Phenoloxidase activity and haemolymph cytology in honeybees challenged with a virus suspension (deformed wings virus DWV) or phosphate buffered suspension (PBS) / Atividade fenoloxidase e citologia de hemolinfa em abelhas infectadas com uma suspensão de vírus (DWV deformed wings) ou suspensão tamponada com fosfato (PBS)

The innate immune system of honeybees mainly consists in antimicrobial peptides, cellular immunity and melanisation. In order to investigate the immune response of honeybees to immune stressors, three stress degrees were tested. Newly emerged bees naturally DWV-infected were collected from a Varroa mite-free apiary and divided into three experimental groups: naturally DWV infected bees, PBS injected bees, and artificially DWV super infected bees. Phenoloxidase activity and haemolymph cellular subtype count were investigated. Phenoloxidase activity was highest (P<0.05) in DWV-superinfected bees, and the haemocyte population differed within the three observed groups. Although, immune responses following DWV infection have still not been completely clarified, this investigation sheds light on the relation between cell immunity and the phenoloxidase activity of DWV-naturally infected honeybees exposed to additional stress such as injury and viral superinfection.(AU)

O sistema imune inato das abelhas consiste principalmente em peptídeos antimicrobianos, imunidade celular e melanização. Para investigar a resposta imune das abelhas a estressores imunológicos, foram testados três graus de estresse. Abelhas recém-emergidas naturalmente infectadas por DWV foram coletadas de um apiário livre de Varroa e divididas em três grupos experimentais: abelhas naturalmente infectadas por DWV, abelhas injetadas com PBS e abelhas superinfectadas artificialmente com DWV. A atividade de fenoloxidase e a contagem de subtipos celulares de hemolinfa foram investigadas. A atividade da fenoloxidase foi maior (P<0,05) nas abelhas super-infectadas com DWV, e a população de hemócitos diferiu entre os três grupos observados. Embora as respostas imunes após a infecção pelo DWV ainda não tenham sido completamente esclarecidas, esta investigação lança luz sobre a relação entre a imunidade celular e a atividade da fenoloxidase das abelhas infectadas naturalmente pelo DWV, expostas a estresse adicional, como lesão e superinfecção viral.(AU)

Pathomorphological and immunohistochemical evaluation of unilateral progressive Mooren’s ulcer in a cat

Background: Mooren’s ulcer is a chronic and painful ulceration of the cornea. It begins progressively in the periphery and spread centrally in cornea. In human, it is seen uniaterally in most of cases. Mooren ulcer has not been reported in any kind of animals up to now. Although its aetiology is not completely enlighted, it has been suspected of the inflammatory reaction against injuries-microbiological and immun mediated effects. Immun response in presence of accumulation of immune complexes into the limbal vessels.As a result of the deficit in the regulatory mechanism because the number of suppressor cells control over B and T lymphocytes, These situations can result in a progressive tendancy to inflammation because the production of autoantibodies and/or lymphokine from cytotoxic T-lymhocytes creates an immune-mediated vasculitis. Numerous immigrant inflammatory cells and proteins are evaded from vessels. After triggering inflammatory cells and releasing of meditors, corneal vascularization, scar tissue and re-epithelization develop. This regenerative-reperative process plays an important role during post-inflammatory process.Case: In this case, it was aimed to detect pathomorphological structure and immunologic relations in progressive Mooren’s ulcer (MU). A 1 year-old mix breed cat was submitted to clinic with complaints of progressive painful and eyesight loss in left eye. There were 1 cm-ulceration, opacification and old haemorrhagic areas at peripheral cornea. Histopathologically, there was wide ulceration including all layers of corneal epithelium and particularly vacuolar degeneration at suprabasal cells. In corneal stroma, numerous neutrophiles and mononuclear cells were infiltrated. Neovascularisation and fibrosis beginning from limbus were also present. This fibrotic progress was confirmed by Masson’s trichrome staining method.[...]

Pathomorphological and immunohistochemical evaluation of unilateral progressive Moorens ulcer in a cat

Background: Moorens ulcer is a chronic and painful ulceration of the cornea. It begins progressively in the periphery and spread centrally in cornea. In human, it is seen uniaterally in most of cases. Mooren ulcer has not been reported in any kind of animals up to now. Although its aetiology is not completely enlighted, it has been suspected of the inflammatory reaction against injuries-microbiological and immun mediated effects. Immun response in presence of accumulation of immune complexes into the limbal vessels.As a result of the deficit in the regulatory mechanism because the number of suppressor cells control over B and T lymphocytes, These situations can result in a progressive tendancy to inflammation because the production of autoantibodies and/or lymphokine from cytotoxic T-lymhocytes creates an immune-mediated vasculitis. Numerous immigrant inflammatory cells and proteins are evaded from vessels. After triggering inflammatory cells and releasing of meditors, corneal vascularization, scar tissue and re-epithelization develop. This regenerative-reperative process plays an important role during post-inflammatory process.Case: In this case, it was aimed to detect pathomorphological structure and immunologic relations in progressive Moorens ulcer (MU). A 1 year-old mix breed cat was submitted to clinic with complaints of progressive painful and eyesight loss in left eye. There were 1 cm-ulceration, opacification and old haemorrhagic areas at peripheral cornea. Histopathologically, there was wide ulceration including all layers of corneal epithelium and particularly vacuolar degeneration at suprabasal cells. In corneal stroma, numerous neutrophiles and mononuclear cells were infiltrated. Neovascularisation and fibrosis beginning from limbus were also present. This fibrotic progress was confirmed by Massons trichrome staining method.[...](AU)

Evaluation of thymic lymphocyte loss of broiler using Digital Analysis of the Lymphoid Depletion System (ADDL) / Avaliação de perda de linfócitos do timo em frangos usando análise digital do sistema de depleção linfóide (ADDL)

