Ocorrência de parasitas gastrintestinais em cães e gatos na rotina do laboratório de enfermidades parasitárias da FMVZ/Unesp-Botucatu, SP.

With the object of determining the occurrence and prevalence of parasites in dogs and cats in the region of Botucatu, a survey was conducted from the results of fecal parasitologic exams feces, processed by the laboratory of parasitic diseases of FMVZ-UNESP/Botucatu from January 2002 to December 2006. 1,012 fecal samples of dogs and cats were evaluated by the technique of Willis Mollay and Faust. In dogs the higher incidence was found for Ancylostoma caninum eggs (38%). In cats, the oocysts of Isospora spp were present in 48.38% of positive samples.

Avaliação de duas técnicas coproparasitológicas convencionais e de um kit comercial na investigação da epidemiologia de parasitas gastrintestinais de cães no Estado de São Paulo

Pós-graduação em Medicina Veterinária - FMVZ

Enteroparasitoses caninas

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Ocorrência de endoparasitos com potencial zoonótico em gatos no município de Andradina, São Paulo, Brasil

Pós-graduação em Ciência Animal - FMVA

Alterações microbiológicas e químicas na silagem de cana-de-açúcar (Saccharum officinarum L.) in natura e queimada, inoculadas ou não, com Lactobacillus buchneri

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Estudos parasitológicos em cães domésticos errantes e carnívoros selvagens generalistas no Parque Nacional do Iguaçu, Foz do Iguaçu

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Lufaxin, a Novel Factor Xa Inhibitor From the Salivary Gland of the Sand Fly Lutzomyia longipalpis Blocks Protease-Activated Receptor 2 Activation and Inhibits Inflammation and Thrombosis In Vivo

Objective-Blood-sucking arthropods' salivary glands contain a remarkable diversity of antihemostatics. The aim of the present study was to identify the unique salivary anticoagulant of the sand fly Lutzomyia longipalpis, which remained elusive for decades. Methods and Results-Several L. longipalpis salivary proteins were expressed in human embryonic kidney 293 cells and screened for inhibition of blood coagulation. A novel 32.4-kDa molecule, named Lufaxin, was identified as a slow, tight, noncompetitive, and reversible inhibitor of factor Xa (FXa). Notably, Lufaxin's primary sequence does not share similarity to any physiological or salivary inhibitors of coagulation reported to date. Lufaxin is specific for FXa and does not interact with FX, Dansyl-Glu-Gly-Arg-FXa, or 15 other enzymes. In addition, Lufaxin blocks prothrombinase and increases both prothrombin time and activated partial thromboplastin time. Surface plasmon resonance experiments revealed that FXa binds Lufaxin with an equilibrium constant approximate to 3 nM, and isothermal titration calorimetry determined a stoichiometry of 1:1. Lufaxin also prevents protease-activated receptor 2 activation by FXa in the MDA-MB-231 cell line and abrogates edema formation triggered by injection of FXa in the paw of mice. Moreover, Lufaxin prevents FeCl3-induced carotid artery thrombus formation and prolongs activated partial thromboplastin time ex vivo, implying that it works as an anticoagulant in vivo. Finally, salivary gland of sand flies was found to inhibit FXa and to interact with the enzyme. Conclusion-Lufaxin belongs to a novel family of slow-tight FXa inhibitors, which display antithrombotic and anti-inflammatory activities. It is a useful tool to understand FXa structural features and its role in prohemostatic and proinflammatory events. (Arterioscler Thromb Vasc Biol. 2012;32:2185-2196.)

Pathogens of Wild Maned Wolves (Chrysocyon brachyurus) in Brazil

The maned wolf, Chrysocyon brachyurus, is an endangered Neotropical canid that survives at low population densities. Diseases are a potential threat for its conservation but to date have been poorly studied. We performed clinical evaluations and investigated the presence of infectious diseases through serology and coprologic tests on maned wolves from Galheiro Natural Private Reserve, Perdizes City, Minas Gerais State, southeastern Brazil. Fifteen wolves were captured between 2003 and 2008. We found high prevalences of antibody to canine distemper virus (CDV; 13/14), canine parvovirus (CPV; 4/14), canine adenovirus type 2 (13/14), canine coronavirus (5/11), canine parainfluenza virus (5/5), and Toxoplasma gondii (6/8), along with Ancylostomidae eggs in all feces samples. Antibodies against Leishmania sp. were found in one of 10 maned wolves, and all samples were negative for Neospora caniman. Evidence of high exposure to these viral agents was also observed in unvaccinated domestic clogs from neighboring farms. High prevalence of viral agents and parasites such as CDV, CPV, and Ancylostomidae indicates that this population faces considerable risk of outbreaks and chronic debilitating parasites. This is the first report of exposure to canine parainfluenza virus in Neotropical free-ranging wild canids. Our findings highlight that canine pathogens pose a serious hazard to the viability of maned wolves and other wild carnivore populations in the area and emphasize the need for monitoring and protecting wildlife health in remaining fragments of the Cerrado biome.

