RCAAP Repository

Glial fibrillary acidic protein (GFAP) belongs to the group of intermediate filaments of the cell cytoskeleton. It is present in astrocytes and in some ependymal cells. Detection of GFAP is useful for the identification of astrocytes in a highly specific manner. In the present study, peroxidase antiperoxidase (PAP) immunohistochemical staining, with a specific antibody against GFAP was employed to determine the distribution and morphology of astrocytes in the hippocampus and dentate gyrus of normal cattle and cattle naturally infected with rabies. Paraffin sections of buffered formalin-fixed tissue were used. Positively brown coloured astrocytes in each region were classified into four levels of intensity. The number of astrocytes per mm2 was determined using a straight scale ocular micrometer. Nuclear area, larger and smaller diameter of astrocytes were also determined with an image analyser system. It was observed that the morphology of astrocytes varied in the différents microregions of hippocampus and dentate gyrus. Cattle with rabies showed astrogliosis represented by strong immunoreaction, enlargement of cell body, cytoplasm and processes. The nuclei were also enlarged and light in colour. Astrocitosis was also present. Neurons showed no alterations. These results represent a contribution to the knowledge of the normal distribution and morphology of astrocytes which permits a better understanding of pathological conditions.

Anticorpos précipitantes contra o antígeno gp51 do vírus da Leucose Enzoótica Bovina foram pesquisados em amostras de soros de vacas leiteiras da região do município de Pitangueiras, no Estado de São Paulo. As amostras sanguíneas eram de animais provenientes de sete propriedades rurais que foram acompanhadas anualmente de 1992 até 1995. Durante o ano de 1992 foram analisados 140 soros, dos quais 24 (17,1%) eram reagentes positivos, 21 (15,0%) suspeitos e 95 (67,9%)negativos. Em 1993, dos 122 soros testados, 25 (20,5%) foram positivos, 8 (6,6%) suspeitos e 89 (72,9%) negativos. No ano de 1994 foram testadas 135 amostras, das quais 45 (33,3%) eram positivas, 19 (14,1%) suspeitas e 71 (52,6%) negativas. Finalmente, em 1995, foram analisadas as amostras de 119 animais sendo que 60 (50,4%) eram positivas, 11 (9,2%) suspeitas e 48 (40,4%) negativas. Os resultados obtidos evidenciam que no decorrer do experimento houve um nítidoaumento na prevalência da Leucose Enzoótica Bovina com o passar do tempo. Sob o prisma da Defesa Sanitária Animal, a crescente disseminação da doença na população estudada serve como alerta sobre o comportamento epidemiológico e sobre a evolução na difusão da enfermidade em nosso meio.

Four rumen cannulated steers were used in a 4 x 4 change over design, to test the effects of the following four treatments: A) cotton seed oil meal, B) soybean oil meal, C) raw soybean and D) toasted soybean, over neutral detergent fiber (NDF) rumen degradability of sugar cane, used as only roughage through nylon bags in situ technique. Sugar cane fiber (NDF)disappearance in the rumen was smallest for whole soybean and highest for cottonseed oil meal (p < 0.05). Soybeans, raw and toasted, showed statistical similarity.

Caprinos e bovinos foram utilizados num delineamento change over 3x3, para avaliar a degradabilidade dos tratamentos: A) farinha de carne e osso com 50% de proteína bruta (FCO 50); B) farinha de carne e osso com 40% de proteína bruta (FCO 40) e C) farelo de soja (FS) pela técnica dos sacos de náilon in situ. Cana-de-açúcar foi o único volumoso ofertado. As taxas de degradabilidade da MS (matéria seca) e PB (proteína bruta) foram semelhantes entre caprinos e bovinos, em todos os tratamentos. Os valores de degradabilidade da FCO-40 e da FCO-50 foram semelhantes, porém menores (p < 0,01) do que o FS a partir de 1,5 hora de incubação. A degradabilidade efetiva da MS foi de 77,4% para o FS; 39,0% para a FCO-40 e 39,1% para a FCO-50. Nos tempos 1,5 h, 3 h, 6 h e 12 h a FCO-50 teve a PB menos degradada que a FCO-40. O FS apresentou sua PB mais degradada dos que as FCO, somente nos tempos de 12 h e 48 h de incubação (p < 0,05). A degradabilidade efetiva da PB foi de 72,4% para o FS; 54,4% para a FCO-40 e 49,8% para a FCO-50. O experimento permitiu concluir que as FCO podem prover maiores quantidades de PB para os intestinos, em relação ao FS, após 12 horas de incubação.

