Construction and preliminary characterization of a river buffalo bacterial artificial chromosome library

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Two-stage rumenostomy in buffaloes

Background: Rumenostomy may be performed for therapeutic and digestibility research purposes in bovines, small ruminants and camelids. Several studies requires romenostomy in buffaloes in order to sample ruminal content for laboratorial assays. However, complications and outcome of rumenostomy was poorly studied in buffaloes. Therefore, the aim of the current study was to describe a two-stage rumenostomy technique in buffaloes, focused on intra and post-operative period.Materials, Methods & Results: Nine Murrah buffaloes were submitted to a 36-h and 12-h of food and water fastening. The animals were given acepromazine and maintained in standing position. Flank local anesthesia was carried out. A circular skin incision was carried out in the center of the left flank, followed by divulsion of the external and internal obliques and transversus abdominus muscles, and incision of the peritoneum. Subsequently, a segment of the dorsal aspect of the rumen was grasped and pulled through the flank incision. The rumen was attached to the peritoneum and skin incision margins in four points (dorsal, ventral, cranial and caudal). Additional simple interrupted sutures attaching the rumen serosa to the skin were applied subsequently. Four additional interrupted horizontal mattress sutures were applied equidistantly, taking bites only in the skin and rumen serosa. Following 12 h, the second stage was carried out. The buffaloes were prepared and restrained as performed for the first stage. A circular flap was excised from the exteriorized rumen and the silicone romenostomy cannula was placed. Clinical parameters, postoperative recovery, weight and behavioral pain scale were assessed. Positioning and anesthesia regimen were adequate for the achievement of the procedure. However, two animals fell in the restraint chute during the first surgical stage. Mild ischemia of the exteriorized rumen segment was observed on the second surgical stage, which resulted in less hemorrhage and enhanced cannula positioning. Complete cicatrization and permanent adhesion of the rumen to the skin were achieved. No ruminal leakage to the abdominal cavity occurred. No signs of pain were reported. There were few cases of laxity of the romenostomy opening leading to drop of cannula, myiasis on the margin of the stoma site and few cases of mild ruminal content leakage on the long-term assessment.Discussion: Restraint in standing position was considered adequate, although lateral recumbence constitutes another option. However, higher risk of contamination and technical difficulties in placing the cannulas are expected if lateral recumbence is considered. In other trials using acepromazine, no accidental recumbence occurred. Xylazine was also indicated for chemical restraint of buffaloes. It is known that flexible cannulas provide better anatomic adjustment and adaptation as well as being effective for sampling ruminal content, as seen in the current study. Ruminal leakage is one of the most frequent complications of romenostomy, which may affect animal's welfare. The animals in the current study presented no variations on the body score, even though on those presenting cannula loosening or ruminal content leakage. Moreover, no significant changes of the ruminal content parameters were noticed. Myiasis was also reported following ruminal surgical interventions, which were mainly attributed to extensive breeding. Loss of the cannula, subcutaneous emphysema and suture dehiscence are common complications of romenostomy. Nonetheless, none of those complications were found on the current study. Thus, romenostomy was feasible and efficient for sampling and performing assays of the ruminal content in buffaloes.

Prevalence and etiology of buffalo mastitis and milk somatic cell countin dry and rainy seasons in a buffalo herd from Analândia, São Paulo State,Brazil

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Padronização de técnica manual para obtenção de plasma rico em plaquetas de bovino

To evaluate the biochemical profile and protein concentration of whey from milk samples of healthy Murrah primiparous and pluriparous buffaloes, 30 female buffaloes were analyzed during a complete lactation. The animals were divided into three groups: G1 = 10 primiparous buffaloes, G2 = 10 pluriparous buffaloes with 2-3 lactations and G3 = 10 pluriparous buffaloes with > 3 lactations. The lactation period was divided into: early stage (I: 1-3 months of lactation), intermediate stage (T: 4-6 months of lactation) and final stage (F: 7-9 months of lactation). Before milk sampling, physical examination of the mammary gland, strip cup test and California Mastitis Test (CMT) were performed. After mammary quarters asepsis, 20mL of milk were collected monthly from each mammary quarter, during a complete lactation, in sterilized plastic bottles without preservative, in order to perform microbiological isolation, biochemical profile and protein electrophoresis in sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE), and 30mL of milk from each mammary quarter were collect, in sterilized plastic bottles containing preservative bronopol to perform the somatic cell count (SCC). A total of 1,042 milk samples were collected from the experimental groups during lactation, of which 923 samples showed negative reaction to CMT and negative microbiological isolation and were selected to biochemical profile analysis and protein electrophoresis in SDS-PAGE. There were influence of parity order and stage of lactation in biochemical profile and protein concentration of healthy Murrah buffaloes'whey. Primiparous buffaloes (G1) showed higher gamma-glutamyltransferase (GGT: 2,346 U/L), alkaline phosphatase (ALP: 181 U/L), phosphorus (P; 56.6mg/dL), potassium (K; 32.0mg/dL) and alpha-lactalbumin (458mg/dL). Buffaloes with 2-3 lactations (G2) showed higher SCC (70,700 cells/mL) and higher concentrations of total protein (1.55g/dL), albumin (100mg/dL), magnesium (Mg; 8.80mg/dL), chlorides (Cl; 176mg/dL), iron (Fe; 10.7 mu g/dL), sodium (Na; 178mMol/L) and lactoferrin (59.5mg/dL). Bufalloes with > 3 lactations (G3) showed higher concentrations of total calcium (Ca; 41.8mg/dL), ionized calcium (iCa; 2.92mMol/L), immunoglobulin A (IgA; 1.32mg/dL), serum albumin (99.1mg/dL), immunoglobulin G (IgG; 49.7mg/dL) and beta-lactoglobulin (1,068mg/dL). During lactation it was observed increase in SCC, GGT, ALP, total protein, albumin, P, Mg, Cl, Na, lactoferrin, serum albumin, IgG and alpha-lactalbumin, as well as decrease in concentrations of Ca, Fe, iCa, K, IgA and beta-lactoglobulin in buffaloes'whey. The results may be used as reference for buffaloes and to support diagnosis and prognosis of diseases common to lactation periods.

