The Influence of porcine parity on colostrum cytokine levels and their passive transfer to piglets / A influência da paridade suína na diferença dos níveis de citocinas no colostro e na transferência passiva para leitões

The limited ability of newborn piglets to produce cytokines may influence lymphocyte development and response to antigen exposure. As a result, colostrum intake is crucial because it contains nutrients that contribute to immune system development in piglets. Our goal was to investigate the effect of sow parity on the transfer of maternal cytokines to nursing piglets. Sixty piglets from nine sows were divided into six groups: piglets from gilts or sows kept with their dams and allowed to suckle normally; piglets from gilts or sows having their dams exchanged and then allowed to suckle normally; piglets from gilts or sows isolated from their dams and bottle-fed a commercial milk replacer formula for pigs. All piglets remained in the diet groups for 24 hours after birth. Concentrations of cytokines in colostrum and serum of gilt/ sows and serum of piglets were then evaluated. The 13 evaluated cytokines had higher concentrations in colostrum and serum of sows than in gilts. Concentrations of GM-CSF, IFNγ, IL-1α, IL-1ß, IL-2, IL-4, IL-6, IL-10, IL-12, IL-18, and TNFα were higher in piglets suckling sows. Piglets that received commercial formula showed higher concentrations of the cytokines IL1-RA and IL-8 than piglets fed colostrum. This outcome can influence piglets' development into adulthood. In short, our findings demonstrated that maternal parity influenced colostrum cytokine composition and its maternal transfer patterns.(AU)

A capacidade limitada dos leitões recém-nascidos de produzir citocinas pode influenciar o desenvolvimento de linfócitos e a resposta à exposição ao antígeno. Portanto, a ingestão de colostro é importante porque contém nutrientes, que contribuem para o desenvolvimento do sistema imunológico do leitão. O objetivo do estudo foi investigar o efeito da paridade da porca na transferência de citocina materna para leitões lactentes. Sessenta leitões de nove porcas foram divididos em seis grupos: leitões de marrãs/porcas mantidas com suas próprias mães e amamentadas normalmente; leitões de marrãs/porcas que foram trocados de mães e amamentados normalmente; leitões de marrãs/porcas que foram isolados das mães e alimentados com mamadeira com substituto do leite para suínos. Os leitões permaneceram nos grupos por 24 horas após o nascimento. Foram avaliadas as concentrações de citocinas no colostro e plasma das marrãs/porcas e no plasma dos leitões. O colostro e o plasma das porcas apresentaram maiores concentrações das 13 citocinas analisadas do que as marrãs. No mesmo sentido, as concentrações de GM-CSF, IFNγ, IL-1α, IL-1ß, IL-2, IL-4, IL-6, IL-10, IL-12, IL-18 e TNFα foram significantemente maiores nos leitões que mamaram o colostro de porcas. Os leitões que receberam fórmula comercial apresentaram, em especial, concentrações das citocinas IL1-RA e IL-8 superiores aos leitões amamentados com colostro. Isso pode influenciar o desenvolvimento até a fase adulta. Portanto, nossos dados demonstraram que a paridade materna influenciou a composição das citocinas do colostro, bem como as características das citocinas na transferência materna.(AU)

Distribution difference of colostrum-derived B and T cells subsets in gilts and sows.

Piglets are highly vulnerable to infections, but colostrum provides them with some protection. The function of colostrum components is unknown, as is if the amount and subsets of leukocytes in colostrum differ between gilts and sows. This study serially characterized leukocyte populations in colostrum for differential leukocyte counts. Differences in humoral and cellular composition of colostrum between 40 gilts and 40 sows (parities orders 3-4) from a commercial herd were examined. Flow cytometry is a useful tool to identify and quantify leukocyte subsets in sow colostrum. Overall, there were no (p ≥ 0.05) parity differences in total macrophages, granulocytes, and T and B cells. However, the sows' colostrum presented significantly higher (p ≤ 0.05) T lymphocyte subsets than gilts, such as central memory CD4+T cells, effector memory CD4+T cells, and central memory CD8+T cells. Among B-lymphocytes, percentages of SWC7+CD5+ cells were significantly higher in sow colostrum than in that of gilts. As expected, IgG concentrations were significantly higher in sows than in gilts. Colostrum from sows had significantly greater mitogenic activity than colostrum from gilts and this fact can be associated with the potential to accelerate the maturation of a newborn's gastrointestinal tract. Our findings suggest that parity order may be one among other factors influencing the cell population and, consequently, the immune adaptive response in piglets that induces neutralizing antibodies and cellular immune responses to antigens.

Detection of sapovirus in brazilian pig farms / Detecção de sapovírus em criações de suínos brasileiras

Sapoviruses (Caliciviridae) are considered important agents of gastroenteritis worldwide affecting animals and humans. In pig farming, the epidemiology is not completely understood because it can affect all stages of production, with symptomatic (diarrhea) or asymptomatic pigs. The aim of our study was to investigate Sapovirus occurrence in Brazilian pig farms. A total of 166 fecal samples of pigs, with different ages, from Minas Gerais, São Paulo, and Mato Grosso States were submitted to RT-PCR reactions and confirmed with nucleotide sequencing of Sapovirus RdRp gene. Six (3.61%) samples were positive and four had partial RdRp gene sequenced, putatively belonging to GVII.1 genogroup, also reported in swine herds in Brazil.

Sapovírus (Caliciviridae) são considerados importantes agentes causadores de gastroenterites em todo o mundo, afetando animais e humanos. Na suinocultura, sua epidemiologia ainda não foi totalmente esclarecida, pois pode afetar todas as fases da produção, com suínos sintomáticos (diarreia) ou assintomáticos. O objetivo do nosso estudo foi investigar a ocorrência de Sapovírus em granjas de suínos brasileiras. Um total de 166 amostras fecais de suínos, com diferentes idades, dos estados de Minas Gerais, São Paulo e Mato Grosso foram submetidas a reações de RT-PCR e confirmadas com sequenciamento de nucleotídeos do gene RdRp do Sapovírus. Seis (3,61%) amostras foram positivas e quatro delas tinham sequenciamento parcial do gene RdRp, supostamente pertencente ao genogrupo GVII.1, previamente relatado em rebanhos suínos no Brasil.

