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Santos P.S.,Federal University of Uberlandia | Nascimento R.,Federal University of Uberlandia | Rodrigues L.P.,Federal University of Uberlandia | Santos F.A.A.,Federal University of Uberlandia | And 5 more authors.
PLoS ONE | Year: 2012

Anaplasmosis, a persistent intraerythrocytic infection of cattle by Anaplasma marginale, causes severe anemia and a higher rate of abortion, resulting in significant loss to both dairy and beef industries. Clinical diagnosis is based on symptoms and confirmatory laboratory tests are required. Currently, all the diagnostic assays have been developed with whole antigens with indirect ELISA based on multiple epitopes. In a pioneer investigation we demonstrated the use of critical motifs of an epitope as biomarkers for immunosensor applications. Mimotopes of the MSP1a protein functional epitope were obtained through Phage Display after three cycles of selection of a 12-mer random peptide library against the neutralizing monoclonal antibody 15D2. Thirty-nine clones were randomly selected, sequenced, translated and aligned with the native sequence. The consensus sequence SxSSQSEASTSSQLGA was obtained, which is located in C-terminal end of the 28-aa repetitive motif of the MSP1a protein, but the alignment and sequences' variation among mimotopes allowed us to map the critical motif STSSxL within the consensus sequence. Based on these results, two peptides were chemically synthesized: one based on the critical motif (STSSQL, Am1) and the other based on the consensus sequence aligned with the native epitope (SEASTSSQLGA, Am2). Sera from 24 infected and 52 healthy animals were tested by ELISA for reactivity against Am1 and Am2, which presented sensitivities of 96% and 100%, respectively. The Am1 peptide was incorporated onto a biolectrode (graphite modified with poly-3-hydroxyphenylacetic acid) and direct serum detection was demonstrated by impedance, differential pulse voltammetry, and atomic force microscopy. The electrochemical sensor system proved to be highly effective in discriminating sera from positive and negative animals. These immunosensors were highly sensitive and selective for positive IgG, contaminants did not affect measurements, and were based on a simple, fast and reproducible electrochemical system. © 2012 Santos et al. Source

Minharro S.,Federal University of Minas Gerais | Minharro S.,Federal University of Tocantins | Silva Mol J.P.,Federal University of Minas Gerais | Dorneles E.M.S.,Federal University of Minas Gerais | And 10 more authors.
PLoS ONE | Year: 2013

Brucellosis is a worldwide distributed zoonosis that causes important economic losses to animal production. In Brazil, information on the distribution of biovars and genotypes of Brucella spp. is scarce or unavailable. This study aimed (i) to biotype and genotype 137 Brazilian cattle isolates (from 1977 to 2008) of B. abortus and (ii) to analyze their distribution. B. abortus biovars 1, 2 and 3 (subgroup 3b) were confirmed and biovars 4 and 6 were first described in Brazil. Genotyping by the panel 1 revealed two groups, one clustering around genotype 40 and another around genotype 28. Panels 2A and 2B disclosed a high diversity among Brazilian B. abortus strains. Eighty-nine genotypes were found by MLVA16. MLVA16 panel 1 and 2 showed geographic clustering of some genotypes. Biotyping and MLVA16 genotyping of Brazilian B. abortus isolates were useful to better understand the epidemiology of bovine brucellosis in the region. © 2013 Minharro et al. Source

Santos T.R.B.,Federal University of Pelotas | Klafke G.M.,Institute Pesquisas Veterinarias Desiderio Finamor | Pappen F.G.,Instituto Federal Catarinense IFC | Nizoli L.Q.,Federal University of Pelotas | And 2 more authors.
Revista Brasileira de Parasitologia Veterinaria | Year: 2013

Field samples of Rhipicephalus (Boophilus) microplus from the state of Rio Grande do Sul, Brazil, were assessed using the following methods: larval packet test (LPT), larval immersion test (LIT) and syringe immersion test (SIT). The following parameters were determined for each population and for the Mozo susceptible reference strain: lethal concentration for 50% (LC50) with its 95% confidence interval (95% CI), regression line slope and resistance ratio (RR). Using the LPT, only one population was susceptible to amitraz, presenting a RR of 1.9. Using the same technique, the other populations presented RRs of between 92.9 and 3445.8 and were considered resistant. The LC50 of the Mozo strain calculated using the LPT, LIT and SIT was 2.9, 27.3, and 52.7 μg/mL, respectively. In general, a good fit to the probit statistical model was only achieved using the LPT. The results obtained in this study impair recommendations for using the LIT and SIT to diagnose amitraz resistance in R. (B.) microplus populations. Additional studies are required to improve the sensitivity of these tests in relation to the LPT. Source

Grisi L.,Federal Rural University of Rio de Janeiro | Leite R.C.,Federal University of Minas Gerais | Martins J.R.S.,Institute Pesquisas Veterinarias Desiderio Finamor | de Barros A.T.M.,Embrapa Gado de Corte | And 5 more authors.
Revista Brasileira de Parasitologia Veterinaria | Year: 2014

The profitability of livestock activities can be diminished significantly by the effects of parasites. Economic losses caused by cattle parasites in Brazil were estimated on an annual basis, considering the total number of animals at risk and the potential detrimental effects of parasitism on cattle productivity. Estimates in U.S. dollars (USD) were based on reported yield losses among untreated animals and reflected some of the effects of parasitic diseases. Relevant parasites that affect cattle productivity in Brazil, and their economic impact in USD billions include: gastrointestinal nematodes - $7.11; cattle tick (Rhipicephalus (Boophilus) microplus) - $3.24; horn fly (Haematobia irritans) - $2.56; cattle grub (Dermatobia hominis) - $0.38; New World screwworm fly (Cochliomyia hominivorax) - $0.34; and stable fly (Stomoxys calcitrans) - $0.34. The combined annual economic loss due to internal and external parasites of cattle in Brazil considered here was estimated to be at least USD 13.96 billion. These findings are discussed in the context of methodologies and research that are required in order to improve the accuracy of these economic impact assessments. This information needs to be taken into consideration when developing sustainable policies for mitigating the impact of parasitism on the profitability of Brazilian cattle producers. Source

Cavalli L.S.,Institute Pesquisas Veterinarias Desiderio Finamor | Batista C.R.,Grande Rio University | Nornberg B.F.S.,Grande Rio University | Mayer F.Q.,Institute Pesquisas Veterinarias Desiderio Finamor | And 4 more authors.
Journal of Invertebrate Pathology | Year: 2013

White spot syndrome virus (WSSV) and Infectious hypodermal and hematopoietic necrosis virus (IHHNV) are two infectious agents associated to economic losses in shrimp aquaculture. As virus spread occurs through vectors and hosts, this study sought to verify the presence of WSSV and IHHNV in Neohelice granulata crab from Lagoa dos Patos estuary in Brazil and nearby shrimp farms. DNA extractions were performed with phenol/chloroform protocol. Molecular diagnosis was carried out by nested PCR for WSSV and one-step PCR for IHHNV. Results showed the presence of WSSV on crabs of both Lagoa dos Patos and farms, while IHHNV was found only on crabs collected in estuary. This is the first study to report IHHNV presence in N. granulata. Moreover, as analyzed crabs had no clinical symptoms or showed in situ mortality, we suggest its use as a bioindicator for virus occurrence in aquatic environments. © 2013 Elsevier Inc. Source

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