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Proteômica para investigar a congelabilidade do semen bovino | Proteomics to investigate bull’s semen freezability
2013
Marcelo Roncoletta | Erika da Silva Carvalho Morani | Lúcia Helena Rodrigues | Paulo Henrique Francheschini
O objetivo do presente estudo foi avaliar se a composição proteica de plasma seminal, verificada por eletroforese bidimensional (2DE), pode interferir na congelabilidade do sêmen de touros, e se existe a possibilidade de predizer tal característica em touros doadores de sêmen. Amostras de 20 touros (diferentes raças) com mínimo de três anos de histórico de produção em Central de Coleta de sêmen foram coletadas. Todos os touros tinham de 2 a 7 anos de idade. A congelabilidade do sêmen foi calculada por no de ejaculados aprovados pós-descongelação / no de ejaculados submetidos à congelação (depois do crivo do padrão de qualidade da empresa para aprovar o ejaculado para ser submetido a congelação). Os touros foram divididos em três grupos: ALTA (=>80% aproveitamento de ejaculados encaminhados à congelação); MÉDIA (>60 a 79% ejaculados aprovados) e BAIXA (=<59% ejaculados aprovados). Sessenta e oito corridas de géis 2DE foram avaliadas nas amostras de plasma seminal, que detectaram 225 spots com quantidade de proteína diferente entre as mesmas (VION). Pela estatística utilizada, comparando-se a congelabilidade do sêmen com quantidade de proteína detectada de cada spot (VION), constatou-se que 2 spots apresentaram-se com quantidade significativa de proteína diferente entre os dois grupos de congelabilidade do sêmen, considerando spots que foram detectados em mais de 75% das amostras corridas. Os touros taurinos apresentaram perfil proteico mais homogêneo quando comparado com os zebuínos, considerando número e frequência de spots. Tais resultados demonstram um potencial para uso da proteômica como ferramenta preditiva da congelabilidade do sêmen de touros, porém com necessidade de maiores estudos, principalmente para melhor compreensão de interações proteicas entre membrana espermática, plasma seminal e diluidores de sêmen utilizados, já que podem interferir no fenótipo proteico avaliado e na congelabilidade do sêmen, seja de forma positiva ou negativa. | The present study was undertaken the protein composition in 2D-electrophoretic pattern (2DE) of the seminal plasma (SP) can interfere in the semen bull freezability, and if we can use that for predicting semen bull freezability. Samples were obtained of 20 bulls (different breeds) with a minimum of 3 years history semen production in commercial semen collection Center. All animals ranged between 2 - 7 years of age. The semen freezability was calculated by # of thawed and approved ejaculates / # of ejaculates submitted to cryopreservation (after semen evaluation and approved to submitted to freeze). The bulls were divided in 3 groups: HIGH (=>80% ejaculates approved); MEDIUM (>60% and <79% ejaculates approved); LOW (=<59% ejaculates approved); the pattern and criteria were the same used in the routine of the commercial semen Center. 68 gels were carried through by 2DE of SP samples indicated 225 detected spots with protein different amount (VION) comparing. Comparing bull´s semen freezability and VION of each spot found difference among 2 spots from High and Low, even considering just spots with % of detection frequency bigger than 75%. The taurine bulls demonstrated more homogeneous profile when comparing with zebu bulls, considering number and frequency of appearance of spots. The results showed that proteomics can be a useful tool to predict the semen freezability, but we´ll need to study better the interactions between sperm membrane, seminal plasma and extender to comprehend better which proteome phenotype interfere positive or negatively in the semen freezability.
