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Teste de ligação espermática à membrana perivitelínica da gema de ovo de galinha como avaliação funcional do sêmen de cães | Sperm-binding to the perivitelline membrane of chicken egg yolk as a functional test for sperm evaluation in dogs Full text
2017
Brito, Maíra Morales | Losano, João Diego Agostini | Angrimani, Daniel de Souza Ramos | Lúcio, Cristina de Fátima | Dalmazzo, Andressa | Nichi, Marcílio | Vannucchi, Camila Infantosi
Teste de ligação espermática à membrana perivitelínica da gema de ovo de galinha como avaliação funcional do sêmen de cães | Sperm-binding to the perivitelline membrane of chicken egg yolk as a functional test for sperm evaluation in dogs Full text
2017
Brito, Maíra Morales | Losano, João Diego Agostini | Angrimani, Daniel de Souza Ramos | Lúcio, Cristina de Fátima | Dalmazzo, Andressa | Nichi, Marcílio | Vannucchi, Camila Infantosi
Durante a fecundação, os espermatozoides interagem com a zona pelúcida (ZP) por meio da ligação entre o acrossomo e as proteínas 2 e 3 (ZP2 e ZP3). A membrana perivitelínica da gema de ovo de galinhas é homóloga à ZP3 de mamíferos, possibilitando a ligação espermática de diversas espécies. Este trabalho padronizou e avaliou a eficiência do teste de ligação espermática à membrana perivitelínica da gema de ovo de galinhas como avaliação funcional do sêmen de cães. Para tal, foram utilizadas nove amostras seminais previamente criopreservadas. Cada amostra foi dividida em duas alíquotas: a primeira foi mantida em banho-maria à 37ºC (vivos) e a segunda submetida a choque térmico com o intuito de induzir dano celular (mortos). As duas alíquotas foram misturadas, correspondendo a 0, 25, 50, 75 e 100% de células viáveis. As amostras foram avaliadas quanto ao número de espermatozoides ligados à membrana perivitelínica por meio da análise computadorizada da motilidade (CASA) ou microscopia convencional. Ademais, as amostras foram avaliadas quanto à motilidade espermática, integridade das membranas acrossomal e plasmática e atividade mitocondrial espermática. O teste de ligação espermática à membrana perivitelínica de ovos de galinha foi considerado um teste de análise seminal exequível tanto para avaliar a capacidade fecundante dos espermatozoides como atributos seminais gerais, especialmente quando realizado em microscopia convencional, expandindo sua praticidade para a maioria dos laboratórios de análise de sêmen canino. | During fertilization, spermatozoa interact with the zona pellucida (ZP) through the binding between the acrosome and proteins 2 and 3 (ZP2 and ZP3). The perivitelline membrane of chicken egg yolk is homologous to the mammalian ZP3, which allows the binding of sperm of several species. The aim of this study was to standardize and evaluate the efficiency of sperm-binding to the perivitelline membrane of chicken eggs as a functional method for canine semen evaluation. For this purpose, nine post-thaw sperm samples were used, which were divided into two aliquots: the first was kept in water bath at 37ºC (live sample) and the second was submitted to cold shock to induce cellular damage (dead sample). The two aliquots were mixed on five proportions, corresponding to 0%, 25%, 50%, 75%, and 100% of viable cells, and the binding test was performed by analyzing the number of spermatozoa bonded to the perivitelline membrane by means of computerized assessment of sperm motility (CASA) or conventional microscopy. Additionally, samples were submitted to sperm motility analysis, evaluation of plasmatic and acrosomal membrane integrity, and sperm mitochondrial activity. The sperm-binding test to the perivitelline membrane of chicken egg yolk was considered a feasible sperm analysis test for both fertilizing capacity and overall sperm attributes evaluation, mainly when the analysis is performed by a conventional microscope, which expands its practicality to the majority of canine reproduction laboratories.
Show more [+] Less [-]Sperm-binding to the perivitelline membrane of chicken egg yolk as a functional test for sperm evaluation in dogs Full text
2017
Maíra Morales Brito | João Diego Agostini Losano | Daniel de Souza Ramos Angrimani | Cristina de Fátima Lúcio | Andressa Dalmazzo | Marcílio Nichi | Camila Infantosi Vannucchi
During fertilization, spermatozoa interact with the zona pellucida (ZP) through the binding between the acrosome and proteins 2 and 3 (ZP2 and ZP3). The perivitelline membrane of chicken egg yolk is homologous to the mammalian ZP3, which allows the binding of sperm of several species. The aim of this study was to standardize and evaluate the efficiency of sperm-binding to the perivitelline membrane of chicken eggs as a functional method for canine semen evaluation. For this purpose, nine post-thaw sperm samples were used, which were divided into two aliquots: the first was kept in water bath at 37ºC (live sample) and the second was submitted to cold shock to induce cellular damage (dead sample). The two aliquots were mixed on five proportions, corresponding to 0%, 25%, 50%, 75%, and 100% of viable cells, and the binding test was performed by analyzing the number of spermatozoa bonded to the perivitelline membrane by means of computerized assessment of sperm motility (CASA) or conventional microscopy. Additionally, samples were submitted to sperm motility analysis, evaluation of plasmatic and acrosomal membrane integrity, and sperm mitochondrial activity. The sperm-binding test to the perivitelline membrane of chicken egg yolk was considered a feasible sperm analysis test for both fertilizing capacity and overall sperm attributes evaluation, mainly when the analysis is performed by a conventional microscope, which expands its practicality to the majority of canine reproduction laboratories.
