Parasites in road-killed wild felines from North of Paraná state, Brazil.

This study aimed to identify the intestinal parasites of road-killed wild felines in the North Central and North, Paraná state, southern Brazil. The animals were monitored by sampling previously established transects. The places where the felines were run over were mapped, the animals were identified, and the gastrointestinal tract was evaluated. The feces were submitted to coproparasitological techniques of spontaneous sedimentation, floating in hypersaturated NaCl solution and centrifugal floating in zinc sulfate. All the parasitic structures detected were photomicrographed. In the coproparasitological analyses were identified oocysts of Cystoisospora spp., eggs of Ancylostomatidae, and Capillaria spp.; eggs of Aelurostrongylus spp., Toxocara spp., Physaloptera spp., Taenia spp., and Spirometra spp.; Aelurostrongylus abstrusus larvae; and eggs and adults of Ancylostoma cati and Taenia spp. One of the cats was parasitized by a flea of Ctenocephalides felis felis. Based on these results, the animals analyzed in this study supplied important samples for the evaluation of parasitic diversity of North of Paraná and suggested that this region may have conditions that allow the maintenance of these parasites life cycles in the environment and among wildlife.

Molecular identification and histological aspects of Renicola sloanei (Digenea: Renicolidae) in Puffinus puffinus (Procellariiformes): a first record / Identificação molecular e aspectos histológicos de Renicola sloanei (Digenea: Renicolidae) em Puffinus puffinus (Procellariiformes): primeiros registros

Abstract Renicolids are parasites that inhabit the renal tubules and ureters of molluscivorous and piscivorous birds. Puffinus puffinus is a migratory seabird that was identified as the definitive host of Renicola spp. Studies focusing on the renicolid species and the resulting renal lesions are valuable for their association with causes of stranding in seabirds. The aim of this study was to identify the renicolid trematodes and evaluate the histological findings in two P. puffinus stranded on the coast of Paraná state, Brazil. The parasites were evaluated by histologic, ultrastructural and molecular assays, while tissue changes were analyzed by histologic methods. The morphological and morphometrical characteristics of the parasites, along with polymerase chain reaction and sequencing assays (ribosomal and mitochondrial regions), identified the species as Renicola sloanei. The results also suggest that this helminth can be the adult form of Cercaria pythionike. The dilation of collecting ducts was the main histological finding in the kidneys. In conclusion, R. sloanei was identified, and for the first time, P. puffinus was described as a host of this digenean inducing mild renal changes.

Resumo Renicolídeos são parasitos que habitam túbulos renais e ureteres de aves que se alimentam de moluscos e peixes. Puffinus puffinus, ave marinha migratória, foi registrada como hospedeiro definitivo de Renicola spp. Estudos relacionados com as espécies de renicolídeos e as lesões renais resultantes são importantes para o entendimento das causas de óbito de aves marinhas. O objetivo deste estudo foi identificar os trematódeos renicolídeos e avaliar os achados histológicos em dois P. puffinus encalhados no litoral do Estado do Paraná, Brasil. Os parasitos foram avaliados por ensaios histológicos, ultraestruturais e moleculares, enquanto as alterações teciduais foram analisadas por métodos histológicos. As características morfológicas e morfométricas dos parasitos, juntamente com a reação em cadeia da polimerase e sequenciamento (regiões ribossomal e mitocondrial), identificaram a espécie como Renicola sloanei. Os resultados também sugerem que este helminto pode ser a forma adulta de Cercaria pythionike. A dilatação dos ductos coletores foi o principal achado histológico renal. Em conclusão, R. sloanei foi identificado, e pela primeira vez P. puffinus foi descrito como hospedeiro deste digenético induzindo alterações renais discretas.

RNAm expression profile of cancer marker genes in HepG2 cells treated with different concentrations of a new indolin-3-one from Pseudomonas aeruginosa.

The present study tested the effects of a newly identified indolin-3-one compound (compound 1), produced by Pseudomonas aeruginosa, on HepG2 cells. The MTT assays demonstrated decreased metabolic activities in HepG2 cells treated with compound 1, with dose- and time-dependent intensifying effect, starting at a concentration of 40 µM. The IC50 after 24, 48, 72, and 96 h treatments were 41.35, 52.7, 92.79 and 66.65 µM of compound 1, respectively. Below 80 µM, no significative damage on erythrocytes membranes was observed by the hemolytic assays. The RT-qPCR revealed that the compound modulated key genes involved in carcinogenesis process, indicating possible indolin-3-one mechanisms of action. The data showed that gene expression alterations promoted by compound 1, in concentrations up to 60 µM after 48 h, led to a decrease in cellular progression and there was no direct cellular damage. In addition, non-cytotoxic concentrations of compound 1 halved the concentration of the chemotherapeutic doxorubicin, maintaining similar therapeutic effect against HepG2 cells. The novelty of the molecule and the biological activities observed in the present study emphasize the potential of the compound 1 in cancer therapy research.

