Avaliação do estresse e do desempenho de suínos na fase de creche, empregando-se técnicas de enriquecimento ambiental / Evaluation of stress and performance of pigs during the nursery phase, using environmental enrichment techniques

O enriquecimento ambiental é uma ferramenta importante dentro dos sistemas de produção, a fim de promover o bem-estar e favorecer a saúde dos animais. Portanto, o objetivo deste trabalho foi avaliar o efeito do enriquecimento ambiental sobre o estresse de suínos na fase de creche. Foram utilizados 32 leitões, alojados em granja experimental, distribuídos em quatro grupos (n= 8): corda, corrente, garrafa PET e controle negativo. Amostras de sangue foram coletadas no início e no final do experimento para contagem de leucócitos circulantes e determinação de antioxidantes não enzimáticos, óxido nítrico, malondialdeído, e de saliva para avaliação do cortisol. Foi aplicado etograma e fez-a ganho médio de peso diário e a conversão alimentar. Os parâmetros avaliados no primeiro dia do experimento não variaram entre os grupos (P>0,05). No último dia do experimento, os valores de neutrófilos e da relação neutrófilo/linfócito foram mais elevados nos leitões do grupo corrente, assim como os valores de cortisol salivar (P<0,05). O ácido úrico apresentou-se mais elevado nos leitões do grupo corrente e o malondialdeído (MDA) nos do grupo garrafa (P< 0,05). Os enriquecimentos ambientais estimularam comportamentos positivos nos leitões, tendo a corda se destacado como o mais atrativo. Por outro lado, a corrente apresentou efeito negativo sobre a fisiologia dos animais, gerando estresse, assim como a garrafa, que induziu a peroxidação lipídica e um menor ganho de peso nos leitões.(AU)

Environmental enrichment is an important tool within production systems to promote welfare and animal health. The aim of this study was to evaluate the effect of enrichment objects on stress of piglets in nursery phase. 32 piglets housed in experimental farm were distributed in 4 groups (n= 8): rope, chain, pet bottle and negative control. Blood samples were collected at the beginning and end of the experiment to count circulating leukocytes and determine non-enzymatic antioxidants, nitric oxide, malondialdehyde, and saliva to evaluate cortisol. At the same time, an etogram and evaluation of mean daily gain and feed conversion were applied. The evaluated parameters on first day of the experiment did not vary between groups (P> 0.05). On the last day, neutrophil and neutrophil /lymphocyte ratios were higher in chain group piglets, as were salivary cortisol values (P< 0.05). Uric acid was higher in chain group and MDA in bottle group (P< 0.05). Environmental enrichment stimulated positive behaviors in piglets, where the rope stood out as the most attractive. On the other hand, chain had a negative effect on animal physiology, generating stress, as well as the bottle that induced lipid peroxidation and a lower weight gain in piglets.(AU)

Histopathological characterization of experimentally induced cutaneous loxoscelism in rabbits inoculated with Loxosceles similis venom

Envenomation by Loxosceles bites is characterized by dermonecrotic and/or systemic features that lead to several clinical signs and symptoms called loxoscelism. Dermonecrotic lesions are preceded by thrombosis of the dermal plexus. Recent studies show that atheromatous plaque is prone to thrombosis due to endothelial cell apoptosis. To the best of our knowledge, there are no reports of microscopic dermal lesion and endothelial cell apoptosis induced by Loxosceles similis venom in the literature. Thus, the aim of the present study is to describe histological lesions induced by L. similis venom in rabbit skin and to elucidate whether apoptosis of endothelial cells is involved in the pathogenesis of loxoscelism. Forty male rabbits were split into two groups: the control group (intradermally injected with 50 µL of PBS) and the experimental group (intradermally injected with 0.5 µg of L. similis crude venom diluted in 50 µL of PBS). After 2, 4, 6 and 8 hours of injection, skin fragments were collected and processed for paraffin or methacrylate embedding. Sections of 5 µm thick were stained by HE, PAS or submitted to TUNEL reaction. Microscopically, severe edema, diffuse heterophilic inflammatory infiltrate, perivascular heterophilic infiltrate, thrombosis, fibrinoid necrosis of arteriolar wall and cutaneous muscle necrosis were observed. Two hours after venom injection, endothelial cells with apoptosis morphology were evidenced in the dermal plexus. Apoptosis was confirmed by TUNEL reaction. It seems that endothelial cell apoptosis and its consequent desquamation is an important factor that induces thrombosis and culminates in dermonecrosis, which is characteristic of cutaneous loxoscelism.