Show more [+] Less [-]Rural area of the Brazilian Pantanal wetlands associated with the occurrence of anti-Leishmania spp. antibodies in dogs | Área rural do Pantanal brasileiro associada à ocorrência de anticorpos anti-Leishmania spp. em cães Full text
2017
Melo, Andréia Lima Tomé | Costa, Andréa Pereira da | Miyazaki, Selma Samiko | Stocco, Matias Bassinello | Semedo, Thiago Borges Fernandes | Pacheco, Thábata dos Anjos | Witter, Rute | Pacheco, Richard de Campos | Labruna, Marcelo Bahia | Marcili, Arlei | Aguiar, Daniel Moura de
Rural area of the Brazilian Pantanal wetlands associated with the occurrence of anti-Leishmania spp. antibodies in dogs | Área rural do Pantanal brasileiro associada à ocorrência de anticorpos anti-Leishmania spp. em cães Full text
2017
Melo, Andréia Lima Tomé | Costa, Andréa Pereira da | Miyazaki, Selma Samiko | Stocco, Matias Bassinello | Semedo, Thiago Borges Fernandes | Pacheco, Thábata dos Anjos | Witter, Rute | Pacheco, Richard de Campos | Labruna, Marcelo Bahia | Marcili, Arlei | Aguiar, Daniel Moura de
Neste trabalho foi realizada uma avaliação sobre a presença de anticorpos anti-Leishmania infantum chagasi em cães domésticos das áreas urbanas e rurais da região do Pantanal brasileiro usando técnicas sorológicas. Um total de 429 cães foram amostrados em três áreas do bioma do Pantanal, incluindo os municípios de Poconé, Santo Antônio de Leverger e Barão de Melgaço, em Mato Grosso, e o município de Corumbá, em Mato Grosso do Sul. A reação de imunofluorescência indireta (RIFI) foi utilizada para detectar anticorpos (ponto de corte de 40) de Leishmania infantum chagasi como antígeno. Devido à possibilidade de reação cruzada entre as espécies do gênero Leishmania, as amostras positivas na RIFI para L. infantum chagasi foram também avaliadas na RIFI utilizando L. amazonensis e L. braziliensis como antígenos. As amostras positivas na RIFI para L. infantum chagasi foram avaliadas utilizando o ensaio de imunoadsorção ligado à enzima (ELISA). Os resultados mostraram a presença de anticorpos contra L. infantum chagasi em 23 (5,36%; IC 95%: 3,50% -8,05%) cães e pelo menos um cão soro-reativo foi encontrado em todos os municípios avaliados neste estudo. Os títulos de anticorpos variaram de 40 a 5.120 e todas as amostras positivas na RIFI foram positivas no ELISA. Entre os 23 cães positivos, nove também reagiram para L. amazonensis e L. braziliensis. A ocorrência de anticorpos anti-L. infantum chagasi em cães foi maior nas áreas rurais (7,06%) do que nas áreas urbanas (2,50%) (P < 0,05). Com base neste estudo, concluímos que cães de áreas rurais do Pantanal tiveram contato com espécies de Leishmania, o que é uma informação relevante, dada a sua importância para a saúde pública. | An evaluation was made of the presence of anti-Leishmania infantum chagasi antibodies in domestic dogs from the urban and rural areas of Brazil’s Pantanal wetland region using serological techniques. A total of 429 dogs were sampled in three areas of the Pantanal biome, including the municipalities of Poconé, Santo Antônio de Leverger, and Barão de Melgaço, in the state of Mato Grosso, and in the municipality of Corumbá, in the state of Mato Grosso do Sul. The immunofluorescence assay (IFA) was used to detect antibodies (cut-off point 40) using Leishmania infantum chagasi antigen. Because of the possibility of cross-reactivity between species of the genus Leishmania, samples that were positive in the IFA against L. infantum chagasi were also tested by IFA in the same conditions, using L. amazonensis and L. braziliensis antigens. IFA-positive samples to L. infantum chagasi were also evaluated using Enzyme-Linked Immunosorbent Assay (ELISA). The results showed the presence of antibodies against L. infantum chagasi in 23 (5.36%; 95% CI: 3.50%-8.05%) dogs and at least one seroreactive dog was found in each of the municipalities evaluated in this study. Antibody titers ranged from 40 to 5,120, and all IFA positive samples were positive in the ELISA. Among the 23 positive dogs, nine were also were seroreactive for L. amazonensis and L. braziliensis antigens. The occurrence of anti- L. infantum chagasi antibodies in dogs was higher in rural areas (7.06%) than in urban areas (2.50%) (P < 0.05). Based on this study, we concluded that dogs from rural areas of the Pantanal wetlands were in contact with Leishmania species, which is relevant information given their importance to public health.
Show more [+] Less [-]Rural area of the Brazilian Pantanal wetlands associated with the occurrence of anti-<i>Leishmania</i> spp. antibodies in dogs Full text
2017
Andréia Lima Tomé Melo | Andréa Pereira da Costa | Selma Samiko Miyazaki | Matias Bassinello Stocco | Thiago Borges Fernandes Semedo | Thábata dos Anjos Pacheco | Rute Witter | Richard de Campos Pacheco | Marcelo Bahia Labruna | Arlei Marcili | Daniel Moura de Aguiar
An evaluation was made of the presence of anti-Leishmania infantum chagasi antibodies in domestic dogs from the urban and rural areas of Brazil’s Pantanal wetland region using serological techniques. A total of 429 dogs were sampled in three areas of the Pantanal biome, including the municipalities of Poconé, Santo Antônio de Leverger, and Barão de Melgaço, in the state of Mato Grosso, and in the municipality of Corumbá, in the state of Mato Grosso do Sul. The immunofluorescence assay (IFA) was used to detect antibodies (cut-off point 40) using Leishmania infantum chagasi antigen. Because of the possibility of cross-reactivity between species of the genus Leishmania, samples that were positive in the IFA against L. infantum chagasi were also tested by IFA in the same conditions, using L. amazonensis and L. braziliensis antigens. IFA-positive samples to L. infantum chagasi were also evaluated using Enzyme-Linked Immunosorbent Assay (ELISA). The results showed the presence of antibodies against L. infantum chagasi in 23 (5.36%; 95% CI: 3.50%-8.05%) dogs and at least one seroreactive dog was found in each of the municipalities evaluated in this study. Antibody titers ranged from 40 to 5,120, and all IFA positive samples were positive in the ELISA. Among the 23 positive dogs, nine were also were seroreactive for L. amazonensis and L. braziliensis antigens. The occurrence of anti- L. infantum chagasi antibodies in dogs was higher in rural areas (7.06%) than in urban areas (2.50%) (P < 0.05). Based on this study, we concluded that dogs from rural areas of the Pantanal wetlands were in contact with Leishmania species, which is relevant information given their importance to public health.
