Search

Made available in DSpace on 2022-04-29T08:30:42Z (GMT). No. of bitstreams: 0 Previous issue date: 2021-01-01 This study aimed to evaluate the correlation of seminal plasma IGF-I and total proteins from peripubertal bulls with semen characteristics and onset of puberty. Sixteen dairy Gyr bulls were separated into early (n = 8) and regular (n = 8) groups, according to the onset of puberty. Semen was monthly collected by electroejaculation from 14 to 26 months of age, and the onset of puberty was retrospectively determined (17.0 ± 1.6 and 19.2 ± 1.5 months, to early and regular group, respectively). Five time points were evaluated (day −60, day −30, day 0, day 30 and day 60 days of puberty), being day 0 considered as beginning of puberty. Scrotal circumference and body weight were also assessed. Semen characteristics were evaluated, the seminal plasma was separated by centrifugation and total protein and IGF-I concentrations were determined. There was no difference between groups to concentration of the seminal plasma total proteins, but we found an interaction between group and age. Seminal plasma IGF-I concentrations were not different between early and regular groups; thus, the data were combined for analysis. Combined IGF-I concentrations were positively correlated with sperm motility and concentration, age, body weight and scrotal circumference. Negative correlation was found between IGF-I concentration and total sperm defects. Increased IGF-I was observed in day −30 and day 0 of puberty in early and regular groups, respectively. Seminal plasma total proteins are involved in peripubertal modifications and IGF-I from Zebus dairy bulls can influence the seminal characteristics and the growth factor increase occurs concomitantly with body growth but cannot be used to define puberty bulls earlier. Departamento de Medicina Veterinária Universidade Federal do Paraná Centro de Ciências Agrárias e da Biodiversidade Universidade Federal do Cariri Universidade Federal Rural da Amazônia Facultad de Ciencias Agropecuarias Universidad de La Salle Departamento de Cirurgia Veterinária e Reprodução Animal FMVZ Universidade Estadual Paulista UNESP Departamento de Salud Animal y Producción de Hato Universidad Nacional de Colombia Universidade Federal de Minas Gerais Escola de Veterinária da UFMG Departamento de Cirurgia Veterinária e Reprodução Animal FMVZ Universidade Estadual Paulista UNESP

Chromatographic profile of seminal plasma from 19 young Guzerat bulls, and their associations with andrologic traits and testosterone were evaluated. Total seminal plasma protein concentrations ranged from 2.86 to 78.82 mg/ml, average of 17.18 ± 20.49 mg/ml. The seminal plasma protein profile by gel filtration showed eight different fractions, being some of them correlated to age, body weight, scrotal circumference, physical (sperm motility and concentration) and morphologic (mojar sperm defects) of the semen. These concentrations were correlated (p

Objetivou-se avaliar caracteristicas quali - quantitativas da carcaca em machos Nelore, submetidos ao bloqueio dos ductos incisivos no periodo pre puberal. Alem disso, objetivou-se avaliar as concentracoes sericas de testosterona e do hormonio do crescimento semelhante a insulina do tipo I (IGF-I), e a arquitetura histologica do orgao vomeronasal (OVN). Trinta e quatro machos, no periodo pre puberal, foram divididos em tres grupos experimentais: inteiros – animais controle sem qualquer procedimento (n = 11); bloqueados - animais com os ductos incisivos bilateralmente obstruidos (n = 10); castrados - animais com orquiepididectomia bilateral (n = 13). O OVN foi obtido no abate, ao final do experimento, para avaliacao histomorfometrica. As coletas de sangue foram realizadas a cada tres meses, em dois turnos (manha e tarde), totalizando cinco avaliacoes. Observaram-se maiores (P 0,05). Registrou-se reducao da altura do epitelio neuro sensitivo do OVN nos bloqueados e castrados (P

