Can intramammary infections change immunoglobulins and acute phase proteins of colostrum and transition milk in dairy goats? / A infecção intramamária pode alterar imunoglobulinas e proteínas de fase aguda do colostro e do leite de transição de cabras leiteiras?

Proteiongram analysis is useful for the early diagnosis of intramammary infections during the period of colostrogenesis. This study aimed to evaluate the profile of total proteins, immunoglobulins, lactoferrin, and gamma-glutamyl transferase (GGT) in the colostrum of dairy goats with intramammary infections. Animals were divided in groups: GI (n=12) of goats without mammary gland infections, and GII (n=8) of goats with mammary gland infections. Intramammary infections were diagnosed using microbiological isolations and somatic cell counts Cs). Total protein was evaluated in the samples using SDS-PAGE shortly after parturition, and 24 and 48 hours after that event. Non-aureus Staphylococcus (NAS) were detected all isolates. At 48 h, GII had high IgG levels and a SCC of 1660.25 x 103/mL. Levels of total protein were high in this group at 24 and 48 h. Albumin levels were high in goats with mastitis at 24 h. Overall, the IgG, lactoferrin, and albumin levels differed between animals with and without intramammary infections at M0. GGT activity was not influenced by the intramammary infection. The results of this study reinforce the importance of the proteinogram as an auxiliary tool in the diagnosis of mastitis in dairy goats.

O proteinograma do colostro/leite pode ser útil para diagnóstico precoce de infecções intramamárias, assim como para a avaliação da intensidade da resposta inflamatória. Este estudo teve como objetivo avaliar o perfil de proteínas, imunoglobulinas, lactoferrina e gamaglutamiltransferase no colostro de cabras leiteiras portadoras s de infecções intramamárias. Os animais foram distribuídos em dois grupos, GI (n 12), composto por cabras sem isolamento microbiológico das glândulas mamárias, e GII (n 8), composto por cabras com resultados positivos na cultura do leite de pelo menos uma das glândulas mamárias. O diagnóstico de infecção intramamária foi realizado logo após o parto, por meio de isolamento microbiológico e de contagem de células somáticas (CCS). Os níveis de proteína total, imunoglobulina A (IgA), imunoglobulina (G) (IgG), lactoferrina, albumina e atividade da gamaglutamiltransferase (GGT) foram avaliados em amostras de colostro/leite de transição, usando-se SDS-PAGE logo após o parto bem como às 24 e às 48 horas após esse evento. Staphylococcus não aureus (NAS) foram encontrados em todos os isolamentos. A concentração de IgG de 186 foi superior no GII apenas às 48h, ao mesmo tempo em que o CCS foi de 1660,25 x 10/mL. No entanto, a proteína total foi maior neste grupo às 24 e às 48h. A albumina foi maior nas cabras com mastite às 24h, e a lactoferrina no momento 0h (M0) e às 48h. Em geral, os valores de IgG, lactoferrina e albumina diferiram entre os animais com e sem infecções intramamárias no M0. A atividade da gamaglutamiltransferase não foi influenciada pela infecção intramamária. Os resultados deste estudo reforçam a importância do proteinograma como uma ferramenta auxiliar no diagnóstico da mastite em cabras leiteiras.

Evaluation of tissue repair using phytotherapeutic gel from Plectranthus neochilus, Schlechter (boldo-gambá) and Cnidoscolus quercifolius Pohl (favela) in Wistar rats / [Avaliação da reparação tecidual do gel fitoterápico de Plectranthus neochilus, Schlechter (Boldo-gambá) e Cnidoscolus quercifolius Pohl (Favela) em ratos Wistar]

The objective of this study was to investigate the effects of a gel formulation from the association of Plectranthus neochilus and Cnidoscolus quercifolius on tissue repair in cutaneous wounds in rats. A surgical wound was induced in 35 Wistar rats and treated according to group: G1 - commercial phytotherapeutic gel; G2 - Carbopol gel 1%; G3, 4, 5 - gel formulation from Boldo-gambá and Favela (FGBF) at 2.5%, 5%, and 10%, respectively. 1ml of the product was applied topically daily, for 14 days. Macroscopic evaluation of the wound showed inflammation, granulation, and epithelization in all groups. The FGBF 2.5% group showed greater angiogenic potential. There was a significant difference between the surgical area of the wounds treated with FGBF 2.5%, 5%, or 10% compared to the group with the commercial phytotherapeutic gel. On histomorphometry of the skin, there were reepithelization of the epidermis and superficial dermis, longitudinal collagen fibers, fibroblasts, and blood vessels, and in the deeper dermis, fibroblasts, transverse and longitudinal collagen fibers, blood vessels, and inflammatory cells. The 2.5% formulation had the greatest increase in fibroblast proliferation and most intense collagenization on day 14 of treatment.(AU)

