Levantamento molecular de micoplasma hemotrópico em mão-pelada (Procyon cancrivorus) do sul do Brasil

Hemoplasmas are non-cultivable bacterial parasites of erythrocytes that infect domestic and wild animals, as well as humans. Their means of transmission and pathogenesis remain contentious issues and difficult to evaluate in wild animals. Procyon cancrivorus is a South American carnivore and occurs in all Brazilian biomes. In this study, we aimed to investigate occurrences of hemoplasmas infecting P. cancrivorus and to identify their 16S rRNA gene, in southern Brazil. DNA was extracted from spleen and blood samples of P. cancrivorus (n = 9) from different locations. Hemoplasma DNA was detected in six samples, based on 16S rRNA gene amplification and phylogenetic analysis. Four of the six sequences belonged to the “Mycoplasma haemofelis group”, which is closely related to genotypes detected in Procyon lotor from the USA; one was within the “Mycoplasma suis group”, closely related to “Candidatus Mycoplasma haemominutum”; and one was within the intermediate group between these clusters. Thus, these sequences showed that the molecular identity of hemoplasmas in the population studied was very variable. In five positive animals, Amblyomma aureolatum ticks and a flea (Ctenocephalides felis felis) were collected. The present study describes the first molecular detection of mycoplasmas in P. cancrivorus.Os micoplasmas hemotrópicos (hemoplasmas) são parasitas bacterianos não-cultiváveis de eritrócitos que infectam tanto animais domésticos e selvagens, como seres humanos. A transmissão e a patogênese são discutíveis e difíceis de avaliar em animais selvagens. O mão pelada (Procyon cancrivorus) é um carnívoro Sul-americano, que ocorre em todos os biomas brasileiros. O objetivo do presente estudo é o de investigar a ocorrência de hemoplasmas infectando P. cancrivorus e identificar seu gene 16S rRNA no Sul do Brasil. O DNA foi extraído do baço e amostras de sangue de P. cancrivorus (n= 9). O DNA de hemoplasma foi detectado em seis amostras, com base na amplificação do gene 16S rRNA e na análise filogenética. Quatro das seis sequências pertencem ao “Grupo Mycoplasma haemofelis”, que estão intimamente relacionadas aos genótipos detectados no Procyon lotor dos EUA; uma dentro do “Grupo Mycoplasma suis”, que está intimamente relacionado ao “Candidatus Mycoplasma haemominutum”, e uma dentro do grupo intermediário entre esses clusters, mostrando assim que há uma diversidade genética de hemoplasmas na população estudada. Em cinco animais positivos, foram coletados carrapatos Amblyomma aureolatum e uma pulga Ctenocephalides felis. O presente estudo traz a primeira detecção molecular de micoplasmas em P. cancrivorus

Pathological findings associated with Dipetalonema spp. (Spirurida, Onchocercidae) infection in two species of Neotropical monkeys from Brazil

Among vector-borne helminths, filarioids of the genus Dipetalonema (Spirurida: Onchocercidae) localize in several tissues and body cavities of several animal species, causing mild to moderate lesions. The pathological findings associated with Dipetalonema spp. infection in Neotropical monkeys from southern Brazil are herein described, along with a fatal case due to filarial polyserositis and entrapment of an intestinal segment. At necropsy, nematodes were observed in abdominal and thoracic cavities, or in the pericardium of 37 (31.3%) out of the 118 individuals examined (i.e., 35 Alouatta guariba clamitans and two Sapajus nigritus). In addition, at histology, 27.0% of positive animals presented microfilarie (inside blood vessels of lung, spleen, liver, and brain) and 8.1% presented adult nematodes in the heart, lung, and liver. In two cases, cross-sections of filarioids were associated with areas of epicardial thickening with intense fibrosis and pyogranulomatous inflammation in the brain, heart, liver, lungs, or spleen. The DNA fragment was amplify using the cox1 gene, sequenced and analyzed to identify the nematode species collected; presence of Wolbachia was assessed in the filarioids using the 16S rRNA gene. At BLAST analysis of the cox1 gene, 10 sequences showed 91.7% nucleotide identity with Dipetalonema gracile, and two with D. gracile (98.5%) and Dipetalonema graciliformis (98.3%). Phylogenetic analyses clustered sequences of the cox1 obtained in this study in two clades corresponding with the host species. Wolbachia sp. endosymbiont was detected in four samples. Data herein reported provide a description of pathological lesions associated with the infection by Dipetalonema spp., suggesting that they may cause disease in Neotropical monkeys. In addition, a better understanding of diversity and biology of Dipetalonema spp. in South America is needed to assess the impact they may cause in native non-human primates from Brazil

The influence of xenobiotics in river sediment on the reproduction and survival of Daphnia magna, 1820, Straus Influência de xenobióticos no sedimento de rio sobre a reprodução e a sobrevivência de Daphnia magna, 1820, Straus