Comparative analysis of the river buffalo somatostatin gene

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Genetic analysis of superoxide dismutase 1 gene in murrah river buffalo

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Prospecting major genes in dairy buffaloes

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Perfil bioquímico sérico de bezerros bubalinos no período neonatal

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Microbiological evaluation of milk samples positive to California Mastitis Test in dairy buffalo cows (Buballus bubalis)

In order to observe the microbiological status of CMT positive samples, 734 apparently health mammary quarters from buffalo cows were submitted to physical evaluation, strip cup test and CMT. After milk samples inoculation in 10% ovine blood agar base media and in MacConkey agar and incubation under aerobic condition for 72 hours at 37 degrees C, identification was proceeded. According to CMT, 227 quarters (30,93%) were positive, among them 73 (32,16%) presented 1+ reaction, 53 (23,35%) were 2+ and 101 (44,49%) were 3+. Microbiological exams of such samples were positive in 147 (64,76%) out of 227 CMT positive samples and among the remaining 72 (31,72%) were negative and 8 (3,52) were contaminated. In the 147 microbiological positive samples 204 bacteria were found in pure or associated growth and the most frequent agents were: Corynebacterium sp (59,25%); Staphylococcus sp (17,65%) among which 86,11% were coagulase negative and 13,89% were coagulase positive; and Micrococcus sp (6,37%). The results revealed that, excluding the eight contaminated samples, 147 (67,12%) quarters out of 219 CMT positive could be considered as bacteria-carrier and that even in a smaller percentage false-positive results can cause problems in a sanitary program for mastitis control in dairy buffalo cows.

Morfologia e morfometria das papilas mamárias de búfalas

Para este estudo, quinze papilas de fêmeas de búfalos e uma de fêmea bovina foram obtidas no matadouro. Estudos macro e microscópico foram realizados por meio de diferentes técnicas: morfológica, histológica e análises, biométrica, histométrica e mesoscópica. As médias foram: papila cranial direita (PCD) 20,95g; papila caudal direita (PCaD) 21,49g; papila cranial esquerda (PCE) 23,52g; papila caudal esquerda (PCaE) 21,35g. O comprimento médio do ducto papilar, do número de pregas e do comprimento das cisternas papilares foi respectivamente: 0,67/5,5 e 2,17cm para a PCD; 0,62/6,0 e 2,26cm para a PCaD; e para o antímero esquerdo, PCD: 0,71/6,0 e 2,26cm e para PCaD: 0,74/5,7 e 2,57cm. A análise de Pearson revelou uma correlação negativa entre o peso do corpo e o peso da glândula mamária. Foi feita a amostragem das três áreas proximal, média e distal e estas foram fixadas em formaldeído a 10%, seguindo-se a investigação mesoscópica em amostras de espessuras de 10, 20 e 40µm coradas pelas técnicas Azan, F3BA Picrosirus e Weigert - Van Gieson. As análises histológicas e morfométricas foram feitas em amostras de 5µm de espessura o que mostrou que a camada muscular estava disposta em dois arranjos, interna circular e externa longitudinal com epitélio cubóide bi-estratificado. A camada muscular bem desenvolvida estava relacionada com a porção média revelando uma espessura de 22,90mm com uma grande quantidade de tecido conectivo (7,10mm).