The present study was conducted to identify the major proteome of the sperm-rich fraction and prostatic fraction of canine seminal plasma. Three semen samples from four healthy dogs were obtained by digital manipulation. The pre-sperm fraction, sperm-rich fraction and prostatic fraction were separated from each ejaculate. Immediately after sperm analysis, a protease inhibitor was added to the sperm-rich fraction and prostatic fraction, and the fractions were separately centrifuged and frozen at −80 °C. The samples were thawed, re-centrifuged, and the total protein concentration was determined. Samples were subjected to 1D SDS-PAGE and Coomassie-blue stained gels, were analyzed by Quantity One 1D Analysis Software. Bands detected in the gels were excised and proteins subjected to digestion with trypsin. Proteins were identified by nano-HPLC-MS and tools of bioinformatics. Tandem mass spectrometry allowed the detection of 268 proteins in the gels of sperm-rich fraction and prostatic fraction of canine ejaculate. A total of 251 proteins were common to the sperm-rich and prostatic fractions, while 17 proteins were present in the sperm-rich fraction and absent in the prostatic fraction. The intensity of the bands detected in range 1 and 2 represented 46.5% of all of the band intensities detected in the 1D gels for proteins of the sperm-rich fraction and 53.0% of all bands in the prostatic fraction. Arginine esterase and lactotransferrin precursor were the protein with the highest intensity observed in the both fractions. Among the proteins present only in the sperm-rich fraction, the proteins UPF0764 protein C16orf89 homolog and epididymal-specific lipocalin-9 were the most abundant. In conclusion, canine sperm-rich fraction and prostatic fraction express a very diverse set of proteins, with unique biochemical properties and functions. Moreover, although most proteins are common to both sperm-rich fraction and prostatic fraction, there are some exclusive proteins in sperm-rich fraction.

The objective of the present study was to describe the relationship of seminal plasma and total sperm cell proteins with the semen freezability parameters of Guzerat bulls. Thirteen bulls were subjected to breeding soundness evaluation. Semen samples were collected, cryopreserved, and then post-thawing sperm kinetics were assessed, where high ( = 7) and low ( = 6) freezability groups were defined. Seminal plasma and total sperm proteins from the 2 groups were separated by 2-dimensional SDS-PAGE, and spots were identified by mass spectrometry. Semen parameters post-cryopreservation were as follows in the high and low freezability groups, respectively: mean total motility, 52.4 ± 20.5 and 13.7 ± 3.9; percentage of normal sperm, 89.0 ± 2.6 and 64.7 ± 14.0; and reactivity of hypo-osmotic swelling test, 38.9 ± 4.7 and 13.6 ± 3.7. Three seminal plasma proteins (osteopontin-K, DNase γ precursor, and DNASE1L3) and 6 proteins from sperm cells (acrosome formation-associated factor isoform 2, annexin A1, disintegrin and metalloproteinase domain-containing protein 2, dihydrolipoyl dehydrogenase, and glyceraldehyde-3-phosphate dehydrogenase) were highly expressed ( < 0.05) in the high freezability group. Another 6 seminal plasma proteins (acrosin inhibitor 1, glutathione peroxidase 3, metalloproteinase inhibitor 2, ephrin-A1, annexin A1, and platelet-activating factor acetylhydrolase) were significantly higher ( < 0.05) in the low freezability group. We described the associations of seminal plasma and sperm cell proteins with post-thawing sperm viability of Guzerat bulls raised in a typical semiarid environment. Such associations indicate that specific seminal plasma proteins more abundant in bulls of low semen freezability may be a response to an early oxidative stress that is not detected by conventional prefreezing semen evaluation. Moreover, specific sperm proteins were more associated with good freezability. The results presented here can serve as guidelines for future research aiming to develop better extenders and/or to improve bull semen selection for cryopreservation.

The present study was conducted to describe the major seminal plasma proteome of rabbits and potential associations between seminal proteins and semen criteria. Semen samples were collected from 18 New Zealand adult rabbits, and seminal plasma proteins were analyzed by 2-D SDS-PAGE and tandem mass spectrometry. Sperm motility, vigor, concentration, morphology and membrane sperm viability were evaluated. Rabbits ejaculated 364 ± 70 million sperm/ml, with 81 ± 6.1% motile cells, 3.8 ± 0.2 vigor and 66.7 ± 2.5% sperm with normal morphology. Based on the viability and acrosome integrity assay, there were 65.8 ± 2.5% live sperm with intact acrosome and most spermatozoa had both intact acrosome and functional membrane. On average, 2-D gels of rabbit seminal plasma had 232 ± 69.5 spots, as determined by PDQuest software (Bio Rad, USA). Mass spectrometry allowed the identification of 137 different proteins. The most abundant proteins in rabbit seminal plasma were hemoglobin subunit zeta-like, annexins, lipocalin, FAM115 protein and albumin. The intensity of the spots associated with these five proteins represented 71.5% of the intensity of all spots detected in the master gel. Multiple regression models were estimated using sperm traits as dependent variables and seminal plasma proteins as independent ones. Also, sperm motility had positive association with beta-nerve growth factor and cysteine-rich secretory protein 1-like and a negative one with galectin-1. The percentage of rabbit sperm with intact membrane was related to seminal plasma protein FAM115 complex and tropomyosin. Then, the population of morphologically normal sperm in rabbit semen was positively linked to carcinoembryonic antigen-related cell adhesion molecule 6-like and down regulated by seminal plasma isocitrate dehydrogenase. Based on another regression model, the variation in the percentage of live sperm with intact acrosome was partially explained by the amount of leukocyte elastase inhibitor and the peptidyl-prolyl cis-trans isomerase A in the rabbit seminal fluid. The current study reports the identification of 137 proteins of rabbit seminal plasma. Major proteins of seminal secretion relate primarily to prevention of damages caused by lipid peroxide radicals and oxidative stress, membrane functionality, transport of lipids to the sperm membrane and temperature regulation. Moreover, finding seminal plasma proteins as indicators of semen parameters will improve assisted reproductive technologies.