AIM: The Taquari River, a tributary of Guaíba Lake, is a major economic and environmental source for Rio Grande do Sul, a state located in the far south of Brazil; METHODS: This study evaluated the interference of sediment from this river in the reproduction and survival of 320 microcrustaceans (80 organisms per site) of the Daphnia magna species. Four sites between the middle course and the river mouth were monitored using eight samples from each site, between June/06 and April/10. Mortality higher than 20% suggested the acute action of the sample on the daphniids, while low reproduction indicated the presence of chronic toxicity. Sometimes an effect on survival was noted, but the number of individuals generated was always low. Semi-static chronic assays (21 day) were developed in 50 mL beakers using sediment and M4 culture medium at a ratio of 1:4 (v:v). Each beakers received a microcrustacean born from lots presenting LC50-24 hours = 0.98 mg K2Cr2O7 ± 0.04 mg totaling ten replicas per sample. The assays were developed under controlled light (16 hours light/day) and temperature (20 ºC ± 2 ºC) conditions; RESULTS: Duncan test showed a variation in reproduction (p < 0.05) with the presence of chronic toxicity. There was less reproductive activity in 91% of the samples, while for survival the reduction attained 9%. Spearman correlation identified the dependence between the rainfall index and reproduction in one of the samples and was absent when the rainfall index was compared to survival; CONCLUSIONS: Taquari River presented episodes of toxicity, and it is necessary to continue studies in the area and to inspect local sources of pollution, as already has been done. Chronic bioassays should be maintained when the purpose is to evaluate environmental quality, since reproduction proved more sensitive than survival.<br>OBJETIVO: O rio Taquari, tributário do Lago Guaíba, constitui importante fonte econômica e ambiental para o Rio Grande do Sul, estado localizado no extremo sul do Brasil; MÉTODOS: O presente estudo avaliou a interferência do sedimento deste rio na reprodução e na sobrevivência de 320 microcrustáceos (80 organismos por local) da espécie Daphnia magna. Foram monitorados quatro locais entre o curso médio e a foz do rio, utilizando oito amostras de cada local, entre junho/06 e abril/10. Mortalidade superior a 20% sugeriu ação aguda da amostra sobre os daphniids, enquanto a baixa reprodução indicou presença de toxicidade crônica. Eventualmente foi observado efeito na sobrevivência, porém a pequena geração de indivíduos foi constante. Ensaios crônicos (21 dias) semi-estáticos foram desenvolvidos em béqueres de 50 mL utilizando sedimento e meio de cultivo M4 na relação de 1:4 (v:v). Cada béquer recebeu um microcrustáceo nascido de lotes apresentando LC50-24 horas = 0,98 mg K2Cr2O7 ± 0,04 mg totalizando dez réplicas por amostra. Os ensaios desenvolveram-se em condições controladas de iluminação (16 horas luz diárias) e temperatura (20 ºC ± 2 ºC) RESULTADOS: O Teste de Duncan mostrou variação na reprodução (p < 0.05) com presença de toxicidade crônica. Ocorreu redução na atividade reprodutiva em 91% das amostras, enquanto que para sobrevivência a redução alcançou 9%. A correlação de Spearman identificou dependência entre índice pluviométrico e reprodução em uma das amostras e esteve ausente quando o índice pluviométrico foi comparado com a sobrevivência CONCLUSÕES: O rio Taquari apresentou episódios de toxicidade sendo necessários estudos na área e fiscalização das fontes poluidoras locais como já vem sendo realizado. Bioensaios crônicos devem ser mantidos quando o objetivo for a avaliação da qualidade ambiental, já que a reprodução foi mais sensível que a sobrevivência

Pathological findings associated with Dipetalonema spp. (Spirurida, Onchocercidae) infection in two species of Neotropical monkeys from Brazil

Among vector-borne helminths, filarioids of the genus Dipetalonema (Spirurida: Onchocercidae) localize in several tissues and body cavities of several animal species, causing mild to moderate lesions. The pathological findings associated with Dipetalonema spp. infection in Neotropical monkeys from southern Brazil are herein described, along with a fatal case due to filarial polyserositis and entrapment of an intestinal segment. At necropsy, nematodes were observed in abdominal and thoracic cavities, or in the pericardium of 37 (31.3%) out of the 118 individuals examined (i.e., 35 Alouatta guariba clamitans and two Sapajus nigritus). In addition, at histology, 27.0% of positive animals presented microfilarie (inside blood vessels of lung, spleen, liver, and brain) and 8.1% presented adult nematodes in the heart, lung, and liver. In two cases, cross-sections of filarioids were associated with areas of epicardial thickening with intense fibrosis and pyogranulomatous inflammation in the brain, heart, liver, lungs, or spleen. The DNA fragment was amplify using the cox1 gene, sequenced and analyzed to identify the nematode species collected; presence of Wolbachia was assessed in the filarioids using the 16S rRNA gene. At BLAST analysis of the cox1 gene, 10 sequences showed 91.7% nucleotide identity with Dipetalonema gracile, and two with D. gracile (98.5%) and Dipetalonema graciliformis (98.3%). Phylogenetic analyses clustered sequences of the cox1 obtained in this study in two clades corresponding with the host species. Wolbachia sp. endosymbiont was detected in four samples. Data herein reported provide a description of pathological lesions associated with the infection by Dipetalonema spp., suggesting that they may cause disease in Neotropical monkeys. In addition, a better understanding of diversity and biology of Dipetalonema spp. in South America is needed to assess the impact they may cause in native non-human primates from Brazil