Seroprevalence of Hepatitis E virus (HEV) in domestic non-commercial pigs reared in small-scale farms and wild boar in South of Brazil

Hepatitis E is a zoonotic emerging disease distributed worldwide. The domestic swine and wild boars (Sus scrofa) are known as important reservoirs of HEV although HEV infections have been detected in other animal species. The southern region of Brazil has the largest swine productions in the country, ranging from highly-specialized commercial swine productions to small-scale non-commercial pig farms. The small-scale farms allow interactions between wild boars and domestic pigs, when occasionally pathogens transmission can occur between these populations. The aim of this study was to determine HEV seroprevalence in non-commercial domestic pigs and wild boars from two southern Brazilian states (RS: Rio Grande do Sul; SC: Santa Catarina), and discuss if the consumption of raw or undercooked meat from these animals is a potential risk to public health. Animals from RS and SC States were sampled. Serum was harvested from wild boar hunted between 2012 and 2016, and from non-commercial small-scale pig farms in 2014. Overall 249 wild boars (56 from RS and 193 from SC) and 382 pigs (261 from RS and 121 from SC) were tested to detect anti-HEV IgG antibodies using a commercial HEV antibody ELISA kit (Thermo fisher), specific for swine. Overall difference was observed (P\u3c0.0001) regarding HEV seroprevalence between wild boar 4.42% (n=249) and non-commercial domestic pigs 46.60% (n=382). In relation to wild boars samples, higher seroprevalence for Hepatitis E was observed in RS (14.29%; n=56) and lower in SC (1.55%; n=193; P\u3c0.0004). In relation to pigs, RS had also higher seroprevalence (53.26%; n=261) than SC (32.23%; n=121; P\u3c0.0002). Although interactions between wild boar and non-commercial domestic pigs are known to occur, the lowest antibody detection in wild boar suggest that these contact may not be sufficient to explain seroprevalence in studied populations. Our results indicate that non-commercial pigs are a more likely source of infection for the human population than wild boar

Description of a Prospective 17DD Yellow Fever Vaccine Cohort in Recife, Brazil

From September 2005 to March 2007, 238 individuals being vaccinated for the first time with the yellow fever (YF) -17DD vaccine were enrolled in a cohort established in Recife, Brazil. A prospective study indicated that, after immunization, anti-YF immunoglobulin M (IgM) and anti-YF IgG were present in 70.6% (IgM) and 98.3% (IgG) of the vaccinated subjects. All vaccinees developed protective immunity, which was detected by the plaque reduction neutralization test (PRNT) with a geometric mean titer of 892. Of the 238 individuals, 86.6% had IgG antibodies to dengue virus; however, the presence of anti-dengue IgG did not interfere significantly with the development of anti-YF neutralizing antibodies. In a separate retrospective study of individuals immunized with the 17DD vaccine, the PRNT values at 5 and 10 years post-vaccination remained positive but showed a significant decrease in neutralization titer (25% with PRNT titers < 100 after 5 years and 35% after 10 years)

Gene expression analysis in anterior pituitary to investigate genetics of swine fertility