The thymus is a lymphoid organ and usually evaluated for the degree of lymphocyte loss with subjective histological techniques. This study aimed to adapt and to apply of the digital analysis of the lymphoid depletion system (ADDL) in the thymus in order to obtain a more accurate analysis. Glucocorticoid was used to induce immunosuppression in 55 broilers at 21 days of age; other 15 broilers were the control group. After euthanasia of the broilers, postmortem examination was made. Both thymic chains were collected and six lobes were selected for histological examination of the degree of lymphocyte depletion (scores 1 to 5) and for submission to all stages of processing by the ADDL system. The artificial constructed neural networks (ANN) obtained 94.03% of correct classifications. In conclusion, it was possible to adopt objective criteria to evaluate thymic lymphoid depletion with the ADDL system.(AU)

O timo é um órgão linfóide, que é normalmente avaliado para o grau de perda de linfócitos a partir de técnicas histológicas subjetivas. Este trabalho teve como objetivo a adaptação e aplicação do sistema de análise digital de depleção linfóide (ADDL) para o timo, a fim de tornar sua análise mais acurada. Glicocorticóides foram utilizados a fim de induzir imunossupressão em 55 aves de 21 dias de idade. Outras 15 aves formaram o grupo controle. Posteriormente, para cada um dos aves, realizou-se a eutanásia e necropsia. Ambas as cadeias do timo foram coletadas e foram selecionadas seis lóbulos para processamento histológico, análise quanto ao grau de depleção linfocitária (escores de 1-5) e submissão a todas as fases do processamento pelo sistema ADDL. Observou-se que a rede neural artificial (RNA) construída obteve 94,03% de classificações corretas. Em conclusão, foi possível adotar critérios objetivos para avaliar a depleção linfóide tímica utilizando o sistema ADDL.(AU)

Evaluation of thymic lymphocyte loss of broiler using Digital Analysis of the Lymphoid Depletion System (ADDL) / Avaliação de perda de linfócitos do timo em frangos usando análise digital do sistema de depleção linfóide (ADDL)

The thymus is a lymphoid organ and usually evaluated for the degree of lymphocyte loss with subjective histological techniques. This study aimed to adapt and to apply of the digital analysis of the lymphoid depletion system (ADDL) in the thymus in order to obtain a more accurate analysis. Glucocorticoid was used to induce immunosuppression in 55 broilers at 21 days of age; other 15 broilers were the control group. After euthanasia of the broilers, postmortem examination was made. Both thymic chains were collected and six lobes were selected for histological examination of the degree of lymphocyte depletion (scores 1 to 5) and for submission to all stages of processing by the ADDL system. The artificial constructed neural networks (ANN) obtained 94.03% of correct classifications. In conclusion, it was possible to adopt objective criteria to evaluate thymic lymphoid depletion with the ADDL system.(AU)

O timo é um órgão linfóide, que é normalmente avaliado para o grau de perda de linfócitos a partir de técnicas histológicas subjetivas. Este trabalho teve como objetivo a adaptação e aplicação do sistema de análise digital de depleção linfóide (ADDL) para o timo, a fim de tornar sua análise mais acurada. Glicocorticóides foram utilizados a fim de induzir imunossupressão em 55 aves de 21 dias de idade. Outras 15 aves formaram o grupo controle. Posteriormente, para cada um dos aves, realizou-se a eutanásia e necropsia. Ambas as cadeias do timo foram coletadas e foram selecionadas seis lóbulos para processamento histológico, análise quanto ao grau de depleção linfocitária (escores de 1-5) e submissão a todas as fases do processamento pelo sistema ADDL. Observou-se que a rede neural artificial (RNA) construída obteve 94,03% de classificações corretas. Em conclusão, foi possível adotar critérios objetivos para avaliar a depleção linfóide tímica utilizando o sistema ADDL.(AU)

The effect of arginine dietary supplementation in broiler breeder hens on offspring humoral and cell-mediated immune responses

The influence of supplementing the diet of broiler breeder hens with arginine (Arg) on their offspring's humoral and cell-mediated immune response was evaluated in two experiments. In experiments I and II, breeder hens were fed diets containing graded levels of Arg (0.943, 1.093, 1.243, 1.393 and 1.543% digestible Arg). In experiment I, the offspring was randomly grouped according to the treatment received by the breeder hens, with five levels of Arg in the maternal diet and six replicates, giving a total 30 experimental units. In experiment II, the offspring were grouped in accordance with the treatment received by the breeder hens; however, Arg was added to the starter diet (1.300, 1.450, 1.600, 1.750 and 1.900% digestible Arg) and also the growing diet (1.150, 1.300, 1.450, 1.600 and 1.750% digestible Arg). Supplementation of the broiler breeder hen diet did not influence (p > 0.05) the development of the lymphoid organs (cloacal bursa, thymus and spleen) of the offspring, whether their diet were supplemented or not. Nevertheless, greater weight and dimensions cloacal bursa were found in the supplemented offspring in comparison with the nonsupplemented offspring. Macrophage phagocytic activity was found to be unaffected (p > 0.05), independently of the Arg supplementation. The offspring fed with supplemented diets showed a linear reduction in the antibody titer against Newcastle Disease (p 0.05) by the breeder hen diet. This study concluded that supplementing the breeder hen diet with arginine is insufficient to improve the humoral and cellular immune response, requiring supplementation of the offspring diet.