Este trabalho objetivou investigar os efeitos de uma formulação em gel da associação da Plectranthus neochilus e da Cnidoscolus quercifolius no processo de reparação tecidual em feridas cutâneas de ratos. Foi induzida uma ferida cirúrgica em 35 ratos Wistar, sendo tratadas de acordo com os grupos: G1 - fitoterápico comercial; G2 - gel de carbopol 1%; G3, G4 e G5 - formulação gel boldo-gambá mais favela (FGBF) 2,5%, 5% e 10%, respectivamente. Aplicou-se 1mL do produto, via tópica, diariamente, durante 14 dias. Na avaliação macroscópica das feridas, verificou-se inflamação, granulação e epitelização em todos os grupos. O grupo FGBF 2,5% apresentou maior potencial angiogênico. Houve diferença significativa entre as áreas cirúrgicas das feridas tratadas com os FGBF 2,5%, 5% ou 10%, comparados ao grupo com o gel fitoterápico comercial. Na histomorfometria da pele, observou-se reepitelização da epiderme e da derme superficial, fibras colágenas longitudinais, fibroblastos e vasos sanguíneos e, na derme profunda, fibroblastos, fibras colágenas transversais e longitudinais, vasos sanguíneos e células inflamatórias. A formulação a 2,5% teve o maior aumento na proliferação de fibroblastos e mais intensa colagenização no dia 14 de tratamento.(AU)

Aquisição de imunidade passiva em cabritos alimentados com colostro de cabras com e sem mastite / Immunity of procurement in passive kids fed colostrum from goats with and without mastitis

O objetivo deste trabalho foi avaliar a transferência de imunidade passiva de cabras, que pariram com mastite, para seus respectivos cabritos. Os animais foram distribuídos em dois grupos, a saber: grupo 1 (GI), constituído por cabritos, filhos de cabras sem isolamento microbiológico em ambas as glândulas mamárias, e grupo 2 (GII), composto por cabritos, filhos de cabras com resultado positivo à lactocultura, em pelo menos uma das glândulas mamárias. Foram coletadas amostras de colostro e sangue à parição, bem como às 24 e às 48 horas após o parto/nascimento. O diagnóstico e o monitoramento da mastite nos animais foram realizados por meio do California Mastitis Test (CMT), contagem de células somáticas e isolamento microbiológico. A proteína total foi mensurada pelo método do biureto, e as concentrações de imunoglobulina A (IgA), imunoglobulina G (IgG), transferrina, albumina e haptoglobina por meio da eletrofoerese em gel de poliacrilamida contendo dodecil sulfato de sódio (SDS-PAGE). Os agentes mais isolados na cultura microbiológica foram os Staphylococcus coagulase negativa. Não houve diferença significativa (P<0,05) entre os valores médios de imunoglobulina G (IgG) nos cabritos provenientes de cabras com mastite quando comparados aos recém-nascidos oriundos de cabras livres de infecções intramamárias. Da mesma forma, a atividade de gamaglutamiltransferase (GGT) não mostrou diferença entre os grupos em todos os momentos avaliados. A ingestão de colostro decorrente de cabras com mastite não causou falha na transferência de imunidade passiva nos respectivos conceptos.(AU)

The aim of this study was to evaluate the transfer of passive immunity goats kidded with mastitis to their kids. The animals were divided into two groups, namely: Group 1 (GI) containing kids, sons of goats without microbiological isolation in both mammary glands, and Group 2 (GII), composed of kids, sons of goats with positive result to lactoculture in at least one of mammary glands. Colostrum samples and blood were collected after delivery, 24 and 48 hours after delivery / birth. The diagnosis and monitoring of mastitis in animals were performed using the California Mastitis Test (CMT), somatic cell count and microbiological isolation. Total protein was measured by the biuret method, and the concentrations of immunoglobulin A (IgA), immunoglobulin G (IgG), transferrin, albumin and haptoglobin through eletrofoerese polyacrylamide gel containing sodium dodecyl sulfate (SDS-PAGE). The agents most isolated in microbiological culture were coagulase-negative Staphylococcus. There was no significant difference (p <0.05) between the acquisition of immunoglobulin G (IgG) in goats from goats with mastitis compared to infants originating free goat mammary infections. Similarly the gamma glutamyl transferase (GGT) was equal in the comparison between groups in all evaluated moments. The colostrum intake resulting from goats with mastitis caused no failure in the passive transfer of immunity in their fetuses.(AU)

