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Toxocariasis is a zoonotic disease of worldwide distribution. The connection between parasitic diseases and conditions that depress the immune system, such as the use of immunosuppressive drugs, has been studied. The purpose of this study was to evaluate the effect of Cyclosporine A (CsA) on the intensity of infection, humoral response and gene transcription of interleukins IL-4, IL-10 and IL-12 in mice experimentally infected with Toxocara canis. To this end, mice were divided into two groups treated with CsA (G1: 10 mg/Kg and G2: 50 mg/kg), the G3 and G4 group received PBS. After the last administration of the drug or PBS (orally every 48 hours for 15 days), groups G1, G2 and G3 were inoculated with 1200 eggs of T. canis. Was collected blood samples on days zero, 15 and 30 days post-inoculation (PI), for ELISA test and the mice were euthanized 30 days PI. The organs and striated muscle tissue were collected for the recovery of larvae. The splenocytes were analyzed by RT-PCR. The intensity of infection in the mice treated with 50 mg/kg of CsA was 65.5% higher than in the control group (p=0.001). An analysis of the kinetics of anti-Toxocara antibody revealed that the groups treated with CsA showed significantly higher mean levels of antibodies on day 15 PI. The transcription of the three tested interleukins showed no statistical difference between G2 and G3 (control). It was concluded that the immunosuppression triggered by CsA (50 mg/Kg) favored the establishment of a larger number of T. canis larvae without, however, altering immunoglobulin production and IL-4, IL-10 and IL-12 transcription on day 30 PI.

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Toxocariasis is a zoonotic disease of worldwide distribution. The connection between parasitic diseases and conditions that depress the immune system, such as the use of immunosuppressive drugs, has been studied. The purpose of this study was to evaluate the effect of Cyclosporine A (CsA) on the intensity of infection, humoral response and gene transcription of interleukins IL-4, IL-10 and IL-12 in mice experimentally infected with Toxocara canis. To this end, mice were divided into two groups treated with CsA (G1: 10 mg/Kg and G2: 50 mg/kg), the G3 and G4 group received PBS. After the last administration of the drug or PBS (orally every 48 hours for 15 days), groups G1, G2 and G3 were inoculated with 1200 eggs of T. canis. Was collected blood samples on days zero, 15 and 30 days post-inoculation (PI), for ELISA test and the mice were euthanized 30 days PI. The organs and striated muscle tissue were collected for the recovery of larvae. The splenocytes were analyzed by RT-PCR. The intensity of infection in the mice treated with 50 mg/kg of CsA was 65.5% higher than in the control group (p=0.001). An analysis of the kinetics of anti-Toxocara antibody revealed that the groups treated with CsA showed significantly higher mean levels of antibodies on day 15 PI. The transcription of the three tested interleukins showed no statistical difference between G2 and G3 (control). It was concluded that the immunosuppression triggered by CsA (50 mg/Kg) favored the establishment of a larger number of T. canis larvae without, however, altering immunoglobulin production and IL-4, IL-10 and IL-12 transcription on day 30 PI.

A toxocaríase é uma zoonose de distribuição mundial. A conexão entre doenças parasitárias e condições que deprimem o sistema imunológico, como o uso de drogas imunossupressoras, tem sido estudada. O objetivo deste estudo foi avaliar o efeito da Ciclosporina A (CsA) na intensidade da infecção, resposta humoral e transcrição gênica das interleucinas IL-4, IL-10 e IL-12 em camundongos experimentalmente infectados com Toxocara canis. Para tanto, os camundongos foram divididos em dois grupos tratados com CsA (G1: 10 mg/Kg e G2: 50 mg/kg), os grupos G3 e G4 receberam PBS. Após a última administração da droga ou PBS (via oral a cada 48 horas por 15 dias), os grupos G1, G2 e G3 foram inoculados com 1200 ovos de T. canis. Foram coletadas amostras de sangue nos dias zero, 15 e 30 dias pós-inoculação (PI), para teste de ELISA e os camundongos foram eutanasiados 30 dias PI. Os órgãos e tecido muscular estriado foram coletados para a recuperação das larvas. Os esplenócitos foram analisados ​​por RT-PCR. A intensidade da infecção nos camundongos tratados com 50 mg/kg de CsA foi 65,5% maior do que no grupo controle (p=0,001). Uma análise da cinética do anticorpo anti-Toxocara revelou que os grupos tratados com CsA apresentaram níveis médios de anticorpos significativamente maiores no dia 15 PI. A transcrição das três interleucinas testadas não apresentou diferença estatística entre G2 e G3 (controle). Concluiu-se que a imunossupressão desencadeada pela CsA (50 mg/Kg) favoreceu o estabelecimento de um maior número de larvas de T. canis sem, no entanto, alterar a produção de imunoglobulinas e a transcrição de IL-4, IL-10 e IL-12 no dia 30 PI.