Besnoitia besnoiti protein disulfide isomerase (BbPDI): molecular characterization, expression and in silico modelling

Besnoitia besnoiti is an apicomplexan parasite responsible for bovine besnoitiosis, a disease with a high prevalence in tropical and subtropical regions and re-emerging in Europe. Despite the great economical losses associated with besnoitiosis, this disease has been underestimated and poorly studied, and neither an effective therapy nor an efficacious vaccine is available. Protein disulfide isomerase (PDI) is an essential enzyme for the acquisition of the correct three-dimensional structure of proteins. Current evidence suggests that in Neosporacaninum and Toxoplasmagondii, which are closely related to B. besnoiti, PDI play an important role in host cell invasion, is a relevant target for the host immune response, and represents a promising drug target and/or vaccine candidate. In this work, we present the nucleotide sequence of the B. besnoiti PDI gene. BbPDI belongs to the thioredoxin-like superfamily (cluster 00388) and is included in the PDI_a family (cluster defined cd02961) and the PDI_a_PDI_a'_c subfamily (cd02995). A 3D theoretical model was built by comparative homology using Swiss-Model server, using as a template the crystallographic deduced model of Tapasin-ERp57 (PDB code 3F8U chain C). Analysis of the phylogenetic tree for PDI within the phylum apicomplexa reinforces the close relationship among B. besnoiti, N. caninum and T. gondii. When subjected to a PDI-assay based on the polymerisation of reduced insulin, recombinant BbPDI expressed in E. coli exhibited enzymatic activity, which was inhibited by bacitracin. Antiserum directed against recombinant BbPDI reacted with PDI in Western blots and by immunofluorescence with B. besnoiti tachyzoites and bradyzoites.

Hookworm aspartic protease, Na-APR-2, cleaves human hemoglobin and serum proteins in a host-specific fashion

Hookworms are voracious blood-feeders. The cloning and functional expression of an aspartic protease, Na-APR-2, from the human hookworm Necator americanus are described here. Na-APR-2 is more similar to a family of nematode-specific, aspartic proteases than it is to cathepsin D or pepsin, and the term nemepsins for members of this family of nematode-specific hydrolases is proposed. Na-apr-2 mRNA was detected in blood-feeding, developmental stages only of N. americanus, and the protease was expressed in the intestinal lumen, amphids, and excretory glands. Recombinant Na-APR-2 cleaved human hemoglobin (Hb) and serum proteins almost twice as efficiently as the orthologous substrates from the nonpermissive dog host. Moreover, only 25% of the Na-APR-2 cleavage sites within human Hb were shared with those generated by the related N. americanus cathepsin D, Na-APR-1. Antiserum against Na-APR-2 inhibited migration of 50% of third-stage N. americanus larvae through skin, which suggests that aspartic proteases might be effective vaccines against human hookworm disease.

An in vitro larval motility assay to determine anthelmintic sensitivity for human hookworm and Strongyloides species

With the implementation of programs to control lymphatic filariasis and soil-transmitted helminths using broad spectrum anthelmintics, including albendazole and ivermectin, there is a need to develop an in vitro assay for detection of drug resistance. This report describes an in vitro assay for measuring the effects of ivermectin and benzimidazoles on the motility of larvae of the hookworm species Ancylostoma ceylanicum, A. caninum, and Necator americanus, and Strongyloides species including Strongyloides stercoralis, and S. ratti. A dose-response relationship was demonstrated with each of the parasite species, with distinct differences observed between the various species. In pilot field testing of the assay with N. americanus larvae recovered from human fecal samples, a dose-response relationship was observed with ivermectin. While the assay has demonstrated the ability to determine drug responsiveness, its usefulness in resistance detection will require correlation with the clinical outcome among individuals infected with parasite strains showing different drug sensitivities.

Rastreio parasitológico de Tritrichomonas foetus em gatos

Dissertação de Mestrado Integrado em Medicina Veterinária