Repositório RCAAP

In order to achieve monolayers and adhesion in cell cultures of hepatocytes from the subtropical fish Metynnis roosevelti (Pisces, Teleostei, Characidae) three different methodologies were used: enzyme cell dissociation by means of a system containing trypsin and versene 0.25%; by means of bovine fetal serum and by the use of blood serum obtained from adult specimens of M. roosevelti. The cell medium used has been a modified F10-199 by the addition of 2 mM L-glutamine, 10 µg/mL insulin, 50 µg/mL fibronectin; and antibiotics. The best results in terms of cell dissociation as well as in terms of adhesion and confluence of the cells monolayer has been achieved by means of the enzymatic cell dissociation.

A pesquisa consta de estudo macroscópico do sistema arterial e venoso do funículo umbilical e da placenta de fetos de gato e caracterização placentária quanto ao tipo, zonária que em 62,5% dos casos apresenta uma fissura em região distal ao funículo umbilical. O funículo é formado por uma porção justafetal, uma porção justaplacentária e um terço médio, encontrando-se: duas artérias umbilicais, uma veia umbilical, dois pedículos vitelíneos e dois pedículos alantoídeos.

Crotalaria spectabilis (crotalária) e a Senna occidentalis (fedegoso) can grow in corn fields and during harvesting this cereal can be contaminated with the seeds of those plants which presents toxicity to animals. The aim of this study was to investigate the erythrocytes morphometric parameters, haematological variables and plasma hormones concentrations of broilers chickens fed rations of two energy levels to which were added 0,1% e 0,5% of Crotalaria spectabilis e a Senna occidentalis seeds, respectively. One hundred and eighty broilers chicks were divided into 6 groups according to a factorial analysis 3 x 2 (control, crotalaria and fedegoso as main treatments and two energy levels, 2,900 and 3,200 kcal ME/kg of ration). The results of this experiment showed that the crotalaria toxin reduced the number of the red blood cells, the haematocrit and mean corpuscular volume (MCV) with no effect on the morphometric studied parameters. These seeds due to its hepatic toxicity induced a high incidence of ascites. The fedegoso seeds were not able to induce ascite in the broilers.

Três vias de fornecimento de dietas pós-operatórias foram comparadas na cicatrização de esofagostomia cervical. Foram utilizados 15 cães, divididos em três grupos (GI, GII e GIII). No GI foi procedida à fluidoterapia durante 48 horas, alimento líquido durante mais 48 horas e pastoso por 72 horas (dieta tradicional), no GII o alimento foi fornecido por sonda faringogástrica, colocada por faringostomia, e no GIII por sonda gástrica, implantada por gastrostomia endoscópica percutânea. Para avaliar a cicatrização, foram realizados exame clínico diário e esofagoscopias semanais, durante seis semanas. À endoscopia perceberam-se algumas ocorrências indesejáveis que retardam a cicatrização da mucosa esofágica: edema entre os pontos na primeira endoscopia pós-operatória (GI) e ulcerações na mucosa esofágica pelo atrito da sonda faringogástrica sobre a cicatriz (GII). A alimentação por sonda gástrica resultou em menor tempo de cicatrização (1,40 ± 0,55 semanas, p < 0,01), em comparação aos alimentados por sonda faringogástrica (4,25 ± 1,50 semanas). No GIII as características clínicas da cicatrização foram de melhor qualidade, provavelmente devido à ausência de movimentos de deglutição e de atrito do bolo alimentar sobre a ferida cirúrgica. Com base nos resultados, recomenda-se o uso de nutrição enteral por sonda gástrica colocada por gastrostomia endoscópica percutânea, durante os primeiros sete dias de pós-operatório, e, na ausência de um endoscópio, a dieta tradicional, que resulta em cicatrização de melhor qualidade do que o uso de sonda por faringostomia.