Prevalence of Eimeria spp. in calves from dairy farms in northern Paraná state, Brazil / Prevalência de Eimeria spp. em bezerros de propriedades leiteiras do norte do estado do Paraná, Brasil

Abstract Bovine coccidiosis is a disease of major importance in cattle herds across the world. The disorder mainly affects young calves, and E. bovis and E. zuernii are considered the most pathogenic species of the genus, however, E. alabamensis have been described in grazing calves. In this study, the prevalence of Eimeria spp. was evaluated in calves on dairy farms in the northern region of the state of Paraná, Brazil. Four hundred calves on 44 dairy farms were tested for the presence of coccidian oocysts. The positives were re-examined and the oocysts were morphometrically analyzed for species identification. All the farms were contaminated and 205 animals (51.25%) presented Eimeria spp. oocysts. Among these, 146 animals (71.22%) were co-infected by two or more species of coccidia. Ten species of Eimeria were identified: E. bovis (in 30.25% of the positive samples), E. alabamensis (26.75%), E. zuernii (22.00%), E. ellipsoidalis (18.50%), E. auburnensis (13.75%), E. canadensis (8.00%), E. cylindrica (7.25%), E. subspherica (5.00%), E. bukidnonensis (3.00%) and E. brasiliensis (0.75%). This study demonstrates the high prevalence of Eimeria spp. in the northern region of Paraná, Brazil, and detection for the first time in our region the pathogenic species E. alabamensis.

Resumo A coccidiose bovina é uma doença de grande importância em rebanhos ao redor do mundo. A desordem afeta principalmente bezerros jovens, e E. bovis e E. zuernii consideradas as espécies mais patogênicas deste gênero, causando grave enterite em animais infectados. No entanto, casos de E. alabamensis foram descritos em bezerros mantidos a pasto. No presente estudo, a prevalência de Eimeria spp. foi avaliada em bezerros de gado leiteiro da região norte do estado do Paraná, Brasil. Quatrocentos bezerros foram amostrados e testados para a presença de oocistos de coccídios. Os positivos foram re-examinados e os oocistos analisados morfologicamente para identificação da espécie. Todas as fazendas estavam contaminadas e 205 (51,25%) animais apresentaram oocistos de Eimeria spp. Destes, 146 (71,22%) animais estava co-infectados por duas ou mais espécies de coccídio. Dez espécies de Eimeria foram identificadas: E. bovis (30,25% de amostras positivas), E. alabamensis (26,75%), E. zuernii (22,00%), E. ellipsoidalis (18,50%), E. auburnensis (13,75%), E. canadensis (8,10%), E. cylindrica (8,00%), E. subspherica (5,00%), E. bukidnonensis (3,00%) e E. brasiliensis (0,75%). Este estudo demonstra a alta prevalência de Eimeria spp. na região norte do estado do Paraná, Brasil, e a detecção, pela primeira vez, de E. alabamensis.

Sensitivity evaluation of a single-step PCR assay using Ehrlichia canis p28 gene as a target and its application in diagnosis of canine ehrlichiosis / Avaliação da sensibilidade da PCR em uma etapa com base no gene p28 de Ehrlichia canis e sua aplicação no diagnóstico da erliquiose canina

The aim of this study was to optimize a PCR assay that amplifies an 843 pb fragment from the p28 gene of Ehrlichia canis and compare it with two other PCR methods used to amplify portions of the 16S rRNA and dsb genes of Ehrlichia. Blood samples were collected from dogs suspected of having a positive diagnosis for canine ehrlichiosis. Amplification of the p28 gene by PCR produced an 843-bp fragment and this assay could detect DNA from one gene copy among 1 billion cells. All positive samples detected by the p28-based PCR were also positive by the 16S rRNA nested-PCR and also by the dsb-based PCR. Among the p28-based PCR negative samples, 55.3 percent were co-negatives, but 27.6 percent were positive in 16S rRNA and dsb based PCR assays. The p28-based PCR seems to be a useful test for the molecular detection of E. canis, however improvements in this PCR sensitivity are desired, so that it can become an important alternative in the diagnosis of canine ehrlichiosis.

O objetivo deste estudo foi aperfeiçoar um ensaio de PCR que amplificasse um fragmento de 843 pares de bases do gene p28 da Ehrlichia canis e compará-lo com outros dois métodos de PCR utilizados para amplificar partes do gene 16S rRNA e dsb do gênero Ehrlichia. Amostras sanguíneas foram colhidas de cães com diagnóstico clínico de erliquiose. A amplificação do gene p28 pela PCR produziu um fragmento de 843pb e esse ensaio permitiu a detecção do DNA de um parasita dentre 1 bilhão de células. Todas as amostras positivas detectadas pela PCR baseada no gene p28 foram também positivas pela nested PCR para detecção do gene 16S rRNA e também pela PCR dsb. Dentre as amostras negativas para a PCR p28, 55,3 por cento foram co-negativas, mas 27,6 por cento foram positivas pela PCR baseada nos genes 16S rRNA e dsb. A PCR p28 parece ser um teste útil para detecção molecular de E. canis, entretanto otimizações na sensibilidade nesta PCR são necessárias, para que esta técnica se torne uma importante alternativa no diagnóstico da erliquiose canina.