The animals were euthanatized according

to Cardoso (2002)

The animals were euthanatized according

to Cardoso (2002). The survey was associated with the project entitled “Description Venetoclax purchase of the Biodiversity of the Helminth Community of Small Mammals in the Pantanal of Mato Grosso do Sul”, sponsored by the Instituto Oswaldo Cruz and the Earthwatch Institute. The capture and necropsy of the rodents were authorized by the Brazilian Institute of Renewable Natural Resources (IBAMA), the federal environmental agency, and were performed according to biosecurity procedures (license numbers CGFAU 009/2002, 197/2002 and 091/2004). Biosecurity techniques and individual safety equipment were used during all procedures. The collection of parasites from T. apereoides captured from the field did not retrieve any males. We, therefore, proceeded with an experimental infection, where both females and males could Autophagy inhibitor chemical structure be obtained. Fourteen gravid females of Trichuris thrichomysi n. sp. recovered from naturally infected T. apereoides specimens captured in Capitão Andrade municipality were cut open with a scalpel and the contents of uterus were emptied into Petri dishes. The eggs were washed twice in dechlorinated water and incubated at 28 ± 2 °C for 60 days. Development

of cultures was observed under a Zeiss ID 02 inverted microscope. After that, 50-μL aliquots were mounted between slides and coverslips to quantify the percentage of embryonated eggs. One thousand below embryonated eggs, in 0.5 mL dechlorinated water, were administered orally to laboratory-bred T. apereoides specimens aged 8–12 weeks. The rodents’ were necropsied after euthanasia in a CO2 chamber and their stools were examined and worms recovered.

The rodents were bred and the parasite life cycle was maintained throughout the identification period in the Laboratorio de Biologia e Parasitologia de Mamíferos Silvestres e Reservatórios – IOC-FIOCRUZ. The whipworms were collected from the cecum of T. apereoides and T. pachyurus, washed in saline solution and fixed by immersion in hot AFA (2% glacial acetic acid, 3% formaldehyde and 95% ethanol). For light microscopy (LM), scanning electron microscopy (SEM) and field emission scanning electron microscopy (FESEM) specimens were prepared following Mafra and Lanfredi (1998), with 10 sec of gold coating for FESEM. Drawings were made with the aid of a camera lucida attached to a Zeiss Standard 20 light microscope. SEM micrographs were taken with a Jeol JSM 5310 and FESEM was performed with a Jeol JSM 6340F. All measurements (mean ± standard deviation) are given in micrometers, except measurements indicated in millimeters (mm).

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