Foram determinados alguns perfis bioquímicos e imunológicos em 16 vacas holandesas com retenção de placenta (RP) e 23 sem retenção (NRP) de 241 partos observados. A alimentação dos animais era baseada em forragens cultivadas no inverno e verão, silagem de milho, feno e concentrado com 18,0% de proteína bruta e 70,0% de nutrientes digestíveis totais. As determinações dos componentes plasmáticos no período peripartal (dias -15, -5, dia do parto, +3 e +5) revelaram maiores concentrações de colesterol (p < 0,01), fósforo (p < 0,01), fosfatase alcalina (p < 0,05), desidrogenase láctica (p < 0,05), aspartatoaminotransferase, uréia, fosfatase ácida e menores níveis de cálcio em vacas RP. As proteínas totais estiveram dentro dos padrões fisiológicos em ambos os grupos. Os níveis de imunoglobulinas demonstraram a incapacidade das vacas RP de produzir IgM, IgG sangüíneas (p < 0,001) e IgA no colostro e no leite (p < 0,01) quando comparadas às vacas NRP. As RP apresentaram, no dia da parturição, quadro de neutrofilia, eosinofilia, monocitose e linfocitose, revelando imunodepressão periférica, quando comparadas com as NRP. Observaram-se complicações nas vacas RP, com 87,5% de endometrite e 62,5% de mastite aguda, demonstrando que estas eram mais susceptíveis. Os resultados reforçam a existência de relação estreiw entre a produção de imunoglobulinas, número de leucócitos, retenção de placenta, inflamação do útero e da glândula mamária, tornando possível a previsão de retenção dos anexos fetais 15 dias antes do parto nas vacas de leite.

The objective of this project was to evaluate the post-intervention pain due to three methods of male sterilization. Twenty seven dogs participated in the study and were distributed in three groups: (OT) dogs submitted to orchiectomy; (ZG) dogs submitted to intratesticular injections of zinc gluconate; and (VT) dogs submitted to vasectomy. Pain was evaluated with the dog post-surgical pain evaluation scale from Melbourne. There was no significant difference amongst median pain scores for the groups. In the group ZG, 44% of the animals needed sedation during the application, and 11% showed secondary complications, with ulceration and swollen testicles. The maximum pain score was higher for the ZG group, requiring post-intervention analgesia for one dog. The results motivate studies with higher number of animals regarding zinc gluconate, suggesting that the dogs be closely monitored during 15 days post-intervention.

The high susceptibility of sperm to the oxidative stress occurs especially due to high content of poly-unsaturated fattyacids (PUFAs) in its plasma membrane. The PUFAs provide the necessary fluidity to the plasma membrane. Howeverdouble bonds present in those fatty acids are more susceptible to oxidative stress. Studies in human indicate thatcryopreservation may lead to damages to the sperm due to oxidative stress. This study aimed to verify if the antioxidantglutathione (GSH) may protect ovine cryopreserved sperm against damages caused by oxidative stress. Semen samplesof four rams were cryopreserved using Tris-egg yolk extender supplemented with different concentrations of reducedglutathione (control, 1, 5 and 10 mM). After thawing, samples were evaluated using conventional (motility and vigor)and functional tests (membrane integrity and mitochondrial activity). Aliquots of each thawed sample were submitted toprotocol of induced lipid peroxidation using ascorbate (20 mM) and ferrous sulphate (4 mM), with further measurementof tiobarbituric acid reactive substances (TBARS), index of oxidative stress. No effect of GSH was observed on variablesassessed by conventional tests. GSH decreased the proportion of intact acrosomes. Samples treated with 5 mM GSHshowed lower percentage of intact membrane cells when compared to control samples and those treated with 10 mM.The percentage of cells with mitochondrial activity was affected by GSH, but no effect on TBARS. Samples from controlgroup were more susceptible to denaturation of chromatin. In conclusion, the addition of Glutathione (GSH) offersprotection to DNA and mitochondrial activity of ovine sperm.