Detection of sapovirus in brazilian pig farms / Detecção de sapovírus em criações de suínos brasileiras

Sapoviruses (Caliciviridae) are considered important agents of gastroenteritis worldwide affecting animals and humans. In pig farming, the epidemiology is not completely understood because it can affect all stages of production, with symptomatic (diarrhea) or asymptomatic pigs. The aim of our study was to investigate Sapovirus occurrence in Brazilian pig farms. A total of 166 fecal samples of pigs, with different ages, from Minas Gerais, São Paulo, and Mato Grosso States were submitted to RT-PCR reactions and confirmed with nucleotide sequencing of Sapovirus RdRp gene. Six (3.61%) samples were positive and four had partial RdRp gene sequenced, putatively belonging to GVII.1 genogroup, also reported in swine herds in Brazil.(AU)

Sapovírus (Caliciviridae) são considerados importantes agentes causadores de gastroenterites em todo o mundo, afetando animais e humanos. Na suinocultura, sua epidemiologia ainda não foi totalmente esclarecida, pois pode afetar todas as fases da produção, com suínos sintomáticos (diarreia) ou assintomáticos. O objetivo do nosso estudo foi investigar a ocorrência de Sapovírus em granjas de suínos brasileiras. Um total de 166 amostras fecais de suínos, com diferentes idades, dos estados de Minas Gerais, São Paulo e Mato Grosso foram submetidas a reações de RT-PCR e confirmadas com sequenciamento de nucleotídeos do gene RdRp do Sapovírus. Seis (3,61%) amostras foram positivas e quatro delas tinham sequenciamento parcial do gene RdRp, supostamente pertencente ao genogrupo GVII.1, previamente relatado em rebanhos suínos no Brasil.(AU)

Effects of crude protein and lactose levels in diets on growth performance, intestinal morphology, and expression of genes related to intestinal integrity and immune system in weaned piglets.

To evaluate the effects of crude protein (CP) and lactose (LAC) for weaned piglets on performance, intestinal morphology, and expression of genes related to intestinal integrity and immune system, 144 piglets with initial weight 7.17 ± 0.97 kg were allotted in a randomized design, in a 2 × 3 factorial arrangement (20.0% and 24.0% CP and 8.0%, 12.0%, and 16.0% LAC) with eight replicates. Piglets fed 20.0% CP had greater weight gain and feed intake. Including 12.0% LAC in the 20.0% CP diet provided higher villous height in the duodenum than 8.0% LAC, and 12.0% or 16.0% LAC in the 24.0% CP diet resulted in higher villous height in the jejunum and ileum, and higher villi/crypt ratio in the ileum than 8.0% LAC. No effects of CP and LAC on interleukin-1ß and tumor necrosis factor-α mRNA were observed. The 16.0% LAC diet provided higher gene expression of transforming-ß1 growth factor. Feeding 20.0% CP resulted in better performance than 24.0% CP. The 12.0% LAC diet promoted greater genetic expression of occludin and zonula occludens. Including 12.0% LAC in the diet may improve intestinal epithelial morphology and integrity, and these improvements are more evident when piglets are fed diets with 24.0% CP.

The role of viral particle integrity in the serological assessment of foot-and-mouth disease virus vaccine-induced immunity in swine.

The efficacy of foot-and-mouth disease virus (FMDV) inactivated vaccines is mainly dependent on the integrity of the whole (146S) viral particles. If the intact capsids disassemble to 12S subunits, antibodies against internal-not protective epitopes, may be induced. Serological correlates with protection may be hampered if antibodies against internal epitopes are measured. Here we compared the performance of different ELISAs with the virus-neutralization test (VNT) that measures antibodies against exposed epitopes. Sera from pigs immunized with one dose of an expired commercial FMDV vaccine were used. This vaccine contained about 50% of O1/Campos and over 90% of A24/Cruzeiro strains total antigen as whole 146S particles. Specific-total antibodies were measured with the standard liquid-phase blocking ELISA (LPBE). We also developed an indirect ELISA (IE) using sucrose gradient purified 146S particles as capture antigen to titrate total antibodies, IgM, IgG1 and IgG2. A good correlation was found between VNT titers and IgG-ELISAs for A24/Cruzeiro, with the lowest correlation coefficient estimated for IgG2 titers. For O1/Campos, however, the presence of antibodies against epitopes different from those of the whole capsid, elicited by the presence of 12S particles in the vaccine, hampered the correlation between LPBE and VNT, which was improved by using purified O1/Campos 146S-particles for the liquid-phase of the LPBE. Interestingly, 146S particles but not 12S were efficiently bound to the ELISA plates, confirming the efficiency of the IE to detect antibodies against exposed epitopes. Our results indicate that any serological test assessing total antibodies or IgG1 against epitopes exposed in intact 146S-particles correlate with the levels of serum neutralizing antibodies in vaccinated pigs, and might potentially replace the VNT, upon validation. We recommend that antigen used for serological assays aimed to measure protective antibodies against FMDV should be controlled to ensure the preservation of 146S viral particles.