Show more [+] Less [-]Toxoplasma gondii in small neotropical wild felids | Toxoplasma gondii em pequenos felinos silvestres neotropicais
2013
William Alberto Cañon-Franco | Flávio Antônio Pacheco de Araújo | Solange Maria Gennari
In the last decade, studies on wildlife worldwide have discovered key epidemiological aspects of the sylvatic cycle of Toxoplasma gondii. However, despite the known role of wild felines as definitive hosts in the transmission and maintenance of this parasite, few studies have focused on the involvement of these animals. Brazil exhibits the largest number of wild felid species in the Americas, all of which have a critical conservation status. However, serological detections, epidemiological studies and some molecular characterizations of T. gondii have primarily used Neotropical felid populations that are maintained in captivity, which does not reflect the disease behavior in free-living conditions. A systematic review of the worldwide scientific literature was conducted focusing on toxoplasmosis in small Neotropical felids. This review covered a number of aspects, including the state of scientific research, parasite transmission in the wild, the genetic characteristics of isolates, the relationship between these genetic characteristics and the pathogenicity of the parasite, and the risk factors linked to conflicts with humans. The present review shows the relevance of studying these felid populations based on their frequent interactions with humans in peri-urban areas and the need for further comprehensive studies to establish the real significance of T. gondii in public and animal health in tropical and temperate regions. | Na ultima década, pesquisas em animais silvestres no mundo todo, tem demonstrado aspectos importantes na epidemiologia do ciclo silvestre de Toxoplasma gondii. No entanto, apesar do papel conhecido dos felinos silvestres como hospedeiros definitivos na transmissão e manutenção desse parasita, há poucos estudos sobre o envolvimento desses animais. O Brasil possui a maior quantidade de espécies de felinos silvestres do continente americano, todas em estado crítico de conservação. Porém, a detecção sorológica, pesquisas epidemiológicas e algumas caracterizações moleculares do T. gondii nestas espécies, utilizaram principalmente populações de felinos neotropicais mantidos em cativeiro, o que não reflete o comportamento da doença em condições de vida livre.Uma revisão sistemática da literatura cientifica mundial foi realizada focando a toxoplasmose em pequenos felinos neotropicais, abrangendo aspectos como o estado da pesquisa cientifica, transmissão do parasita na vida silvestre, características genéticas dos isolados e sua relação com a patogenicidade, além dos fatores de risco ligados aos conflitos com o homem. Esta revisão mostra a importância do estudo dessas populações de felinos, em função das frequentes interações com o homem em áreas peri-urbanas e a necessidade de estudos mais abrangentes, que estabeleçam a real importância do T. gondii no tangente á saúde pública e saúde animal nas regiões tropicais e temperadas.
Show more [+] Less [-]Presença de Toxocara spp. e ancilostomídeos em áreas de peridomicílios de uma comunidade pesqueira no Centro-Oeste do Brasil | Presence of Toxocara spp. and hookworms in peridomiciliary areas of a fishing community in the Mid-west region of Brazil
2013
Andreia Fernandes Brilhante | Vânia Lúcia Brandão Nunes | Maria Elizabeth Cavalheiros Dorval
O objetivo do estudo foi verificar a contaminação do solo de peridomicílios de uma comunidade pesqueira localizada no município de Bonito, Mato Grosso do Sul, Brasil, onde é frequente a ocorrência de casos de Larva migrans cutânea. Utilizando-se das técnicas de centrifugo-flutuação, Baermann-Moraes e centrifugo- sedimentação, amostras de solo e fezes de cães foram analisadas e os resultados evidenciaram altos índices de contaminação por ovos de Toxocara spp., Ancilostomídeos e larvas de nematoides. Estes achados podem estar associados aos casos de Larvas migrans cutânea e às elevadas taxas de eosinofilia em escolares daquela comunidade. | The aim of this study was to investigate soil contamination in peridomiciliary areas from a fishing community in the municipality of Bonito, Mato Grosso do Sul state, Brazil, where is often found the occurrence of cases of cutaneous larva migrans. Using the techniques of zinc sulphate centrifugal flotation, Baermann-Moraes and centrifugal sedimentation, soil samples and feces of dogs were analyzed and the results showed high levels of contamination by Toxocara spp., hookworm and larvae of nematodes. These findings may be associated with cases of cutaneous larva migrans and high rates of eosinophilia in school children of that community.