The present study was conducted to identify the major seminal plasma protein profile of boars and its associations with semen criteria. Semen samples were collected from 12 adult boars and subjected to evaluation of sperm parameters (motility, morphology, vitality, and percent of cells with intact acrosome). Seminal plasma was obtained by centrifugation, analyzed by two-dimensional SDS-PAGE, and proteins identified by mass spectrometry (electrospray ionization quadrupole time-of-flight). We tested regression models using spot intensities related to the same proteins as independent variables and semen parameters as dependent variables (P ≤ 0.05). One hundred twelve spots were identified in the boar seminal plasma gels, equivalent to 39 different proteins. Spermadhesin porcine seminal protein (PSP)-I and PSP-II, as well as spermadhesins AQN-1, AQN-3 and AWN-1 represented 45.2 ± 8% of the total intensity of all spots. Other proteins expressed in the boar seminal plasma included albumin, complement proteins (complement factor H precursor, complement C3 precursor and adipsin/complement factor D), immunoglobulins (IgG heavy chain precursor, IgG delta heavy chain membrane bound form, IgG gamma-chain, Ig lambda chain V-C region PLC3, and CH4 and secreted domains of swine IgM), IgG-binding proteins, epididymal-specific lipocalin 5, epididymal secretory protein E1 precursor, epididymal secretory glutathione peroxidase precursor, transferrin, lactotransferrin and fibronectin type 1 (FN1). On the basis of the regression analysis, the percentage of sperm with midpiece defects was related to the amount of CH4 and secreted domains of swine IgM and FN1 (r² = 0.58, P = 0.006), IgG-binding protein (r² = 0.41, P = 0.024), complement factor H precursor (r² = 0.61, P = 0.014) and lactadherin (r² = 0.45, P = 0.033). The percentage of sperm with tail defects was also related to CH4 and secreted domains of swine IgM and FN1 (r² = 0.40, P = 0.034), IgG-binding protein (r² = 0.35, P = 0.043) and lactadherin (r² = 0.74, P = 0.001). Sperm motility, in turn, had association with the intensities of spots identified as lactadherin (r² = 0.48, P = 0.027). In conclusion, we presently describe the major proteome of boar seminal plasma and significant associations between specific seminal plasma proteins and semen parameters. Such relationships will serve as the basis for determination of molecular markers of sperm function in the swine species.

The Saanen is a highly productive breed, and for this reason, it has been raised in Brazil, but mostly under climate conditions completely different from where the breed originated. The objective of this study was to investigate variations in semen parameters and sperm membrane proteins from Saanen bucks (n = 7) raised in Northeastern Brazil, during dry season (September, October, and November) and rainy season (March, April, and May). We showed that during the dry season, sperm motility, concentration, and the percentage of normal sperm decreased as compared to the rainy season. Rectal temperatures of bucks had no significant (p > 0.05) variations during the dry and rainy seasons. However, temperatures of left and right skin testis were higher (p < 0.05) during the dry as compared to the rainy season. Expression of three proteins (lysine-specific demethylase 5D, adenosine triphosphate (ATP) synthase subunit d, and radial spoke head protein 9 homolog) in sperm membrane were more intense in rainy season and only one protein (cytosol aminopeptidase) had greater expression in the dry season of the year. Our results show that mechanisms of testicular thermoregulation of Saanen bucks did not prevent a decrease in seminal parameters during the dry season. This deterioration may be related to reduced expression of proteins associated with important functions in sperm membrane.