The research looked upon the influence of dietary nitrogen level or equivalent ruminal degradable protein (RDP) on semen biochemistry characteristics and freezability of sheep spermatozoa. Eighteen young rams, were allocated in tree blocs of six animals each, according to scrotal circumference. The rams were randomly distributed to three treatments: A (control) a maintenance diet, B and C, maintenance diet added 20 g and 40 g of urea (75% and 150% of RDP), respectively. Three weeks after dietary adaptation, blood and semen samples were collected every week for nine weeks, to analyze blood plasma and semen levels of urea, transaminases (AST and ALT), acid phosphatases, frutose and citric acid. Once a week for six weeks the motility percent, motility rates, abnormal acrosome percent and abnormal total percent between pre-freezing and post-thaw of cryoprotected semen samples were freezing, evaluated and calculated to mathematical difference. There was a linear regression (p < 0.01) between levels of UP 18.26 mg%, 34.42 mg% and 49.77 mg%, and levels of semen urea 32.47 mg%, 51.44 mg% e 60.46 mg%, for treatments A, B and C respectively. There was no effect of treatments on fructose, citric acid or enzymes acid phosphatase and transaminases levels, and of motility percent, motility rating, abnormal acrosome percent and abnormal total percent differences. The excess of nitrogen or RDP dietary levels elevates urea in blood plasma and semen, but did not affect biochemistry characteristics and freezability of rams spermatozoa.

Avaliaram-se comparativamente as provas de soroaglutinação rápida, soroaglutinação em tubo, 2-mercaptoetanol e antígeno tamponado acidificado no diagnóstico da brucelose bovina. Todas as provas apresentaram boa concordância quando considerada a interpretação preconizada pelo Ministério da Agricultura do Brasil. As provas do antígeno tamponado acidificado e do 2-mercaptoetanol apresentaram alta concordância. Neste sentido, o presente estudo propõe o uso da prova do antígeno tamponado acidificado como triagem para o diagnóstico da brucelose bovina.

Uma cadela, Dachshund, quatro anos, foi encaminhada ao H.V. da FMVZ-UNESP-Botucatu-SP com histórico de prostração, emagrecimento progressivo, anorexia, cansaço e tosse seca. Ao exame clínico observou-se apatia, secreção nasal serosa bilateral e dispnéia intensa. Radiografia torácica demonstrou efusão pleural, edema pulmonar acentuado, desvio dorsal da traquéia e possível massa mediastinal. A ultra-sonografia, após drenagem torácica, confirmou a presença de massa mediastinal, sendo classificada como neoplasia maligna de origem epitelial pela citologia aspirativa por agulha fina. O animal veio a óbito logo após o diagnóstico de neoplasia. À necropsia constatou-se massa mediastinal encapsulada, consistência macia e superfície de corte com coloração branco-acinzentada e áreas necrótico-hemorrágicas, localizada na região ântero-ventral do tórax. O exame histopatológico demonstrou células epiteliais neoplásicas, células linfóides (timócitos) com morfologia normal, vasos sangüíneos de pequeno e médio calibre, formações císticas com conteúdo eosinofílico e corpúsculos de Hassall. A imunohistoquímica apresentou positividade para citoqueratina AE1/AE3 e UCHL, confirmando o diagnóstico de timoma.

Twenty one adult horses, males and females, were pretreated with 0.14 mg/kg of hyoscine-N-butylbromide intravenously and injected 5 minutes later with 0.02 mg/kg, iv of detomidine (group A, n = 9) or saline (group B, n = 12). Mean arterial pressure measurements and electrocardiography were performed during 65 minutes. After hyoscine injection the heart rate was increased by 43% and 65% in A and B groups, respectively. Heart rate remained increased after injection of detomidine, returning to baseline values after 15 minutes. No increase in the mean arterial pressure (MAP) was noticed after hyoscine but the MAP was increased by 62% after detomidine, returning to basal measurements until the end of observation time. No additional increase was noted in the group B horses. Hyoscine shortened PR and QT intervals in both groups, but after detomidine, PR and QT intervals enlarged significantly at the end of the experiment. The second degree atrioventricular block occurred in 3 horses after 40 minutes only in group B. It was concluded that hyoscine prevented detomidine induced bradycardia and may be an useful drug combination against the bradycardia induced by this alpha-2 agonist, in horses.

A completely randomized trial was conducted in order to evaluate the hydrogen ion concentration (pH), total titratable acidity and methylene blue reduction time test in rumen fluid of cross bred and raced goats, grazing on grasslands and rangelands of semi arid regions of Brazil, during rainy and dry season of the year. A 2³ factorial arrangement was used to analyse the experiment (two types of bread, pasture and season). Significant effects were obtained for type of pasture on total titratable acidity. The pH was influenced by season and type of pasture. The microbial activity, mensured through methylene blue reduction time, was more intensive on rainy season and rangelands than dry season and grasslands, respectively. In the interpretation of the results of these tests for goats, should be considered mainly the season and type of pasture, without despising the breed.

