Somatic cell nuclear transfer is associated with altered expression of angiogenic factor systems in bovine placentomes at term.

Low efficiency of somatic cell cloning by nuclear transfer has been associated with alterations of placental vascular architecture. Placental growth and function depend on the growth of blood vessels; VEGF-A and bFGF are the most important factors controlling neovascularization and vascular permeability in the placenta. We hypothesize that the VEGF-A and bFGF systems are disrupted in placentomes from cloned animals, contributing to the placental abnormalities that are common in these clones. We determined mRNA expression and protein tissue localization of VEGF-A, bFGF, and their receptors in placentomes from cloned and non-cloned bovine fetuses at term. Real-time RT-PCR revealed that VEGFR-2 mRNA was increased in cloned male-derived placentomes, while mRNA of bFGF and its receptors were decreased in placentomes of cloned females. VEGF-A system proteins were found to be located in placentomal endothelial, maternal and fetal epithelial and stromal cells; there was a variable pattern of cellular distribution of these proteins in both cloned and non-cloned animals. Alterations in the expression of VEGF-A and bFGF systems suggest that angiogenic factors are involved in abnormal placental development in cloned gestations, contributing to impaired fetal development and poor survival rates.

Simultaneous presence of bovine papillomavirus and bovine leukemia virus in different bovine tissues: in situ hybridization and cytogenetic analysis.

Bovine papillomavirus (BPV) DNA sequences were detected in different tissues, in addition to epithelium. Cytogenetic abnormalities were observed in blood lymphocytes. The presence of more than one virus in a single tissue is a difficult aspect to evaluate, especially when the DNA sequences are detected in tissues that are not specifically targeted by the virus. BPV and bovine leukemia virus (BLV) are clastogenic, causing chromosome aberrations in peripheral blood lymphocytes. In the present study, we investigated the simultaneous presence of DNA sequences of both viruses and the possibility of vertical transmission and compared the types of chromosome aberrations related to viral action. BPV 1, 2, and 4 DNA sequences were found in three females of the herd and in their offspring. BLV DNA sequences were not detected in their progeny. A newborn calf that was negative for BLV infection showed specific chromosome rearrangements possibly related to the effect of infection with BPV.

Can established cultured papilloma cells harbor bovine papillomavirus?

Papillomaviruses have been reported to be very difficult to grow in cell culture. Also, there are no descriptions of cell cultures from lesions of bovine cutaneous papillomatosis, with identification of different bovine papilloma virus (BPV) DNA sequences. In the present report, we describe primary cell cultures from samples of cutaneous lesions (warts). We investigated the simultaneous presence of different BPV DNA sequences, comparing the original lesion to different passages of the cell cultures and to peripheral blood. BPV 1, 2 and 4 DNA sequences were found in lesion samples, and respective cell cultures and peripheral blood, supporting our previous hypothesis of the possible activity of these sequences in different samples and now also showing how they can be maintained in different passages of cell cultures.

Toxoplasma gondii infection in Brazilian domestic outpatient cats.

The occurrence of Toxoplasma antibodies in domestic outpatient cats in the city of São Paulo was evaluated using the indirect immunofluorescence assay. A total of 248 blood samples obtained from male and female cats seen at the Veterinary Teaching Hospital at the University of São Paulo between February 1996 and January 1997 were tested. Of these, 17.7% were positive, with a 64 titer being detected in most animals. The frequence of Toxoplasma antibodies was significantly higher in older cats, those fed raw meat and those with free access to the outdoor environment. There was no significant difference in reactivity between males and females. We conclude that diet and free access to the outdoor environment were equally important as predisposing factors to the risk of Toxoplasma infection.

[The preliminary stage of labor in cattle in relation to the clinical signs of labor and the course of progesterone secretion for the prediction of the calving time]. / Das Vorbereitungsstadium der Geburt beim Rind unter Berücksichtigung der klinischen Geburtsanzeichen und des Progesteronverlaufs für die Vorhersage des Geburtstermins.