Foram avaliados 24 touros da raça Guzerá, de 24 a 34 meses de idade, com o objetivo de estudar o perfil da concentração sérica de testosterona e sua variação relativa à idade, peso corporal e características andrológicas. A concentração média de testosterona foi de 1,36 ng/mL, com influência do horário de coleta nos níveis circulantes. A equação de regressão da concentração sérica de testosterona em função do peso corporal dos animais apresentou efeito linear, sugerindo que, para cada quilograma a mais no peso corporal, ocorre acréscimo de 0,0082 ng/mL nas concentrações de testosterona. A regressão da concentração sérica de testosterona e a circunferência escrotal (CE) também apresentou efeito linear indicando que, para cada centímetro a mais na CE nos touros Guzerá, ocorreu aumento de 0,2319 ng/mL na concentração de testosterona. A regressão da concentração espermática do sêmen em função da concentração sérica de testosterona apresentou efeito linear, os resultados mostram que, para cada 0,1 ng/mL a mais na concentração de testosterona circulante, ocorreu aumento de 9,29 x 10(6) espermatozoides/mL de sêmen. Os resultados indicam que os níveis circulantes de testosterona podem auxiliar na identificação e seleção de touros Guzerá, com maior potencial reprodutivo. Twenty-four Guzerat bulls were evaluated from 24 to 34 months of age, aiming to determinate the bulls serum testosterone concentration profile and their variation according to age, body weight and andrologic traits. The average of testosterone concentration was 1.36 ng/mL, with effect of time at harvest on circulating levels. The regression equation of testosterone according to body weight showed linear effect, suggesting that for each kilogram added to the body weight, an increase of 0.0082 ng/mL on serum testosterone concentrations occurs; and the regression of testosterone according to scrotal circumference (SC) also showed a linear effect. The results show that for each centimeter added to SC in Guzerat bulls, an increase of 0.2319 ng/mL on testosterone concentrations occurs. The regression of sperm concentration according to serum testosterone showed a linear effect, the results show that for each 0.1 ng/mL added to serum testosterone concentration, an increase of 9.29x10(6) spermatozoa/mL of semen occurs. The results indicate that circulating levels of testosterone may help on identification and selection of Guzerat bulls with major reproductive potential.

Recently, comprehensive studies were conducted regarding the reproductive development, age at puberty, spermatogenesis and the protein profile in the seminal plasma of Santa Ines rams. Despite the abundant information obtained from these studies regarding these tropically adapted rams in Brazil, it is still unclear how sperm parameters relate to the expression of molecular components of the reproductive tract. In this regard, the present study was conducted to determine if sperm parameters were empirically associated with the seminal plasma proteins described in two-dimensional electrophoresis maps. Seminal plasma proteins were separated by two-dimensional electrophoresis and the respective maps electronically analyzed. Protein spots associated with higher or lower sperm motility were then identified by tandem mass spectrometry. Based on sperm motility, the ejaculates were divided into two groups: those containing up to 80% motile cells (G1; n = 11) and those with more than 80% motile sperm (G2; n = 10). On average, 236 spots per gel were detected. Eleven spots were significantly more intense in gels from animals with ejaculates characterized by higher semen motility scores (G2). The intensity of three other spots was higher in gels from the G1 group. All spots differentially expressed for G1 and G2, were present in at least 90% of the gels. From the 13 spots differentially expressed in G1 and G2, four spots were identified by tandem mass spectrometry. Spots expressed with more intensity in the ejaculates with higher sperm motility (G2) were identified as arylsulfatase A and zinc-alpha-2-glycoprotein. On the other hand, two spots associated with G1 were identified as ram seminal vesicles protein 22 kDa (RSVP-22) and bodhesin-2. Knowledge of these identities represents a crucial step toward the comprehension of how specific seminal plasma proteins are related to sperm motility.

This study aimed to define sperm membrane protein markers of semen freezability of boars with the aid of a proteomic approach. Semen from fourteen adult boars were subjected to slow freezing and rapid thawing. After thawing, sperm vigor and motility were analyzed, and based on these results, animals were separated into two groups: good (GFEs) and poor freezability (PFEs). Sperm membrane proteins were extracted and subjected to two-dimensional electrophoresis. Stained gels were analyzed by computerized resources to indicate differentially expressed protein spots, that were identified by mass spectrometry. Six animals showed good freezability with average sperm vigor and motility of 2.2±0.8 and 41.8±22.9, respectively, whereas eight boars showed poor freezability, with 1.9±0.6 and 26.8±17.5 of sperm vigor sperm motility, respectively. An average of 263±62.2 spots per gel and 234.2±54.6 of spots consistently present in all gels were detected. The intensities of five spots were significantly different between groups. Fc fragment of IgG binding protein and lactadherin were more intense in the PFE group, while Arylsulfatase A and F-actin capping protein subunit alpha 1 were more expressed in the GEF group. Based on their functions and interactions with other proteins, we conclude that these four sperm membrane proteins may act as potential markers of boar semen freezability.