Associação de alterações anatomopatológicas com o Helicobacter spp. por meio de análise imuno-histoquímica em estômagos de leitões

O objetivo deste trabalho foi relacionar os achados anatomopatológicos das lesões gástricas subclínicas de ocorrência natural em leitões com a presença, ou não, de Helicobacter spp. por meio da imuno-histoquímica. Foram utilizados 48 leitões de linhagem genética comercial. Os animais foram adquiridos em uma granja comercial, com peso médio de 34 Kg e idade média de 79 dias; após o abate, seus estômagos foram coletados e avaliados. Avaliações histopatológicas e imuno-histoquímicas foram realizadas em amostras das regiões anatômicas aglandular e glandular. Macroscopicamente, 34 (70,83%) leitões apresentaram lesões na região aglandular, enquanto que em 14 animais (29,17%) não foram encontradas alterações nesta região. Dos estômagos com lesão, 14 foram classificados como grau 1, seis como grau 2 e 14 como grau 3. Microscopicamente, 44 amostras (91,66%) apresentaram paraqueratose. Deste total, 22 apresentaram a forma discreta, 20 a moderada e dois a acentuada. Na avaliação macroscópica da porção glandular, 41 (85,4%) animais apresentaram alteração em pelo menos uma das três regiões, e em somente sete (14,6%) não foram encontradas lesões em nenhuma delas. Em 14 deles, houve aumento da atividade mucípara, em dois, houve erosão e, em cinco, hiperemia.  As lesões na região glandular do estômago foram mais extensas no antro e no cárdia, seguidas do fundo. Em relação à análise imuno

The aim of this study was to relate the anatomopathological findings of naturally occurring subclinical gastric lesions in piglets, with or without Helicobacter spp. through immunohistochemistry. Forty-eight piglets of commercial genetic lineage were used. The animals were acquired in a commercial farm, with an average weight of 34 kg and an average age of 79 days, and after slaughter, their stomachs were collected and evaluated. Samples from the glandular and aglandular anatomical regions were evaluated. Macroscopically, 34 (70.83%) samples had lesions on aglandular region, while 14 (29.17%) nothing had. Of the injured stomachs, 14 were classified as grade 1, six as grade 2 and 14 as grade 3. Microscopically, 44 samples (91.66%) showed parakeratosis. Of these, 22 showed a discreet manner, 20 moderate and two severe. In the glandular region, in 41 (85.4%) samples there was a change in at least one of the three regions, and only seven animals (14.6%) showed no change in any of the three. Fourteen samples showed increased muciparous activity, two showed erosion and five hyperemia. The lesions were higher in antral regions and cardic, followed the fundus. In relation to immunohistochemistry, 21(43.8%) samples were negative in all areas, 24 (50%) were positive in at least one, and none were positive in all. The anatomopathological findings showed a statistical relationship with the bacteria, and its immunostaining, not associated with gastric lesions in certain regions, demonstrates its saprophytic and opportunistic character.

Occurrence of antibodies against Neospora caninum in sows and factors associated with infection in commercial herds in two regions of the state of Santa Catarina, Brazil / Ocorrência de anticorpos contra Neospora caninum em porcas e fatores associados à infecção em rebanhos comerciais de duas regiões de Santa Catarina, Brasil

Neosporosis is a disease that affects ruminants, horses, swine, and wild animals, causing reproductive disorders and economic losses. The importance of the infection in pigs has not been fully elucidated. The objective of the present study was to verify the occurrence of antibodies against Neospora caninum and the possible factors associated with infection in swine herds from two regions of the state of Santa Catarina, Brazil. A total of 498 blood samples were collected from sows of western (five properties) and southern (four properties) regions of the state, and an epidemiological questionnaire was administered. Sera obtained from the blood were tested by indirect fluorescent antibody test (IFAT) (cutoff point 1:50) using tachyzoites of N. caninum NC1 strain as antigen. Positive samples were two-fold serially diluted until maximum reactivity titer. The results were analyzed using Fisher and Chi-square tests to determine correlation between serology and associated factors. The overall prevalence was 18.9% (94/498) with titers of 50 (27), 100 (26), 200 (22), 400 (10), and 800 (9). Eighty of 256 samples (31.3%) from the western region were positive, while 14 out of 242 samples (5.8%) from the southern region were positive. A correlation (p < 0.01) was observed among the seroprevalence and contact with dogs (OR = 3.97, IC 2.64-6.43), presence of rodents (OR = 7.37 IC 4.06-13.50), and water source (OR = 2.38 IC 1.44-3.91). The use of anti-bird nets was identified as a protective factor against infection (OR = 0.36 IC 0.07-0.25). These results demonstrate the high prevalence of antibodies against N. caninum in commercial swine herds in the state of Santa Catarina compared to that in other regions of Brazil.(AU)

Neosporose é uma doença que afeta ruminantes, equinos, suínos e animais selvagens, causando distúrbios reprodutivos e perdas econômicas. A importância da infecção em suínos não está totalmente elucidada. O objetivo do presente estudo foi verificar a ocorrência de anticorpos contra Neospora caninum e os possíveis fatores associados à infecção em rebanhos suínos de duas regiões do estado de Santa Catarina. Foram coletadas 498 amostras de sangue de porcas nas regiões oeste (cinco propriedades) e sul (quatro propriedades) do estado, e foi aplicado um questionário epidemiológico. Os soros obtidos foram avaliados pela Reação de Imunofluorescência Indireta (RIFI, 1:50) usando taquizoítos da cepa NC1 de N. caninum como antígeno. As amostras positivas foram analisadas em diluições seriadas de base dois até a titulação máxima reativa. Os resultados foram analisados por meio dos testes de Fisher e do qui-quadrado para determinar a correlação entre sorologia e fatores de risco. A prevalência geral foi de 18,9% (94/498), com títulos de 50 (27), 100 (26), 200 (22), 400 (10) e 800 (9). Das 256 amostras da região oeste, 80 (31,3%) foram positivas, enquanto 14 (5,8%) das 242 amostras da região sul foram positivas. Observou-se correlação (p < 0,01) entre soroprevalência e contato com cães (OR = 3,97; IC 2,64-6,43), presença de roedores (OR = 7,37 IC 4,06-13,50) e fonte de água (OR = 2,38 IC 1,44- 3,91). O uso de redes antipassáros foi identificado como fator de proteção contra a infecção (OR = 0,36 IC 0,07-0,25). Esses resultados demonstram a alta prevalência de anticorpos contra N. caninum em rebanhos suínos comerciais no estado de Santa Catarina em comparação com outras regiões do Brasil.(AU)