The effect of arginine dietary supplementation in broiler breeder hens on offspring humoral and cell-mediated immune responses

The influence of supplementing the diet of broiler breeder hens with arginine (Arg) on their offspring's humoral and cell-mediated immune response was evaluated in two experiments. In experiments I and II, breeder hens were fed diets containing graded levels of Arg (0.943, 1.093, 1.243, 1.393 and 1.543% digestible Arg). In experiment I, the offspring was randomly grouped according to the treatment received by the breeder hens, with five levels of Arg in the maternal diet and six replicates, giving a total 30 experimental units. In experiment II, the offspring were grouped in accordance with the treatment received by the breeder hens; however, Arg was added to the starter diet (1.300, 1.450, 1.600, 1.750 and 1.900% digestible Arg) and also the growing diet (1.150, 1.300, 1.450, 1.600 and 1.750% digestible Arg). Supplementation of the broiler breeder hen diet did not influence (p > 0.05) the development of the lymphoid organs (cloacal bursa, thymus and spleen) of the offspring, whether their diet were supplemented or not. Nevertheless, greater weight and dimensions cloacal bursa were found in the supplemented offspring in comparison with the nonsupplemented offspring. Macrophage phagocytic activity was found to be unaffected (p > 0.05), independently of the Arg supplementation. The offspring fed with supplemented diets showed a linear reduction in the antibody titer against Newcastle Disease (p < 0.05) at seven days of age, and a quadratic response (p < 0.05) at 28 days of age. The antibody titer in the non-supplemented offspring was not influenced (p > 0.05) by the breeder hen diet. This study concluded that supplementing the breeder hen diet with arginine is insufficient to improve the humoral and cellular immune response, requiring supplementation of the offspring diet.(AU)

Fontes de óleo na dieta e sua influência no desempenho e na imunidade de frangos de corte / Sources of oil in the diet and its influence on the performance and the immunity of broilers

[...] O objetivo deste estudo foi avaliar o efeito do consumo de óleos ricos em ácidos graxos poli-insaturados ômega-6 (PUFAs n-6) e ômega-3 (PUFAs n-3) sobre o desempenho e a resposta imunológica de frangos de corte frente a um desafio antigênico. Foram comparadas dietas formuladas com 7% de óleo de soja (OS), linhaça (OL) ou sardinha (OP), fornecidas a 240 frangos da linhagem Cobb, divididos em 24 grupos de 10 aves cada, num arranjo experimental 3x2 (3 tipos de óleo e aves vacinadas ou não vacinadas) e 4 repetições. O óleo de soja é rico em ácido linoleico, um PUFA n-6, o óleo de linhaça é fonte de ácido alfa-linolênico, um PUFA n-3, e o óleo de sardinha, de outros PUFAs n-3, como os ácidos eicosapentaenoico e docosahexaenoico. O consumo de ração, o ganho de peso e a conversão alimentar foram avaliados aos 21, 35 e 42 dias. Aos 7 e aos 21 dias de idade, metade das aves recebeu vacina contra doença de Newcastle. Quinze dias após a imunização, avaliou-se a produção de anticorpos pelo método de ELISA, expressa pela densidade óptica a 450 nm (D.O. 450nm). Apenas as aves alimentadas com ração contendo OS apresentaram maior imunidade humoral (P<0,05) após a vacinação. A resposta linfoproliferativa das aves, que expressa a imunidade celular, foi maior entre as aves vacinadas, em comparação às aves não vacinadas (P<0,05), independentemente do óleo utilizado. A fonte de óleo da ração ou a vacinação não influenciaram o ganho de peso das aves (P>0,05). Entre as aves que receberam dieta com OS, as aves vacinadas apresentaram pior conversão alimentar (P<0,05). Nos grupos que consumiram ração com OL ou OP, a vacinação não influenciou a conversão alimentar (P>0,05), considerando todo o período experimental. A utilização de óleo rico em PUFA n-6 na dieta de frangos de corte aumentou a resposta humoral, mas não influenciou a resposta celular frente a um desafio antigênico.(AU)

[...] The objective of this study was to evaluate the effect of the consumption of oils rich in omega-6 (n-6 PUFA) and omega-3 (n-3 PUFA) polyunsaturated fatty acids on the performance and the immune response of broilers submitted to an antigenic challenge. Diets were formulated with either 7% soybean oil (SO), linseed oil (LO) or sardine oil (PO) and provided to 240 Cobb broilers which were divided into 24 groups of 10 birds each, following a 3x2 experimental arrangement (3 types of oil and vaccinated or non-vaccinated birds) and four replications. Soybean oil is rich in linoleic acid (n-6 PUFA), linseed oil a source of alfa-linolenic acid (n-3 PUFA) and the sardine oil is a source of eicosapentaenoic and docosahexaenoic acids (other n-3 PUFA). Feed intake, weight gain and feed conversion were evaluated at 21, 35 and 42 days. Half of the birds were vaccinated against Newcastle disease at 7 and 21 days. Fifteen days after the immunization, the production of antibodies was evaluated by ELISA and expressed by optical density at 450 nm (O.D. 450 nm). Only the birds fed ration containing SO presented higher humoral immune response (p<0.05) after vaccination. The lymphoproliferative response, which expresses the cellular immunity, was higher in vaccinated than in the unvaccinated birds (P<0.05), regardless of the oil used. Neither the oil source in the ration nor the vaccination influenced birds' weight gain (P>0.05). The vaccination impaired the feed conversion of the birds fed diet containing SO (P<0.05) but did not influence feed conversion of the birds fed rations with LO or PO (P>0.05). The use of oil rich in n-6 PUFA in broilers' diet increased humoral response, but did not influence the cellular response against an antigenic challenge.(AU)

Proteínas séricas de potros da raça Puro Sangue Árabe recém-desmamados ou com mais de trinta dias de desmame / Serum protein profile in Arabian foals recently weaned or at more than thirty days after weaning