Fourteen Holstein-Zebu calves were used to evaluate the effects of probiotic with Lactobacillus acidophilus, Streptococcus faecium and Sacharomyces cerevisae in diet on performance and meat quality of veal calves. Calves were randomly assigned in two treatments: whole milk and whole milk with probiotic. Calves were weight at birth and every fifteen days until slaughter, between 160 and 190 kg body weight. Body weight daily gain, dry matter intake, feed efficiency, dressing percentage of carcass, fecal score and sensorial analyses in Longissimus dorsi muscle were evaluated in a completely randomized block design. Data were interpreted by variance analysis and means compared by F test. Weekly fecal scores were interpreted by non-parametric variance analysis, by Wilcoxon test. Any differences were observed in sensorial analysis of meat samples, and data indicate a quite clear coloration, with compatible intensity of red color wanted by market. Average values of body weight daily gain, feed efficiency, weight and dressing percentage of carcass for both treatments were 892 and 945 g, 1.41 and 1.30, 90.9 and 98.8 kg, 57.1 and 58.3%, respectively, for calves fed whole milk and whole milk with probiotic (p &gt;; 0.05).

Vinte bezerros machos Holandeses, desaleitados, com cerca de 100 dias de idade e 109 kg de peso, foram alimentados com proteína texturizada de soja (PTS) na forma sólida ou líquida.Os animais foram manejados de forma a manter-se a formação de suas goteiras esofageanas, após o desaleitamento, e receberam quatro tratamentos, dentro de um delineamento em blocos ao acaso: A) 100% PTS na forma sólida, mais 4 kg de água; B) 66% PTS na forma sólida, e 33% PTS dissolvida em 4 kg de água; C) 33% PTS na forma sólida, e 66% PTS dissolvida em 4 kg de água; D) 100% PTS dissolvida em 4 kg de água. As porções líquidas das dietas foram fornecidas em biberões e à temperatura de 35 graus centígrados. Desempenhos em ganhos de peso, ingestões de matéria seca e conversões alimentares não mostraram diferenças entre tratamentos.

The celiac ganglion plays a major role in the innervation of the stomach, intestines, liver and pancreas and also contributes to the innervation of the spleen and is therefore essential for the control of gastrointestinal motility. Clinicians and surgeons should be familiar with the nervous supply to these organs, specially in regard to digestive atonies, hemorrhagic gastroenteritis, gastric torsion and intestinal invaginations. We studied the gross and microscopic anatomy of the celiac, celiacmesenteric and cranial mesenteric ganglia. We carried out 30 adults domestic cats, 10 males and 20 females. Red neoprene latex was injected into the thoracic aorta. Following this procedure, the animals were frozen for at least 48 hours. After unfreezing, fixation was concluded in 10% aqueous formaldehyde solution. Light microscopic studies were conducted using hematoxylin-eosin, Masson’s stain, the reticulin method and acid hematoxylin fosfotungstic. Located in periarterial position we observed 7 celiac ganglions, 4 to the right and 3 to the left. Most of them (23.3%) were elliptic. We also observed 24 celiac mesenteric ganglions. In two cases, the celiac mesenteric ganglion was composed of two portions: a right and a left portion, which join caudally to the cranial mesenteric artery. In 11 samples we noticed a right celiac mesenteric ganglion with a left cranial mesenteric portion surrounded the left face of the cranial mesenteric artery, thus contributing to its asymmetric semilunar form. These findings suggest that the fusion of the celiac and the mesenteric ganglions prevails. The result of this fusion is the celiac mesenteric ganglion that was formed of the neurons immersed in an abundant connective tissue matrix and are involved by a capsule which contains elastic, collagens and reticular fibers and continues at the ganglion’s fusion sites.

