Paulo César da Silva

Avaliação de populações de maracujazeiro-doce (Passiflora alata Curtis) obtidas de polinização aberta

Fourteen horticultural characteristics of five sweet passion fruit (Passiflora alata Curtis) populations were evaluated in Jaboticabal-SP, from April 2000 to March 2001. A considerably high variability among plants and low among populations was observed for these characters. The number of fruits per plant (NF), number of seeds/fruit (NS), fruit mass (MF) and production (PRO) were the characters with the highest levels of variability among plants, thus, allowing then for the selection of superior plants for these characters. Averages of production (PRO), number of fruits/plant (NF), rind thickness (EC), number of seeds/fruit (NS) and pulp yield (%P), compared by the Tukey test at 5% probability, did not present significant differences among the populations.

Serum protein concentrations, including acute phase proteins, in calves experimentally infected with Salmonella Dublin

The aim of this study was to evaluate serum protein concentrations in calves experimentally inoculated with Salmonella Dublin. Twelve healthy 10 to 15-day-old Holstein calves were randomly allotted into two groups, control and infected with 108 CFU of Salmonella Dublin orally. The calves were subjected to physical evaluation and blood samples were collected shortly before administration of the bacteria and also 24, 48, 72, 96, 120 and 168 hours post-infection. The concentration of serum proteins was determined through sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE). Thirty serum proteins ranging from molecular weight of 24,000 Da to molecular weight of 236,000 Da were detected. Serum concentrations of ceruloplasmin (125,000 Da), haptoglobin (45,000 Da), acid glycoprotein (40,000 Da) and a 34,000 Da protein were significantly increased in the experimentally infected calves, when compared with their concentrations in the control animals. Therefore, this study showed that S. Dublin infection could lead to the increase of certain serum proteins in calves.

Acute phase proteins: a potential approach for diagnosing chronic infection by Trypanosoma vivax

The present study aimed to assess potential changes in acute phase proteins in sheep experimentally infected with Trypanosoma vivax. There were studied eight male sheep, four used as controls and four infected with 10(5) T. vivax trypomastigotes. Blood samples were collected at two points times before infection and then at 5,7, 9, 11, 13, 15, 20, 30, 45, 60, 75, 90, 105 and 120 days post-infection (dpi). Blood samples were centrifuged and allotted, and acute phase proteins were then separated by electrophoresis on acrylamide gel containing sodium dodecyl sulfate. Protein concentrations were determined by computer-assisted densitometry. Total protein was determined by colorimetric biuret method. Trypanosomes were counted daily using a 5 mL aliquot of blood smear on a glass slide under a 22 × 22 mm coverslip. Parasites were counted in 100 microscopic fields (40× magnification), and then multiplied by a correction factor. The results were expressed as parasites per mL of blood. For statistical analyses, we used the Wilcoxon test at 5% significance level. There was found a reduction in several acute phase proteins and increase in antitrypsin and transferrin. This finding can be used for the diagnosis of T. vivax infection, especially in chronic infection.

Influence of parity on concentrations of enzymes, proteins, and minerals in the milk of cows

Avaliou-se a influencia do numero de paricoes nos valores de alguns parâmetros bioquimicos e do perfil eletroforetico do soro lacteo de vacas de corte. Trinta e cinco vacas da raca Canchim foram alocadas em cinco grupos: vacas de primeira lactacao, segunda lactacao, terceira e quarta lactacoes, quinta lactacao e sexta lactacao. As amostras de secrecao lactea foram coletadas imediatamente apos (dia 0) e 1, 2, 7, 15 e 30 dias apos o parto. As concentracoes de gamaglutamiltranferase (GGT), proteina total, calcio, fosforo, magnesio e calcio ionizado foram avaliadas. A separacao eletroforetica das proteinas foi realizada em matriz de gel de poliacrilamida (SDS-PAGE). A atividade de GGT e as concentracoes de imunoglobulina G, calcio e fosforo nao foram influenciadas pelo numero de paricoes. As concentracoes de proteina total, calcio ionizado, magnesio imunoglobulina A, lactoferrina, β - lactoglobulina e α - lactoalbumina, foram influenciadas pelo numero de partos das vacas. A excecao dos teores de fosforo e α - lactoalbumina em poucos grupos, a concentracao d a s demais caracteristicas decresceu no decorrer do periodo de lactacao.