Show more [+] Less [-]Measurement of serum lactate values in domestic cats (Felis catus) submitted to physical and chemical restraint | Mensuração dos valores de lactato sérico em felinos domésticos (Felis catus) submetidos a contenção física e química Full text
2017
Siragusi, Rafael Henrique de Souza | Barros, Valeska Tallia Mendes de | Fiorini, Emiliana Andrade | Bergamo, Tiago Montalvão | Siqueira, Rafael Cerântola | Manhoso, Fábio Fernando Ribeiro | Franco, Rodrigo Prevedello
Measurement of serum lactate values in domestic cats (Felis catus) submitted to physical and chemical restraint | Mensuração dos valores de lactato sérico em felinos domésticos (Felis catus) submetidos a contenção física e química Full text
2017
Siragusi, Rafael Henrique de Souza | Barros, Valeska Tallia Mendes de | Fiorini, Emiliana Andrade | Bergamo, Tiago Montalvão | Siqueira, Rafael Cerântola | Manhoso, Fábio Fernando Ribeiro | Franco, Rodrigo Prevedello
A utilização do lactato sérico é cada vez mais comum na Medicina Veterinária, por ser um excelente marcador de oxigenação tecidual, elevando-se em situações de hipóxia tecidual e realizando a glicólise anaeróbica como fonte de energia. Pesquisas científicas em cães o apontam como fator prognóstico em diversas afecções, sendo pouco utilizado em felinos domésticos devido ao elevado limiar de estresse da espécie, podendo interferir em seus valores. Assim, este trabalho avaliou os valores de lactato sérico em felinos domésticos durante contenção física e após contenção química, utilizando a associação de 8 mg/kg de cetamina-S, 0,4 mg/kg de midazolam e 2 mg/kg de cloridrato de tramadol. Foram estudados 20 felinos, fêmeas, sem raça definida, de seis meses a cinco anos de idade, clinicamente saudáveis, divididos em dois grupos, com GPR (n = 10) representado por felinos que passaram apenas por contenção física e com GCR por felinos que passaram por contenção física (TPR) e após 30 horas por contenção química (TCR). Foi observado em ambos os grupos (GPR e GCR) e durante a contenção física a hiperlactatemia sérica (GPR = 6.23 ± 0.9 mmol / L – GCR – TPR = 6.66 ± 1.59 mmol / L) com valores médios de 4,42 ± 1,28 mmol/L durante a contenção química (GCR – TCR). Conclui-se que a contenção física interfere nos valores do lactato sérico, com redução dos valores na utilização da contenção química na espécie felina. | The use of serum lactate has become increasingly common in Veterinary Medicine, since it is an excellent marker of tissue oxygenation, elevated in situations of tissue hypoxia, where anaerobic glycolysis is used as an energy source. Studies point out its use in dogs as a prognostic factor in several affections; however it is not commonly used in domestic felines due to the high stress threshold of the species, which may interfere with their values. The aim of the present study was to evaluate serum lactate values in domestic cats during physical restraint and after chemical restraint using the combination of 8mg/kg ketamine-S, 0.4 mg/kg midazolam, and 2 mg/kg Tramadol. To this end, 20 clinically healthy female cats without defined race, from six months to five years of age, were divided into two groups, GPR (n = 10) represented by felines that had only physical restraint and the GCR (n = 10) by felines that underwent physical restraint (TPR) and after 30 hours by chemical restraint (TCR). It was observed that in both groups (GPR and CPR) during physical restraint presented hyperlactatemia (GPR = 6.23 ± 0.9 mmol/L – GCR – Tpr = 6.66 ± 1.59 mmol/L) and mean values of 4.42 ± 1.28 mmol/L during chemical restraint (GCR – Tcr). It was concluded that physical restraint interferes with serum lactate values, with the reduction of such values with the use of chemical restraint in the feline species.
Show more [+] Less [-]Measurement of serum lactate values in domestic cats (<i>Felis catus</i>) submitted to physical and chemical restraint Full text
2017
Rafael Henrique de Souza Siragusi | Valeska Tallia Mendes de Barros | Emiliana Andrade Fiorini | Tiago Montalvão Bergamo | Rafael Cerântola Siqueira | Fábio Fernando Ribeiro Manhoso | Rodrigo Prevedello Franco
The use of serum lactate has become increasingly common in Veterinary Medicine, since it is an excellent marker of tissue oxygenation, elevated in situations of tissue hypoxia, where anaerobic glycolysis is used as an energy source. Studies point out its use in dogs as a prognostic factor in several affections; however it is not commonly used in domestic felines due to the high stress threshold of the species, which may interfere with their values. The aim of the present study was to evaluate serum lactate values in domestic cats during physical restraint and after chemical restraint using the combination of 8mg/kg ketamine-S, 0.4 mg/kg midazolam, and 2 mg/kg Tramadol. To this end, 20 clinically healthy female cats without defined race, from six months to five years of age, were divided into two groups, GPR (n = 10) represented by felines that had only physical restraint and the GCR (n = 10) by felines that underwent physical restraint (TPR) and after 30 hours by chemical restraint (TCR). It was observed that in both groups (GPR and CPR) during physical restraint presented hyperlactatemia (GPR = 6.23 ± 0.9 mmol/L – GCR – Tpr = 6.66 ± 1.59 mmol/L) and mean values of 4.42 ± 1.28 mmol/L during chemical restraint (GCR – Tcr). It was concluded that physical restraint interferes with serum lactate values, with the reduction of such values with the use of chemical restraint in the feline species.