To evaluate changes in the externally recognizable indications of parturition, the body temperature and progesteroneprofile were examined in 9 Holstein-Friesian heifers and 14 Holstein-Friesian cows in the preparatory phase of parturition. A special external obstetric examination was performed on these animals in the last 263 hours of pregnancy. Furthermore, the vaginal and rectal temperature was measured, and blood samples were taken. Of the blood samples the plasma progesterone concentration was determined. The time of birth could be predicted to be in the next 22 hours on the basis of the relaxation of the broad pelvic ligaments in 52.5% of the cases. A prediction was possible in 43.5% of the cases on the basis of the drop in temperature, and in 91.3% of the animals using the reduction in progesterone levels. Considering the clinical signs of parturition "relaxation of the pelvic ligaments" and a "decrease in temperature", predicting the time of parturition was possible in 65.2% of the animals. In 8.7% of the cases, in which no reduction in the progesterone level below 1.5 ng/ml blood plasma could be seen, the time of delivery could be determined by considering the relaxation of the broad ligaments and the reduction in temperature. A prediction of the time of parturition to be in the next 22 hours was possible for all animals the indicators of parturition.

[Changes in calcium concentration in bovine udder secretions during the preparatory stages of labor in relation to prediction of the time of calving]. / Veränderungen des Kalziumgehalts im bovinen Eutersekret während des Vorbereitungsstadiums der Geburt mit Berücksichtigung der Vorhersage des Abkalbetermins.

Nine heifers and 10 cows of the breed 'German Black Pied' were used in order to evaluate the practicability of measurement of changes in concentration of calcium in udder secretions during the preparatory stage of labour for the prediction of the time of calving. A gradual increase in the concentration of calcium in udder secretions ante partum (a.p.) was determined. Eleven days a.p., the average concentration of calcium was 14.2 mmol/l. Thereafter, the calcium content gradually increased and reached a mean value of 52.1 mmol/l sub partu (secretions of all udder quarters), and 52.4 mmol/l and 51.8 mmol/l in secretions of fore and hind quarters, respectively. Despite the increase in calcium, the time of calving in individual animals could not be predicted reliably, or with a high probability, because the increase in calcium content did not occur in all cows, the variation of the values was high, and the differences between values during the last 48 h a.p. were only small.

Chromosome aberrations in cattle raised on bracken fern pasture.

Thirteen cows maintained on natural bracken fern (Pteridium aquilinum) were analyzed cytogenetically. The frequency of structural chromosome aberrations detected in peripheral blood cells was significantly higher when compared to that detected in animals raised on pasture containing no bracken fern. We discuss the clastogenic action of fern and its synergistic action with infection by type 2 and 4 papilloma virus in the same animals.

Laboratory and clinical features of experimental feline leptospirosis.

In order to assess the clinical, laboratorial and epidemiological aspects of feline leptospirosis, ten female and male adult cats were experimentally inoculated with pathogenic and autochthonous field isolate of Leptospira interrogans. Five of them were inoculated subcutaneously with serovar icterohaemorrhagiae (R-192) and the others five with serovar canicola. No clinical and laboratorial alterations were found in these animals. Antileptospiral agglutinins were detected in 90% of the infected cats, shortly after the 1st week after inoculation. The leptospiral agglutinins were detected for 8 to 12 weeks and the elimination of leptospires through urine was observed only in animals infected with serovar canicola, beginning 2 to 4 weeks after inoculation and lasting for 2 to 8 weeks. Isolations of leptospires from blood and kidneys were unsuccessful.

Bovine papillomavirus transmission and chromosomal aberrations: an experimental model.