Occurrence of antibodies against Neospora caninum in sows and factors associated with infection in commercial herds in two regions of the state of Santa Catarina, Brazil / Ocorrência de anticorpos contra Neospora caninum em porcas e fatores associados à infecção em rebanhos comerciais de duas regiões de Santa Catarina, Brasil

Neosporosis is a disease that affects ruminants, horses, swine, and wild animals, causing reproductive disorders and economic losses. The importance of the infection in pigs has not been fully elucidated. The objective of the present study was to verify the occurrence of antibodies against Neospora caninum and the possible factors associated with infection in swine herds from two regions of the state of Santa Catarina, Brazil. A total of 498 blood samples were collected from sows of western (five properties) and southern (four properties) regions of the state, and an epidemiological questionnaire was administered. Sera obtained from the blood were tested by indirect fluorescent antibody test (IFAT) (cutoff point 1:50) using tachyzoites of N. caninum NC1 strain as antigen. Positive samples were two-fold serially diluted until maximum reactivity titer. The results were analyzed using Fisher and Chi-square tests to determine correlation between serology and associated factors. The overall prevalence was 18.9% (94/498) with titers of 50 (27), 100 (26), 200 (22), 400 (10), and 800 (9). Eighty of 256 samples (31.3%) from the western region were positive, while 14 out of 242 samples (5.8%) from the southern region were positive. A correlation (p < 0.01) was observed among the seroprevalence and contact with dogs (OR = 3.97, IC 2.64-6.43), presence of rodents (OR = 7.37 IC 4.06-13.50), and water source (OR = 2.38 IC 1.44-3.91). The use of anti-bird nets was identified as a protective factor against infection (OR = 0.36 IC 0.07-0.25). These results demonstrate the high prevalence of antibodies against N. caninum in commercial swine herds in the state of Santa Catarina compared to that in other regions of Brazil.

Neosporose é uma doença que afeta ruminantes, equinos, suínos e animais selvagens, causando distúrbios reprodutivos e perdas econômicas. A importância da infecção em suínos não está totalmente elucidada. O objetivo do presente estudo foi verificar a ocorrência de anticorpos contra Neospora caninum e os possíveis fatores associados à infecção em rebanhos suínos de duas regiões do estado de Santa Catarina. Foram coletadas 498 amostras de sangue de porcas nas regiões oeste (cinco propriedades) e sul (quatro propriedades) do estado, e foi aplicado um questionário epidemiológico. Os soros obtidos foram avaliados pela Reação de Imunofluorescência Indireta (RIFI, 1:50) usando taquizoítos da cepa NC1 de N. caninum como antígeno. As amostras positivas foram analisadas em diluições seriadas de base dois até a titulação máxima reativa. Os resultados foram analisados por meio dos testes de Fisher e do qui-quadrado para determinar a correlação entre sorologia e fatores de risco. A prevalência geral foi de 18,9% (94/498), com títulos de 50 (27), 100 (26), 200 (22), 400 (10) e 800 (9). Das 256 amostras da região oeste, 80 (31,3%) foram positivas, enquanto 14 (5,8%) das 242 amostras da região sul foram positivas. Observou-se correlação (p < 0,01) entre soroprevalência e contato com cães (OR = 3,97; IC 2,64-6,43), presença de roedores (OR = 7,37 IC 4,06-13,50) e fonte de água (OR = 2,38 IC 1,44- 3,91). O uso de redes antipassáros foi identificado como fator de proteção contra a infecção (OR = 0,36 IC 0,07-0,25). Esses resultados demonstram a alta prevalência de anticorpos contra N. caninum em rebanhos suínos comerciais no estado de Santa Catarina em comparação com outras regiões do Brasil.

Protection against Toxoplasma gondii cysts in pigs immunized with rROP2 plus Iscomatrix / Proteção contra cistos de Toxoplasma gondii em suínos imunizados com rROP2 associado à Iscomatrix

This study aimed to evaluate the humoral immune response in pigs immunized intranasally and intramuscularly with recombinant Toxoplasma gondii rROP2 protein in combination with the adjuvant Iscomatrix. Twelve mixed breed pigs divided into three groups (n=4) were used, G1 received recombinant ROP2 proteins (200 µg/dose) plus Iscomatrix, G2 received PBS plus Iscomatrix, and G3 as the control group. The intranasal (IN) and intramuscular (IM) routes were used. Animals were challenged orally with VEG strain oocysts and treated on day three after challenge. Fever, anorexia, and prostration were the clinical signs observed in all animals. All the G1 animals produced antibodies above the cut-off on the day of the challenge, while the G2 and G3 remained below the cut-off. Better partial protection against parasitemia and cyst tissue formation was observed in G1 than G3. The protection factors against tissue cyst formation were 40.0% and 6.1% for G1 and G2, respectively, compared to G3. In conclusion, there were not systemic antibody responses in pigs with IN immunization with rROP2+Iscomatrix; however, after IM immunization, those animals produced higher titers than animal controls. We associated these results with partial protection obtained against parasitemia and tissue cysts formation.(AU)