The arterial blood supply of the sinoatrial node has been studied in 30 hearts of females, of Piètrain swine breed. The hearts had, after being detached and adequately cleaned, their coronary arteries injected with colored Neoprene latex 450 solution, and fixed later in formalin solution at 10% for consecutive dissection. In 27 (90% ± 5.47) hearts the sinus node is nourished by collateral branches of the right coronary artery, more exactly, of the right proximal atrial branch exclusively in 18 (60% ± 8.94) organs or associated: with the right intermedio atrial branch in 7 (23.33% ± 7.72) hearts and with the right distal atrial branch in 2 (6.66% ± 4.55). In 3 (10% ± 5.47) organs, the sinus node is nourished by collaterals coming from the right and left coronary arteries, always by the right proximal atrial branch associated with the left proximal atrial branch in 2 (6.66% ± 4.55) or with right intermedio and left proximal atrials branches in 1 (3.33% ± 3.27) organ. Arterial anastomosis involving branches related to the nourishment of the sinus node in swine of Piètrain breed were observed.

Estudou-se a vascularização arterial em nove fêmeas adultas de cateto, incluindo uma jovem e uma gestante, e em seis fêmeas de queixada (três jovens e três adultas). Para este estudo as artérias foram injetadas com solução de látex Neoprene 650 corado e após fixação de formol foram dissecadas. O material foi obtido na Universidade Federal do Paraná - Campus Palotina e na Universidade Estadual Paulista - Campus de Ilha Solteira. As artérias responsáveis pela irrigação sangüínea dos ovários, tubas uterinas e útero são as artérias útero-ováricas, que emitem as artérias uterinas e ováricas, e as artérias vaginais, estando presentes em todas as amostras, porém variando suas origens. As artérias ováricas emitiram ramos para os ovários, tubas uterinas (ramos tubáricos) e extremidades craniais dos cornos uterinos (ramos tubo-uterinos) e as artérias uterinas enviaram ramos para os cornos uterinos, corpos uterinos e porções craniais da cérvix uterina. As artérias vaginais enviaram ramos para a cérvix e a vagina.

Foram eviscerados os globos oculares esquerdos de 32 ratos, linhagem Wistar, divididos em quatro grupos (A, B, C, D) constituídos, cada um, de cinco testemunhas e três controles. Nos animais-testemunha introduziu-se, dentro da capa córneo-escleral, uma esfera de resina acrílica (metilmetacrilato), previamente confeccionada e esterilizada por autoclavagem, ao passo que nos controles a cavidade eviscerada foi mantida sem prótese. Os ratos dos grupos A, B, C e D foram sacrificados respectivamente aos 7, 15, 30 e 90 dias de pós-operatório, quando os conteúdos orbitários esquerdos foram exenterados e preparados para o exame histopatológico. Observou-se que os animais-testemunha tiveram resposta inflamatória do tipo tecido de granulação ao redor da prótese de cavidade, com edema inflamatório da córnea especialmente nos grupos A e B, quando se iniciou a regressão da inflamação aguda. A cavidade orbitária manteve o tamanho em todos os grupos nos animais-testemunha e houve contração significativa nos animais-controle. Com estas observações, foi possível concluir que a esfera de resina acrílica é uma opção, de baixo custo e fácil confecção, para correção de defeito estético causado pela perda do globo ocular.

Bovine karyotyping has become an important diagnostic tool in animal breeding. In the prenatal period it can diagnose several chromosomal abnormalities such as Robertsonian translocations, testicle feminization syndrome, gonadal dysgenesis and Klinefelter’s syndrome. An important cell source for karyotype analysis is the amniotic fluid. It has been extensively used in humans but in bovine, however, this is not the case despite its diagnostic value. Since a small percentage of cells is viable, cells and their growth conditions as well as the handling of the material should be optimal to insure a successful analysis. For this, we have compared the growth efficiency for bovine amniocytes in two media, employing cells from 10 to 14 weeks of gestation. Amniocytes were cultured in the Amniomax (Gibco-BRL/ Life Technologies, Rockville, MD USA) medium during eleven days and in the RPMI 1640 (Gibco-BRL) medium during sixteen days at 37ºC and 5% CO2, then fixed and GTG banded. All the cultures with RPMI showed a poor cell growth, regardless the gestational age. Out of the samples cultured in Amniomax one presented 100% of cellular confluence at day 11 (10 weeks of gestation) and the others resulted in an increased proliferation compared with those that were cultured in RPMI. To ensure a successful karyotyping, amniotic fluid from cows with gestational ages of 10-12 weeks should be used and care should be taken for critical steps in preparation of spread metaphases - hypotonic and trypsin treatments.