Show more [+] Less [-]Atualização sobre a epidemiologia de Dirofilaria immitis na América do Sul e no México: revisão de literatura | Update on Dirofilaria immitis epidemiology in South America and Mexico: literature review Full text
2017
Bendas, Alexandre José Rodrigues | Mendes-de-Almeida, Flavya | Guerrero, Jorge | Labarthe, Norma
Atualização sobre a epidemiologia de Dirofilaria immitis na América do Sul e no México: revisão de literatura | Update on Dirofilaria immitis epidemiology in South America and Mexico: literature review Full text
2017
Bendas, Alexandre José Rodrigues | Mendes-de-Almeida, Flavya | Guerrero, Jorge | Labarthe, Norma
Dirofilaria immitis é um nematoide de ampla distribuição geográfica, que ocorre com maior frequência em áreas quentes e úmidas do planeta. O primeiro registro de sua ocorrência na América do Sul foi realizado em 1878, no Brasil. Naquela época os registros eram poucos e raramente de fácil obtenção, razão pela qual reuni-los facilitará a recuperação da memória ao longo dos anos. Quatro bases de dados (Scopus, MEDLINE, LILACS e PubMed) foram estudadas utilizando-se as palavras-chave “Dirofilaria” ou “heartworm”, os nomes dos países da América do Sul e o México. Nenhum registro foi encontrado para quatro países (Bolívia, Equador, Guiana Francesa e Uruguai) e para outros três (Suriname, Guiana e Paraguai) os registros eram antigos. Apenas o Chile é o território onde houve estudos registrados com ausência do parasita. Os outros países (México, Peru, Colômbia, Venezuela, Argentina e Brasil) apresentam registros com frequência variável no tempo ou no espaço. Assim, as informações reunidas indicam que infecções por D. immitis ocorrem na maior parte da América do Sul e no México e que os médicos veterinários devem instituir programas preventivos para garantir cuidados médicos de qualidade aos pacientes e para proteger a saúde destes e de suas famílias. | Dirofilaria immitis (Leidy, 1856; Raillet & Henry 1911) is a parasite that is widely disseminated around the globe, with a higher prevalence in warm, humid climates. The first report of its occurrence in South America is from 1878 in Brazil. At that time, reports were scarce and difficult to retrieve – therefore, gathering them will facilitate record-keeping over time. Four databases were searched (Scopus, MEDLINE, LILACS, and PubMed) and the search keywords were “Dirofilaria” or “heartworm” and the countries’ names. Four countries lacked reports (Bolivia, Ecuador, French Guiana, and Uruguay) and other three (Suriname, Guyana, and Paraguay) had only old reports. Chile was the only country in which studies were conducted over time, and no infected dogs were registered. For the other six countries (Mexico, Peru, Colombia, Venezuela, Argentina, and Brazil), reports showed that the infection frequency varied over time and with the surveyed area. Therefore, the information indicates that D. immitis is established, and veterinarians must institute preventive programs to optimally care for their patients and protect the health of their families.
Show more [+] Less [-]Atualização sobre a epidemiologia de<i> Dirofilaria immitis </i>na América do Sul e no México: revisão de literatura Full text
2017
Alexandre José Rodrigues Bendas | Flavya Mendes-de-Almeida | Jorge Guerrero | Norma Labarthe
Dirofilaria immitis é um nematoide de ampla distribuição geográfica, que ocorre com maior frequência em áreas quentes e úmidas do planeta. O primeiro registro de sua ocorrência na América do Sul foi realizado em 1878, no Brasil. Naquela época os registros eram poucos e raramente de fácil obtenção, razão pela qual reuni-los facilitará a recuperação da memória ao longo dos anos. Quatro bases de dados (Scopus, MEDLINE, LILACS e PubMed) foram estudadas utilizando-se as palavras-chave “Dirofilaria” ou “heartworm”, os nomes dos países da América do Sul e o México. Nenhum registro foi encontrado para quatro países (Bolívia, Equador, Guiana Francesa e Uruguai) e para outros três (Suriname, Guiana e Paraguai) os registros eram antigos. Apenas o Chile é o território onde houve estudos registrados com ausência do parasita. Os outros países (México, Peru, Colômbia, Venezuela, Argentina e Brasil) apresentam registros com frequência variável no tempo ou no espaço. Assim, as informações reunidas indicam que infecções por D. immitis ocorrem na maior parte da América do Sul e no México e que os médicos veterinários devem instituir programas preventivos para garantir cuidados médicos de qualidade aos pacientes e para proteger a saúde destes e de suas famílias.
Show more [+] Less [-]Influência da suplementação do fator de crescimento epidermal (EGF) em diferentes momentos da maturação in vitro de oócitos caninos | Influence of epidermal growth factor (EGF) supplementation at different times of in vitro maturation of canine oocytes Full text
2017
Pereira, Leda Maria Costa | Bersano, Paulo Ricardo Oliveira | Lopes, Maria Denise
Influência da suplementação do fator de crescimento epidermal (EGF) em diferentes momentos da maturação in vitro de oócitos caninos | Influence of epidermal growth factor (EGF) supplementation at different times of in vitro maturation of canine oocytes Full text
2017
Pereira, Leda Maria Costa | Bersano, Paulo Ricardo Oliveira | Lopes, Maria Denise