O objetivo deste estudo foi avaliar a resposta imune humoral em suínos imunizados pelas vias intranasal e intramuscular com proteínas recombinantes rROP2 do Toxoplasma gondii associadas ao adjuvante Iscomatrix. Doze suínos cruzados divididos em 3 grupos (n=4) foram utilizados. O G1 recebeu proteína recombinante ROP2 (200mg/dose) associada ao adjuvante Iscomatrix; o G2 recebeu PBS associado ao Iscomatrix; e o G3 foi o grupo controle. As vias intranasal (IN) e intramuscular (IM) foram utilizadas. Os animais foram desafiados por via oral com a cepa VEG e tratados no dia três após o desafio. Febre, anorexia e prostração foram os sinais clínicos observados em todos os animais. Todos os animais do G1 produziram anticorpos acima do ponto de corte no dia do desafio, enquanto os animais do G2 e G3 permaneceram abaixo do ponto de corte no desafio. Proteção parcial contra parasitemia e formação de cistos teciduais foram observadas nos suínos do G1 comparados ao G3. Os fatores de proteção contra a formação de cistos teciduais foram 40,0% e 6,1% no G1 e G2, respectivamente, comparados com o G3. Como conclusão, não houve estimulação da resposta imune humoral sistêmica nos suínos após as imunizações IN com rROP2+Iscomatrix. Estes animais, porém, após a imunização IM, produziram títulos de anticorpos mais altos que os animais controles. Esses resultados foram associados a uma proteção parcial contra a parasitemia e formação de cistos teciduais.(AU)

Associação de alterações anatomopatológicas com o Helicobacter spp. por meio de análise imuno-histoquímica em estômagos de leitões

O objetivo deste trabalho foi relacionar os achados anatomopatológicos das lesões gástricas subclínicas de ocorrência natural em leitões com a presença, ou não, de Helicobacter spp. por meio da imuno-histoquímica. Foram utilizados 48 leitões de linhagem genética comercial. Os animais foram adquiridos em uma granja comercial, com peso médio de 34 Kg e idade média de 79 dias; após o abate, seus estômagos foram coletados e avaliados. Avaliações histopatológicas e imuno-histoquímicas foram realizadas em amostras das regiões anatômicas aglandular e glandular. Macroscopicamente, 34 (70,83%) leitões apresentaram lesões na região aglandular, enquanto que em 14 animais (29,17%) não foram encontradas alterações nesta região. Dos estômagos com lesão, 14 foram classificados como grau 1, seis como grau 2 e 14 como grau 3. Microscopicamente, 44 amostras (91,66%) apresentaram paraqueratose. Deste total, 22 apresentaram a forma discreta, 20 a moderada e dois a acentuada. Na avaliação macroscópica da porção glandular, 41 (85,4%) animais apresentaram alteração em pelo menos uma das três regiões, e em somente sete (14,6%) não foram encontradas lesões em nenhuma delas. Em 14 deles, houve aumento da atividade mucípara, em dois, houve erosão e, em cinco, hiperemia.  As lesões na região glandular do estômago foram mais extensas no antro e no cárdia, seguidas do fundo. Em relação à análise imuno(AU)

The aim of this study was to relate the anatomopathological findings of naturally occurring subclinical gastric lesions in piglets, with or without Helicobacter spp. through immunohistochemistry. Forty-eight piglets of commercial genetic lineage were used. The animals were acquired in a commercial farm, with an average weight of 34 kg and an average age of 79 days, and after slaughter, their stomachs were collected and evaluated. Samples from the glandular and aglandular anatomical regions were evaluated. Macroscopically, 34 (70.83%) samples had lesions on aglandular region, while 14 (29.17%) nothing had. Of the injured stomachs, 14 were classified as grade 1, six as grade 2 and 14 as grade 3. Microscopically, 44 samples (91.66%) showed parakeratosis. Of these, 22 showed a discreet manner, 20 moderate and two severe. In the glandular region, in 41 (85.4%) samples there was a change in at least one of the three regions, and only seven animals (14.6%) showed no change in any of the three. Fourteen samples showed increased muciparous activity, two showed erosion and five hyperemia. The lesions were higher in antral regions and cardic, followed the fundus. In relation to immunohistochemistry, 21(43.8%) samples were negative in all areas, 24 (50%) were positive in at least one, and none were positive in all. The anatomopathological findings showed a statistical relationship with the bacteria, and its immunostaining, not associated with gastric lesions in certain regions, demonstrates its saprophytic and opportunistic character.(AU)

Serum concentration of acute phase proteins and cytokines in vaccinated pigs challenged with foot-and-mouth disease virus serotype O

The present study aimed to investigate the kinetics of acute phase proteins and cytokines in the serum of immunized or naive pigs following infection with a field isolate of foot-and-mouth disease (FMD) virus. Five-month-old SPF pigs were intramuscularly immunized with or without the 146S antigen (7.5 µg per head) purified from FMD virus (O/Andong/SKR/2010) and were subsequently challenged 30 days post-vaccination (dpv) with FMD virus on each footpad. Serum samples were obtained at 28 dpv and on days 1, 3, and 5 post-infection (dpi). At 28 dpv, the serum concentrations of C-reactive protein and tumor necrosis factor alpha (TNF-α) were higher in the immunized pigs compared with the non-immunized control pigs. Upon challenge, the levels of all measured acute phase proteins (i.e., C-reactive protein, major acute phase protein, S-amyloid A, and haptoglobin) plateaued at 1 dpi in the immunized pigs whereas they gradually increased up to 5 dpi in the non-immunized control pigs. Levels of serum TNF-α remained higher in the immunized challenged pigs compared with the non-immunized challenged pigs. The serum concentration of interleukin (IL)-6 in the immunized challenged pigs was undetectable at all time points, but moderately increased upon challenge in the non-immunized challenged pigs. IL-1ß was not detected in any of the pigs at any of the observed time points. Collectively, these findings show that the immunized pigs exhibit a rapid serum acute phase protein response following subsequent challenge with the field FMD virus compared with the non-immunized pigs.(AU)

Serological prevalence of Brucella spp. in feral pigs and sympatric cattle in the Pantanal of Mato Grosso do Sul, Brazil / Prevalência sorológica de Brucella spp. em porcos ferais e bovinos em simpatria no Pantanal do Mato Grosso do Sul, Brasil