In order to determine the profile of leptin and insulin values in seminal plasma and their correlations with sperm patterns during the peripuberty, a total of 16 crossbred Gyr dairy bulls were monitored from 60 days before to 60 days after puberty, separated into two groups, early and regular, according to the onset of puberty. Hormone values were determined by radioimmunoassay after gel filtration in fast performance liquid chromatography and semen patterns, according to the Brazilian College of Animal Reproduction. Insulin concentrations in seminal plasma did not differ between groups, nor between the ages, with the pooling of the data showing average of 35.15±16.93, 36.60±26.21, 26.82±09.81, and 43.56±31.71 32.24±16.71 U/mL for the pubertal period of -60, -30, 0, +30 and +60 days, respectively. These values were not correlated with sperm parameters, but differed in the group of animals with the highest percentage of major defects (31.08±18.58 U/mL) compared with those with lower percentage of these sperm defects (40.01±25.37 U/mL). Leptin was not correlated with sperm parameters and did not differ between groups or stage of sexual development, with concentrations of 23.10±10.43, 24.35±9.42, 22.41±9.90, 23.76±9.69 and 24.99±11.42 ng/mL for the periods of puberty aforementioned, respectively, after being grouped. These results demonstrated the binding of insulin to the structural quality of the sperm cell, and this hormone is indicated in andrologycal evaluation of future breeding soundness.

Semen of twelve Tabapuã young bulls, which were according to patterns established by CBRA (1998) after cryopreservation, was submitted to fast and slow thermoresistence tests, hypoosmotic swelling test, induced acrosome reaction by heparin, sperm viability test by Trypanblue post-freezing and post-incubation with heparin, and pregnancy rates at first artificial insemination of Nellore heifers. Continuous variable averages of the mentioned tests were compared by Student–Newman–Keuls test and subjective variables were compared by Kruskal-Wallis nonparametric test (p0.05) to preview pregnancy rate, even if using the multiple regression model that included all variables. One can conclude that the model proposed in this study was not effective to predict pregnancy rate. Hypoosmotic Swelling Test can be suggested to evaluate functional and structural integrities of plasma membrane and the capacity of spermatozoa going through acrosome reaction.

É possível que a expressividade de alguns elementos do plasma seminal dos bovinos, como proteínas e hormônios, possa servir como marcadores para sêmende alta ou baixa fertilidade. Vários estudos têm demonstrado a associação de proteínas do plasma seminalcom a fertilidade de touros. Dentre as mais estudadas, destacam-se aquelas com afinidade à heparina, que exercem importantes papéis na capacitaçãoespermática e na reação acrossômica. Alguns fatoresendócrinos e/ou locais, podem estar associados à expressividadee/ou função destas proteínas, auxiliandonas condições espermáticas favoráveis à fecundação. Dentre estes, destacam-se a insulina, a leptina e o fatorde crescimento semelhante à insulina do tipo I. Assim sendo, evidenciam diferenças entre animais,estando associados à estrutura e as condições metabólicasda célula espermática, auxiliando na determinação da qualidade do plasma seminal. Desta maneira,o estudo das proteínas do plasma seminal associadoà condição metabólica destes hormônios, presentesneste meio, pode servir como importante parâmetrode avaliação da condição reprodutiva do macho.

The objectives of the present study were to investigate the effects of three equilibration times (0, 2, and 4h) and two extenders (TRIS or Bioxcell) for cryopreservation of bull semen. Semen from 12 Gyr bulls was cryopreserved using an automated freezing machine. There were significant interactions between equilibration times and extenders for sperm motility and membrane integrity. The control treatment (0h equilibration) had the lowest values (P0.05) for total (MOT) and progressive motilities (PROG), and percentage of sperm with intact plasma and acrosomal membranes (IPIA), with no significant differences between extenders. Extender TRIS had greater cryoprotective action than Bioxcell, with greater MOT, PROG, IPIA at 2 and 4h, as well as the lowest proportion of damaged plasma membrane (DPM, 72.2% vs. 85.8%) for all times. Equilibration for 4h yielded the most desirable (P0.05) for MOT, PROG, and IPIA, and the least DPM percentage (86.5, 78.0, and 72.6% for 0, 2, and 4h, respectively). Overall, the combination of TRIS and 4h of equilibration was the most desirable semen cryopreservation method, with greatest MOT, PROG, and IPIA (TRIS-T4=26.8%; BIO-T4=18.3%) and the least DPM. In conclusion, based on objective analyses, equilibration during cryopreservation was essential for maintaining motility and integrity of sperm membranes; equilibration for 4h yielded the greatest sperm survival, independent of the extender used.