In the present study we investigated the effect of selection for fertility traits on gene expression in anterior pituitary (AP) of NE selection lines. The objective of this study was to identify differentially expressed genes in the anterior pituitary of sows with different fertility phenotype. A White Composite base population originated two selection lines, one control line and another selected line for prolificacy. Selection was conducted based on an index, in which ovulation rate and embryo survival were included as parameters. Sows from both lines were slaughtered during follicular development phase. Differential display PCR (DD-PCR) was used to search for differences between lines in AP gene expression. Northern analysis and microarray were used to confirm DD-PCR results. Microarray was constructed using swine anterior pituitary clones (from the present study) and follicle clones from a concurrent DD-PCR project. In the expression study, three genes were identified as differentially expressed in the same direction of DD-PCR using Northern hybridizations. Using microarray analysis, 13 genes were differently expressed (P \u3c 0.05) for the two lines. At D2 nine genes were differently expressed, and at D4, the expression of six genes differed between lines. In general, genes differentially expressed are involved in cell growth, iron pool regulation, and energy production. In conclusion, long-term selection for fertility plays an important role in changing gene expression patterns in the anterior pituitary. Selected transcripts isolated from DD-PCR were mapped. Eleven markers were mapped using somatic cell hybrid panel (SCHP). Eight markers were mapped using a radiation hybrid panel (RH). SCHP mapping allows regional assignment of markers to chromosomes, and RH mapping refines mapping location and defines marker order not resolved in the linkage maps of the pig, which is important in finding genes in QTL region, in applying marker-assisted selection, and in increasing gene density in the porcine genomic map. Furthermore, they are useful for development of the porcine transcript map and the high-resolution BAC contig map

Gene expression analysis in anterior pituitary to investigate genetics of swine fertility

In the present study we investigated the effect of selection for fertility traits on gene expression in anterior pituitary (AP) of NE selection lines. The objective of this study was to identify differentially expressed genes in the anterior pituitary of sows with different fertility phenotype. A White Composite base population originated two selection lines, one control line and another selected line for prolificacy. Selection was conducted based on an index, in which ovulation rate and embryo survival were included as parameters. Sows from both lines were slaughtered during follicular development phase. Differential display PCR (DD-PCR) was used to search for differences between lines in AP gene expression. Northern analysis and microarray were used to confirm DD-PCR results. Microarray was constructed using swine anterior pituitary clones (from the present study) and follicle clones from a concurrent DD-PCR project. In the expression study, three genes were identified as differentially expressed in the same direction of DD-PCR using Northern hybridizations. Using microarray analysis, 13 genes were differently expressed (P \u3c 0.05) for the two lines. At D2 nine genes were differently expressed, and at D4, the expression of six genes differed between lines. In general, genes differentially expressed are involved in cell growth, iron pool regulation, and energy production. In conclusion, long-term selection for fertility plays an important role in changing gene expression patterns in the anterior pituitary. Selected transcripts isolated from DD-PCR were mapped. Eleven markers were mapped using somatic cell hybrid panel (SCHP). Eight markers were mapped using a radiation hybrid panel (RH). SCHP mapping allows regional assignment of markers to chromosomes, and RH mapping refines mapping location and defines marker order not resolved in the linkage maps of the pig, which is important in finding genes in QTL region, in applying marker-assisted selection, and in increasing gene density in the porcine genomic map. Furthermore, they are useful for development of the porcine transcript map and the high-resolution BAC contig map

Indicadores do bem-estar em suínos Welfare indicators in swine

A busca por produtos de qualidade exige do produtor mudanças no sistema de produção de suíno que priorizem, em particular, o bem-estar do animal. As mudanças são necessárias para atender à demanda da sociedade e ampliar os mercados internos e externos. O bem-estar na espécie suína pode ser avaliado por meio das respostas comportamentais, fisiológicas, ligadas à sanidade e à produção. Em função do exposto, o objetivo deste trabalho é revisar a literatura em relação aos critérios científicos utilizados para indicar o bem-estar da espécie suína nos sistemas de produção.In order to obtain good quality products, the swine production system should prioritize the animals welfare. Changes are necessary to meet society's demand and expand domestic and foreign markets. The swine's welfare can be assessed by behavioral and physiological traits related to health and production. Thus, our goal is to review in the literature what are the scientific criteria that are used to indicate the swine welfare in the production systems