Cryptococcus: isolamento ambiental e caracterização bioquímica / Cryptococcus: environmental isolation and biochemical characterization

O gênero Cryptococcus caracteriza-se por ser uma levedura responsável por infecção sistêmica, causada pelas espécies Cryptococcus neoformans e Cryptococcus gattii. O fungo é encontrado em substratos de origem animal e vegetal, e a infecção ocorre com a inalação de basidiósporos ou leveduras desidratadas infectantes presentes no ambiente. O presente trabalho teve por objetivo pesquisar a existência de microfocos de Cryptococcussp.em amostras ambientais da cidade de Araçatuba, São Paulo, com a finalidade de minimizar os riscos de contaminação do homem e dos animais, buscando o conhecimento da ecoepidemiologia do Cryptococcus. Foram colhidas 50 amostras oriundas de ocos e troncos de árvores (Cassiasp., Ficussp., Caesalpinea peltophorides) de 10 locais representativos do perímetro urbano, as quais foram encaminhadas ao Laboratório de Bacteriologia e Micologia da Faculdade de Medicina Veterinária de Araçatuba-Unesp, onde foram processadas e semeadas em placas de Petri contendo ágar semente de Níger e Sabouraud dextrose com clorafenicol e incubadas à temperatura de 30ºC, por um período não inferior a cinco dias. Posteriormente, foram submetidas às provas bioquímicas: produção de urease, termotolerância a 37ºC e quimiotipagem em ágar CGB (L-canavanina-glicina-azul de bromotimol). A análise dos resultados revelaram que 17 (34%) dos cultivos foram positivos para o gênero Cryptococcus, sendo nove (18%) para Cryptococcus gattiie oito (16%) para Cryptococcus neoformans. Outras leveduras correlacionadas, como Rhodotorula sp. e Candida sp., também foram isoladas. Conclui-se que os basidiósporos de Cryptococcusencontram-se dispersos na natureza, constituindo microfocos ambientais, não vinculados necessariamente a um único hospedeiro.(AU)

Cryptococcosis is an opportunistic fungal infection caused by Cryptococcusyeasts, especially C. neoformans and Cryptococcus gattii. The fungus is found in substrates of animal and vegetable origin, and infection occurs through inhalation and seedlings present in the environment. The present study aimed to investigate the existence of microfocus Cryptococcus sp. from the environmental samples of Araçatuba city, São Paulo, featuring new niches, by decoupling the direct relationship between fungus and host in order to minimize the risk of contamination of man and animals, understanding the ecoepidemiology of Cryptococcus. Fifty samples from hollows and tree trunks were harvested (Cassia sp., Ficus sp., Caesalpinea peltophorides) from ten representatives in the urban perimeter. The samples were immediately sent to the Laboratory of Bacteriology and Mycology, Faculty of Veterinary Medicine Araçatuba - Unesp where they were processed and plated on Petri dishes containing agar seed Niger and Sabouraud dextrose agar with chloramphenicol, incubated at 30ºC for a period of no less than 5 days. Afterwards they were subimitted to biochemical tests: urease production, thermotolerance at 37°C and quimiotipagem in CGB agar (L- Canavanine-Glycine-Bromothymol blue). The results showed that 17 (34%) cultures were positive for Cryptococcus, 9 (18%) for Cryptococcus gattii and 8 (16%) for Cryptococcus neoformans. Other yeast correlated as Rhodotorula sp. and Candida sp. were isolated. We conclude that the infectious propagules of Cryptococcus are dispersed in nature and constitute an environmental microfocus, not necessarily being bound to a single host.(AU)

Caracterização bioquímica e molecular de Cryptococcus spp. isolados de excretas ambientais de pombos (Columba livia domestica) / Biochemical and molecular characterization of Cryptococcus spp. isolates from environmental feces of pigeons (Columba livia domestic)