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AIM: The aim of the present study was to examine the vaccine immune response in ewes supplemented with Bacillus toyonensis BCT-7112T during a period of 5-day supplementation before vaccination against a recombinant Clostridium perfringens epsilon toxin (rETX). METHODS AND RESULTS: Ewes were vaccinated with 200 µg of rETX adjuvanted with 10% aluminium hydroxide. The treat group was orally supplemented with B. toyonensis BCT-7112T (3 × 108 viable spores) for 5 days prior to the first and second vaccination. Ewes supplemented with B. toyonensis BCT-7112T showed higher neutralizing antibody titres than the non-supplemented ewes (P < 0·05), with an increase in serum levels for total IgG anti-rETX by 3·2-fold (P < 0·0001), and for both IgG isotypes IgG1 and IgG2 by 2·1-fold and 2·3-fold (P < 0·01), respectively, compared with the control group. The peripheral blood mononuclear cells of ewes in the supplemented group had a higher (P < 0·05) cytokine mRNA transcription levels for IL-2 (6·4-fold increase), IFN-γ (2·9-fold increase) and transcription factor Bcl6 (2·3-fold increase) compared with the control group. CONCLUSION: We conclude that a 5 days of supplementation with B. toyonensis BCT-7112T prior vaccination is sufficient to significantly improve the humoral immune response of ewes against C. perfringens recombinant ETX vaccine. SIGNIFICANCE AND IMPACT OF THE STUDY: These findings open a new perspective in the utilization of B. toyonensis BCT-7112T as an immunomodulator since a 5 days period of probiotic supplementation is sufficient to improve the vaccine immune response.

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O herpesvírus equídeo 1 (EHV-1) apresenta distribuição mundial e causa graves prejuízos à equideocultura. É agente de surtos de doença respiratória, reprodutiva e neurológica, em equídeos jovens e adultos. A glicoproteína D (gD) do envelope viral é essencial para ligação e penetração em células permissivas e direcionamento do sistema imunológico do hospedeiro, induz respostas imunes humorais e celulares, sendo um antígeno apropriado para ser utilizado em vacinas e imunodiagnóstico. O objetivo deste trabalho foi expressar e caracterizar a gD do EHV-1 em Pichia pastoris para posterior utilização como antígeno em técnicas de imunodiagnóstico e formulação de vacinas recombinantes. Uma sequência de DNA que codifica uma forma truncada da gDEHV-1 foi clonada no vetor pPICZαA de expressão em P. pastoris. Obteve-se uma proteína de ~41 kDa, como esperado. A proteína apresentou glicosilação entre 4 kDa e 16 kDa, demonstrada por deglicosilação enzimática. A proteína recombinante foi caracterizada antigenicamente e imunogenicamente por Western blot, utilizando-se anticorpos policlonais equinos anti-EHV-1, e por ELISA indireto em modelo murino, demonstrando que a gD recombinante manteve epítopos similares aos da proteína nativa. Esses resultados sugerem que a gDEHV-1 é um antígeno promissor para uso como imunobiológico no controle do EHV-1.(AU)

Equine herpesvirus 1 (EHV-1) has a worldwide distribution and causes serious damage to horse breeding. It is an agent of respiratory, reproductive and neurological disease outbreaks in young and adult equids. Viral envelope glycoprotein D (gD) is essential for binding and penetration into permissive cells and targeting the host immune system, inducing humoral and cellular immune responses, and is an appropriate antigen for use in vaccines and immunodiagnostics. The objective of this work was to express in Pichia pastoris and to characterize EHV-1 gD for later use as an antigen in immunodiagnostic techniques and formulation of recombinant vaccines. A DNA sequence encoding a truncated form of gDEHV-1 has been cloned into the P. pastoris expression vector pPICZαA. A protein of ~41 kDa was obtained as expected. The protein presented glycosylation between 4 kDa and 16 kDa, demonstrated by enzymatic deglycosylation. The recombinant protein was antigenically and immunogenically characterized by Western blot using equine polyclonal anti-EHV-1 antibodies, and by indirect ELISA in a murine model, demonstrating that the recombinant gD maintained epitopes similar to those of the native protein. These results suggest that gDEHV-1 is a promising antigen for use as an immunobiological in the control of EHV-1.(AU)