Proteinograma do soro lácteo de ovelhas da raça Santa Inês em diferentes fases de lactação

This study aimed to evaluate the influence of lactation phases on the proteinogram of whey protein in Santa Ines ewes. Ewes were accompanied in a semi-intensive system using the same sanitary and nutritional management evaluated at 15, 30, 60 and 90 days postpartum (end of weaning and lactation). Clinical examination of the mammary gland was carried out through and bacteriological culture. The screening of the material resulted in 44 milk samples of healthy glands concurrent negative by CMT and bacteriological culture exam. For obtaining the whey protein renin solution was used. The whey was fractionated into aliquots and kept in the -80C freezer to later separation of protein fractions. For determination of total protein of whey protein was employed the biuret, observing the linearity of the test. Separation of protein fractions was performed, using polyacrylamide gel containing sodium dodecyl sulfate (SDS-PAGE). Eigth protein were observed including lactoferrin, serum albumin, IgA, IgG (heavy chain IgG (IgG CP), light chain IgG (IgG CL), β-lactoglobulin, α-lactalbumin and proteins identified as PM 15000 and PM 29000. No significant difference was observed at different stages of lactation in the following protein: IgA (P>0.3895), lactoferrin (P>0.1611), PM 29000 (P>0.4879), α-lactalbumin (P>0.0799) and PM15000 (P>0.4494). In total protein (P<0.0022), albumin protein (P<0.0377) and IgG (P<0.0354) it was observed a significant variation in the first moments of observations, in the β-lactoglobulin protein (P<0.0005) there was significant variation with reduction of 15 to 30 days postpartum with progressive elevation until the last stage of lactation (90 days postpartum). The SDS-PAGE technique allowed the quantification of eigth whey proteins in health ewes. The protein fractions identified reflect the profile of whey to ovine species, with influence of stages of lactation in albumin, IgG and β-lactoglobulin.

Proteinograma sérico, com ênfase em proteínas de fase aguda, de bovinos sadios e bovinos portadores de enfermidade aguda de ocorrência natural

Over the last few decades acute phase proteins (APP) have become the biomarkers of choice in human medicine to identify and monitor inflammation and infection. There is no reason to suppose that clinical investigations in veterinary medicine would not be equally assisted by APP assays. Aiming to verify the importance of APPs as biomarkers of inflammatory diseases in domestic cattle, serum protein profiles, especially APPs, were determined through the SDS-PAGE electrophoresis technique. Thirty animals were allotted in two groups: 15 healthy cattle and 15 clinically ill cattle (5 with staphylococcal mastitis, 5 with photosensitization and 5 with onphalophlebitis). All animals were submitted to daily blood sampling during 7 days, while interned in the Veterinarian Hospital from UNESP, Jaboticabal campus. Ceruloplasmin and haptoglobin were significantly elevated in animals with mastitis, photosensitization and onphalophlebitis (275.17% and 343.71%; 175.17% and 230.19%; 114.47% and 144.47%, respectively). α1-acid glycoprotein behaved as a good biomarker only in animals with mastitis and photosensitization, elevating respectively 198.14% and 145.89% of their serum levels. Fibrinogen was a reliable indicator only in animals undergoing mastitis, with a raise of 146.5%. The diverse responsiveness of different APP under distinct inflammatory stimuli was clear. Ceruloplasmin and haptoglobin were more sensible and, therefore, reliable biomarkers to the diseases studied in this species.