Os pombos possuem uma rica diversidade de microrganismo, entre eles fungos sapróbios, como do gênero Cryptococcus, que podem atuar como agentes patogênicos para o homem e animais. Objetivou-se o isolamento, a caracterização bioquímica e a molecular de amostras de Cryptococcus spp. de excretas ambientas de pombos. Foram colhidas 100 amostras ambientais de pontos equidistantes e representativos da área da cidade de Araçatuba, São Paulo. As amostras foram rasteladas do solo de vias públicas, armazenadas em frasco coletor e encaminhadas para o Laboratório de Bacteriologia e Micologia da FMVA, onde foram processadas e cultivadas em duplicata, em placas de Petri contendo ágar Sabouraud dextrose a 4% e Niger. Em seguida, foram incubadas à temperatura ambiente e a 30ºC, respectivamente, por um período não inferior a 15 dias. Após a observação diária, as colônias sugestivas para levedura foram reisoladas em ágar Niger e submetidas a testes bioquímicos para posterior caracterização molecular pela técnica da PCR. Como resultado, a caracterização bioquímica e a molecular isolaram 32 colônias leveduriformes, sendo 8% dos cultivos positivos para Cryptococcus neoformans var. neoformans, 17% para Rhodotorula rubidae e 7% Candida albicans. Pelo exposto, concluiu-se que excretas ambientais de pombos constituem um microfoco para Cryptococcus neoformans var. neoformans e outras leveduras com potencial patogênico, representando um risco à saúde pública, sendo necessárias medidas preventivas, como a higienização com a correta remoção das excretas, a fim de minimizar os riscos de exposição ambiental(AU)

Pigeons have a rich diversity of microrganisms, including fungi saprobes such as the genus Cryptococcus that can act as a pathogen for humans and animals. The aim of this was their isolation, biochemical and molecular characterization of samples of Cryptococcus from ambientas avian excreta. One hundred environmental samples,representative of the area equidistant from Araçatuba, São Paulo points were collected. The samples were removed and stored in collection bottles, sent to the Laboratory of Bacteriology and Mycology of FMVA, where they were processed and cultivated in duplicate on Petri dishes containing Sabouraud dextrose 4% and Niger and incubated at room temperature 30° C, respectively, for a period of no less than 15 days. Colonies after daily observation which were suggestive for yeast growth were re-isolated in Niger agar and subjected to biochemical analisis for further molecular characterization by PCR tests. As results, the biochemical and molecular characterization of 32 yeast colonies revealed 8% of cultures positive for Cryptococcus neoformans var. neoformans, 17% Rhodotorula rubidae and 7% Candida albicans. It was concluded that environmental avian excreta are active sites for replication of Cryptococcus neoformans var. neoformans and other yeasts with pathogenic potential. These constitute a public health risk, and preventive measures are necessary, such as cleaning and complete removal of excreta in order to minimize the risk of environmental exposure(AU)

Infecção congênita em cabritos por Mycoplasma agalactiae / Congenital infection by Mycoplasma agalactiae in goat kids

This study aimed to report three cases of contagious agalactia (CA) by Mycoplasma agalactiae in goat kids born with polyarthritis. The nanny goats belonging to two different herds presented clinical signs of CA during pregnancy and in parturition they were apparently healthy. The carpal articulations of the three goat kids, the tarsus articulation in one, and thigh-femoral articulation in another showed swelling, pain and impairment of the flexion-extension movements. The articular liquid was collected from two goat kids at birth and revealed a content which varied from transparent to fibrinopurulent, presenting a yellow coloring. The samples were plated on modified Hayflick. The colonies had the appearance of "fried egg" and were confirmed as being M. agalactiae by biochemical tests and 16S rRNA PCR. Blood was collected from three animals soon after birth and submitted to the indirect ELISA test for the determination of the titration of the anti- M. agalactiae antibodies. The results confirmed that the goat kids were infected during pregnancy by M. agalactiae and resulted in the birth of an offspring with clinical signs of CA being immune tolerant.(AU)

TNF- and IL-6 immunohistochemistry in rat renal tissue experimentaly infected with Leptospira interrogans serovar Canicola

Leptospirosis is a public health problem worldwide and its etiology remains unclear. Its pathogenesis involves a complex interaction between host and infecting microorganism. The inflammatory reaction that controls the infection process also underscores many pathophysiological events occurring in leptospirosis. We investigated the presence of tumor necrosis factor- (TNF-) and interleukin-6 (IL-6) in renal tissues by immunohistochemical and histopathological examination in animals experimentally inoculated with Leptospira serovar Canicola. All the tests were carried out 2, 7, 14, 21 or 28 days after inoculation. Although TNF- and IL-6 had been detected in tissues throughout the observation period, these cytokines appeared more intensely during the initial phase of infection. Therefore, both TNF- and IL-6 were associated with the immunopathogenesis of leptospirosis. This profile suggests a high immunocellular response throughout the early infection stages followed by subsequent humoral response.