Several authors have emphasized the importance of monitoring estrous cycle in bitches and mentioned technique examples of how it can be done. The aim of this study was to evaluate the salivary progesterone quantification technique in order to identify ovulation in this species. To compound the experimental group, 13 bitches of different breeds (no specific breed, English Bulldog, Bernesse Mountain Dog)and different ages (from 11 months to 9 years old) were used. Blood and saliva samples were collected simultaneously in all animals, starting about the first day of proestrus clinical signs. Salivary samples were collected with a specific commercial device (Salivette®). This method was effective, since it has become possible to obtain enough volume in almost all samples to quantify salivary progesterone (100 μLfor duplicate quantification). Serum progesterone was quantified by radioimmunoassay and salivary progesterone by enzyme immunoassay, both of them with commercial kits. There was an increasing, linear and positive correlation between salivary and serum progesterone (r=0.704; p<0.0001) in bitches. One of the animals had an anovulatory cycle, in which, for 13 days, seric progesterone levels were maintained between 1.67 and 3.76 ng/mL and salivary progesterone levels were maintained between 19.55 and 236.77 pg/mL. It was concluded that salivary progesterone levels measured by enzyme immunoassay is a technique that can be used to evaluate the presence or absence of ovulation in bitches.

The present study was conducted to verify if the elevation of plasma concentrations of estradiol during superovulatory treatments affects the oocyte transport in buffalo females, as well as if the inferior quality of buffalo oocytes and/or some functional difference on the oviduct of these animals is responsible for the low embryo recovery rate in superovulated uffaloes when compared to cows subjected to the same treatment. Oviducts of 10 buffaloes and 15 of cows, treated to induce a single ovulation were used. The oviducts were placed on Petri dishes and received the following treatments: 5 buffalo oocytes with no E2 (G-BufBuf and G-BovBuf), 5 bovine oocytes with no E2 (G-BufBov and G-BovBov), 5 buffalo oocytes with E2 (G-BufE2Buf and G-BovE2Buf) and 5 bovine oocytes with E2 (G-BufE2Bov and G-BovE2Bov; factorial 2x2x2). Oocytes were incubated for 24h. Subsequently, oviducts were washed and oocytes were recovered and counted. Since no interactions were found between E2 treatment, oviducts and oocytes species, main effects were analyzed separately. Recovery rate and number of oocytes was higher on cattle compared to buffaloes (35.0+8.6% and 1.4+0.3 vs. 10.0±4.6% and 0.5±0.2, respectively; p<0.05); no effect of E2 treatment was observed on recovery rate and number of oocytes (29.8±9.0% and 1.3±0.4 vs. 16.9±6.1% and 0.7±0.2, respectively; p>0.05); the number of buffaloes and bovine oocytes recovered were similar (1.4±0.4 and 0.6±0.2, respectively; p>0.05). Oocytes recovery rate showed a trend (P=0.07) to be higher when buffalo oocytes were implanted when compared to bovine oocytes (35.2±9.2% vs. 12.9±5.4%). Present results suggest that oocyte transport by the oviduct of buffaloes and bovine was not dependent on oocytes species or E2 supplementation to the culture medium.