Enzootic haematuria and urinary bladder cancer in cattle are associated with feeding on bracken fern and bovine papillomavirus (BPV) infection. An increased rate of chromosomal aberrations in peripheral blood lymphocytes from chronically affected haematuric cows raised in bracken fern pastures has been reported, suggesting the presence of BPV in the peripheral blood of afflicted animals. The purpose of the present investigation was to examine the role of peripheral blood as a potential BPV-transmitting agent and search for clastogenic effects in experimentally infected animals kept on a bracken fern-free diet. Healthy cows were inoculated with blood samples of haematuric animals every two weeks for 18 months. Recipient cows, their offspring, donor animals and a control group were kept on a bracken fern-free diet throughout the experiment. Clinical and molecular analyses for detection of BPV infection were carried out periodically in all groups. Short-term lymphocyte cultures were performed to assess chromosomal aberration levels. The donor cows, the recipient cows and their offspring presented increased levels of chromosomal aberrations. BPV-2 DNA was identified by Southern blotting, PCR and cycle-sequencing of PCR products in peripheral blood of donor and recipient animals and in the progeny of recipient animals. Data support both the concept that BPV can be transmitted through blood and the hypothesis that infection with the virus causes the clastogenic alterations observed in the present experimental model. The presence of BPV-2 DNA and chromosomal alterations in peripheral blood of offspring at the moment of birth is evidence for vertical transmission of BPV.

Possibility of vertical transmission of caprine arthritis-encephalitis virus in neonate kids

Estudou-se a capacidade do vírus da artrite-encefalite dos caprinos (CAEV) infectar o feto ou o cabrito neonato pela via de transmissão transplacentária ou no momento do parto. Foram utilizados 26 cabritos recém-nascidos, filhos de cabras sororreagentes aos antígenos do CAEV e que nasceram de partos eutócicos. Na pesquisa de anticorpos séricos anti-CAEV, foi utilizada a técnica de imunodifusão em gel de ágar. Nenhum cabrito nasceu sororreagente aos antígenos do vírus, indicando que a possibilidade de transmissão vertical transplacentária da infecção foi menor do que 3,8 por cento (< 1:26).

Aspectos clínicos da artrite-encefalite dos caprinos / Clinical features of caprine arthritis-encephalitis infections

Caprinos de 14 plantéis localizados no estado de São Paulo, naturalmente infectados pelo vírus da artriteencefalite dos caprinos, foram clinicamente avaliados. Demonstrou-se que 17,1 % (64/374) dos caprinos sororreagentes apresentavam a forma clínica articular da enfermidade e que 6,6% (17/249) das cabras sororreagentes apresentavam a forma mamária.

Variaçäo da concentraçäo de imunoglobulinas G e M, de proteína total e suas fraçöes eletroforéticas e da atividade da gamaglutamiltransferase no soro sangüíneo de vacas holandesas, antes e após o parto / Variation of G and M immunoglobulins, total protein and eletrophoretic fractions and activiTy of gama glutamyltransferase in Holstein cow blood sera before and after parturition

Com o objetivo de determinar a dinâmica do proteinograma sangüíneo e da atividade enzimática da gamaglutamiltransferase sérica de vacas antes e após o parto, foram utilizadas 26 vacas do período pré-parto até 180 dias após o parto, todas da raça Holandesa. Os resultados permitiram concluir que as vacas, no momento do parto, apresentam menores concentraçöes séricas de proteína total, das fraçöes albumina e betaglobulina, bem como de imunoglobulinas G e M, do que nos períodos ante e pós-parto, apresentando, a partir dos nove dias pós-parto, elevaçäo gradativa desses constituintes até os 180 dias após o parto. Näo houve diferença na atividade da enzima gamaglutamiltransferase nos momentos estudados

Valores de referência da atividade enzimática da aspartato-aminotransferase e da gama-glutamiltransferase em bovinos da raça Jersey. Influência dos fatores etários, sexuais e da infecçäo pelo vírus da leucose dos bovinos / Reference values of the enzymatic activities of the aspartate aminotransferase and gamma glutamyltransferase of Jersey breed. The influence of age and sexual factors, and of the infection by the bovine leukosis virus