Avaliação dos meios PBS e whitten no cultivo de morulas de Mus musculus

A fim de estabelecer um parâmetro para a avaliação da viabilidade após o descongelamento, os resultados do cultivo de embriões frescos de camundongos, no meio PBS Modificado sem adição de C02, foram comparados aos resultados obtidos após o cultivo no meio de Whitten Modificado em atmosfera com 5% de C02. Foram coletadas 1458 mórulas provenientes de fêmeas Mus musculus da cepa Suíço Albina CF1, previamente superovuladas com 6-8UI de ECG e 6-8UI de HCG, com intervalo de 48 horas. Destas, 1051 foram cultivadas em PBS Modificado acrescido de 20% de SFB, a 37ºC e 95% de umidade. As restantes 407 foram cultivadas no meio de Whitten Modificado, acrescido de 3% de BSA, a 37ºC, de 95% de umidade e 5% de CO2. AS avaliações foram realizadas com 24h de cultivo, sendo considerados os embriões que atingiram os estágios de blastocisto inicial (Bi), blastocisto (Bl), blastocisto expandido (Bx), blastocisto em eclosão (BH) ou blastocisto eclodido (Be), e após 48h de cultivo os que atingiram pelo menos o estágio de blastocisto expandido. Os resultados obtidos após 24h (87,9 e 98,4%) e após 48h de cultivo (95,1 e 93,8%) em PBS e Whitten respectivamente, não apresentaram diferenças estatisticamente significativas (p< 0,05)

Avaliação dos meios PBS e whitten no cultivo de morulas de Mus musculus

A fim de estabelecer um parâmetro para a avaliação da viabilidade após o descongelamento, os resultados do cultivo de embriões frescos de camundongos, no meio PBS Modificado sem adição de C02, foram comparados aos resultados obtidos após o cultivo no meio de Whitten Modificado em atmosfera com 5% de C02. Foram coletadas 1458 mórulas provenientes de fêmeas Mus musculus da cepa Suíço Albina CF1, previamente superovuladas com 6-8UI de ECG e 6-8UI de HCG, com intervalo de 48 horas. Destas, 1051 foram cultivadas em PBS Modificado acrescido de 20% de SFB, a 37ºC e 95% de umidade. As restantes 407 foram cultivadas no meio de Whitten Modificado, acrescido de 3% de BSA, a 37ºC, de 95% de umidade e 5% de CO2. AS avaliações foram realizadas com 24h de cultivo, sendo considerados os embriões que atingiram os estágios de blastocisto inicial (Bi), blastocisto (Bl), blastocisto expandido (Bx), blastocisto em eclosão (BH) ou blastocisto eclodido (Be), e após 48h de cultivo os que atingiram pelo menos o estágio de blastocisto expandido. Os resultados obtidos após 24h (87,9 e 98,4%) e após 48h de cultivo (95,1 e 93,8%) em PBS e Whitten respectivamente, não apresentaram diferenças estatisticamente significativas (p< 0,05)

A two-plasmid strategy for engineering a dengue virus type 3 infectious clone from primary Brazilian isolate

Dengue infections represent one of the most prevalent arthropod-borne diseases worldwide, causing a wide spectrum of clinical outcomes. Engineered infectious clone is an important tool to study Dengue virus (DENV) biology. Functional full-length cDNA clones have been constructed for many positive-strand RNA viruses and have provided valuable tools for studying the molecular mechanisms involved in viral genome replication, virion assembly, virus pathogenesis and vaccine development. We report herein the successful development of an infectious clone from a primary Brazilian isolate of dengue virus 3 (DENV3) of the genotype III. Using a two-plasmid strategy, DENV3 genome was divided in two parts and cloned separately into a yeast-bacteria shuttle vector. All plasmids were assembled in yeast by homologous recombination technique and a full-length template for transcription was obtained by in vitro ligation of the two parts of the genome. Transcript-derived DENV3 is infectious upon transfection into BHK-21 cells and in vitro characterization confirmed its identity. Growth kinetics of transcript-derived DENV3 was indistinguishable from wild type DENV3. This system is a powerful tool that will help shed light on molecular features of DENV biology, as the relationship of specific mutations and DENV pathogenesis