O objetivo deste estudo foi avaliar a influência do fator de crescimento epidermal (EGF) em diferentes momentos da maturação in vitro de oócitos caninos. Os ovários foram coletados de 55 cadelas consideradas sadias e isolados assepticamente, imersos em solução fisiológica e transportados refrigerados. Os complexos cumulus-oócito (COCs) grau 1 foram selecionados e divididos em dois grupos, denominados grupo controle (GC) e grupo tratamento (GT). No GC, 698 COCs grau I foram cultivados em placas de quatro poços contendo meio TCM-199 suplementado com 25 mM de HEPES, 100 UI/mL de penicilina, 100 mg/mL de estreptomicina, 26 mM de bicarbonato de sódio, 1,5 mM de piruvato de sódio, 2,9 mM de lactato de sódio penta hidratado, 0,6 mM de cisteína, 0,03 UI/mL de hCG, 0,5 µg/mL de FSH, 20 µg/mL de estrógeno em estufa úmida a 38ºC, 5% de CO2 nos períodos de 24h, 48 h e 72 h . Já no GT, 547 COCs receberam o mesmo meio de maturação acrescido de 10 ηg/mL do EGF. Modelos de regressão logística foram elaborados para estimar as chances do oócito ser observado nos estágios de maturação nuclear em diferentes tempos de cultivo. Com base nos resultados encontrados, o meio suplementado com EGF demonstrou 2,56 vezes mais chances de ter um oócito no estágio de metáfase I (M-I) do que o meio sem EGF (p < 0,0001). Os resultados desse estudo demonstraram também que o tempo de 72 h mostrou 5,88 vezes mais chances de ter um oócito no estágio de metáfase II (M-II) do que o tempo de 2 h (p = 0,0001) e 7,69 vezes mais chance do que o tempo de 48h (p = 0,0001). As chances de se encontrar um oócito em M-II também foram 9,09 vezes maiores no meio suplementado com EGF do que no meio sem EGF (p = 0,0001). Dessa forma, estes resultados demonstraram a importância essencial do EGF em diferentes momentos da maturação oocitária, sendo componente chave para a aquisição da competência meiótica nas cadelas, aumentando os índices de M-I e M-II. | The aim of this study was to evaluate the influence of epidermal growth factor (EGF) on in vitro maturation of canine oocytes at different times of the process. Ovaries were collected from 55 bitches considered healthy and aseptically isolated, immersed in physiological solution (0.9% NaCl) and transported under refrigeration. Grade 1 cumulus-oocyte complexes (COCs) were selected and divided into two groups: control group (CG) and treatment group (TG). In CG 698 grade I COCs were placed in 4-well plates containing TCM-199 medium supplemented with 25 mM HEPES, 100 IU/mL penicillin, 100 mg/mL streptomycin, 26 mM sodium bicarbonate, 1.5 mM sodium pyruvate, 2.9 mM sodium lactate pentahydrate, 0.6 mM cysteine, 0.03 IU/mL hCG, 0.5 μg/mL FSH, 20 μg/mL estrogen at 38.5ºC in a humidified atmosphere of 5% CO2 in times of 24 h, 48 h, and 72 h. In TG 547 COCs received the same maturation medium plus 10 ηg/mL EGF. Logistic regression models (SAS, 2011) were constructed in order to estimate the chances of oocytes being observed at nuclear maturation stages in different culture times (24 h, 48 h, and 72 h). Based on the results found EGF-supplemented medium showed 2.56 times more chances of having an oocyte at metaphase I (M-I) than medium without EGF (p < 0.0001). The results of this study demonstrated that the time of 72 h showed 5.88 times more chances of having an oocyte at metaphase II (M-II) compared to time of 24 h (p = 0.0001) and 7.69 times more chance than time of 48 h (p = 0.0001). The chances of finding an oocyte at M-II were also 9.09 times higher in medium supplemented with EGF than in medium without EGF (p = 0.0001). Thus, these results demonstrated the essential importance of EGF at different moments of oocyte maturation, being a key component for the acquisition of meiotic competence in bitches, increasing the M-I and M-II rates.
Show more [+] Less [-]Influence of epidermal growth factor (EGF) supplementation at different times of <i>in vitro</i> maturation of canine oocytes Full text
2017
Leda Maria Costa Pereira | Paulo Ricardo Oliveira Bersano | Maria Denise Lopes
The aim of this study was to evaluate the influence of epidermal growth factor (EGF) on in vitro maturation of canine oocytes at different times of the process. Ovaries were collected from 55 bitches considered healthy and aseptically isolated, immersed in physiological solution (0.9% NaCl) and transported under refrigeration. Grade 1 cumulus-oocyte complexes (COCs) were selected and divided into two groups: control group (CG) and treatment group (TG). In CG 698 grade I COCs were placed in 4-well plates containing TCM-199 medium supplemented with 25 mM HEPES, 100 IU/mL penicillin, 100 mg/mL streptomycin, 26 mM sodium bicarbonate, 1.5 mM sodium pyruvate, 2.9 mM sodium lactate pentahydrate, 0.6 mM cysteine, 0.03 IU/mL hCG, 0.5 μg/mL FSH, 20 μg/mL estrogen at 38.5ºC in a humidified atmosphere of 5% CO2 in times of 24 h, 48 h, and 72 h. In TG 547 COCs received the same maturation medium plus 10 ηg/mL EGF. Logistic regression models (SAS, 2011) were constructed in order to estimate the chances of oocytes being observed at nuclear maturation stages in different culture times (24 h, 48 h, and 72 h). Based on the results found EGF-supplemented medium showed 2.56 times more chances of having an oocyte at metaphase I (M-I) than medium without EGF (p < 0.0001). The results of this study demonstrated that the time of 72 h showed 5.88 times more chances of having an oocyte at metaphase II (M-II) compared to time of 24 h (p = 0.0001) and 7.69 times more chance than time of 48 h (p = 0.0001). The chances of finding an oocyte at M-II were also 9.09 times higher in medium supplemented with EGF than in medium without EGF (p = 0.0001). Thus, these results demonstrated the essential importance of EGF at different moments of oocyte maturation, being a key component for the acquisition of meiotic competence in bitches, increasing the M-I and M-II rates.