The objective of this study was to determine the prevalence of anti-Brucella antibodies in feral pigs and bovines simpatrics in the Pantanal subregions of Paiaguás and Nhecolândia. The study was conducted in the municipality of Corumbá, Mato Grosso do Sul State, Brazil. A total of 105 feral pigs and 256 cattle were sampled in 12 farms, in all animals blood samples were collected for the serological diagnosis with Rose Bengal Test (RBT) for screening, 2-Mercaptoethanol (2-ME) confirmatory test and comparative test with Fluorescence Polarization Assay (FPA). The prevalence of positive feral pigs were 1% (1/105) in the RBT and FPA and no positive AAT results were confirmed in the 2-ME test. The prevalence of positive cattle sampled was 11.32%, 4.3% and 7.42% in the RBT, 2-ME and FPA tests respectively. The degree of agreement obtained between the serological tests used in cattle was Kappa = 0.506 (p 0.001), 95% CI (0.282 - 0.729). The results of the serological tests demonstrated that brucellosis is widespread in bovine herds of the region studied, but the same type of exposure to the agent did not occur in feral pigs according to the diagnostic tests used.(AU)

O objetivo deste estudo foi estimar a prevalência de anticorpos anti-Brucella em porcos ferais e bovinos simpátricos nas sub-regiões pantaneiras do Paiaguás e Nhecolândia. O estudo foi conduzido no município de Corumbá, Estado de Mato Grosso do Sul, Brasil. Foram amostrados 105 porcos ferais e 256 bovinos em 12 propriedades, em todos os animais foram coletadas amostras de sangue para o diagnóstico sorológico com Antígeno Acidificado Tamponado (AAT) para triagem, teste confirmatório 2-Mercaptoetanol (2-ME) e comparativo com Teste de Polarização Fluorescente (TPF). A prevalência de porcos ferais positivos foi de 1% (1/105) no AAT e TPF, não tendo sido confirmado nenhum resultado positivo do AAT no 2-ME. A prevalência de bovinos positivos amostrados foi de 11,32% (29/256), 4,3% (10/256) e de 7,42% (19/256) nos testes AAT, 2-ME e no TPF, respectivamente. O grau de concordância obtido entre os testes sorológicos utilizados nos bovinos foi de Kappa = 0,506 (p 0.001), 95% CI (0.282 0.729). Os resultados dos testes sorológicos demonstraram que a brucelose está disseminada nos rebanhos bovinos da região estudada, porém o mesmo tipo de exposição ao agente não ocorreu nos porcos ferais de acordo com os testes de diagnóstico utilizados.(AU)

IgG Induced by Vaccination With Ascaris suum Extracts Is Protective Against Infection.

Human ascariasis has a global and cosmopolitan distribution, and has been characterized as the most prevalent neglected tropical disease worldwide. The development of a preventive vaccine is highly desirable to complement current measures required for this parasitic infection control and to reduce chronic childhood morbidities. In the present study, we describe the mechanism of protection elicited by a preventive vaccine against ascariasis. Vaccine efficacy was evaluated after immunization with three different Ascaris suum antigen extracts formulated with monophosphoryl lipid A (MPLA) as an adjuvant: crude extract of adult worm (ExAD); crude extract of adult worm cuticle (CUT); and crude extract of infective larvae (L3) (ExL3). Immunogenicity elicited by immunization was assessed by measuring antibody responses, cytokine production, and influx of tissue inflammatory cells. Vaccine efficacy was evaluated by measuring the reductions in the numbers of larvae in the lungs of immunized BALB/c mice that were challenged with A. suum eggs. Moreover, lung physiology and functionality were tested by spirometry to determine clinical efficacy. Finally, the role of host antibody mediated protection was determined by passive transfer of serum from immunized mice. Significant reductions in the total number of migrating larvae were observed in mice immunized with ExL3 61% (p < 0.001), CUT 59% (p < 0.001), and ExAD 51% (p < 0.01) antigens in comparison with non-immunized mice. For the Ascaris antigen-specific IgG antibody levels, a significant and progressive increase was observed with each round of immunization, in association with a marked increase of IgG1 and IgG3 subclasses. Moreover, a significant increase in concentration of IL-5 and IL-10 (pre-challenge) in the blood and IL-10 in the lung tissue (post-challenge) was induced by CUT immunization. Finally, ExL3 and CUT-immunized mice showed a marked improvement in lung pathology and tissue fibrosis as well as reduced pulmonary dysfunction induced by Ascaris challenge, when compared to non-immunized mice. Moreover, the passive transfer of specific IgG antibodies from ExL3, CUT, and ExAD elicited a protective response in naïve mice, with significant reductions in parasite burdens in lungs of 65, 64, and 64%, respectively. Taken together, these studies indicated that IgG antibodies contribute to protective immunity.

Efficacy of energy supplementation on growth performance and immune response of suckling pigs.

Three studies were performed to determine the effects of oral energy sources on the performance, immune status, and intestinal morphology of piglets. In Exp. 1, 50 litters were selected based on genotype and parity order to determine the optimum amount of supplemental energy for neonatal pigs. They were distributed according to a 5 × 5 Latin square design, in which columns were represented by 5 sows and rows were represented by 5 newborn weight categories. Treatments consisted of 2 oral doses of 0, 2, 4, 8, or 16 mL of rice bran oil (RBO). There was a positive response of RBO on BW with a numerical difference (P = 0.12) and a significant linear effect (P = 0.04) at weaning. In Exp. 2, litters (n = 340) were randomly allotted to a 2 × 2 factorial arrangement of treatments: RBO supplemented either through oral doses (without supplementation or with 2 mL of RBO) or through prestarter feed (without supplementation or with 2% of RBO replacing 2% of soybean oil). Piglets orally receiving RBO showed a numerical increase in BW at weaning (P = 0.101) compared with the control group. Moreover, piglets supplemented by prestarter feed tended to increase feed intake (P = 0.084). In Exp. 3, litters (n = 245) were selected based on genotype and parity order and they were distributed according to a 7 × 7 Latin square design planed in the same way as in Exp. 1. Each neonatal pig within a litter was randomly assigned to 1 of 7 treatments: 1) control (no supplementation), 2) 2 mL of RBO enriched with omega-3 fatty acids, 3) 2.33 mL of pure glycerin, 4) 1.3 mL of soybean oil, 5) 1.4 mL of linseed oil, 6) 1.68 mL of coconut oil, and 7) 2 mL of RBO. Animal performance was analyzed using 2 data sets: all data (average initial BW of 1.479 kg) and low birth weight (LBW) piglet data (≤1.220 kg initial BW with average of 0.985 kg). Body weight or BW gain (BWG) were not different among treatments (P > 0.10) when all data were analyzed. However, LBW piglets fed coconut oil tended to increase BW (P = 0.099) during the first week. In conclusion, the oral use of RBO may increase BW at weaning and the supplementation by prestarter feed may enhance the feed intake of piglets. In addition, coconut oil may increase BW of LBW piglets in the first week of life without affecting mortality, immune response, or gut morphology. The routine practice of oral supplementation of energy for newborn pigs could be an important tool in swine production.