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O herpesvírus equídeo 1 (EHV-1) apresenta distribuição mundial e causa graves prejuízos à equideocultura. É agente de surtos de doença respiratória, reprodutiva e neurológica, em equídeos jovens e adultos. A glicoproteína D (gD) do envelope viral é essencial para ligação e penetração em células permissivas e direcionamento do sistema imunológico do hospedeiro, induz respostas imunes humorais e celulares, sendo um antígeno apropriado para ser utilizado em vacinas e imunodiagnóstico. O objetivo deste trabalho foi expressar e caracterizar a gD do EHV-1 em Pichia pastoris para posterior utilização como antígeno em técnicas de imunodiagnóstico e formulação de vacinas recombinantes. Uma sequência de DNA que codifica uma forma truncada da gDEHV-1 foi clonada no vetor pPICZαA de expressão em P. pastoris. Obteve-se uma proteína de ~41 kDa, como esperado. A proteína apresentou glicosilação entre 4 kDa e 16 kDa, demonstrada por deglicosilação enzimática. A proteína recombinante foi caracterizada antigenicamente e imunogenicamente por Western blot, utilizando-se anticorpos policlonais equinos anti-EHV-1, e por ELISA indireto em modelo murino, demonstrando que a gD recombinante manteve epítopos similares aos da proteína nativa. Esses resultados sugerem que a gDEHV-1 é um antígeno promissor para uso como imunobiológico no controle do EHV-1.(AU)

Equine herpesvirus 1 (EHV-1) has a worldwide distribution and causes serious damage to horse breeding. It is an agent of respiratory, reproductive and neurological disease outbreaks in young and adult equids. Viral envelope glycoprotein D (gD) is essential for binding and penetration into permissive cells and targeting the host immune system, inducing humoral and cellular immune responses, and is an appropriate antigen for use in vaccines and immunodiagnostics. The objective of this work was to express in Pichia pastoris and to characterize EHV-1 gD for later use as an antigen in immunodiagnostic techniques and formulation of recombinant vaccines. A DNA sequence encoding a truncated form of gDEHV-1 has been cloned into the P. pastoris expression vector pPICZαA. A protein of ~41 kDa was obtained as expected. The protein presented glycosylation between 4 kDa and 16 kDa, demonstrated by enzymatic deglycosylation. The recombinant protein was antigenically and immunogenically characterized by Western blot using equine polyclonal anti-EHV-1 antibodies, and by indirect ELISA in a murine model, demonstrating that the recombinant gD maintained epitopes similar to those of the native protein. These results suggest that gDEHV-1 is a promising antigen for use as an immunobiological in the control of EHV-1.(AU)

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Probiotics modulate the immune response and can increase the effectiveness of vaccines. Bacillus toyonensis is widely used as a probiotic in animal feed. The aim of this study was to assess the effects of B. toyonensis administration on the immune response to an experimental recombinant vaccine against bovine herpesvirus type 5 (BoHV-5) in mice. Mice were vaccinated with BoHV-5 recombinant glycoprotein D and supplemented with the probiotic B. toyonensis in two regimes: one group received the probiotic only during seven days prior to the initial vaccination while the second group was given the probiotic throughout the experimental period of seven weeks. Animals supplemented with probiotic B. toyonensis in two regimes showed an increase in total immunoglobulin (Ig)G, IgG1 and IgG2a levels in serum, in addition to higher titres of antibodies capable of neutralising the BoHV-5 virus than non-supplemented animals (P<0.05). Splenocytes from the supplemented mice had higher mRNA transcription levels of cytokines interleukin (IL)-4 and IL-12. These results show that the use of this probiotic may significantly contribute to the response elicited by recombinant vaccines, especially those that rely on increasing antibody and cell-mediated immune responses for efficacy. Further, the data support an immunomodulatory effect for probiotic B. toyonensis and imply that enhance effect on the immune response against a BoHV-5 recombinant vaccine in mice.