Com o objetivo de estabelecer os valores de referência de parâmetros bioquímicos que avaliem a funçäo hepática, por meio da determinaçäo da atividade enzimática da aspartato-aminotransferase (AST) e da gama-glutamiltransferase (GGT), de bovinos da raça Jersey, criados no Estado de Säo Paulo, bem como avaliar a influência dos fatores etários, sexuais e da infecçäo pelo vírus da leucose dos bovinos sobre os referidos parâmetros bioquímicos, foram colhidas e examinadas amostras de soro sangüíneo de 170 animais, sendo 106 fêmeas e 17 machos, considerados clinicamente sadios e näo reagentes ao antígeno glicoprotéico (gp51) da cápsula do vírus da leucose dos bovinos e 47 fêmeas soro-reagentes ao mencionado vírus. Os valores de referência obtidos para a AST foram iguais a 33,91ñ10,99U/l enquanto para a GGT obtiveram-se valores médios de 13,21ñ12,7U/l. A avaliaçäo dos resultados demonstrou que as transaminases sofreram influência dos fatores etários, pois comprovou-se os teores séricos da AST aumentaram significativamente com o evoluir da idade, sendo o valor mínimo observado nas bezerras com até 3 meses de idade (26,39ñ22,17U/l) e valor máximo em vacas com idade entre 48 e 72 meses de idade (38,60ñ11,24U/l). Para os níveis séricos da GGT observou-se que nas bezerras com até 3 meses de idade os níveis séricos (31,15ñ38,70U/l) foram maiores do que os observados nos animais mais velhos (entre 8,70ñ2,40U/l e 14,24ñ10,37U/l). Nesta pesquisa näo foram observadas influências dos fatores sexuais e da infecçäo pelo vírus da leucose dos bovinos sobre a atividade enzimática das transaminases

Prevalência de anticorpos antivírus da arterite dos eqüinos em cavalos criados no Estado de Säo Paulo / Prevalence of equine arteritis virus specific antibodies in horses of Säo Paulo State, Brazil

Utilizou-se a prova de soroneutralização em microplacas para detecção de anticorpos antivírus da arterite dos eqüinos em 659 amostras de soro sangüíneo de animais criados no Estado de São Paulo. A prevalência de anticorpos na população estudada foi igual a 18,2 por cento. A raça Mangalarga foi a que apresentou maior taxa de prevalência, 33,3 por cento. Animais na faixa etária de 6 a 24 meses de idade apresentaram a maior taxa de prevalência, 30,4 por cento, e as fêmeas apresentaram prevalência de 22, 9 por cento, mais alta do que nos machos

With the purpose of studying the prevalence of equine viral arteritis in horses raised in São Paulo State, Brazil, by the standard microtiter serum neutralization test, 659 serum samples were investigated. The prevalence of antibodies in the horse population was 18.2 per cent, which was significantly higher in Mangalarga horses (33.3 per cent) than in any other breed (Thoroughbred, Arab, Quarter Horse, mixed breeds and others). The distribuition of horses by age showed that horses between 6 to 24 months of age (30.4 per cent) had a higher prevalence (30.4 per cent) rate than others. The female horses prevalence rate of 22.9 per cent was significantly higher than in male horses.

Influência da forma de administraçäo e da quantidade fornecida de colostro sobre a concentraçäo de proteína total e de suas fraçöes eletroforéticas no soro sangüíneo de bezerros da raça Holandesa / Influence of volume and method of administration of colostrum on total protein and eletrophoretic serum fractions in Holstein calves

This study was carried out to evaluate the effect of the amount of colostrum (two and four liters) and methods of colostrum feeding (direct suckling from the cow, feeding colostrum using nipple bottle, and no feeding colostrum) on different blood traits. Thirty two Holstein calves (four groups of eight animals each) were randomly allotted to four treatments: treatment 1 - calves were hand fed four liters of colostrum; treatment 2 - calves were hand fed two liters of colostrum; treatment 3 - calves were fed colostrum directly from the cow, and treatment 4 - calves were not fed colostrum. Higher values of total serum protein, betaglobulins and gamaglobulins were observed in the calves after 24 hours of colostrum feeding. No differences among animals fed colostrum directly from the cow and hand fed two or four liters of colostrum were observed for total protein and eletrophoretic fractions. However lower values of total and eletrophoretic fractions were observed for calves that were not fed colostrum