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An adult free-ranged female maned wolf was rescued from a periurban area subject to anthropogenic disturbances in the Minas Gerais, Brazil. The animal presented poor body condition and anemia. The clinical condition rapidly deteriorated culminating in dead and a necropsy was performed. The main gross lesions were marked anemia and blood content in the intestines accompanied by many types of parasites. The protozoa Rangelia vitalii was identified by histopathological analysis predominantly within the cytoplasm of endothelial cells of capillaries of the small intestine. The lymph nodes, spleen, bone marrow, dermis, lungs and kidney had similar protozoal forms but with mild or moderate intensity. Rangelia vitalii was confirmed by molecular assays. Dec 21, 2017 - Recover My Files Crack is a data recovery program that uses the principle of File carving to retrieve your lost data from unallocated places. Connectify Hotspot Pro 2018 Crack Plus License key [Latest]. 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These findings emphasize the fragility of Brazilian ecosystems, especially in disturbed areas, reinforcing the necessity of efforts to preserve these areas and wild carnivores, some of which are threatened with extinction, such as the maned wolf. History In November 2013, one adult maned wolf female weighting 18 kg was rescued from a periurban house located in the Rio Acima municipality (20°5′44.732″ S and 43°47′24.263″ W) in the state of Minas Gerais (MG), southeast of Brazil. The animal had physical debilitation and pain in the right foreleg. After its rescue by the Brazilian Federal Environmental Agency (IBAMA), it was transported to a veterinary clinic. Clinical evaluation showed right elbow luxation and tick infestation. Fipronil spray 0.25% was administered. Blood sample were collected in an ethylene diamine tetraacetic acid (EDTA) tube for further molecular analyses and for preparation of blood smears. Eleven days later, the animal died and necropsy was performed at the Pathology Sector, School of Veterinary Medicine, Universidade Federal de Minas Gerais (UFMG). The study was approved by Ethics Committee for Animal Research of the UFMG under protocol 332/2013 and by the Brazilian Institute for Environment and Natural Renewable Resources (IBAMA, Belo Horizonte, MG, Brazil) under license 34633-5. Necropsy and sampling Necropsy was performed and the animal was grossly evaluated. Samples of brain, lung, trachea, heart, esophagus, stomach, intestine, kidney, bone marrow, spleen, and lymph nodes were collected in duplicate: one portion of each tissue was fixed in 10% neutral-buffered formalin for histopathology; and the other portion of the same tissue was frozen at −20 °C and stored for subsequent DNA extraction. For search for the parasites in the intestines, anatomical segments were separated by double ligatures, the contents removed and the mucosae scraped. The collected material was washed in strainers and the solid portion was fixed and preserved in acetic formaldehyde. For histopathological analysis, tissues were dehydrated using a series of increasing ethanol concentrations, cleared in xylene, and embedded in paraffin. Sections of 4 μm were obtained and stained with hematoxylin and eosin (HE) for routine histological evaluation under light microscopy (). Parasite identification in histological sections was performed according to. DNA extraction and PCR amplification DNA was extracted from helminths, organs and blood samples using Wizard ® Genomic DNA Purification Kit (Promega, Madison, WI, USA), according to the manufacturer's instructions for animal tissue and whole blood, respectively. Atlas De Parasitologia Veterinaria Pdf Reader DownloadIn all PCR assays, the reaction mixture in the first round contained 7.5 μL of GoTaq ®Green Master Mix (Promega, Madison, WI, USA), 0.6 μL of a solution containing the mixed primers (10 mM) and 5.4 μL of nuclease free water. A 1.5 μL of total DNA was added to the reaction mixture to obtain a final volume of 15 μL. The reaction mixtures in the second round assays were similar, except the templates were the products from the first round PCR reactions (1.5 μL). Sets of primers used to detect the parasite species and amplifications were performed using a touchdown PCR programmed as previously (,,, ) ().
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