Cell proliferation markers in the transplanted canine transmissible venereal tumor / Marcadores de proliferação celular no tumor venéreo transmissível canino transplantado

Adult male mongrel dogs were subcutaneously transplanted with the canine transmissible venereal tumor (TVT) on the hypogastric region. Twelve specimens of tumors were collected, half during the proliferative phase and the other half during the regressive phase. Fragments of the tumor were fixed in 10 percent buffered formalin and routinely processed for light microscopy. Sections of 4µm were stained by Schorr or AgNOR or either immunostained for MIB1 (Ki67). Schorr stain, AgNOR and MIB1 showed an increased proliferative activity through mitotic index, nuclear argyrophilic protein stain and cycling tumoral cells in the growing tumors, respectively. All of the three cell proliferation markers were able to distinguish the TVT in both evolution phases. MIB1 monoclonal antibody was the best in the morphologic evaluation of growth and regression of TVT. This resulted in higher values than AgNORs counting and mitotic index. MIB1 immunostaining was the most effective parameter of the proliferative activity of TVT. However, a significant correlation has been detected only between mitosis counting and AgNORs.

Cães machos, adultos, mestiços, foram transplantados com células do tumor venéreo transmissível canino (TVTC), na região hipogástrica. Foram coletados doze espécimes do TVTC, sendo metade durante a fase proliferativa e metade durante a fase regressiva. Fragmentos do tumor foram fixados em formol a 10 por cento, tamponado e processado rotineiramente para microscopia de luz. Secções de 4µm foram coradas pelo Shorr, ou pela AgNOR, ou ainda, imunocorado para MIB 1 (Ki67). As colorações pelo Shorr, AgNOR, ou MIB 1 mostraram um aumento do índice mitótico, coloração da proteína argirofílica nuclear e células tumorais ciclando em tumores em crescimento, respectivamente. Todos os três marcadores de proliferação celular foram capazes de distinguir o TVTC em ambas as fases de evolução. O anticorpo monoclonal MIB 1 foi o melhor na avaliação morfológica, de crescimento e regressão do TVTC. Isto resultou em um valor maior que a contagem de AgNOR e do índice mitótico. A imunomarcação com MIB1 foi o parâmetro mais efetivo da atividade proliferativa. No entanto, só foi observada uma correlação positiva entre a contagem de mitose e a AgNOR.

Síndrome de Kartagener em um cão (Canis lupus familiaris) da raça Cocker Spaniel Inglês / Kartagener's syndrome in an English Cocker Spaniel dog (Canis lupus familiaris)

Kartagener is syndrome is a rare disorder described in dogs determined by the triad situs inversus totalis, chronic sinusitis, and bronchial disease. The clinical signs of chronic respiratory and other less common presentations (such as hydrocephalus and subfertility) are due to a concomitant ciliary dysfunction. This article describes the clinical, radiographic, and computed tomography findings and treatment in a dog affected by Kartagener is syndrome since early months of life. This is the first case report of this syndrome in a domestic dog in Brazil and the only known scientific report in the English Cocker Spaniel.

Apoptosis in the transplanted canine transmissible venereal tumor during growth and regression phases / Apoptose no tumor venéreo transmissível canino durante as fases de crescimento e regressão

Twelve male, mongrel, adult dogs were subcutaneously transplanted with cells originated from two canine transmissible venereal tumors (TVT). The aim was to demonstrate and to quantify the occurrence of apoptosis in the TVT regression. After six months of transplantation, a tumor sample was obtained from each dog, being six dogs with TVT in the growing phase and six in the regression phase as verified by daily measurements. Samples were processed for histological and ultrastructural purposes as well as for DNA extraction. Sections of 4µm were stained by HE, Shorr, methyl green pyronine, Van Gieson, TUNEL reaction and immunostained for P53. The Shorr stained sections went through morphometry that demonstrated an increase of the apoptotic cells per field in the regressive tumors. It was also confirmed by transmission electron microscopy, which showed cells with typical morphology of apoptosis and by the TUNEL reaction that detected in situ the 3'OH nick end labeling mainly in the regressive tumors. The regressive TVTs also showed an intensified immunostaining for P53 besides a more intense genomic DNA fragmentation detected by the agarose gel electrophoresis. In conclusion, apoptosis has an important role in the regression of the experimental TVT in a way that is P53-dependent.