Show more [+] Less [-]Development of the omasum of Nelore cattle (Bos taurus indicus) during the prenatal period | Desenvolvimento do omaso de bovinos da raça Nelore (Bos taurus indicus) durante o período pré-natal
2013
Gabriela Mansano do Nascimento | Silvia Helena Venturoli Perri | Roberto Gameiro de Carvalho | Cristina Maria Rodrigues Monteiro
<!--[if gte mso 9]><xml> <o:OfficeDocumentSettings> <o:AllowPNG/> </o:OfficeDocumentSettings> </xml><![endif]--><p class="MsoNormal"><span style="mso-ansi-language: EN-US;" lang="EN-US">The development of the stomach of domestic mammals is a complex process, especially in ruminants. The aim of this study was to analyze this organ during prenatal period. Omasum samples of Nelore fetuses (Bos taurus indicus) were divided into five groups of six samples each: 1- fetuses from 9 to 15 weeks (8 to 21cm) of gestation, 2 - fetuses from 16 to 22 weeks (23 to37cm), 3 - fetuses from 23 to 29 weeks (40 to 58cm), 4 - fetuses from 30 to 36 weeks (61 to 77cm) and 5 - fetuses from 37 to43 weeks (79 to 88cm). The histological sections were stained with Hematoxyl in and Eosin and Picrossirius and examined by light microscopy. In fetuses of 11 cm it was observed mucosa with uniform primary, secondary and tertiary blades and all layers present. The fetus with 13.5 cm presented quaternary blades emergence, papillae on primary and secondary blades and formation of the muscularis mucosae. At 28 cm it were observed papillae in development, as well as the blades, especially the quaternary. In 42 cm fetuses was early epithelial keratinization and at 56 cm it was fully keratinized. In 60 cm fetuses the blades presented large papillae with tapered apex and at 65 cm there was decreased epithelium and increase of the muscle layer. In the fetus of 83 cm it were observed thin sections, discrete muscularis mucosa and papillae over the entire organ surface. It was concluded that the blades and papillae were structures that presented more changes during fetal development.</span></p> | <!--[if gte mso 9]><xml> <o:OfficeDocumentSettings> <o:AllowPNG/> </o:OfficeDocumentSettings> </xml><![endif]--><p class="MsoNormal">O desenvolvimento do estômago de mamíferos domésticos é um processo complexo, especialmente em ruminantes. Essa pesquisa analisou o desenvolvimento desse órgão durante o período pré-natal. Foram utilizadas amostras de omaso de fetos da raça Nelore (Bos taurus indicus) divididos em cinco grupos com seis amostras cada: 1º – fetos com 9 a 15 semanas (8 a 21cm) de gestação, 2º – fetos com 16 a 22 semanas (23 a 37cm), 3º – fetos com 23 a 29 semanas (40 a 58cm), 4º – fetos com 30 a 36 semanas (61 a 77cm) e 5º– fetos com 37 a 43 semanas (79 a 88cm). Os cortes histológicos foram corados com Hematoxilina e Eosina e Picrossirius e examinados ao microscópio de luz. No feto de 11 cm, foi observado mucosa com lâminas primárias, secundárias e terciárias uniformes e demais camadas presentes. No feto de 13,5 cm surgiram as lâminas quaternárias, papilas nas lâminas primárias e secundárias e a formação da muscular da mucosa. Aos 28 cm, as papilas estavam em pleno desenvolvimento, assim como as lâminas, principalmente as quaternárias. No feto de 42 cm houve início de queratinização epitelial e, no de 56 cm, estava totalmente queratinizado. No feto de 60 cm, as lâminas possuíam papilas grandes e afiladas no ápice e, aos 65 cm, houve diminuição do epitélio e aumento da camada muscular. O feto de 83 cm apresentou lâminas delgadas, muscular da mucosa discreta e papilas por toda a superfície. Concluiu-se que as lâminas e papilas foram as estruturas que mais apresentaram modificações durante o desenvolvimento fetal.</p>