Analysis of the Stayability in Milk Buffaloes using Survival Models

In this study the trait Stayability (SA) was evaluated according to the year of cull after first calvin, i.e., SA 1 to 6 for 1 to 6 years from first calving in lactating females from bubaline milk herds spread in nine farms located in São Paulo state. Informations were used regarding 1027 lactating Murrah breed buffaloes. The statistical analyses were made using LIFEREG (SAS, 1999) procedure. The SA was evaluated using the fixed effects: farm production, birth year, calving season (Season 1- April to September and Season 2 October - March) and class of milk yield at 270 days. The age at first calving (AFC) was considered as a random effect. The mean observed for total milk yield was 1458.75Kg. Calving Season 2 encloses 65.6% of births. The means of cull age, in months, and the percentage of SA were, respectively: 10.69 e 69% (SA1), 19.30 e 63% (SA2), 26.4 e 54% (SA3), 33.15 e 42% (SA4), 38.53 e 36% (SA5) e 42.65 e 26% (SA6). It is verified that most of culls happens after the first lactation, among the sixth and eleventh month after first calving. It was observed that the factors: farm production, birth year and class of milk yield at 270 days affected significantly all SAs. Factors like calving season and the age at first calving (AFC) were only significant for SAL Being significant the factor AFC in level of 1% and factor time in 10%. For other SAs these factors were not statistically significant.

Factors affecting somatic cell counts and their relations with milk and milk constituent yield in buffaloes

Data concerning daily milk yield (MY), percentage of milk fat (%F), protein (%P), lactose (%LT), and total solids (%TS), and somatic cell counts (SCC) for a herd of 222 Murrah buffalo reared in the state of São Paulo, Brazil, were collected monthly from 1997 to 2000 in order to study the factors affecting SCC and their relation to milk production and constituents during lactation. SCC decreased in the second month of lactation and increased thereafter, up to the ninth month of lactation. The interaction of month of lactation x order of calving was significant. Mean MY observed during the first month of lactation was 6.87 kg, which increased to 7.65 kg during the second month, and then decreased until the ninth month of lactation (3.83 kg). During the different months of lactation, %F, %P, %LT, and %TS ranged from 6.28 to 8.38%, 4.05 to 4.59%, 4.96 to 5.34%, and 16.94 to 18.55%, respectively. Calving year, calving order, and order of month of lactation significantly affected MY, %F, %P, %LT, and %TS. The regression coefficients of transformed SCC on MY and %LT were negative and significant during all months of lactation, showing that milk and lactose yield decreased with increased transformed SCC, causing losses to buffalo milk producers.

Growth hormone mRNA expression in the pituitary of Bos indicus and Bos taurus x Bos indicus crossbred young bulls treated with recombinant bovine somatotropin

The effects of breed and of recombinant bovine somatotropin (rbST) treatment on growth hormone gene expression were studied in young bulls. The experiment was completely randomized in a [2 × 2]-factorial arrangement, using two levels of rbst (0 or 250 mg/animal/14 days), and two breed groups (Nelore and Simmental x Nelore crossbred). A CDNA encoding Bos indicus growth hormone was cloned and sequenced for use as a probe in Northern and dot blot analyses. Compared to the Bos taurus structural gene, the Bos indicus CDNA was found to begin 21 bases downstream from the transcription initiation site and had only two discrepancies (C to T at position 144-His and T to C at position 354-Phe), without changes in the polypeptide sequence. However, two amino acid substitutions were found for Bubalus spp., which belong to the same tribe. The rbst treatment did not change any of the characteristics evaluated (body and pituitary gland weights, growth hormone MRNA expression level). Crossbred animals had significantly higher body weight and heavier pituitaries than Nelore cattle. Pituitary weight was proportional to body weight in both breed groups. Growth hormone MRNA expression in the pituitary was similar (P>0.075) for both breed and hormonal treatment groups, but was 31.9% higher in the pure Nelore group, suggesting that growth hormone gene transcription regulation differs among these breeds.

Relación entre el recuento de células somáticas y la producción de leche de búfalas en el estado de São Paulo, Brasil

Mastitis is an inflammatory process in the udder that can affect the quality and quantity of milk produced causing economic losses and risks for health. Considering the somatic cell count (SCCs) as indicator of udder health and the milk yield (MY) of buffaloes from São Paulo State, this study aimed to quantify the related losses in milk due to somatic cells count (SCC). 9404 sources of information from 2198 lactations that occurred between 1997 and 2004 were analysed. There was no relation between MY and the SCCs in the buffaloes at first parity. For the second parity in the months 1, 2, 5, 6 and 7 of lactation, there was a negative and significant relationship between SCCs and MY. For parities of three or more there was a significant and negative regression coefficient during every month of lactation betrween MY and SCC. The average losses varied from 0,18 to 2,2 milk liters per unit of SCCs. The results indicated large losses observed in the miltiparous buffaloes and that this category needs received special attention in terms of udder health. The effect of farm, parity and year, must be considered comparison between animals.