Human Breast-Milk Feeding Enhances the Humoral and Cell-Mediated Immune Response in Neonatal Piglets.

Background: The benefits of breastfeeding infants are well characterized, including those on the immune system. However, determining the mechanism by which human breast milk (HBM) elicits effects on immune response requires investigation in an appropriate animal model. Objective: The primary aim of this study was to develop a novel porcine model and to determine the differential effects of feeding HBM and a commercial milk formula (MF) on immune response and gastrointestinal microbial colonization in a controlled environment. Methods: Male piglets were fed HBM (n = 26) or MF (n = 26) from day 2 through day 21. Piglets were vaccinated (n = 9/diet group) with cholera toxin and cholera toxin subunit B (CTB) and tetanus toxoid at 21 d or were fed placebo (n = 6/diet group) and then weaned to a standard solid diet at the age of 21 d. Humoral and cell-mediated immune responses were assessed from blood on days 35 and 48. Immune response was further examined from tissues, including mesenteric lymph nodes (MLNs), Peyer's patches (PPs), and spleen. The colonization of gut microbiota was characterized from feces on days 16 and 49. Results: Serum antibody titers in piglets fed HBM were 4-fold higher (P < 0.05) to CTB and 3-fold higher (P < 0.05) to tetanus toxoid compared with piglets fed MF on day 48. Compared with MF, the numbers of immunoglobulin A antibody-producing cells to CTB were 13-fold higher (P < 0.05) in MLNs and 11-fold higher (P < 0.05) in PPs in the HBM diet group on day 51. In addition, significantly increased T cell proliferation was observed in the HBM group relative to the MF group. Furthermore, microbial diversity in the HBM group was lower (P < 0.05) than in the MF group. Conclusions: This porcine model appears to be valid for studying the effects of early postnatal diet on immune responses and the gastrointestinal microbiome. Our results lay the groundwork for future studies defining the role of infant diet on microbiota and immune function.

Efficacy of leptospiral commercial vaccines on the protection against an autochtonous strain recovered in Brazil

In swine and bovines, leptospirosis prevention and control is carried out via vaccination of susceptible animals using bacterins. However, the efficiency of leptospirosis vaccines has been questioned. This work aimed to investigate the potency of five leptospirosis vaccines sold commercially in Brazil, challenging the animals with one autochthonous strain of Leptospira, Canicola serovar, denoted LO4, isolated from swine. The standard protocol was followed, and renal carriers of Leptospira were identified among the surviving animals by culture and PCR. Of the five vaccines tested, only two proved effective. None of the surviving animals was positive by culture; however, one animal was positive by PCR. Three of the five vaccines sold commercially in Brazil for the immunization of swine or bovines failed the test of the efficacy to protect the vaccinated animals following challenge with an autochthonous Leptospira strain, Canicola serovar. The two vaccines provided protection against the renal carrier state in the surviving animals. The criteria used to produce leptospirosis bacterins sold commercially in Brazil must be reviewed. The industry should support researches on leptospiral vaccinology to improve the quality of the present vaccines and discover new immunogenic strains, because it is known that vaccination is one of the most important tools to increase the reproduction rates in livestock.(AU)

Characterization and expression of DEC205 in the cDC1 and cDC2 subsets of porcine dendritic cells from spleen, tonsil, and submaxillary and mesenteric lymph nodes.

Conventional dendritic cells (cDCs) are divided into the following different subtypes: cDC1, which promotes a Th1 response, and cDC2, which stimulates a Th2 and Th17 response. These cells have not been characterized in porcine lymphoid tissues. DEC205 is a receptor that increases antigen presentation and allows DCs to cross-present antigens. The objectives of this work were to characterize cDCs subsets in the tonsil, submaxillary and mesenteric lymph nodes and spleen lymphoid tissues and to determine their expression of DEC205 by flow cytometry. The cDC1 (MHCIIhighCADM1highCD172a-/low) and cDC2 (MHCIIhighCADM1highCD172a+) phenotypes were confirmed by the expression of characteristic cDC1 and cDC2 transcripts (FLT3, XCR1 and FCER1α). Among all lymphoid tissues, the spleen had the highest frequency of total cDCs. The cDC1:cDC2 ratio showed that all lymph tissues had higher levels of cDC1 than levels of cDC2. DEC205+ cDCs were found in all analyzed tissues, albeit with different frequencies. Our research will facilitate the study on the function of these cells and the investigation of the strategies for DEC205 targeting and functional studies.