O presente estudo teve por objetivo correlacionar o número de bactérias espiraladas e as alterações histológicas da mucosa gástrica em cães de vida livre. Foram analisadas biopsias gástricas endoscópicas de 28 cães assintomáticos. Para análise histológica, foi realizada avaliação qualitativa, onde foram atribuídos escores de 0 a 3, considerando a densidade de bactérias espiraladas por campo (400x), a presença de células inflamatórias, o número de agregados linfoides e a existência de alteração degenerativa glandular. A prevalência de Helicobacter spp, identificado pela histologia (Carbol-Fucscina) e positividade no teste da urease, foi de 100%. Dos 28 cães, 18 (64,3%) receberam escore 3 e 10 (35,7%) o escore 2 para a densidade de bactérias. O infiltrado inflamatório predominantemente linfoplasmocitário foi de grau leve (escore 1) em 17 (60,7%) cães e moderado em 6 (21,4%) cães. Dos 28 cães, 14 (50%) receberam escore 1 para degeneração glandular e 9 (32,1%), o escore 0. As regiões do corpo e antro apresentaram maior número de resultados positivos à histopatologia. Apesar do número elevado de bactérias encontrado nas amostras analisadas, as alterações histológicas foram classificadas como de grau leve na maioria dos animais. A presença do Helicobacter spp. não parece estar relacionado com sintomatologia de gastrite.

O óleo de copaíba é uma resina extraída pela perfuração do tronco de árvores do gênero Copaifera sp., usado como cicatrizante em várias regiões do Brasil. São comprovados seus efeitos antimicrobianos e antiinflamatórios, sem relatos de atividade sobre a proliferação celular. No presente trabalho foi observada a dinâmica da proliferação de células MDBK (Madin Darby Bovine Kidney) mantidas em meio de cultivo adicionado de diferentes concentrações do óleo de copaíba, utilizando-se como controle células mantidas em meio sem adição do oleoresina (grupo M) e células no meio com aplicação do solvente tween 80, na diluição 10-3 (grupo MT). Diluições decimais de 10-1 até 10-3 mostraram-se tóxicas e, portanto, os estudos de proliferação partiram da diluição 10-4 até 10-7. Observou-se que houve crescimento mais acelerado em todos os grupos adicionados do óleo-resina nas primeiras 24 horas, com destaque para a diluição 10-5, que teve sua taxa de proliferação 5,47 vezes maior que a do grupo M. Concluiu-se que o óleo-resina de copaíba se mostrou estimulante da multiplicação celular, o que pode ser um dos mecanismos de seu efeito positivo sobre a cicatrização, somado àqueles previamente comprovados na literatura.

The following study describes meniscus ruptures associated to cranial cruciate ruptures, in 34 dogs of different breeds, ages and weights. Before surgery the animals underwent clinical and radiographic examinations. All animals presented either total or partial cranial cruciate ruptures: 21 (24.14%) of the animals didn’t present any meniscus lesions, and the rest (75.86%) presented only a medial meniscus lesion. The lesions found in the medial meniscus were the following: 33 stifles (37.93%) presented with eversion of the caudal pole (Type 1), 15 (17.24%) showed a bucked handle lesion (Type 6), 3 (3.45%) presented with fibrillation lesion (Type 4), 3 (3.45%) multiple fibrillation lesion (Type 3), 3 (3.45%) longitudinal lesion (Type 2), 1 (1.15%) lesion type 7 and 10 (11.49%) presented multiple lesion. Surgical procedure for cranial cruciate rupture included: tibial tuberosity advancement (TTA) (49 stifle), tibial plateau leveling osteotomies (TPLO) (15 stifle), closing wedge osteotomy (CWO) (14 stifle), extracapsular (4 stifle) and meniscectomy alone (5 stifle), and all these techniques guaranteed weight baring and return to function in the first week after surgery, with no complications. Through this study we could demonstrate that meniscus tear is highly associated to cranial cruciate rupture and that the most common is type 1 (eversion of the caudal pole) and that cronicity of the lesion increases the probability of meniscus tear.