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ABSTRACT The research aimed to study the histology of ovaries, lateral oviducts, common oviducts and spermatheca, and morphometry of the ovaries of Tropidacris collaris (Stoll, 1813) (Orthoptera: Romaleidae), reared under the photoperiods 10:14, 12:12 and 14:10 (L:D). Morphometry was conducted using a binocular magnifying glass adapted with a milimetric ocular, being registered the length and the width of the ovaries. Subsequently the ovarioles were separated and counted. For the histological analysis, the organs were fixated in alcoholic Boüin, included in paraplast and stained by H.E., Mallorys trichromic and P.A.S. Photoperiods studied did not influence the morphometry of the ovaries, number of ovarioles and the histology of the organs. The averages of the ovarioles number were of 195,62, 202,62 and 208,25 for the fotoperíodos of 10L:14D, 12L:12D and 14L:10D, respectively, being more numerous in the left-side ovary. Each ovarioles presented tubular morphology with distinct regions (terminal filament, germarium and vitellarium). The lateral oviducts are internally covered by simple cubic epithelial tissue with numerous folds, sustained on the connective tissue and, externally, by a striated muscular tissue layer. The common oviduct presents the same histological constitution of the lateral oviduct, except by the presence of the epithelium with cuticular intima and a well developed muscular layer. The spermatheca is constituted by columnar pseudo-stratified epithelial tissue with cuticular intima and striated muscular tissue associated to connective tissue.

RESUMO A pesquisa teve o objetivo de descrever a histologia dos ovários, ovidutos laterais, oviduto comum e espermateca, e realizar a morfometria dos ovários de Tropidacris collaris (Stoll, 1813) (Orthoptera: Romaleidae), submetido aos fotoperíodos de 10L:14E, 12L:12E e 14L:10E. Para morfometria utilizou-se uma lupa binocular adaptada com uma ocular milimétrica, sendo registrado o comprimento e as larguras dos ovários. Posteriormente, os ovaríolos foram separados e contados. Para análise histológica, os órgãos foram fixados em Boüin alcoólico, incluídos em paraplast e corados pela H.E., tricrômico de Mallory e P.A.S (Ácido Periódico de Schiff). Os resultados mostraram que não houve influência dos fotoperíodos sobre a morfometria dos ovários, número de ovaríolos e histologia dos órgãos. Foram encontrados ovários préreprodutivos e reprodutivos. As médias do número de ovaríolos foram de 195,62, 202,62 e 208,25 para os fotoperíodos de 10L:14E, 12L:12E e 14L:10E, respectivamente, sendo mais numerosos no ovário esquerdo. Cada ovaríolo apresentou morfologia tubular com regiões bem distintas (filamento terminal, germário e vitelário). O oviduto lateral é revestido internamente por tecido epitelial simples cúbico com numerosas dobras, apoiado no tecido conjuntivo e externamente por uma camada de tecido muscular estriado. O oviduto comum apresenta a mesma constituição histológica do oviduto lateral, exceto pela presença de epitélio com íntima cuticular e uma camada muscular bem desenvolvida. A espermateca é constituída por tecido epitelial pseudo-estratificado colunar com íntima cuticular e tecido muscular estriado associado a tecido conjuntivo.

RESUMO

Although trans fatty acids are known to influence essential fatty acid (EFA) metabolism and serum levels of lipids and lipoproteins, little is known about their effects on the metabolism of mammary glands (MGs) during lactation. In this study, 5 groups of lactating Wistar rats were fed semisynthetic diets containing 7% soy oil (control); 7% partially hydrogenated vegetable oil (7% PHVO); 13% PHVO +7% soy oil (13% PHVO); 5% PHVO +2% soy oil (5% PHVO), and 3.5% PHVO +3.5% soy oil (3.5% PHVO). Dams were killed on the 12th day of lactation. Weight, lipid content (LC), in vivo lipogenesis rate (LR) and activity of the lipogenic (ATP-citrate lyase and malic) enzymes were evaluated in the MGs. Maternal food intake, weight gain, and total MG weight were not found to be significantly different between the groups. The groups with 13, 7 and 5% PHVO presented a lower (p < 0.05) LR in MGs when compared to controls, whereas MG LC was higher in the 7 and 13% groups than in controls. The observed decrease in MG lipogenesis was accompanied by a diminution (p < 0.05) in the activities of ATP-citrate lyase and malic enzymes. These data indicate a potential impairment of lipid metabolism in the MG by trans isomers in lactating rats.

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