Com o intuito de fornecer subsídios para programas de conservação, seleção e expansão da raça Gir no Nordeste do Brasil, objetivou-se avaliar o histórico do rebanho Gir registrado no nordeste brasileiro, com base na sua estrutura populacional e no progresso genético e fenotípico de características de desenvolvimento ponderal. Foram utilizadas informações de pedigree de animais nascidos no período de 1964 a 2006 e dados dos pesos ajustados aos 205 e 365 dias de idade de bovinos nascidos de 1978 a 2006. O progresso genético para a raça no Nordeste foi ocasionado exclusivamente pelo melhoramento ambiental. O tamanho efetivo da população reduziu a partir da década de 90, e o coeficiente de endogamia médio aumentou durante o período estudado. É imprescindível que o tamanho efetivo da raça Gir do nordeste seja ampliado e que haja maior controle de acasalamentos entre indivíduos aparentados, para prevenção da endogamia e conservação da variabilidade genética e viabilidade da raça. This paper provides an evaluation of the population structure, phenotype and genetic trends of registered Gyr herd cattle in northeast Brazil. The study provides important baseline information for the management, conservation and potential population expansion of this economically and culturally important cattle breed. Pedigree data were analyzed for individuals born between 1964 and 2006. Body weight values were adjusted to 205 and 365 days of age for animals born between 1978 and 2006. Phenotypic change of zebu Gyr in northeast Brazil is solely due to environmental improvement. However, there is potential for artificial selection for weight gain in young cattle. Effective population size decreased during the 1990s and the average inbreeding coefficient increased during the studied period. An increase of the effective population size of Gyr in northeast Brazil is strongly recommended, along with an increase in the management of the mating process to prevent inbreeding and to maintain the genetic variability of the breed.

O objetivo deste trabalho foi estimar correlações genéticas e fenotípicas de características reprodutivas e ponderais em 579 touros Nelore, em criação extensiva. As características reprodutivas estudadas foram: circunferência escrotal, consistência, volume e forma testiculares, comprimento dos testículos esquerdo e direito, largura dos testículos esquerdo e direito, motilidade e vigor espermáticos, defeitos espermáticos maiores, menores e totais e classificação andrológica por pontos. As características foram analisadas pelo método de máxima verossimilhança restrita, com algoritmos livres de derivadas, sob modelo animal, com inclusão da matriz de numeradores dos coeficientes de parentesco entre os animais e seus ascendentes. As correlações genéticas entre circunferência escrotal e as características peso corporal, volume testicular, motilidade espermática, vigor espermático, defeitos espermáticos menores, defeitos espermáticos totais e classificação andrológica por pontos foram, respectivamente, 0,72, 0,99, 0,72, 0,60, -0,67, -0,12 e 0,64. As maiores correlações fenotípicas encontradas entre peso e circunferência escrotal, características físicas e morfológicas do sêmen, quando comparadas às correlações entre idade e as mesmas características, são indicativas de que o peso tem maior influência na condição reprodutiva. As correlações genéticas entre classificação andrológica por pontos e as características: peso, circunferência escrotal, volume testicular, defeitos espermáticos maiores e defeitos espermáticos totais foram, respectivamente, 0,19, 0,64, 0,71, -0,47 e -0,58.