A evolução morfológica do ducto epididimário de cobaio (Cavia porcellus, L.) foi avaliada nas faixas etárias de 10, 20, 30, 45,60, 70, 90 e 100 dias após o nascimento, mostrando-se complexa, o que é decorrente da própria diferenciação pós-natal do epitélio epididimário. Assim sendo, o segmento inicial epididimário mostrou um aumento de altura epitelial correspondente aos valores médios de alturas das células principais do epitélio tubular, crescente após 45 dias de idade pós-natal. As alturas epiteliais obtidas em nível do segmento médio foram maiores em idades juvenis (de 10 a 45 dias) do que na idade pré-puberal (60 dias), tendo diminuído entre os 70 e 100 dias de idade. No segmento terminal epididimário, verificou-se um decréscimo marcante da altura epitelial tubular a partir da idade puberal (70 dias) até a idade adulta (a partir de 90 dias). Além disso, este segmento apresentou ondulações epiteliais que cessaram aos 70 dias de idade, momento em que a luz tubular se preenche com espermatozoides e esfoliações celulares. Os tipos celulares do epitélio tubular epididimário, células principais, basais e apicais, foram observados em todas as idades. Em epidídimos de animais muito jovens, observou-se a predominância de células colunares indiferenciadas. A partir de 20 dias de idade, houve prevalência natural das células principais sobre os demais tipos celulares, como é peculiar ao epitélio epididimário de mamíferos, de modo geral.

The effects of swim bladder injection with thioglycolate, Escherichia coli lipopolysaccharide (LPS) and heat-inactivated Aeromonas hydrophila were assessed on hematological responses in pacu, Piaractus mesopotamicus (Characidae). A quantitative assessment was done on erythrocytes, thrombocytes e leucocytes at 6, 24, and 48 h pos-injection of the inflammatory agents and compared with fish injected with saline solution (control). Fish injected with inactivated A.hydrophila showed a reduction of erythrocytes and hemoglobin, whereas the hematocrit increased 6 h pos-injection.The results show that thioglycolate and LPS also induced a reduction on hemoglobin and an increase on the hematocrit. The thrombocytes count decreased 6 h post A. hydrophila injection, whereas increased 48 hours post LPS injection. The leukocytes count increased after 6 h post A. hydrophila injection, while the lymphocytes and PAS-positive granularleukocytes (PAS-LG) count decreased after 24 h post injection. In fish injected with thioglycolate or with LPS showed an increase in the LG-PAS counts when compared to A. hydrophila or control groups. The monocytes count was not affected by the different inflammatory agents.

Bovine tuberculosis, persists in several countries as a threat to health of the herds despite the efforts given to the control and eradication of the disease. In order to detect immunogenic proteins with a potential to be used in a bovine tuberculosis diagnosis test, serum samples obtained from reactors and no reactors to intradermal tuberculin test, were evaluated for reactivity to Mycobacterium bovis antigens from a bacilli originated in Goiás – Brazil. The proteins immunogenicity from M. bovis was obtained by Western blot according to the molecular weight profile. The majority of the PPD positive bovine (67.92%) recognized a protein with 26kDa, suggesting the use of this protein in a serological test for bovine tuberculosis.

Atopic dermatitis (AD) is an inflammatory, pruritic and chronic allergic skin disease. It is recognized as the second most common allergic skin disease of dogs, after flea allergy. Pruritus is the predominant sign of canine AD, and it affects a variety of areas of the body, leading to intense suffering in both the animal and its owner. The long-term use of glucocorticoid therapy can be devastating because of its numerous side effects and secondary diseases. Cyclosporine (CsA) has been considered a good therapeutic option to treat canine AD. CsA inhibits the activation of cells that initiate the cutaneous immune response (Langerhans cells and lymphocytes) and that mediate allergic reactions (mast cells and eosinophils). It also decreases the release of histamine and other cytokines. The objective of this study was to analyze the efficacy of CsA (5 mg/kg, SID for 60 days) to reduce skin lesions and pruritus in 21 atopic dogs using CADESI-03 and two scales to quantify the levels of body itching. This immunomodulatory therapy was considered to be an effective treatment for atopic dogs because it reduced skin lesions by 70% after 60 days of therapy. During that period, there was a 52.6% reduction of body itching as assessed via a verbal numeric scale, and there was a significant reduction of body itching on a qualitative scale, as the maximal levels of pruritus (“three” and “four”) were hardly observed after immunomodulatory therapy. CsA was effective and safe in the treatment of canine atopic dermatitis.