A fase perinatal do desenvolvimento constitui um dos períodos de vida mais desafiadores para o sistema imunológico dos potros. O objetivo do presente estudo foi verificar o perfil protéico sérico de parâmetros relacionados à imunidade de equinos jovens no período perinatal, verificando-se a transferência de imunidade passiva. Oito animais desmamados há um dia, formaram o Grupo 1 (G1), enquanto vinte animais desmamados há mais de trinta dias formaram o Grupo 2 (G2). A concentração sérica de proteína total foi determinada por refratometria. Para o fracionamento das proteínas, utilizou-se eletroforese em gel de acrilamida. Os resultados obtidos foram submetidos à análise de medidas repetidas e ao teste Tukey (p<0,05) para comparação das médias. As concentrações de IgA apresentaram diferença (p<0,05) entre os grupos, porém os valores observados encontravam-se dentro do considerado normal para equinos adultos. Não houve diferença (p>0,05) nas concentrações de IgG. O estabelecimento adequado da imunidade celular ocorre durante a fase neonatal, nos animais que ingerem adequadamente o colostro e o leite. O presente estudo determinou diferenças no perfil protéico sérico de parâmetros relacionados à imunidade de equinos jovens no período imediato ao desmame, comparados com animais desmamados há mais de 30 dias. De acordo com os valores observados, concluiu-se que os animais, mesmo desmamados precocemente, obtiveram transferência adequada de imunidade passiva.(AU)

The perinatal phase of foal development is one of the most challenger period for the immune system. The present study has analyzed serum protein profile, considering variables related to immunity in foals at the perinatal phase, verifying passive immunity transfer. The group 1 (G1) contained eight foals evaluated one day after weaning, and group 2 (G2) included twenty foals at more than thirty days after weaning. Total protein concentration was determined by means of refractometry. The concentration of serum proteins was determined through sodium dodecyl sulphatepolyacrylamide gel electrophoresis. Results were submitted to analysis of variance and Tukey test (P<0.05). IgA concentration showed difference (P<0.05) between the two studied groups, however data were within adult healthy horses normal values. IgG didn't show statistical difference (P>0.05). The cellular immunity establishment occurs in the neonatal phase, in foals that suckled colostrum and milk properly. The present study showed differences in serum protein profile, considering variables related to immunity, in foals immediately after weaning comparing to foals at more than 30 days after weaning. According to the observed values, we conclude that foals, even early weaned, showed proper passive immunity tranfer.(AU)

Absorção de IgG via colostro em leitões biológicos e adotados após a uniformização da leitegada / Absorption of IgG via colostrum in biological piglets and adopted piglets after crossfostering

Nove fêmeas de quinto parto (OP5) foram imunizadas com 4mg e 2mg de albumina sérica bovina (BSA) aos 70 e 100 dias de gestação, respectivamente. A uniformização da leitegada foi realizada 4,9±1,9h após o nascimento, antes de os leitões efetuarem a primeira mamada. As leitegadas foram compostas por cinco leitões biológicos (LB) e cinco leitões adotados (LA), com pesos semelhantes ao nascimento. Foram coletadas amostras de sangue dos leitões ao nascimento e 24h após, das fêmeas ao pós-parto e de colostro de cada grupo de tetos ao parto e 24h após. As amostras de soro e colostro foram quantificadas para IgG pelo ELISA indireto. A densidade ótica de IgG anti-BSA (DOIgG-BSA) dos leitões (24h de vida) foi correlacionada com a das fêmeas. A DOIgG-BSA entre LB e LA foi semelhante, assim como entre os grupos de tetos, ao parto e 24h após. Entretanto, ocorreu redução na DOIgG-BSA do parto até 24h após. LB e LA absorveram a mesma quantidade de IgG via colostro, quando a uniformização foi realizada até 5h pós-parto, independentemente do teto em que os leitões mamaram, uma vez que esses possuem a mesma concentração de IgG.(AU)

Nine sows of fifth parity (PO5) were immunized with 4mg and 2mg of bovine serum albumin (BSA) at 70 and 100d of gestation, respectively. Cross fostering was performed 4.9±1.9h after birth, before piglets had their first suckling. Litters were composed of five biological piglets (BP) and five adopted piglets (AP), with similar weight at birth. Blood samples were collected from piglets (at birth and at 24h of life) and from females (after farrowing) and colostrum from each group of teats (at farrowing time and after 24h). Samples of serum and colostrum were quantified to IgG by indirect ELISA. Optical density of IgG anti-BSA (ODIgG-BSA) from piglets (24h of life) was correlated with dams. ODIgG-BSA was similar among BP and AP, as well as among pairs of teats (at farrowing time and after 24h). However, there was a decrease in ODIgG-BSA from farrowing up to 24h after birth. BP and AP absorbed the same amount of IgG via colostrum, when cross fostering was evaluated 5h after farrowing, regardless of the teat suckled, since these have the same concentration of IgG.(AU)

Cell-mediated immune response to Leishmania chagasi experimental infection of BALB/c immunosuppressed mice

Leishmaniasis, a zoonosis of worldwide distribution, presents a significant impact on immunosupressed patients. This study aimed to evaluate Leishmania chagasi infection in BALB/c mice immunosuppressed with dexamethasone. Spleen cells stimulated or not with L. chagasi were cultured for cytokine quantification (IFN-gama, IL-2, IL-4 and IL-10) by sandwich ELISA. Parasite loads in the spleen and liver were determined by means of culture microtitration. Immunosuppressed groups showed statistically lower spleen weight and CD4-cell percentage in blood on the day of infection and produced Th1 and Th2 cytokines on other days of the study. The other infected groups, weather immunosupressed or not, also produced Th1 and Th2 cytokines. Parasite loads in the spleen and liver were not statistically different among the groups. It was concluded that L. chagasi infection was not affected by dexamethasone-induced immunosuppression, probably due the reversible effect of the treatment.(AU)

Tuberculin skin testing in HIV-infected patients in Campo Grande, Mato Grosso do Sul State, Brazil