Show more [+] Less [-]Clinical and epidemiological characterization of leishmaniasis in dogs in São Paulo State | Caracterização clínica e epidemiológica das leishmanioses em cães no Estado de São Paulo
2013
Cauê Pereira Toscano | Claudio Nazaretian Rossi | Vítor Márcio Ribeiro | Marcia Dalastra Laurenti | Carlos Eduardo Larsson
<!--[if gte mso 9]><xml> <o:OfficeDocumentSettings> <o:AllowPNG/> </o:OfficeDocumentSettings> </xml><![endif]--><p class="MsoNormal"><span style="mso-ansi-language: EN-US;" lang="EN-US">The occurrence of visceral or cutaneous canine leishmaniasis cases among dermatophatic dogs admitted to the veterinary teaching hospital School of Veterinary Medicine and Animal Husbandry - University of São Paulo from 1997 to 2008 was retrospectively characterized. For the diagnosis of 427 animals, serological (Enzyme-Linked Immunosorbent Assay, Indirect Immunofluorescence and/or Complement Fixation), parasitological and molecular (Polymerase Chain Reaction) tests were performed, which resulted in 117 (27.4%) confirmed positive cases. Of these, 96 (82.1%) were pure breed dogs and the others 21 (17.9%) were cross breed, being 57 (48.7%) females and 60 (51.3%) males. In terms of age, 26 (22.2%) were younger than 36 months, 48 (41%) were between 36 and 84 months, and 43 (36.8%) were older than seven years old. Sixty-four (54.7%) dogs were considered oligosymptomatic, 44 (37.6%) symptomatic, and the remaining nine (7.7%) asymptomatic. The main clinical findings were, in descending order, dermal lesions, lymphadenopathy, anemia, dysorexia, prostration and splenomegaly. As for hematological disorders, relative and absolute lymphopenia was the most important findings. Ninety-six (82.1%) of the positive animals correspond to allochthonus cases from São Paulo municipality.</span></p> | <!--[if gte mso 9]><xml> <o:OfficeDocumentSettings> <o:AllowPNG/> </o:OfficeDocumentSettings> </xml><![endif]--><p class="MsoNormal">Caracterizou-se retrospectivamente, a partir do ano de 1997 até março de 2008, a ocorrência de casos de leishmaniose, visceral ou tegumentar, dentre os cães dermatopatas atendidos em serviço especializado de hospital-escola veterinário de Faculdade de Medicina Veterinária e Zootecnia da Universidade de São Paulo. Para o diagnóstico em 427 animais, foram utilizados testes sorológicos (Ensaio Imunoenzimático Indireto, Imunofluorescência Indireta e/ou Fixação de Complemento), parasitológicos diretos e moleculares (Reação em Cadeia da Polimerase), que resultaram na confirmação de 117 (27,4%) casos positivos. Da amostragem, 96 (82,1%) eram cães de raça definida, os demais 21 (17,9%) não dispunham de plena definição racial, sendo 57 (48,7%) fêmeas e 60 (51,3%) machos. Quanto à faixa etária, 26 (22,2%) apresentavam idade inferior a 36 meses, 48 (41%) entre 36 e 84 meses e 43 (36,8%) ultrapassavam o sétimo ano de vida. Considerou-se 64 (54,7%) cães como oligossintomáticos, 44 (37,6%) sintomáticos e os nove (7,7%) remanescentes como assintomáticos. As principais alterações clínicas observadas foram, em ordem decrescente, lesões tegumentares, linfonodomegalia, anemia, disorexia, prostração e esplenomegalia. Quanto às alterações hematológicas, a mais importante foi a linfopenia relativa e absoluta. Dos animais positivos para a enfermidade, 96 (82,1%) correspondeu a casuística alóctone ao Município de São Paulo.