Doze cães, adultos, machos e sem raça definida foram transplantados subcutaneamente, na região hipogástrica, com células originadas de dois tumores venéreos transmissíveis caninos (TVT). O objetivo do estudo foi demonstrar e quantificar a ocorrência de apoptose na regressão do TVT. Após seis meses, foi obtido um tumor de cada animal, totalizando seis em crescimento e seis em regressão. Fragmentos dos tumores foram processados para avaliação histológica, ultra-estrutural e também para extração de DNA. Cortes de 4µm foram corados em HE, Shorr, verde de metila pironina e Van Gieson e alguns foram submetidos à reação do TUNEL e à imunoistoquímica para P53. Secções coradas pelo Shorr, submetidas à morfometria, demonstram maior índice apoptótico nos tumores em regressão. Esse achado foi confirmado pela microscopia eletrônica de transmissão que evidenciou células com morfologia típica de apoptose e pela reação de TUNEL que marcou mais células nos tumores em regressão que naqueles em crescimento. A imunomarcação para P53 foi mais intensa nos tumores em regressão, assim como a fragmentação internucleossômica do genoma mostrada pela eletroforese em gel de agarose. Concluiu-se que a apoptose tem importante papel na regressão do TVT transplantado experimentalmente, sendo, nesse caso, dependente de P53 para a sua execução.

Virulence insights from the Paracoccidioides brasiliensis transcriptome

Paracoccidioides brasiliensis, the etiologic agent of paracoccidioidomycosis, is a dimorphic fungus, which is found as mycelia at 22-26 degrees C and as yeasts at 37 degrees C. A remarkable feature common to several pathogenic fungi is their ability to differentiate from mycelium to yeast morphologies, or vice-versa. Although P. brasiliensis is a recognized pathogen for humans, little is known about its virulence genes. In this sense, we performed a search for putative virulence genes in the P. brasiliensis transcriptome. BLAST comparative analyses were done among P. brasilienses assembled expressed sequence tags (PbAESTs) and the sequences deposited in GenBank. As a result, the putative virulence PbAESTs were grouped into five classes, metabolism-, cell wall-, detoxification-related, secreted factors, and other determinants. Among these, we have identified orthologs of the glyoxylate cycle enzymes, a metabolic pathway involved in the virulence of bacteria and fungi. Besides the previously described alpha- and beta-glucan synthases, orthologs to chitin synthase and mannosyl transferases, also important in cell wall synthesis and stabilization, were identified. With respect to the enzymes involved in the intracellular survival of P. brasiliensis, orthologs to superoxide dismutase, thiol peroxidase and an alternative oxidase were also found. Among the secreted factors, we were able to find phospholipase and urease orthologs in P. brasiliensis transcriptome. Collectively, our results suggest that this organism may possess a vast arsenal of putative virulence genes, allowing the survival in the different host environments.

Paracoccidioides brasiliensis translation and protein fate machineries revealed by functional genome analysis

The translational and post-translational modification machineries of Paracoccidioides brasiliensis were assessed by means of comparative analyses of PbAESTs (P. brasiliensis assembled expressed sequence tags) with sequences deposited on different databases. Of the 79 sequences corresponding to cytosolic ribosomal proteins, we were able to find 78 in the P. brasiliensis transcriptome. Nineteen of the 27 Saccharomyces cerevisiae genes related to translation initiation were also found. All eukaryotic elongation factors were detected in P. brasiliensis transcriptome, with eEF1A as one of the most expressed genes. Translation termination is performed, in eukaryotes, by factors 1 and 3 (eRF1, eRF3). In P. brasiliensis transcriptome it was possible to identify eRF3, but not eRF1. Sixteen PbAESTs showing aminoacyl-tRNA synthetase-predicted activities were found in our analyses, but no cysteinyl-, leucyl-, asparagyl- and arginyl-tRNA synthetases were detected. Among the mitochondrial ribosomal proteins, we have found 20 and 18 orthologs to S. cerevisiae large and small ribosomal subunit proteins, respectively. We have also found three PbAESTs similar to Neurospora crassa mitochondrial ribosomal genes, with no similarity with S. cerevisiae genes. Although orthologs to S. cerevisiae mitochondrial EF-Tu, EF-G and RF1 have been found in P. brasiliensis transcriptome, no sequences corresponding to functional EF-Ts were detected. In addition, 64 and 28 PbAESTs associated to protein modification and degradation, respectively, were found. These results suggest that these machineries are well conserved in P. brasiliensis, when compared to other organisms.