A cross-sectional study on HIV/AIDS was carried out in 108 outpatients from the university hospital of the Federal University of Mato Grosso do Sul, Campo Grande, Brazil, from July to December 2008, to investigate latent tuberculosis infection using the tuberculin skin test (TST). The prevalence of positive results was 13.9 percent. The CD4+ T cell count (p = 0.091) and the diagnosis time (p = 0.010) were statistically significant when compared with TST positivity. In the cohort of HIV/AIDS patients who had latent tuberculosis infection, the median diagnosis time was eight years. Undetectable viral load presented significant association (p = 0.046) with tuberculosis infection. The fact that numerous individuals with HIV/AIDS infection presented a negative reaction to the tuberculin skin test is probably related to alterations in the cellular immune response induced by HIV infection. The tuberculin test is a useful tool for the detection of latent tuberculosis infection and should be performed in all HIV/AIDS individuals at the time of the diagnosis and on a yearly basis, if negative. Both the early identification of the tuberculosis infection by the tuberculin skin test at the moment of immunological restoration and chemoprophylaxis in infected individuals are mechanisms to control HIV/AIDS and tuberculosis coinfection.(AU)

Toxoplasma gondii: humoral and cellular immune response of BALB/c mice immunized via intranasal route with rTgROP2 / Toxoplasma gondii: avaliação da resposta imune humoral e celular de camundongos BALB/c imunizados pela via nasal com rTgROP2

TgROP2 is an intracellular protein associated with rhoptries of Toxoplama gondii and an antigen component of a candidate vaccine for toxoplasmosis. he purpose of the present study was to evaluate the eficacy of rTgROP2 to stimulate humoral and cellular immune responses in BALB/c mice via intranasal injection. TgROP2 partial coding sequence was (196-561) amplified by PCR from genomic T. gondii RH strain DNA and cloned into the pTrcHis expression vector. Escherichia coli Rosetta 2 cells transformed with pTrcHis-TgROP2 showed high levels (1 mg.mL potentiation -1) of recombinant protein after 4 hours of IPTG induction. Recombinant TgROP2 exhibited an apparent Mr equal to 54 kDa. In order to test immunogenicity of the recombinant protein, 10 BALB/c mice received 10 µg of rROP2 protein + 10 µg of Quil-A via intranasal injection. Doses were administered at days 0, 21, and 42. Three animals were euthanized and used to evaluate cellular immune response on day 62. Five (50%) and two (20%) out of ten animals produced IgG (DO mean = 0.307; cut-of = 0.240) and IgA (DO mean = 0.133, cut-of = 0.101), respectively, by ELISA on day 62. he proliferation of splenocytes revealed high stimulation index (SI) when co-cultured with 5, 10 and 15 µg.mL potentiation -1 of rTgROP2. These results indicate that intranasal immunization with recombinant protein ROP2 plus Quil-A can elicit both cellular and humoral immune responses in BALB/c mice. (AU)

TgROP2 é uma proteína localizada nas roptrias do Toxoplasma gondii, sendo um antígeno candidato a componente de uma vacina contra a toxoplasmose. O objetivo do presente estudo foi avaliar a eficácia da TgROP2 recombinante em estimular a resposta imune celular e humoral de camundongos BALB/c após estímulo intranasal. A sequência da TgROP2 foi amplificada pela PCR a partir da cepa RH e clonada em vetor de expressão pTrc-His. Após a transformação em Escherichia coli- Rosetta 2, a pTrcHis-TgROP2 exibiu alto nível de expressão após 4 horas de indução com IPTG. A proteína recombinante apresentou uma massa molecular aparente de aproximadamente 54 kDa. Para avaliar a imunogenicidade dessa proteína recombinante, 10 camundongos receberam, pela via intranasal, 10 µg da rROP2 associado a 10 µg de Quil-A. Três doses foram realizadas nos dias 0, 21 e 42. No dia 62 do experimento, três animais foram eutanasiados para avaliar as respostas imune celular e humoral. Cinco (50%) e dois (20%) dos 10 animais apresentaram níveis de IgG (DO média = 0,307; ponto de corte = 0,240) e IgA (DO média = 0,133; ponto de corte = 0,101) acima do ponto de corte no ELISA no dia 62. A proliferação de esplenócitos revelou altos Índices de Estimulação (SI), quando as células foram cultivadas com 5, 10 e 15 µg.mL elevado a -1 de rTgROP2. Os resultados obtidos indicam que a via nasal pode estimular tanto a resposta imune celular como a humoral. (AU)

Níveis crescentes de parede de levedura sobre a resposta imune celular e perfil hematológico de frangos de corte / Cellular immune response and hematological parameters of broilers from different age broiler breeders, fed with a sorghum meal with different yeast wall levels

Um experimento foi conduzido para avaliar o efeito de níveis crescentes de parede de levedura e idade das matrizes reprodutoras sobre o peso dos órgãos linfóides, a resposta imune celular e o perfil hematológico de frangos de corte. Foram utilizados 3.360 pintos de corte da linhagem Cobb, distribuídos em delineamento inteiramente casualisado, em esquema fatorial 2x5, mais dois controles, sendo duas idades de matrizes (34 e 57 semanas de idade) e cinco níveis de suplementação de parede de levedura (zero, um, dois, três e quatro kg de parede de levedura/tonelada de ração). A idade das matrizes influenciou a resposta de todas as variáveis. A inclusão de 3kg de parede de levedura/tonelada de ração promoveu, na progênie de reprodutoras de 57 semanas, reação inflamatória mais intensa quando comparada a dieta controle, no entanto não houve aumento significativo no número de heterófilos e linfócitos circulantes. Conclui-se que a utilização da parede de levedura associada ao sorgo ou não em rações de frangos de corte ainda necessita de estudos complementares, que incluam, por exemplo, os componentes purificados da parede de levedura (MOS e ß-glucano).(AU)