Neste trabalho investigou-se em camundongos BALB/c, machos e fêmeas, os efeitos comportamentais da exposição aguda a uma formulação comercial do glifosato (GF) em uma dose duas vezes maior que a dose sem efeito observado(NOAEL). A neurotoxicidade aguda ao GF foi determinada por meio da análise da atividade geral, de parâmetros sensoriais, psicomotores, do sistema nervoso central e autônomo em machos e fêmeas. Os efeitos exploratório, de ansiedade e depressão induzidos pelo GF foram observados no campo aberto, labirinto em cruz elevado e no teste da suspensão da cauda, respectivamente. O GF promoveu poucos sinais de neurotoxicidade. A capacidade exploratória de fêmeas foi reduzida no campo aberto. Nenhum sinal de ansiedade foi detectado tanto em machos como em fêmeas no labirinto em cruz elevado, porém, notou-se redução da exploratória neste aparelho. No teste de suspensão da cauda tanto as fêmeas como machos mostraram aumento no tempo de imobilidade. Não foi observado neste caso interação entre sexo e tratamento. Concluiu-se que a exposição ao dobro da dose da NOAEL do GF induziu poucos sinais de neurotoxicidade e poucos efeitos sexualmente dimórficos em camundongos machos e fêmeas.

Considering the limitations of current methods of anaplasmosis control, the development of a more effective vaccine is required. Previous studies, using proteomic and genomic approaches, have identified new membrane proteinsin Anaplasma marginale that may be vaccine candidates. These include VirB9, VirB10 and elongation factor-Tu (EFTu).Although the role of these proteins in the membrane of A. marginale has not been properly characterized, production of the recombinant proteins rVirB9, rVirB10, and rEF-Tu has been achieved. However, these recombinant proteins have not yet been exploited in vaccine formulations. The present study describes the use of rVirB9, rVirB10and rEF-Tu, emulsificated in adjuvant Montanide, in mice. A strong humoral immune response was induced under these conditions, with both IgG1 and IgG2a production. The IgG2a/IgG1 ratios revealed a predominance of IgG1.However, splenocytes of the animals that received rVirB9 or rVirB10 produced high levels of gamma interferon after in vitro stimulation, indicating a specific cellular immune response to these proteins. Therefore, further studies are required to adjust the profile of the immune response in order to perform tests of protection against A. marginale in cattle.

Avian infectious bronchitis virus (IBV) (Nidovirales: Coronaviridae) is a chicken Gammacoronavirus with the highestevolution rate in the genus and, despite the recently reported proofreading activity of its polymerase, intra and interhost diversity is a well-documented phenomenon. This study aimed to assess the genetic variation of serial passages of a variant genotype IBV strain in vitro. Strain CRG-BETA, propagated in chicken embryos, was inoculated in VERO cells monolayers up to the 4th passage and each passage was monitored with an RT-PCR targeted to the S1 gene (nt 705to 1094) and an RT-PCR to the protein 5a mRNA. All passages were positive to RT-PCRs to S1 and passages 1 to 3 to5a mRNA; S1 sequences showed no polymorphism. The finding of IBV mRNA in the cell cultures demonstrates that the CRG-BETA IBV strain is replicating in the VERO cells and regarding S1 sequence analysis, the lack of nucleotide mutations shows that CRG-BETA might have reached a fixed status. As a conclusion, different genotypes of IBV present different evolutionary patterns not only in vivo as previously known, but also in vitro, as described herein.

The aim of this study was to verify the quantitative alterations of the neurons bodies in the brain cortex of malnourished dogs. Seven brains of mongrel male dogs that were previously diagnosticated as malnourished were collected. The dog shad similar constitutionalist characteristics of cranium (mesaquicefalus). It was chosen different gyrus of the braincortex to been study; those gyrus were sampled, prepared according to conventional histological technique and stainedby modified cresil violet, for becoming evident the neurons bodies. The slides were analyzed with the 20x ocular. Our results indicate a expressive reduction in the number of neuron bodies in malnourished dogs (10,8), when compared with dogs in normal nutrition conditions (16,35). In conclusion, it’s possible that the malnutrition leads to an reduction of neurons bodies in brain cortex of dogs, when compared to those with normal condition of nutrition.