Response activity of alveolar macrophages in pulmonary dysfunction caused by Leptospira infection

Leptopspirosis is a syndrome with different clinical manifestations including the most severe and often fatal forms of pulmonary disease of unknown etiology. Pulmonary injury during the inflammatory process has been associated with the excessive number of alveolar macrophages (AMs) and polymorphonuclear leukocytes stimulated in the lungs and with the production of reactive oxygen and nitrogen intermediates and other inflammatory mediators. The aim of the present work was to evaluate the cellular immune response of AMs or inflammatory cells of hamsters during leptospirosis. The activity of AMs was determined by measuring nitric oxide (NO) and protein production as well as inflammatory cell infiltration in bronchoalveolar lavage (BAL) fluid. Pulmonary activity during infection was monitored by measuring pH, pressure of oxygen (PaO2), and pressure of carbon dioxide (PaCO2) in blood samples. Cellular immune response and its role in the genesis of leptospirosis have been incriminated as the main causes of tissue and pulmonary injuries, which consequently lead to the pulmonary dysfunction in severe cases of leptospirosis. The present results show a low production of NO in both supernatant of alveolar macrophage culture and BAL. In the latter, protein production was high and constant, especially during acute infection. Total and differential cell count values were 2.5X10(6) on day 4; 7.3X10(6) on day 21; and 2.3X10(6) on day 28 after infection, with lymphocytes (84.04%) predominating over neutrophils (11.88%) and monocytes (4.07%). Arterial blood gas analysis showed pulmonary compromising along with the infectious process, as observed in parameter values (mean±SD) evidenced in the infected versus control group: PaO2 (60.47mmHg±8.7 vs. 90.09mmHg±9.18), PaCO2 (57.01mmHg±7.87 vs. 47.39mmHg±4.5) and pH (7.39±0.03 vs. 6.8±1.3). Results indicated that Leptospira infection in hamsters is a good experimental model to study leptospirosis. However, some of the immune parameters showed variations which might be associated with the animal species.

Cytokine and antibody production during murine leptospirosis

The aim of the present study was to investigate the kinetics of humoral and cellular responses during leptospirosis. We observed that the presence of tumor necrosis factor-alpha (TNF-alpha) and interleukin-6 (IL-6) was associated with antibody production and bacterial recovery, and the compromising of both TNF-alpha and IL-6 in the immunopathogenesis of leptospirosis during an experimental infection of BALB/c mice inoculated with Leptospira interrogans serovar Canicola was verified. Results showed higher levels of TNF-alpha and IL-6 in the initial phase of infection, in which the greatest bacterial clearance was observed. However, when the bacterial recovery was compared with the kinetics of the production of antibodies, the results revealed a kinetics proportionally inverted to antibody production. This fact may be related to some inhibitory factor which could be responsible for the selective suppression of the cellular immune response. We concluded that during leptospirosis there was a greater mobilization of the cellular immune response activity, mainly in the initial phase of the infectious process, for posterior involvement of the humoral response, and that both TNF-alpha and IL-6 could be associated with the immunopathogenesis of the disease.

Role of cytokines, NO, and H2O2 on the immunopathology of Leptospirosis in genetically selected mice

Immune response to leptospirosis is mainly humorally mediated, and involves opsonization of leptospires for phagocytosis by macrophages and neutrophils. However, some aspects are still unknown. For a more detailed analysis of the cellular immune response to leptospirosis infection, trials were carried out in order to determine the hydrogen peroxide and nitric oxide (H2O2 and NO) production stimulated or not by Interferon-gamma. The participation of some specific cytokines, such as Tumor Necrosis Factor-alpha (TNF-alpha); Interferon-gamma (IFN-gamma); Interleukin-6 (IL-6); and Interleukin-4 (IL-4), in the immunopathology of this infection was also investigated. For this purpose, we analyzed the supernatant from peritoneal macrophage cell culture and the splenic cells of mice genetically selected as High (H) and Low (L) antibody producers, and inbred Balb/c mice infected with Leptospira interrogans serovar icterohaemorrhagiae. The IL-6 production varied from release peaks to inhibition in H, L, and Balb/c mice. Similar behavior was observed for IL-4, produced only by H and Balb/c mice. The three strains presented constant and elevated production of TNF-alpha until day 14, suggesting its effective participation in the initial phase of the infection. Meanwhile, all the three strains presented a constant and irregular IFN-gamma production, with release peaks between the 7th and 14th days in L mice. The H and Balb/c mice strains presented a higher tendency to Th2 response pattern, whereas L mice tended towards Th1 response.