An experiment was carried out to evaluate the effect increasing levels of yeast wall and broiler breeders' age levels on lymphoid organs weight, cellular immune response and hematological parameters in broilers. A total of 3,360 Cobb broilers were allotted, in a completely randomized design and a 2x5 factorial arrangement, and two controls, compound of two broiler breeders age (34 and 57 weeks of age) and five yeast wall levels (zero, one, two, three and four kg of yeast wall/ton of diet). Broiler breeders' age affected all studied variables. The inclusion of 3kg of yeast wall/ton of meal increased, at 57 weeks age broiler breeders, more intense inflammatory reaction when compared to control diet; however the circulated heterophils and lymphocytes numbers were not increased. In conclusion, the use of yeast wall, associated or not to sorghum on broilers diet still needs complementary studies, for example, purified components of yeast wall (MOS and ß-glucano).(AU)

Estudo retrospectivo do perfil clínico e epidemiológico de cães com sarna demodécica / Retrospective study of the clinical-epidemiological profile of dog with demodicosis

A demodiciose canina é uma doença parasitária, inflamatória, caracterizada pela presença de umaumento na contagem da população de Demodex canis que ocorre por falha na imunidade celular, geneticamente programada ou por doenças que comprometam a imunocompetência do paciente. Avaliou-se o perfil de pacientes com sarna demodécica, tratados com moxidectina e atendidos em uma clínica particular especializada em dermatologia, na cidade de São Paulo, entre 2008 e 2013.Foram atendidos 51 pacientes, dentre esses 54,9%, eram fêmeas e 45,1% machos. Os cães de raça definida totalizaram 88,2% dos animais e, destes, 50,98% possuíam pelame longo. Grande parte dos animais (85%) apresentou prurido e infecção bacteriana secundária concomitante. A ocorrência de efeitos colaterais à moxidectina foi relatada por 23,5% dos proprietários, sendo os sintomas gastroentéricos os mais frequentes. (AU)

Canine demodicosis is an inflammatory parasitic disease, characterized by the presence of an increased population of Demodex canis. The disease may be due to the lymphocytes failure in react to theparasite infection, to genetic disorders or to secondary. It was evaluated 51 dogs treated with moxidectin in a Veterinary Clinic, localizated in São Paulo in the period 2008-2013. 54.9% were females and 45.1% males. The pure breed dogs totaled 88.2% of the animals, of these, 50.98% had long haircoat. The occurrence of side effects reported by owners was 23.5% of cases, gastroenteric symptoms being the most common. (AU)

Estudo retrospectivo do perfil clínico e epidemiológico de cães com sarna demodécica / Retrospective study of the clinical-epidemiological profile of dog with demodicosis

A demodiciose canina é uma doença parasitária, inflamatória, caracterizada pela presença de umaumento na contagem da população de Demodex canis que ocorre por falha na imunidade celular, geneticamente programada ou por doenças que comprometam a imunocompetência do paciente. Avaliou-se o perfil de pacientes com sarna demodécica, tratados com moxidectina e atendidos em uma clínica particular especializada em dermatologia, na cidade de São Paulo, entre 2008 e 2013.Foram atendidos 51 pacientes, dentre esses 54,9%, eram fêmeas e 45,1% machos. Os cães de raça definida totalizaram 88,2% dos animais e, destes, 50,98% possuíam pelame longo. Grande parte dos animais (85%) apresentou prurido e infecção bacteriana secundária concomitante. A ocorrência de efeitos colaterais à moxidectina foi relatada por 23,5% dos proprietários, sendo os sintomas gastroentéricos os mais frequentes.

Canine demodicosis is an inflammatory parasitic disease, characterized by the presence of an increased population of Demodex canis. The disease may be due to the lymphocytes failure in react to theparasite infection, to genetic disorders or to secondary. It was evaluated 51 dogs treated with moxidectin in a Veterinary Clinic, localizated in São Paulo in the period 2008-2013. 54.9% were females and 45.1% males. The pure breed dogs totaled 88.2% of the animals, of these, 50.98% had long haircoat. The occurrence of side effects reported by owners was 23.5% of cases, gastroenteric symptoms being the most common.

Avaliação da imunidade celular na pele de cães naturalmente infectados com Leishmania (Leishmania) infantum chagasi e sua correlação com transmissibilidade ao vetor / Evaluation of cellular immunity in the skin of naturally infected dogs with Leishmania (Leishmania) infantum chagasi and its relationship with the vector transmissibility

Foram selecionados aleatoriamente 38 cães naturalmente infectados por Leishmania (Leishmania) infantum chagasi oriundos de Araçatuba, São Paulo, área endêmica para leishmaniose visceral, e os quais foram distribuídos em dois grupos: o primeiro com 24 cães sintomáticos e, o outro, composto por 14 animais assintomáticos. Correlacionou-se a caracterização clínica (assintomáticos ou sintomáticos) com os: potencial em infectar o inseto vetor, padrão inflamatório da pele, perfil de imunidade celular e parasitismo tegumentar desses cães. Quanto ao número de formas amastigotas/mm2 no tegumento, não houve diferença estatisticamente significativa entre os grupos (p = 0,1584), sendo que a densidade de parasitos na pele mostrou uma correlação positiva moderada com os títulos de anticorpos anti-Leishmania (p = 0,042). Quanto à infectividade ao vetor, evidenciada pelo xenodiagnóstico, 16 (66,7%) cães sintomáticos e 13 (93%) assintomáticos foram capazes em transmitir Leishmania aos flebotomíneos, dentre estes, respectivamente, seis (37,5%) e oito (61,5%) animais não apresentavam parasitismo cutâneo, embora todos, de ambos os grupos, tenham sido positivos quando da pesquisa de parasitos no linfonodo poplíteo. Observou-se um maior percentual de transmissibilidade para o vetor a partir de cães assintomáticos (p = 0,0494). As alterações histológicas cutâneas foram similares em ambos os grupos e se caracterizaram, de modo geral, por um infiltrado inflamatório, ora focal ora difuso, na derme, constituído, principalmente, por células mononucleares (macrófagos, linfócitos e plasmócitos), variando de discreto a intenso. Quanto à caracterização da resposta imune celular no tegumento dos animais, somente a densidade (células/mm2) de células iNOS+ foi significantemente maior na derme dos cães sintomáticos quando comparado aos assintomáticos (p = 0,0368). Observou-se correlação positiva moderada entre a densidade de parasitos na pele e a densidade de macrófagos (p = 0,031), de células T CD4+ (p = 0,015) e CD8+ (p = 0,023), e, também, naquela de células iNOS+ relativamente a de células T CD3+ (p = 0,005), CD4+ (p = 0,001) e CD8+ (p = 0,0001). Verificou-se, ainda, correlação negativa moderada com o número de manifestações clínicas e/ou gravidade da enfermidade (p = 0,028), confirmando, assim, maior transmissibilidade de parasitos ao vetor, a partir de animais assintomáticos, demonstrando-se, dessa forma, que tais cães são fontes de infecção em potencial para insetos vetores em áreas endêmicas para leishmaniose visceral canina.