The present study evaluated functional aspects of binder of sperm 1 (BSP1) in the bovine species. In a first experiment, cumulus-oocyte complexes (n = 1274) were incubated with frozen-thawed ejaculated sperm (18 hours) in Fert-TALP medium containing: heparin, 10, 20, or 40 μg/mL BSP1. Heparin followed by gelatin affinity chromatography was used for purification of BSP1 from bovine seminal vesicle fluid. With ejaculated sperm, cleavage rates were similar when Fert-TALP medium was incubated with heparin (74.1 ± 2.7%), 10 μg/mL BSP1 (77.8 ± 3.1%), or 20 μg/mL BSP1 (74 ± 2.0%). Day-7 blastocyst rates were equivalent after incubations with heparin (40.8 ± 5.0%) and 10 μg/mL BSP1 (34.1 ± 4.4%), but reduced after 20 μg/mL BSP1 (22.4 ± 2.9%) and 40 μg/mL BSP1 (19.3 ± 4.1%; P < 0.05). In the second experiment, cumulus-oocyte complexes (n = 1213) were incubated with frozen-thawed cauda epididymal sperm (18 hours) in Fert-TALP medium containing: no heparin, heparin, 10, 20, or 40 μg/mL. Cleavage and blastocyst rates were similar after treatments with heparin (68.5 ± 1.3% and 24.7 ± 3.2%, respectively) or without heparin (65.5 ± 1.8% and 27.3 ± 1.6%, respectively). Cleavage was higher after treatment with any BSP1 concentrations (74.2 ± 2.7%-79.0 ± 1.1%) than without heparin (P < 0.05). Also, cleavage was better after Fert-TALP medium incubation with 40 μg/mL BSP1 (79.0 ± 1.1%) than with heparin (68.5 ± 1.3%; P < 0.05). Embryo development was higher (P < 0.05) after treatment with 20 μg/mL BSP1 (35.6 ± 2.5%) and 40 μg/mL (41.1 ± 2%) than after incubations with heparin (24.7 ± 3.2%) or without heparin (27.3 ± 1.6%). Interestingly, BSP1 did not cause reductions in blastocyst rates after fertilization with epididymal sperm, as observed with ejaculated sperm. On the basis of immunocytochemistry, there was BSP1 binding to frozen-thawed ejaculated but not to epididymal sperm. Also, anti-BSP1 reaction remained on ejaculated sperm (as expected) and appeared on epididymal sperm after incubation with purified BSP1. Acrosome reaction of ejaculated and epididymal sperm was induced after incubation with purified BSP1 as well, indicating an effect of BSP1 on capacitation. In conclusion, purified BSP1 from bull seminal vesicles was able to bind to and induce capacitation of ejaculated and epididymal sperm. Also, BSP1 added to fertilization media and allowed proper cleavage and embryo development, with the effects being modulated by previous exposure or not of spermatozoa to seminal plasma.

The present study evaluated the effects of heat stress on the ram seminal plasma proteome. Six Morada Nova rams were scrotal insulated for 8 days. Scrotal circumference, sperm parameters, and seminal fluid proteins were evaluated before (Day 0) and twice during scrotal insulation (Days 4 and 8), and weekly until semen parameters returned to preinsulation values (normal). Seminal proteins were analyzed by two-dimensional SDS-PAGE and mass spectrometry. Scrotal circumference decreased from 30 ± 0.4 cm on Day 0 to 22.6 ± 0.6 cm on Day 36 (P

Gummosis is an aggressive disease caused by the necrotrophic fungus Lasiodiplodia theobromae (Pat.) Griffon & Maubl that threatens commercial cashew orchads in Brazil. To study the molecular mechanisms involved in the cashew response to L. theobromae , a proteomic analysis of stems from the commercial cashew clone BRS 226 (resistant) was conducted at early times post-artificial infection. In addition, changes in the stem proteome profiles of gummosis resistant and susceptible cashew plants grown under field condition and naturally exposed to pathogen were also compared. After two-dimensional gel electrophoresis (2D-PAGE), 73 proteins showed statistically significant differences in spot abundance. Of these, 31 spots were identified in BRS 226 stems compared with mock-inoculated controls and 32 in stems collected from field-grown resistant and susceptible cashew plants. L. theobromae -responsive proteins were mainly involved in energy metabolism pathways, stress and defense, cell signaling and protein metabolism indicating modulation of various cellular functions upon fungal infection. As stress-inducing factors seem to be important for susceptibility to disease, the change in the abundance relative these proteins may possibly indicate an attempt to maintain cellular homeostasis, as resistance determinant factor, related with a possible role in the regulation of oxidative burst. These findings provide the first information about the cellular mechanisms acting in the Anacardium occidentale genotypes associated with the pathophysiological state of infection with L. theobromae . Biological significance Gummosis caused by Lasiodiplodia theobromae , a necrotrophic fungus, is the major disease of cashew plants in the semi-arid conditions of northeastern Brazil. Although various studies were carried out on this pathosystem, there is no information available on the molecular mechanisms of plant defense related to the incompatible interaction of cashew with L. theobromae . Therefore, this original study comprises a differential proteomic analysis of cashew stems from: (i) resistant dwarf clone BRS 226 mock-inoculated (control) and artificially inoculated with L. theobromae ; and (ii) cashew plants bearing resistant and susceptible traits to gummosis, originated from open pollination of BRS 226 in a commercial orchard with high disease incidence. The contribution of the reprogrammed proteins to molecular events triggered in cashew plants challenged by L. theobromae has a great relevance in the identification of the host candidate proteins linked to biological pathways that respond to L. theobromae infection. Furthermore this study may contribute to improve breeding programs aimed at selecting resistant/tolerant cashew clones toward this pathogen.