A model of chronic IgE-mediated food allergy in ovalbumin-sensitized mice

Food allergy is most frequently the result of IgE-mediated hypersensitivity reactions. Here, we describe a chronic model in which some of the intestinal and systemic consequences of continuous egg white solution ingestion by ovalbumin-sensitized eight-week-old BALB/c mice, 6 animals per group, of both sexes, were investigated. There was a 20 percent loss of body weight that began one week after antigen exposure and persisted throughout the experiment (3 weeks). The sensitization procedure induced the production of anti-ovalbumin IgG1 and IgE, which were enhanced by oral antigen exposure (129 percent for IgG1 and 164 percent for IgE, compared to sensitization values). Intestinal changes were determined by jejunum edema at 6 h (45 percent Evans blue extravasation) and by a significant eosinophil infiltration with a peak at 48 h. By day 21 of continuous antigen exposure, histological findings were mild, with mast cell hyperplasia (100 percent) and increased mucus production (483 percent). Altogether, our data clearly demonstrate that, although immune stimulation was persistently occurring in response to continuous oral antigen exposure, regulatory mechanisms were occurring in the intestinal mucosa, preventing overt pathology. The experimental model described here reproduces the clinical and pathological changes of mild chronic food allergy and may be useful for mechanistic studies of this common clinical condition.

Immune and hormonal activity in adults suffering from depression

An association between depression and altered immune and hormonal systems has been suggested by the results of many studies. In the present study we carried out immune and hormonal measurements in 40 non-medicated, ambulatory adult patients with depression determined by CID-10 criteria and compared with 34 healthy nondepressed subjects. The severity of the condition was determined with the Hamilton Depression Rating Scale. Of 40 depressed patients, 31 had very severe and 9 severe or moderate depression, 29 (72.5 percent) were females and 11 (27.5 percent) were males (2.6:1 ratio). The results revealed a significant reduction of albumin and elevation of alpha-1, alpha-2 and beta-globulins, and soluble IL-2 receptor in patients with depression compared to the values obtained for nondepressed subjects (P<0.05). The decrease lymphocyte proliferation in response to a mitogen was significantly lower in severely or moderately depressed patients when compared to control (P<0.05). These data confirm the immunological disturbance of acute phase proteins and cellular immune response in patients with depression. Other results may be explained by a variety of interacting factors such as number of patients, age, sex, and the nature, severity and/or duration of depression. Thus, the data obtained should be interpreted with caution and the precise clinical relevance of these findings requires further investigation

Autotransplante de cordas tendíneas: nova técnica para o tratamento cirúrgico da insuficiência mitral por rotura de cordas tendíneas da cúspide anterior / Autotransplantation of chordae tendineae: new technique for the surgical approach of mitral insufficiency

Uma das causas mais freqüentes de prolapso valvar mitral, com conseqüente regurgitaçao, é a rotura de cordas tendíneas. Várias técnicas têm sido empregadas em tal situaçao. Entretanto, a substituiçao valvar mitral tem sido o procedimento mais utilizado. É apresentada uma nova técnica cirúrgica empregada em 4 pacientes com insuficiência valvar mitral severa devido a rotura de cordas tendíneas da cúspide anterior da valva mitral. Um autotransplante de cordas da vliva tricúspide para a valva mitral é realizado. Em 2 casos, um retalho é retirado da cúspide anterior da valva tricúspide com cordas tendíneas e parte do músculo papilar correspondente. Nos outros 2 casos a cúspide posterior da valva tricúspide foi retirada, inteiramente, com cordas e músculo papilar. Na primeira variante técnica, a cúspide anterior da valva tricúspide foi reparada e, na segunda, através, da plicatura do anel, a valva tricúspide foi transformada em bicúspide. A peça retirada é transplantada para a valva mitral, sendo suturada a cúspide doadora com a cúspide anterior da valva mitral e o músculo papilar doador com o topo do músculo papilar posterior da valva mitral em 3 casos e com o músculo papilar anterior e l caso. Os 4 pacientes foram reestudados clínica e laboratorialmente aos 15, ll, 2 e l mês de pós-operatório, respectivamente, com execelente evoluçao. Apesar da limitada experiência, acreditamos ser esta nova técnica uma boa alternativa para o tratamento cirúrgico da insuficiência mitral por rotura de cordas tendíneas da cúspide anterior.

Mucosa intestinal com localizacao bucal. / Intestinal mucosa with oral localization

O presente trabalho relata o achado de tecido epitelial intestinal na mucosa alveolar de um paciente de 18 anos. A entidade manifestou-se apos a extracao do dente da area e recebeu as hipoteses clinicas de lesao de celulas gigantes perifericas e granuloma piogenico. A localizacao da heterotopia, pela primeira vez relatada na literatura, e a idade do paciente levaram os autores a questionar a provavel origem e natureza da entidade em questao