Thirty-eight dogs naturally infected by Leishmania (Leishmania) infantum chagasi were randomly selected from Araçatuba of São Paulo state (Brazil), an endemic area for visceral leishmaniasis. The subjects were distributed into two groups: the first comprising 24 symptomatic dogs and the second consisting of 14 asymptomatic dogs. Possible correlations of clinical characterization (in both symptomatic and asymptomatic subjects) with potential to infect the insect vector, skin inflammatory pattern, cutaneous cellular immunity profile and cutaneous parasitism were investigated. Regarding to the number of cutaneous leishmania amastigotes/mm2, there was not a significant difference between the groups (p = 0.1584); density of skin parasites showed a moderate positive correlation with anti-Leishmania antibody titers (p = 0.042). Concerning to the infectivity to the insect vector, as evidenced by xenodiagnosis, 16 (66.7%) symptomatic and 13 (93%) asymptomatic dogs were able to transmit Leishmania to phlebotomines. Among these, six (37.5%) and eight (61.5%) dogs, respectivelly, did not show cutaneous parasitism, although all participant dogs were found positive when searching for parasites in the popliteal lymph nodes. Asymptomatic dogs showed higher transmissibility to the vector than symptomatic ones (p = 0.0494). Histological features in the skin were similar in both groups and were generally characterized by an inflammatory infiltrate, either diffuse or focal, in the dermis, mainly consisted of mononuclear cells (macrophages, lymphocytes and plasma cells), varying from mild to intense. Concerning characterization of the cutaneous cellular immune response, only iNOS+ cells density (cells/mm2) was significantly higher in the dermis of the symptomatic dogs compared to the asymptomatic ones (p = 0.0368). Moderate positive correlation between the cutaneous parasites density and the macrophages density (p = 0.031), the CD4+ T cells (p = 0.015) and CD8+ T cells (p = 0.023) were observed. Furthermore, density of iNOS+ cells in relation to CD3+ T cells (p = 0.005), CD4+ T cells (p = 0.001) and CD8+ T cells (p = 0.0001) were found positively correlated at a moderate level. It was also found a moderate negative correlation between cutaneous parasites density and the number of clinical signs and/or disease severity (p = 0.028), confirming grater parasites transmission to the vector from asymptomatic animals. It was therefore demonstrated that these dogs are potential sources of infection to insect vectors in endemic areas for canine visceral leishmaniasis.

Avaliação da imunidade celular na pele de cães naturalmente infectados com Leishmania (Leishmania) infantum chagasi e sua correlação com transmissibilidade ao vetor / Evaluation of cellular immunity in the skin of naturally infected dogs with Leishmania (Leishmania) infantum chagasi and its relationship with the vector transmissibility

Foram selecionados aleatoriamente 38 cães naturalmente infectados por Leishmania (Leishmania) infantum chagasi oriundos de Araçatuba, São Paulo, área endêmica para leishmaniose visceral, e os quais foram distribuídos em dois grupos: o primeiro com 24 cães sintomáticos e, o outro, composto por 14 animais assintomáticos. Correlacionou-se a caracterização clínica (assintomáticos ou sintomáticos) com os: potencial em infectar o inseto vetor, padrão inflamatório da pele, perfil de imunidade celular e parasitismo tegumentar desses cães. Quanto ao número de formas amastigotas/mm2 no tegumento, não houve diferença estatisticamente significativa entre os grupos (p = 0,1584), sendo que a densidade de parasitos na pele mostrou uma correlação positiva moderada com os títulos de anticorpos anti-Leishmania (p = 0,042). Quanto à infectividade ao vetor, evidenciada pelo xenodiagnóstico, 16 (66,7%) cães sintomáticos e 13 (93%) assintomáticos foram capazes em transmitir Leishmania aos flebotomíneos, dentre estes, respectivamente, seis (37,5%) e oito (61,5%) animais não apresentavam parasitismo cutâneo, embora todos, de ambos os grupos, tenham sido positivos quando da pesquisa de parasitos no linfonodo poplíteo. Observou-se um maior percentual de transmissibilidade para o vetor a partir de cães assintomáticos (p = 0,0494). As alterações histológicas cutâneas foram similares em ambos os grupos e se caracterizaram, de modo geral, por um infiltrado inflamatório, ora focal ora difuso, na derme, constituído, principalmente, por células mononucleares (macrófagos, linfócitos e plasmócitos), variando de discreto a intenso. Quanto à caracterização da resposta imune celular no tegumento dos animais, somente a densidade (células/mm2) de células iNOS+ foi significantemente maior na derme dos cães sintomáticos quando comparado aos assintomáticos (p = 0,0368). Observou-se correlação positiva moderada entre a densidade de parasitos na pele e a densidade de macrófagos (p = 0,031), de células T CD4+ (p = 0,015) e CD8+ (p = 0,023), e, também, naquela de células iNOS+ relativamente a de células T CD3+ (p = 0,005), CD4+ (p = 0,001) e CD8+ (p = 0,0001). Verificou-se, ainda, correlação negativa moderada com o número de manifestações clínicas e/ou gravidade da enfermidade (p = 0,028), confirmando, assim, maior transmissibilidade de parasitos ao vetor, a partir de animais assintomáticos, demonstrando-se, dessa forma, que tais cães são fontes de infecção em potencial para insetos vetores em áreas endêmicas para leishmaniose visceral canina. (AU)