Show more [+] Less [-]Composição nutricional e avaliação de diferentes métodos de determinação de gordura em alimentos úmidos para cães e gatos | Nutritional composition and evaluation of different methodologies for fat determination in wet feed for dogs and cats Full text
2017
Urrego, Maria Isabel Gonzalez | Ernandes, Mariane Ceschin | Matheus, Laura Fantucci de Oliveira | Santos, Karine de Melo | Oba, Patricia Massae | Silva, Cláudio Galeno Piantino | Vendramini, Thiago Henrique Annibale | Pedrinelli, Vivian | Brunetto, Márcio Antonio
Composição nutricional e avaliação de diferentes métodos de determinação de gordura em alimentos úmidos para cães e gatos | Nutritional composition and evaluation of different methodologies for fat determination in wet feed for dogs and cats Full text
2017
Urrego, Maria Isabel Gonzalez | Ernandes, Mariane Ceschin | Matheus, Laura Fantucci de Oliveira | Santos, Karine de Melo | Oba, Patricia Massae | Silva, Cláudio Galeno Piantino | Vendramini, Thiago Henrique Annibale | Pedrinelli, Vivian | Brunetto, Márcio Antonio
Devido ao aumento do número de cães e gatos domiciliados, a indústria de alimentos para animais de estimação tem expandido a gama de produtos existentes no mercado de pet food. Para obter informações mais relevantes sobre o segmento de alimentos úmidos para cães e gatos, este trabalho determinou a composição nutricional, avaliou as informações declaradas nos rótulos e comparou a composição com as recomendações da Fediaf de proteína e gordura. Também foram comparadas três metodologias diferentes de análise de gordura: extrato etéreo (CFa), extrato etéreo após hidrólise ácida (CFAH) e teor de gordura obtido no analisador Ankom XT15 (ANKOM) para determinar o método mais adequado de avaliação de gordura em alimentos úmidos. Foram avaliadas 25 marcas de alimentos úmidos, 13 para cães e 12 para gatos. As análises de composição centesimal obtidas neste estudo foram comparadas com a informação nutricional declarada nos rótulos e com as necessidades mínimas recomendadas pela Fediaf para cada espécie. Os resultados da composição nutricional, os valores descritos no rótulo e a avaliação dos três métodos para determinação da gordura foram comparados com o emprego do teste t e modelo misto com medidas repetidas nas mesmas amostras, respectivamente (p < 0,05) no programa SAS. Já a avaliação da adequação nutricional de proteína e do teor de gordura foram analisados por cálculos matemáticos de diferença e proporção. Para a maioria dos alimentos avaliados não foi observada diferença entre a composição nutricional dos alimentos úmidos e os valores declarados em rótulo, e houve predominância de produtos que excederam as recomendações mínimas de proteína e gordura da Fediaf para ambas as espécies. Quanto às metodologias de extração de gordura, não foi observada diferença entre os três métodos avaliados (p = 0,68). Concluiu-se que os alimentos úmidos avaliados atendem às informações declaradas pelos fabricantes e também às recomendações nutricionais da Fediaf com base na ingestão energética recomendada. Em relação às metodologias avaliadas para determinação de gordura nestes alimentos, a similaridade entre tais resultados justifica o uso da técnica de maior facilidade ou de menor custo. | As a consequence of the increasing number of dog and cat owners, the pet food industry is expanding the range of pet food products in the market. In order to obtain more necessary information about the wet food segment for dogs and cats, the aim of this study was to determine the nutritional composition, to evaluate the information declared on the labels, and to compare the composition with the FEDIAF recommendations for protein and fat. Furthermore, three different methodologies of fat analysis were compared: crude fat (CFa), crude fat after acid hydrolysis (CFAH), and fat content obtained with Ankom XT15 (ANKOM) to determine the most adequate method for fat determination in wet foods. Twenty-five wet food products were evaluated, 13 wet foods for dogs and 12 for cats. Centesimal composition analyses obtained in this study were compared with guaranteed analysis declared on the label and with FEDIAF minimum recommended requirements for each species. The results of the nutritional composition and the values described on the label and the evaluation of the three fat determination methods were compared using the mixed model test with repeated measurements in the same samples, respectively (p < 0.05) in the SAS program, evaluation of protein adequacy and fat content were analyzed by mathematical calculations of difference and proportion. No difference was observed between nutritional composition of wet foods and the values declared on the labels for the majority of the diets analyzed, and there was a predominance of products that exceeded FEDIAF minimum recommendations of protein and fat for both species. No difference was observed between the three methods of fat content evaluation (p = 0.68). It was concluded that wet foods evaluated in this study match the label information and FEDIAF nutrient requirement recommendations, considering recommended calorie intake. All three fat determination methodologies evaluated were similar, justifying the choice of the easiest or cheapest method.
Show more [+] Less [-]Nutritional composition and evaluation of different methodologies for fat determination in wet feed for dogs and cats Full text
2017
Maria Isabel Gonzalez Urrego | Mariane Ceschin Ernandes | Laura Fantucci de Oliveira Matheus | Karine de Melo Santos | Patricia Massae Oba | Cláudio Galeno Piantino Silva | Thiago Henrique Annibale Vendramini | Vivian Pedrinelli | Márcio Antonio Brunetto
As a consequence of the increasing number of dog and cat owners, the pet food industry is expanding the range of pet food products in the market. In order to obtain more necessary information about the wet food segment for dogs and cats, the aim of this study was to determine the nutritional composition, to evaluate the information declared on the labels, and to compare the composition with the FEDIAF recommendations for protein and fat. Furthermore, three different methodologies of fat analysis were compared: crude fat (CFa), crude fat after acid hydrolysis (CFAH), and fat content obtained with Ankom XT15 (ANKOM) to determine the most adequate method for fat determination in wet foods. Twenty-five wet food products were evaluated, 13 wet foods for dogs and 12 for cats. Centesimal composition analyses obtained in this study were compared with guaranteed analysis declared on the label and with FEDIAF minimum recommended requirements for each species. The results of the nutritional composition and the values described on the label and the evaluation of the three fat determination methods were compared using the mixed model test with repeated measurements in the same samples, respectively (p < 0.05) in the SAS program, evaluation of protein adequacy and fat content were analyzed by mathematical calculations of difference and proportion. No difference was observed between nutritional composition of wet foods and the values declared on the labels for the majority of the diets analyzed, and there was a predominance of products that exceeded FEDIAF minimum recommendations of protein and fat for both species. No difference was observed between the three methods of fat content evaluation (p = 0.68). It was concluded that wet foods evaluated in this study match the label information and FEDIAF nutrient requirement recommendations, considering recommended calorie intake. All three fat determination methodologies evaluated were similar, justifying the choice of the easiest or cheapest method.