Perfil eletroforético do colostro de ovelhas suplementadas com propileno glicol e cobalto associado à vitamina B12 no final da gestação

This study aimed to evaluate the proteinogram of the colostrum of ewes submitted to administration of propylene glycol and cobalt associated with vitamin B12 in late pregnancy. Eighteen pregnant Santa Ines ewes 18 months to 5 years old were distributed by probabilistic sampling into three experimental groups, about 30 days before the expected delivery date. In group 1 (G1/n=6), daily oral doses of 30ml propylene glycol PA were administered; Group 2 (G2/n=2) received a daily oral dosage of 1mg cobalt chloride in 1% solution and 2mg of vitamin B12 intramuscularly once a week, and Group 3 (G3/n=6) was the control group. Soon after delivery 30mL of colostrum was harvested from each ewe, which were stored in appropriate containers and sent to the laboratory. After homogenization, we added to each 1000μL of colostrum 75μL solution of rennin, which was kept in a water bath at 37°C for about 20 minutes and centrifuged at 21.000G for 10 minutes in a refrigerated centrifuge. Later, the intermediate fraction, corresponding to colostrum whey, was aliquoted and kept in a -80oC ultrafreezer for subsequent determination of proteins. The determination of the total colostral protein whey was performed using a commercial reagent, observing the linearity test for colostrum. The separation of proteins was performed using the technique of electrophoresis on polyacrylamide gel containing sodium dodecyl sulfate (SDS-PAGE). Lactoferrin, IgA, albumin, IgG heavy chain (IgGCP), β-casein, IgG light chain (IgGCL), β-lactoglobulin and α-lactalbumin proteins were identified. There was no influence of the administration of supplements during late pregnancy on the concentration of proteins identified in the colostrum of the ewes.

Effects of intramammary infection on whey proteinograms of sheep during lactation

The study aimed to identify potential biomarkers of mammary gland infection in Santa Ines sheep. Commercial flocks of sheep provided the same hygiene, sanitary, and nutritional management under semi-intensive production systems were monitored during the lactation stage-and assessed 15, 30, 60, and 90 days after delivery (through the end of lactation and weaning). The California Mastitis Test (CMT) was performed on the mammary glands. Milk was collected for bacterial examination and protein analysis. Bacterial culture and biochemical characterization of the samples were performed. Forty-two milk samples from healthy glands (negative CMT and bacterial testing) and 43 milk samples from infected glands (positive CMT and bacterial testing) taken at the predefined time points were assessed. A rennin solution was used to obtain the whey. The proteins analysis was performed using sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), which allowed for the quantification of nine whey proteins produced in healthy glands: serum albumin, lactoferrin, IgA, IgG heavy-chain (IgG HC), IgG light-chain (IgG LC), total IgG (IgG HC + IgG LC), α-lactalbumin, β-lactoglobulin, protein with MW 15.000 Da, protein with MW 29.000 Da and eleven whey proteins secreted by infected glands, including haptoglobin and α-1-acid glycoprotein. A comparison of whey proteins between healthy and infected glands showed increases (P<0.05) in the secreted and total contents of all proteins, except for IgG LC and α-lactoalbumin. The most significant changes were observed in α-1-acid glycoprotein, lactoferrin and haptoglobin, which showed three-, five-, and seven-fold increases in secretion, respectively. This study showed that haptoglobin, α-1-acid glycoprotein, lactoferrin, albumin, and the IgA and IgG immunoglobulins may serve as potential biomarkers for mammary gland infection in sheep.

Avaliação dos parâmetros genéticos em progênies de meio-irmãos de uma sub-população de milho composto Flint para resistência à lagarta-da-espiga Helicoverpa zea (Bod.)

One hundred of half-sib families of a subpopulation of maize (Zea mays L.)Flint Composite were evaluated in the field, to verify the genotype resistance to the corn earworm Helicoverpa zea (Bod.). For this subpopulation, the obtained value of corn earworm damage mean was 1.14 cm of length in the ear determined by the Widstrom scale and the experimental variance coefficient (CVE) was 23.4 %. Of the evaluated genetic parameters the estimates of heritability was 6%, genetic variance was 0,0015 cm2 and phenotypic variance was 0.025 cm2 for H. zea damage. However the length of the husk top and husk compactation the obtained values of the heritability was 75% and 72% respectively. Therefore, this subpopulation of the maize present genetic variability sufficient for utility in the breeding programs, though the resistance to the corn earworm can be increased, through the breeding of morphologic characters directly related to the insect, such as the husk compactation and length.