Thirty-eight dogs naturally infected by Leishmania (Leishmania) infantum chagasi were randomly selected from Araçatuba of São Paulo state (Brazil), an endemic area for visceral leishmaniasis. The subjects were distributed into two groups: the first comprising 24 symptomatic dogs and the second consisting of 14 asymptomatic dogs. Possible correlations of clinical characterization (in both symptomatic and asymptomatic subjects) with potential to infect the insect vector, skin inflammatory pattern, cutaneous cellular immunity profile and cutaneous parasitism were investigated. Regarding to the number of cutaneous leishmania amastigotes/mm2, there was not a significant difference between the groups (p = 0.1584); density of skin parasites showed a moderate positive correlation with anti-Leishmania antibody titers (p = 0.042). Concerning to the infectivity to the insect vector, as evidenced by xenodiagnosis, 16 (66.7%) symptomatic and 13 (93%) asymptomatic dogs were able to transmit Leishmania to phlebotomines. Among these, six (37.5%) and eight (61.5%) dogs, respectivelly, did not show cutaneous parasitism, although all participant dogs were found positive when searching for parasites in the popliteal lymph nodes. Asymptomatic dogs showed higher transmissibility to the vector than symptomatic ones (p = 0.0494). Histological features in the skin were similar in both groups and were generally characterized by an inflammatory infiltrate, either diffuse or focal, in the dermis, mainly consisted of mononuclear cells (macrophages, lymphocytes and plasma cells), varying from mild to intense. Concerning characterization of the cutaneous cellular immune response, only iNOS+ cells density (cells/mm2) was significantly higher in the dermis of the symptomatic dogs compared to the asymptomatic ones (p = 0.0368). Moderate positive correlation between the cutaneous parasites density and the macrophages density (p = 0.031), the CD4+ T cells (p = 0.015) and CD8+ T cells (p = 0.023) were observed. Furthermore, density of iNOS+ cells in relation to CD3+ T cells (p = 0.005), CD4+ T cells (p = 0.001) and CD8+ T cells (p = 0.0001) were found positively correlated at a moderate level. It was also found a moderate negative correlation between cutaneous parasites density and the number of clinical signs and/or disease severity (p = 0.028), confirming grater parasites transmission to the vector from asymptomatic animals. It was therefore demonstrated that these dogs are potential sources of infection to insect vectors in endemic areas for canine visceral leishmaniasis. (AU)

Efeito do dimetildioctadecilamônio na resposta imune humoral e celular de cobaios inoculados com o herpesvírus bovino-1 inativados / Effect of dimethyl dioctadecyl ammonium chloride on the humoral and cellular response to inactivated bovine herpesvirus-1 in guinea pigs

Avaliou-se o potencial adjuvante do cloreto de dimetildioctadecilamônio (DDA cloreto) em estimular as respostas imune humoral e celular, do tipo hipersensibilidade cutânea tardia (DTH), em cobaios que receberam preparações de antígeno contendo o herpesvírus bovino tipo 1 (BHV-1) inativado. Os animais foram vacinados com o BHV-1 em cinco diferentes formulações, representadas pelos grupos: A- adjuvante completo / incompleto de Freund, B- hidróxido de alumínio (Al(OH)3), C- DDA cloreto, D- associação Al(OH)3 / DDA cloreto e E- BHV-1 em solução aquosa sem adjuvante. Os animais do grupo F, grupo-controle negativo, foram inoculados apenas com meio de cultivo celular. Os maiores títulos de anticorpos neutralizantes do BHV-1, expressos em log2, foram obtidos nos grupos D, A e C, com títulos de 6,25, 6,0 e 5,25, respectivamente. Na avaliação da DTH, os maiores aumentos na espessura da dobra da pele foram observados nos grupos A (2,4mm), C (1,8mm) e D (1,1mm). O DDA cloreto, utilizado tanto de forma isolada quanto em associação, determinou a potencialização das respostas imunológicas humoral e celular de cobaios imunizados com o BHV-1 inativado.(AU)

The adjuvanticity of dimethyl dioctadecyl ammonium chloride (DDA chloride) to induce humoral and cell mediated (delayed type hypersensitivity DTH –) immune responses was assessed in guinea pigs that received antigen preparations containing inactivated bovine herpesvirus type 1 (BHV-1). The animals were vaccinated with five different formulations, containing BHV-1 represented by groups: A- complete/incomplete Freund adjuvant, B- aluminum hydroxide (Al(OH)3), C- DDA chloride, D- association Al(OH)3/DDA and E- BHV-1 in aqueous solution without adjuvant. Group F was the negative control group and the animals received only cell culture medium. The higher titers of BHV-1 neutralizing antibodies, expressed in log2, were obtained in groups D, A and C with values of 6.25, 6.0 and 5.25, respectively. In the DTH assessment, the higher increase in skin thickness was observed in group A (2.4mm), C (1.8mm) and D (1.1mm). These results showed that DDA chloride used alone or in association to other substance boosted the humoral and cell mediated immune responses in guinea pigs immunized with inactivated BHV-1.(AU)