Show more [+] Less [-]Caracterização genética do gene da hemaglutinina em vírus da cinomose canina de cães naturalmente infectados no Brasil | Genetic characterization of the haemagglutinin gene in canine distemper virus strains from naturally infected dogs in Brazil Full text
2017
Cortez, Adriana | Heinemann, Marcos Bryan | Fonseca Junior, Antônio Augusto | Costa, Luciana Fachini da | Souza, Vanessa Aparecida Feijó de | Megid, Jane
Caracterização genética do gene da hemaglutinina em vírus da cinomose canina de cães naturalmente infectados no Brasil | Genetic characterization of the haemagglutinin gene in canine distemper virus strains from naturally infected dogs in Brazil Full text
2017
Cortez, Adriana | Heinemann, Marcos Bryan | Fonseca Junior, Antônio Augusto | Costa, Luciana Fachini da | Souza, Vanessa Aparecida Feijó de | Megid, Jane
A cinomose canina é uma das principais doenças infecciosas em cães e animais selvagens, resultando em alta morbidade e mortalidade. O gene H tem uma das maiores variabilidades genéticas entre os genes codificados pelo vírus da cinomose canina (CDV), e tem sido utilizado para caracterizar as estirpes de CDV, permitindo a identificação de linhagens específicas. A variação no gene H pode permitir que o vírus evite o reconhecimento por anticorpos induzidos pela vacina, resultando em falha vacinal. O objetivo deste estudo foi caracterizar o gene H em estirpes de CDV de cães infectados naturalmente no estado de São Paulo. A análise filogenética revelou que as estirpes de CDV brasileiras estão geneticamente relacionadas as estirpes circulantes no Uruguai, na Argentina e na Europa. Não foi encontrada nenhuma evidência da circulação no estado de São Paulo das linhagens América do Sul 2 e 3. O grau de divergência genética entre linhagens selvagens de CDV brasileiras e as estirpes vacinais podem sugerir a possibilidade de falhas vacinais e consequentemente a ocorrência de surtos de cinomose canina. | Canine distemper is one of the major infectious diseases in dogs and wild animals, resulting in high morbidity and mortality. The H gene has the greatest genetic variability among the genes encoded by the canine distemper virus (CDV) genome, and it has been used to characterise field samples, allowing the identification of specific lineages. Variation in the H gene can allow the virus to evade recognition by vaccine-induced antibodies, resulting in vaccine failure. The purpose of this study was to characterise H gene in CDV strains from naturally infected dogs in the state of São Paulo. The phylogenetic analysis revealed that Brazilian CDV strains were genetically related to the circulating CDV strains in Uruguay, Argentina, and Europe. We found no evidence of South America 2 and 3 CDV lineages circulating in Brazilian dogs. The degree of genetic divergence between wild Brazilian CDV strains and vaccine strains may suggest the possibility of vaccine failures and consequently the occurrence of canine distemper outbreaks.
Show more [+] Less [-]Caracterização genética do gene da hemaglutinina em vírus da cinomose canina de cães naturalmente infectados no Brasil Full text
2017
Adriana Cortez | Marcos Bryan Heinemann | Antônio Augusto Fonseca Junior | Luciana Fachini da Costa | Vanessa Aparecida Feijó de Souza | Jane Megid
A cinomose canina é uma das principais doenças infecciosas em cães e animais selvagens, resultando em alta morbidade e mortalidade. O gene H tem uma das maiores variabilidades genéticas entre os genes codificados pelo vírus da cinomose canina (CDV), e tem sido utilizado para caracterizar as estirpes de CDV, permitindo a identificação de linhagens específicas. A variação no gene H pode permitir que o vírus evite o reconhecimento por anticorpos induzidos pela vacina, resultando em falha vacinal. O objetivo deste estudo foi caracterizar o gene H em estirpes de CDV de cães infectados naturalmente no estado de São Paulo. A análise filogenética revelou que as estirpes de CDV brasileiras estão geneticamente relacionadas as estirpes circulantes no Uruguai, na Argentina e na Europa. Não foi encontrada nenhuma evidência da circulação no estado de São Paulo das linhagens América do Sul 2 e 3. O grau de divergência genética entre linhagens selvagens de CDV brasileiras e as estirpes vacinais podem sugerir a possibilidade de falhas vacinais e consequentemente a ocorrência de surtos de cinomose canina.
Show more [+] Less [-]Esporotricose - gato como fonte de infecção em foco epidêmico familiar em Guarulhos, São Paulo: relato de caso | Sporotrichosis - cat as the source of familiar infection outbreak in Guarulhos, São Paulo: a case report Full text
2017
Galati, Leandro Haroutune Hassesian | Tomimori, Jane | Taborda, Carlos Pelleschi | Michalany, Nilceo Schwery | Larsson Júnior, Carlos Eduardo | Larsson, Carlos Eduardo
Esporotricose - gato como fonte de infecção em foco epidêmico familiar em Guarulhos, São Paulo: relato de caso | Sporotrichosis - cat as the source of familiar infection outbreak in Guarulhos, São Paulo: a case report Full text
2017
Galati, Leandro Haroutune Hassesian | Tomimori, Jane | Taborda, Carlos Pelleschi | Michalany, Nilceo Schwery | Larsson Júnior, Carlos Eduardo | Larsson, Carlos Eduardo
A esporotricose é uma dermatozoonose que tem como agentes etiológicos fungos do gênero Sporothrix. É responsável por quadros micóticos ditos de implantação. Essa micose, de características antropo e saprozoonóticas e cujas principais fontes de infecção são os felinos domésticos, o solo e os vegetais, tem sido responsável por surtos epizoóticos e epidêmicos no sul e sudeste brasileiro. Este relato apresenta o caso de um felino diagnosticado e tratado para esporotricose pelo Serviço de Dermatologia do Hospital Veterinário da FMVZ/USP, após ter sido encaminhado pelo Departamento de Dermatologia da EPM/Unifesp, onde os proprietários do animal estavam sendo tratados para esporotricose humana. | Sporotrichosis is a dermatozoonosis caused by fungi of the Sporothrix genus. It is classified as an implantation mycosis. This fungal infection, with anthropozoonotic and saprozoonotic characteristics and which has domestic felines, soil, and vegetables as main sources of infection, has been responsible for epizootic and epidemic outbreaks in southern and southeastern Brazil. This report presents the case of a feline diagnosed and treated for sporotrichosis at Dermatology Service of FMVZ/USP, after referral from the Dermatology Department of EPM/Unifesp, where the owners of the cat were being treated for human sporotrichosis.