We analyzed the chromosomes of 117 cows of different breeds for the identification of centric fusion and the chromosomes of 100 young Andaluz and Brasileiro de Hipismo mares for the identification of 63,X lines, using the technique of X chromosome identification based on interstitial heterochromatin on the long arm.

Foram investigadas as células dendríticas (CD) na pele normal de cinco bezerros das raças Nelore, cinco da raça Holandesa Preta e Branca e cinco animais mestiços por meio da imunomarcação pela proteína S-100. Os animais mestiços foram infestados experimentalmente com 100 larvas de primeiro estádio de Dermatobia hominis e deles foram colhidas biópsias de pele parasitada às 24, 48, 72 e 168 horas após a infestação. Biópsias de pele destes animais, colhidas antes da infestação, foram utilizadas como controle. A imunomarcação das CDs foi feita empregando-se anticorpos de coelhos antiproteína S-100 e a técnica da avidina-biotina-peroxidase. Além das CDs, melanócitos, nervos e células endoteliais apresentaram imunomarcação pela proteína S-100. As DCs foram observadas exclusivamente na derme superficial, próximas à camada basal, tanto nos animais infestados como nos não-infestados. Não se detectou diferença significativa no número de CDs que pudesse ser atribuída à raça dos animais. Nos animais parasitados por D. hominis, as CDs apresentavam-se mais intensamente coradas e com os prolongamentos mais espessos do que nos controles não-parasitados. Além disso, nos animais parasitados observou-se um decréscimo significativo no número de CDs a partir de 24 horas após a infestação.

Sexual maturation involves restraint of gonadal activity from birth to puberty, while somatic development continues. Fetal gonadal steroidogenesis is very important for the maintenance of pregnancy and foaling in mares. The present study was conducted to evaluate gonadal steroid serum levels in foals after birth. Five fillies born at Equília Stud Farm, Avaré, SP, Brazil, were studied. Blood samples were collected daily in the morning during the first week of life. Progesterone serum levels were measured by a commercial radioimmunoassay (RIA) kit and estradiol serum levels with a sensitized non-commercial RIA. At birth, both progesterone and estradiol serum levels were high (13.46 ± 5.5 nmol/l and 7.95 ± 1.5 nmol/l) and decreased to undetectable levels at the end of the first week of life. A negative correlation was found between fillie age and gonadal serum steroid concentration. Results show that fetal gonadal activity persists until birth, and decreases gradual and slowly during the first week of life, as reflected by steroid concentrations in the blood.

Compacted mouse morulae were frozen at 0.3ºC/min. or 0.5ºC/min. from -6ºC to -24ºC or -32ºC in 10% of glycerol plus different sucrose concentrations with or without 0.1% of honeybee royal jelly. Embryos were thawed in water bath at 22ºC for 20 seconds and cryoprotectant dilution was done in three steps. Embryos were cultured in Whitten’s medium for 24, 48 and 72 hours at 37ºC, 5% of CO2 and 100% of humidity. The in vitro development ranged from 56.6% to 100% after 72 hours. Expanded blastocysts were transferred to pseudopregnant recipients on the third day of the estrous cycle. Viable fetuses rates for embryos frozen to -24 or -32ºC at 0.3ºC/minute in 10% glycerol + 10% sucrose, 10% glycerol + 10% sucrose + 0.1% honeybee royal jelly, 10% glycerol + 0.1% honeybee royal jelly or 10% glycerol were respectively: 28.1% and 13.6%, 48.7% and 31.9%, 28.6% and 13.2%, 20.0% and 42.4%. Viable fetuses for embryos frozen to -24ºC or -32ºC at 0.5ºC/minute in 10% glycerol + 10% sucrose or 10% glycerol + 10% sucrose + 0.1% honeybee royal jelly were respectively 29.0% and 15.3%, 48.8% and 32.0%. We can conclude that addition of 10% sucrose to 10% glycerol was efficient for embryo freezing at 0.3 or 0.5ºC/minute and plunged in liquid nitrogen at -24ºC. The honeybee royal jelly addition provided higher viable fetuses rates when embryos were cooled at 0.3 or 0.5ºC/minute and plunged in liquid nitrogen at -24ºC.