Serological prevalence of Brucella spp. in feral pigs and sympatric cattle in the Pantanal of Mato Grosso do Sul, Brazil / Prevalência sorológica de Brucella spp. em porcos ferais e bovinos em simpatria no Pantanal do Mato Grosso do Sul, Brasil

The objective of this study was to determine the prevalence of anti-Brucella antibodies in feral pigs and bovines simpatrics in the Pantanal subregions of Paiaguás and Nhecolândia. The study was conducted in the municipality of Corumbá, Mato Grosso do Sul State, Brazil. A total of 105 feral pigs and 256 cattle were sampled in 12 farms, in all animals blood samples were collected for the serological diagnosis with Rose Bengal Test (RBT) for screening, 2-Mercaptoethanol (2-ME) confirmatory test and comparative test with Fluorescence Polarization Assay (FPA). The prevalence of positive feral pigs were 1% (1/105) in the RBT and FPA and no positive AAT results were confirmed in the 2-ME test. The prevalence of positive cattle sampled was 11.32%, 4.3% and 7.42% in the RBT, 2-ME and FPA tests respectively. The degree of agreement obtained between the serological tests used in cattle was Kappa = 0.506 (p 0.001), 95% CI (0.282 - 0.729). The results of the serological tests demonstrated that brucellosis is widespread in bovine herds of the region studied, but the same type of exposure to the agent did not occur in feral pigs according to the diagnostic tests used.

O objetivo deste estudo foi estimar a prevalência de anticorpos anti-Brucella em porcos ferais e bovinos simpátricos nas sub-regiões pantaneiras do Paiaguás e Nhecolândia. O estudo foi conduzido no município de Corumbá, Estado de Mato Grosso do Sul, Brasil. Foram amostrados 105 porcos ferais e 256 bovinos em 12 propriedades, em todos os animais foram coletadas amostras de sangue para o diagnóstico sorológico com Antígeno Acidificado Tamponado (AAT) para triagem, teste confirmatório 2-Mercaptoetanol (2-ME) e comparativo com Teste de Polarização Fluorescente (TPF). A prevalência de porcos ferais positivos foi de 1% (1/105) no AAT e TPF, não tendo sido confirmado nenhum resultado positivo do AAT no 2-ME. A prevalência de bovinos positivos amostrados foi de 11,32% (29/256), 4,3% (10/256) e de 7,42% (19/256) nos testes AAT, 2-ME e no TPF, respectivamente. O grau de concordância obtido entre os testes sorológicos utilizados nos bovinos foi de Kappa = 0,506 (p 0.001), 95% CI (0.282 0.729). Os resultados dos testes sorológicos demonstraram que a brucelose está disseminada nos rebanhos bovinos da região estudada, porém o mesmo tipo de exposição ao agente não ocorreu nos porcos ferais de acordo com os testes de diagnóstico utilizados.

Chito-oligosaccharide as growth promoter replacement for weaned piglets: performance, morphometry, and immune system / Quitooligossacarídeo como substituto aos promotores de crescimento para leitões desmamados: desempenho, morfometria e imunidade

The objective of this study was to evaluate the chito-oligosaccharide (COS) against two growth promoter antibiotics, colistin and lincomycin, with respect to growth performance, incidence of diarrhea, visceral characteristics, morphometry, and serum immunoglobulin levels (IgA, IgG, and IgM). A total of 96 Pen Ar Lan® piglets (48 barrows and 48 females), weaned at 17 days and with body weight (BW) 5.33 ± 0.37 kg, were subjected to the evaluation of growth performance and serum. Twenty-four animals, females, 35-day-old and with BW 6.86 ± 0.64 kg, were used for the assessment of histology and visceral organ weight. The three treatments were a basic diet formulation supplemented with COS (100 mg kg-1), colistin (40 mg kg-1), or lincomycin (4.4 mg kg-1). The antibiotic treatments showed higher average daily gain (ADG) than COS treatment during the period of 49 to 63 days; whereas the feed conversion ratio (FCR) was higher and incidence of diarrhea was lower for the colistin treatment than for other treatments. The spleen weight and the small intestinal length were higher and duodenal pH was lower for COS than for antibiotics. Morphometry indicated greater villus height and higher ratio of villus height to crypt depth with colistin than with COS and a lower lesion score compared with other treatments. The serum IgA concentration was higher for COS in 35-day-old piglets. According to the results, COS wasnot efficient to replace colistin as a growth promoter for piglets weaned at 17 days; however, the resultsrelated to the immune system suggested that COS is a potentially promising product during weaning.(AU)

Realizou-se este trabalho com o objetivo de avaliar o quitooligossacarídeo (QOS) frente a dois antibióticos utilizados como promotores de crescimento, a colistina e lincomicina, quanto ao desempenho, frequência de diarreia, características viscerais, morfometria e imunoglobulinas séricas, IgA, IgG e IgM. Foram utilizados 96 leitões de genética Pen Ar Lan®, desmamados com idade média de 17 dias e peso de 5,33 ± 0,369 kg para avaliação de desempenho e imunoglobulinas séricas. Para a avaliação histológica e peso dos órgãos, foram utilizados 24 animais com idade de 35 dias e peso de 6,86 ± 0,64 kg. Os três tratamentos consistiram em uma dieta basal acrescida de QOS (100 mg Kg -1), dieta basal com colistina 40 mg Kg -1 e dieta basal com lincomicina 4,4 mg Kg -1. Para o desempenho, os antibióticos apresentaram melhor ganho diário de peso (GPD) dos 49 aos 63 dias de idade em relação ao QOS e a conversão alimentar (CA) foi melhor para a colistina em comparação com os demais tratamentos, além disso apresentou menor incidência de diarreia aquosa quando comparada com os outros tratamentos. Para as vísceras, observou-se aumento do peso relativo do baço, maior comprimento do intestino delgado e menor pH duodenal para os leitões que receberam QOS em relação aos antibióticos. A morfometria indicou maior altura de vilosidades e melhor relação vilos/cripta para a colistina em relação ao QOS e menor escore lesional em relação aos demais tratamentos. A concentração sérica de imunoglobulinas apontou uma maior concentração de IgA para o QOS aos 35 dias. Frente aos resultados observados, embora o QOS tenha promovido estímulos sobre o sistema imune do animal, este não foi eficiente parasubstituir a colistina como promotor de crescimento para leitões desmamados com 17 dias de idade.(AU)