</p><!--[if gte mso 9]><xml> <w:WordDocument> <w:View>Normal</w:View> <w:Zoom>0</w:Zoom> <w:TrackMoves/> <w:TrackFormatting/> <w:HyphenationZone>21</w:HyphenationZone> <w:PunctuationKerning/> <w:ValidateAgainstSchemas/> <w:SaveIfXMLInvalid>false</w:SaveIfXMLInvalid> <w:IgnoreMixedContent>false</w:IgnoreMixedContent> <w:AlwaysShowPlaceholderText>false</w:AlwaysShowPlaceholderText> <w:DoNotPromoteQF/> <w:LidThemeOther>PT-BR</w:LidThemeOther> <w:LidThemeAsian>X-NONE</w:LidThemeAsian> <w:LidThemeComplexScript>X-NONE</w:LidThemeComplexScript> <w:Compatibility> <w:BreakWrappedTables/> <w:SnapToGridInCell/> <w:WrapTextWithPunct/> <w:UseAsianBreakRules/> <w:DontGrowAutofit/> <w:SplitPgBreakAndParaMark/> <w:EnableOpenTypeKerning/> <w:DontFlipMirrorIndents/> <w:OverrideTableStyleHps/> </w:Compatibility> <m:mathPr> <m:mathFont m:val="Cambria Math"/> <m:brkBin m:val="before"/> 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Show more [+] Less [-]Molecular epidemiology of Listeria monocytogenes isolated from different sources in Brazil | Epidemiologia molecular de Listeria monocytogenes isoladas de diferentes fontes no Brasil
2013
Andrea Micke Moreno | Renata Paixão | Luisa Zanolli Moreno | Débora Dirani Sena de Gobbi | Daniele Cristine Raimundo | Thais Sebastiana Porfida Ferreira | Ernesto Hofer | Maria Helena Matte | Marina Moreno
<!--[if gte mso 9]><xml> <o:OfficeDocumentSettings> <o:AllowPNG/> </o:OfficeDocumentSettings> </xml><![endif]--><p class="MsoNormal"><span style="mso-ansi-language: EN-US;" lang="EN-US">Listeria monocytogenes is an important foodborne pathogen that primarily affects pregnant women, neonates, the elderly and immune-compromised individuals, and it may cause abortion, septicemia, and meningitis. From the 13 capsular groups described, serotypes 4b, 1/2b and 1/2a are most closely related to human infection. For this reason, serotyping has limited value as an epidemiological tool; thus, improved discriminatory typing methods are required to enhance knowledge of L. monocytogenes contamination and infection. The aim of this study was to characterize the genetic diversity of L. monocytogenes isolates in the pork processing industry in Sao Paulo, Brazil and human infection isolates by ERICPCR and single enzyme AFLP. Serotypes 1/2c and 4b were frequent among isolates from pork and slaughterhouse/market environments, whereas serotypes 4b and 1/2a were observed among human isolates. ERIC-PCR and AFLP revealed 34 and 31 distinct profiles, respectively, which had tendencies of separation according to serogroup and isolate origin. The genetic profiles from slaughterhouse and market environments suggest the possibility of different sources of Listeria contamination in the environment, although in certain cases, continuous contamination caused by the persistence of clonal strains is also a possibility.</span></p><!