Show more [+] Less [-]Esporotricose - gato como fonte de infecção em foco epidêmico familiar em Guarulhos, São Paulo: relato de caso Full text
2017
Leandro Haroutune Hassesian Galati | Jane Tomimori | Carlos Pelleschi Taborda | Nilceo Schwery Michalany | Carlos Eduardo Larsson Júnior | Carlos Eduardo Larsson
A esporotricose é uma dermatozoonose que tem como agentes etiológicos fungos do gênero Sporothrix. É responsável por quadros micóticos ditos de implantação. Essa micose, de características antropo e saprozoonóticas e cujas principais fontes de infecção são os felinos domésticos, o solo e os vegetais, tem sido responsável por surtos epizoóticos e epidêmicos no sul e sudeste brasileiro. Este relato apresenta o caso de um felino diagnosticado e tratado para esporotricose pelo Serviço de Dermatologia do Hospital Veterinário da FMVZ/USP, após ter sido encaminhado pelo Departamento de Dermatologia da EPM/Unifesp, onde os proprietários do animal estavam sendo tratados para esporotricose humana.
Show more [+] Less [-]Analytical strategy for determination of chloramphenicol in different biological matrices by liquid chromatography - mass spectrometry Full text
2017
Śniegocki Tomasz | Gbylik-Sikorska Małgorzata | Posyniak Andrzej
Analytical strategy for determination of chloramphenicol in different biological matrices by liquid chromatography - mass spectrometry Full text
2017
Śniegocki Tomasz | Gbylik-Sikorska Małgorzata | Posyniak Andrzej
Introduction: The main problem in determination of chloramphenicol in food of animal origin is a large number of matrices. The main target of this study was to create a method for determination and confirmation of chloramphenicol in products and food of animal origin. Material and Methods: Each 5 g matrix sample was mixed with 5 mL of water and 10 mL of acetonitrile/ethyl acetate, homogenised, and centrifuged. The organic layer was evaporated and redissolved in 6 mL of 4% NaCl. The extract was cleaned up by SPE technique. Chloramphenicol was analysed by LC-MS/MS in electrospray mode. Results: The procedure was validated according to the Commission Decision No. 2002/657/EC. The apparent recoveries were in the range of 92.1% to 107.1% with a repeatability less than 11.0% (4.4%-11.0%) and within-laboratory reproducibility below 13.6% (4.7%-13.6%). Conclusion: The method was successfully validated and proved to be efficient, precise, and useful for quantification of chloramphenicol in more than 20 different matrices.
Show more [+] Less [-]Analytical strategy for determination of chloramphenicol in different biological matrices by liquid chromatography - mass spectrometry Full text
2017
Śniegocki, Tomasz | Gbylik-Sikorska, Małgorzata | Posyniak, Andrzej
Introduction: The main problem in determination of chloramphenicol in food of animal origin is a large number of matrices. The main target of this study was to create a method for determination and confirmation of chloramphenicol in products and food of animal origin. Material and Methods: Each 5 g matrix sample was mixed with 5 mL of water and 10 mL of acetonitrile/ethyl acetate, homogenised, and centrifuged. The organic layer was evaporated and redissolved in 6 mL of 4% NaCl. The extract was cleaned up by SPE technique. Chloramphenicol was analysed by LC-MS/MS in electrospray mode. Results: The procedure was validated according to the Commission Decision No. 2002/657/EC. The apparent recoveries were in the range of 92.1% to 107.1% with a repeatability less than 11.0% (4.4%-11.0%) and within-laboratory reproducibility below 13.6% (4.7%-13.6%). Conclusion: The method was successfully validated and proved to be efficient, precise, and useful for quantification of chloramphenicol in more than 20 different matrices.
Show more [+] Less [-]Dogs’ gastrointestinal parasites and their association with public health in Iran Full text
2017
Kohansal Mohammad Hasan | Fazaeli Asghar | Nourian Abbasali | Haniloo Ali | Kamali Koorosh
Dogs’ gastrointestinal parasites and their association with public health in Iran Full text
2017
Kohansal Mohammad Hasan | Fazaeli Asghar | Nourian Abbasali | Haniloo Ali | Kamali Koorosh
Introduction: Dogs harbour zoonotic parasites that cause serious infections in humans, such as visceral larva migrans, ocular larva migrans, cystic echinococcosis, and alveolar echinococcosis. Studies on dogs’ gastrointestinal parasites in different geographical locations are required to increase knowledge of the risk of canine zoonoses in human populations.
Show more [+] Less [-]Dogs’ gastrointestinal parasites and their association with public health in Iran Full text
2017
Kohansal, Mohammad Hasan | Fazaeli, Asghar | Nourian, Abbasali | Haniloo, Ali | Kamali, Koorosh
Introduction: Dogs harbour zoonotic parasites that cause serious infections in humans, such as visceral larva migrans, ocular larva migrans, cystic echinococcosis, and alveolar echinococcosis. Studies on dogs’ gastrointestinal parasites in different geographical locations are required to increase knowledge of the risk of canine zoonoses in human populations.Material and Methods: The presence of parasites was examined in 450 faecal samples collected from eight zones of Zanjan province, northwest Iran from June to November 2015. The samples were examined using the sedimentation concentration method and modified Ziehl-Neelsen staining.Results: Gastrointestinal parasites were found in 86 (19.1%) faecal samples. Sarcocystis spp. (7.3%), Taenia/Echinococcus spp. (5.6%), Toxocara spp. (1.8%), and Cystoisospora spp. (1.6%) were the most common parasites observed. The other detected parasites consisted of Dicrocoelium dendriticum (0.7%), Eimeria spp. (0.7%), Cryptosporidium spp. (0.4%), Physaloptera spp. (0.4%), Giardia spp. (1.3%), and Spirocerca lupi (1.3%). The lowest parasite infection rates belonged to Trichuris vulpis and Acanthocephalans (0.2% each).Conclusion: This study provides current information on the infection rates in dog populations in Zanjan Province. Furthermore, the study shows a high prevalence of gastrointestinal parasitic infections, including zoonotic ones and particularly Taenia/Echinococcus spp., potentially transmissible to humans and thus relevant to public health.
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