The objective of the present study was to evaluate the effect of different hormonal stimulation treatments on the antral follicular population of naturalized Canindé goats. Adult goats (n=17) having estrous cycles at regular intervals were treated with intra-vaginal sponges containing 60 mg medroxyprogesterone acetate for 11 days, combined with an application of 50 μg d-cloprostenol on the Day 8 of treatment. For ovarian stimulation, goats were distributed into the following experimental groups: (a) multiple doses (MD), with a total of 120 mg NIH-FSH P1 in five intramuscular injections (30/30; 20/20 and 20 mg) at 12-h intervals; (b) three doses (TD), with a total of 120 mg NIH-FSH P1 in three intramuscular injections (60; 40 and 20 mg) at 24 h intervals; (c) one dose (OD), which consisted of the use of 70 mg NIH-FSH P1 combined with 200 IU eCG administered intramuscularly 36 h before sponge removal. In the MD and TD groups, FSH injections were begun on the Day 8 of progestagen treatment. The ovaries of all animals were observed by transrectal real time ultrasonography (TRU) during the follicular stimulation protocols. All follicles ≥2 mm were counted, measured and classified according to greatest diameter. The ultrasonographic assessment of the ovaries provided for well-defined ovarian structures. At the time of insertion of the sponges (Day 0), significant differences were observed (P0.05) for the mean number of large follicles between the treated groups. Meanwhile, on Day 11, the three treatments did not differ (P0.05), regardless of the follicular category. The diameter of small follicles was similar in MD, TD and OD during the whole period of the study. In the TD group, diameter of the large follicles was less (P0.05) on Day 10 when compared to MD and OD. However, these differences were not observed on Day 11. In conclusion, the three treatments produced comparable distribution of the follicular populations. However, the single dose treatment can be preferred because of its simplicity and efficacious follicular response.

Um total de 12 touros da raça Guzerá, de 24 a 34 meses de idade, avaliados andrologicamente, foram submetidos a teste da libido com o objetivo de estudar o comportamento sexual em curral, por cinco, 10 e 15 minutos de observação; e associar com características andrológicas (aspectos físicos: motilidade, vigor, turbilhonamento e concentração espermáticos, e volume do sêmen; e morfológicos: defeitos espermáticos maiores, menores e totais, do sêmen), circunferência escrotal (CE), classificação andrológica por pontos (CAP), peso corporal, idade e concentrações séricas de testosterona. Observou-se, de forma geral, baixa resposta dos animais aos testes, uma vez que menos da metade dos animais foi classificada com libido média ou alta. Verificou-se pequena variação na pontuação entre os tempos de observação, com escore 3,0 em cinco minutos e 3,3 em 15 minutos de avaliação (p>0,05). A associação entre a libido e as concentrações séricas de testosterona (0,78), reforçam o achado de influência desta característica na expressão do interesse sexual em touros jovens Guzerá.

A raça Gir Leiteiro é uma excelente opção para os rebanhos leiteiros no Brasil pelas características apresentadas por seus animais, como rusticidade, adaptabilidade e produção de leite expressiva. A hipoplasia testicular causa problemas reprodutivos nos touros podendo levar à subfertilidade ou, em casos mais graves, à infertilidade. No Brasil, a incidência dessa patologia varia de 5 a 15%. Devido à grande importância que animais da raça Gir Leiteiro têm na pecuária brasileira e aos impactos negativo, tanto em termos de fertilidade quanto economicamente, causados pela hipoplasia testicular, este trabalho objetivou traçar o perfil proteico diferencialmente expresso entre animais com esta patologia e clinicamente normais. Amostras de sêmen de 18 animais (9 hipoplásicos e 9 normais) foram coletadas e o plasma seminal foi centrifugado e submetido à eletroforese bidimensional (2-DE). Os géis foram corados com Coomassie Blue G-250, digitalizados e analisados pelo programa PDQuest 2-D Analysis Software para identificação dos spots. Foram identificados 33 spots diferencialmente expressos entre os dois grupos de animais, sendo 26 mais intensamente expressos em animais hipoplásicos. Os spots foram cortados dos géis e submetidos à espectrometria de massa sendo identificadas 38 proteínas e, dentre estas, o grupo de proteínas conhecidas como Binder of Sperm Proteins (BSP) e a platelet-activating fator acetylhydrolase (PAF AH) foram as mais presentes nos spots identificados nos animais hipoplásicos. Os animais também foram submetidos à avaliação clinico-andrológica e houve diferenças estatísticas quanto ao comprimento, largura e volume testicular, turbilhonamento e defeitos espermáticos (p