--[if gte mso 9]><xml> <w:WordDocument> <w:View>Normal</w:View> <w:Zoom>0</w:Zoom> <w:TrackMoves/> <w:TrackFormatting/> <w:HyphenationZone>21</w:HyphenationZone> <w:PunctuationKerning/> <w:ValidateAgainstSchemas/> <w:SaveIfXMLInvalid>false</w:SaveIfXMLInvalid> <w:IgnoreMixedContent>false</w:IgnoreMixedContent> <w:AlwaysShowPlaceholderText>false</w:AlwaysShowPlaceholderText> <w:DoNotPromoteQF/> <w:LidThemeOther>PT-BR</w:LidThemeOther> <w:LidThemeAsian>X-NONE</w:LidThemeAsian> <w:LidThemeComplexScript>X-NONE</w:LidThemeComplexScript> <w:Compatibility> <w:BreakWrappedTables/> <w:SnapToGridInCell/> <w:WrapTextWithPunct/> <w:UseAsianBreakRules/> <w:DontGrowAutofit/> <w:SplitPgBreakAndParaMark/> 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mso-style-priority:99; mso-style-parent:""; mso-padding-alt:0cm 5.4pt 0cm 5.4pt; mso-para-margin-top:0cm; mso-para-margin-right:0cm; mso-para-margin-bottom:10.0pt; mso-para-margin-left:0cm; line-height:115%; mso-pagination:widow-orphan; font-size:11.0pt; font-family:"Calibri","sans-serif"; mso-ascii-font-family:Calibri; mso-ascii-theme-font:minor-latin; mso-hansi-font-family:Calibri; mso-hansi-theme-font:minor-latin; mso-fareast-language:EN-US;} </style> <![endif]--> | <!--[if gte mso 9]><xml> <o:OfficeDocumentSettings> <o:AllowPNG/> </o:OfficeDocumentSettings> </xml><![endif]--><p class="MsoNormal">Listeria monocytogenes é um importante patógeno de origem alimentar que afeta principalmente grávidas, neonatos, idosos e indivíduos imunocomprometidos, e pode causar abortamento, septicemia e meningite. Dos 13 grupos capsulares descritos, os sorotipos 4b, 1/2b e 1/2a são os mais relacionados à infecção humana. Por esta razão, a sorotipagem possui valor limitado como ferramenta epidemiológica e, dessa forma, métodos mais discriminatórios são necessários para melhorar o conhecimento sobre a contaminação e a infecção por L. monocytogenes. O objetivo deste estudo foi caracterizar a diversidade genética de isolados de L. monocytogenes da indústria de processamento de carne suína no Estado de São Paulo, Brasil, e compará-los a isolados de casos de infecção humana através do ERIC-PCR e AFLP com uma única enzima. Os sorotipos 1/2c e 4b foram frequentes em carne suína e ambientes de abatedouros e mercados, enquanto os sorotipos 4b e 1/2a foram observados nos isolados de humanos. ERIC-PCR e AFLP resultaram em 34 e 31 perfis distintos, respectivamente, com uma tendência a separar de acordo com o sorogrupo e a origem do isolado. Os perfis genéticos de ambiente dos abatedouros e mercados sugerem a possibilidade de diferentes origens de contaminação por Listeria nos ambientes estudados, porém, em alguns casos, é possível que ocorra a persistência de cepas clonais causando contaminação contínua.</p><!--[if gte mso 9]><xml> <w:WordDocument> <w:View>Normal</w:View> <w:Zoom>0</w:Zoom> <w:TrackMoves/> <w:TrackFormatting/> <w:HyphenationZone>21</w:HyphenationZone> <w:PunctuationKerning/> <w:ValidateAgainstSchemas/> <w:SaveIfXMLInvalid>false</w:SaveIfXMLInvalid> <w:IgnoreMixedContent>false</w:IgnoreMixedContent> <w:AlwaysShowPlaceholderText>false</w:AlwaysShowPlaceholderText> <w:DoNotPromoteQF/> <w:LidThemeOther>PT-BR</w:LidThemeOther> <w:LidThemeAsian>X-NONE</w:LidThemeAsian> <w:LidThemeComplexScript>X-NONE</w:LidThemeComplexScript> <w:Compatibility> <w:BreakWrappedTables/> <w:SnapToGridInCell/> <w:WrapTextWithPunct/> <w:UseAsianBreakRules/> <w:DontGrowAutofit/> <w:SplitPgBreakAndParaMark/> <w:EnableOpenTypeKerning/> 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