%), Amblyomma cajennense sensu stricto (4,55%) e Rhipicephalus (Boophilus) microplus (1,82%). Estes carrapatos foram submetidos à extração de DNA e testados pela reação em cadeia pela polimerase (PCR) alvejando os genes citrato sintase (gltA) e o de proteína de membrana externa de 190kDa (ompA). Três espécimes de A. cajennense s.s. foram positivos para Rickettsia amblyommatis. As sequências dos nucleotídeos obtidas a partir dos produtos de PCR mostraram 99-100% de identidade com as sequências correspondentes de R. amblyommatis quando analisadas pelo BLAST. Desta forma, os resultados indicam a circulação de R. amblyommatis nos carrapatos e/ou uma cepa muito próxima circulando na microrregião.

Molecular and serological detection of Theileria equi, Babesia caballi and Anaplasma phagocytophilum in horses and ticks in Maranhão, Brazil / Detecção molecular e serológica de Theileria equi, Babesia caballi e Anaplasma phagocytophilum em equinos e carrapatos no Maranhão

Equine piroplasmosis is a tick-borne disease caused by the intraeytrhocytic protozoans Babesia caballi and Theileria equi. It has been reported as a main equine parasitic disease. In addition, Anaplasma phagocytophilum, the causative agent of granulocytic ehrlichiosis, causes a seasonal disease in horses. Both diseases, can be detrimental to animal health. In this sense, blood samples and ticks were collected from 97 horses raised in the microregion of Baixada Maranhense, Maranhão State, Brazil. Serum samples were subjected to Indirect Fluorescence Antibody Test (IFAT) and blood samples and ticks to Polymerase Chain Reaction (PCR) to evaluate the infection by Theileria equi, Babesia caballi and Anaplasma phagocytophilum. The overall seroprevalence was 38.14%, 18.55% and 11.34% for T. equi, B. caballi and A. phagocytophilum, respectively. The results of PCR from blood samples showed 13.40% and 3.09% positive samples to T. equi and B. caballi, respectively. A total of 170 tick specimens were collected and identified as Dermacentor nitens, Amblyomma cajennense sensu lato and Rhipicephalus (Boophilus) microplus. It was detected 2.35% (4/170) and 0.59% (1/170) positive tick samples by PCR for T. equi and B. caballi, respectively. All samples were negative to A. phagocytophilum. No statically difference (p>0.05) was observed when gender, age, use of ectoparasiticide and tick presence were analyzed. A BLASTn analysis of the sequenced samples indicated 97 to 100% similarity with T. equi 18S rRNA gene sequences in GenBank and 98 to 100% with B. caballi. Genetic analysis classified the obtained sequences as T. equi and B. caballi cluster, respectively. It can be concluded that these pathogens occur and are circulating in the studied area.(AU)

A piroplasmose equina é uma doença transmitida por carrapatos causada pelos protozoários intraeritrocitários Babesia caballi e Theileria equi. É relatada como uma doença parasitária comum em equinos. Além disso, Anaplasma phagocytophilum, o agente causal da ehrlichiose granulocítica, causa uma doença sazonal em equinos. Ambas as doenças, podem ser prejudiciais para a saúde animal. Nesse sentido, amostras de sangue e carrapatos foram coletadas de 97 cavalos criados na microrregião da Baixada Maranhense, estado do Maranhão, Brasil. As amostras de soro foram submetidas ao Teste de Imunofluorescência Indireta (RIFI) e amostras de sangue e os carrapatos a Reação da Polimerase em Cadeia (PCR) para avaliar a infecção por Theileria equi, Babesia caballi e Anaplasma phagocytophilum. A prevalência foi de 38,14%, 18,55% e 11,34% para T. equi, B. caballi e A. phagocytophilum, respectivamente. Os resultados da PCR para as amostras de sangue demonstraram 13,40% e 3,09% de positividade para T. equi e B. caballi, respectivamente. Um total de 170 specimens de carrapatos foi coletado e foram identificados Dermacentor nitens, Amblyomma cajennense sensu lato and Rhipicephalus (Boophilus) microplus. Obteve-se 2,35% (4/170) e 0,59% (1/170) positivos por PCR para T. equi e B. caballi, respectivamente. Todas as amostras foram negativas para A. phagocytophilum. Não houve diferença estatística significativa (p>0.05) em relação ao sexo, idade, uso de ectoparasiticida e presença de carrapatos. A análise BLASTn das amostras sequenciadas para gene 18S rRNA indicaram 97 a 100% de similaridade com T. equi e 98-100% com B. caballi no GenBank. Análises genéticas classificaram as sequencias obtidas no mesmo clado que T. equi e B. caballi, respectivamente. Podemos concluir que estes patógenos estão circulando na área de estudo.(AU)

Comparative growth of spotted fever group Rickettsia spp. strains in Vero cells.

In Brazil, the spotted fever group (SFG) Rickettsia rickettsii and Rickettsia parkeri related species are the etiological agents of spotted fever rickettsiosis. However, the SFG, Rickettsia rhipicephali, that infects humans, has never been reported. The study of growth dynamics can be useful for understanding the infective and invasive capacity of these pathogens. Here, the growth rates of the Brazilian isolates R. rickettsii str. Taiaçu, R. parkeri str. At#24, and R. rhipicephali HJ#5, were evaluated in Vero cells by quantitative polymerase chain reaction. R. rhipicephali showed different kinetic growth compared to R. rickettsii and R. parkeri.

Comparative growth of spotted fever group Rickettsia spp. strains in Vero cells

In Brazil, the spotted fever group (SFG) Rickettsia rickettsii and Rickettsia parkeri related species are the etiological agents of spotted fever rickettsiosis. However, the SFG, Rickettsia rhipicephali, that infects humans, has never been reported. The study of growth dynamics can be useful for understanding the infective and invasive capacity of these pathogens. Here, the growth rates of the Brazilian isolates R. rickettsii str. Taiaçu, R. parkeri str. At#24, and R. rhipicephali HJ#5, were evaluated in Vero cells by quantitative polymerase chain reaction. R. rhipicephali showed different kinetic growth compared to R. rickettsii and R. parkeri.