Melittin peptide kills Trypanosoma cruzi parasites by inducing different cell death pathways.

Antimicrobial peptides (AMPs) are components of the innate immune response that represent desirable alternatives to conventional pharmaceuticals, as they have a fast mode of action, a low likelihood of resistance development and can act in conjunction with existing drug regimens. AMPs exhibit strong inhibitory activity against both Gram-positive and Gram-negative bacteria, fungi, viruses, metazoans and other parasites, such as the protozoan Leishmania. Melittin is a naturally occurring AMP, which comprises 40-50% of the dry weight of Apis mellifera venom. Our group has recently shown that crude A. mellifera venom is lethal to Trypanosoma cruzi, the Chagas disease etiologic agent, and generates a variety of cell death phenotypes among treated parasites. Here, we demonstrate that the melittin affected all of T. cruzi developmental forms, including the intracellular amastigotes. The ultrastructural changes induced by melittin suggested the occurrence of different programmed cell death pathways, as was observed in A. mellifera-treated parasites. Autophagic cell death appeared to be the main death mechanism in epimastigotes. In contrast, melittin-treated trypomastigotes appeared to be dying via an apoptotic mechanism. Our findings confirm the great potential of AMPs, including melittin, as a potential source of new drugs for the treatment of neglected diseases, such as Chagas disease.

Objective assessment of the cryoprotective effects of dimethylformamide for freezing goat semen.

The aim of this work was to assess the cryoprotective effects of dimethylformamide (DMF) for freezing goat semen, using an objective analysis by computer-assisted sperm analysis (CASA). Twenty-one ejaculates (seven per animal) were collected from three stud bucks with the aid of an artificial vagina and immediately evaluated for gross and microscopic characteristics. The semen was diluted in two steps with a Tris-egg yolk extender containing 6% glycerol or 6% DMF, frozen in 0.50-mL straws, and stored in liquid nitrogen. Samples were accessed for sperm morphology, sperm membrane structural and functional integrity, and by CASA, immediately after thawing. There were differences (P<0.05) between glycerol and DMF with regard to subjective progressive motility (23.9±2.2% vs. 16.6±2.0%), objective progressive motility (3.5±0.4% vs. 1.8±0.3%), linearity (53.9±1.6% vs. 48.1±1.4%) and amplitude of lateral head (2.3±0.1 vs. 2.9±0.1 mm), which confirmed the efficiency of glycerol. In conclusion, dimethylformamide could be used as an alternative cryoprotectant for goat semen freezing. However it was showed that no benefits were derived by using dimethylformamide to replace glycerol at an equal 6% concentration.

Trypanosoma vivax nos tecidos testicular e epididimário de ovinos experimentalmente infectados / Trypanosoma vivax in testicular and epidydimal tissues of experimentally infected sheep

Quatro ovinos machos, com cerca de 12 meses de idade (Ovinos 1-4), foram infectados por via intravenosa com aproximadamente 1,25x10(5) tripomastigotas de Trypanosoma vivax, outros quatro ovinos (Ovinos 5-8) destinaram-se ao grupo controle. Após a infecção, exames clínicos visando avaliar temperatura retal, freqüências cardíaca e respiratória e parasitemia foram realizados diariamente por 30 dias, tempo estabelecido para o término do experimento. A avaliação do hematócrito foi realizada a cada cinco dias. Ao final do período experimental, os animais foram castrados e os testículos e epidídimos submetidos ao exame anatomopatológico. Amostras destes órgãos dos Ovinos 1, 4 e 5 foram tomadas para a realização da reação em cadeia da polimerase (PCR). Os parâmetros clínicos (hipertermia, aumento das freqüências cardíaca e respiratória, aumento de volume dos linfonodos e palidez das mucosas) mantiveram-se para o grupo infectado acima dos valores mostrados pelo grupo controle durante todo o período experimental. A parasitemia foi observada a partir do 3º dia pós-infecção (dpi) com picos nos 6-10os dpi e nos 15-18os dpi. Os Ovinos 1 e 4 apresentaram, a partir do 25º dpi, anemia acentuada. Macroscopicamente, todos os testículos dos animais do grupo infectado apresentaram-se flácidos e com coloração pálida. Microscopicamente, observaram-se degeneração testicular moderada a acentuada, epididimite multifocal e hiperplasia do epitélio epididimário. A análise por PCR de T. vivax nos tecidos testicular e epididimário resultou em 100% de positividade para ovinos infectados experimentalmente. As lesões epididimárias e testiculares associadas à presença do parasita nesses órgãos, detectada por PCR, sugerem a participação do parasita no mecanismo etiopatogênico de danos reprodutivos.(AU)

Four adult sheep (number 1, 2, 3 and 4), all males, were inoculated intravenously with 1ml of blood containing 1.25x10(5) trypomastigotes of Trypanosoma vivax, and Sheep 5, 6, 7 and 8 were used as control. After infection, clinical exams considering rectal temperature, respiratory and cardiac frequencies, and parasitaemia were recorded daily for a 30-day experiment period. Blood samples were obtained for 5-day intervals to hematocrit analysis. At the end of the experimental period, the sheep were orquiectomized. Testes and epididymides from these animals were studied anatomopathologically. Samples from these tissues of Sheep 1, 4 and 5 were taken to polymerase chain reaction (PCR). Clinical parameters remained for the infected group above the values observed in the control group during the experimental period. Parasitaemia was observed on day 3 post-infection, and the highest values occurred between day 6 and 10, and day 15 and 18 post-infection. Sheep 1 and 4 showed severe anemia on day 25 post-infection. All sheep of the infected group showed flabby and palid testes. Histologically, moderate to severe testicular degeneration, multifocal epididymitis and hyperplasia of epididymal epithelium were observed. The result of T. vivax PCR analysis in the testes and epididymal tissues was positive in 100% of the samples of the experimentally infected sheep. Epididymal and testicular lesions associated with the presence of the parasite in these tissues, shown by PCR, suggest the participation of T. vivax in the pathophysiological mechanism of reproductive damage.(AU)