A comparison of immunological, chemical and surgical castration of Nelore bulls.

The objective was to compare effects of immunological, chemical and surgical castrations on testicular characteristics, scrotal surface temperatures, sperm quality, and serum testosterone concentrations in Bos indicus bulls. Eighty Nelore bulls (∼20 mo) were grazed on pasture, fed a supplement and slaughtered at ∼480 kg. Bulls were allocated into four groups (n = 20/group). The control group (CON) was non-castrated and there were three castration methods: immune (IMM - Bopriva®), injected on D-30, D30 and D60; chemical (CHE), an intratesticular injection of 40% CaCl2 + 0.5% dimethylsulfoxide on D0; and surgical (SUR) on D0. The CHE group were surgically castrated on D60, due to testicular swelling and necrosis of scrotal skin. Most scrotal surface temperatures (infrared thermography) were significantly higher on D15 in CHE and SUR compared to CON, due to inflammation. All bulls were subjected to a breeding soundness evaluation on D-7 and slaughtered on D220. Scrotal circumference and testicular volume did not differ among groups (P > 0.05) at D0, but at D15, both end points were highest (P < 0.05) in the CHE group (due to swelling), although both end points were smaller for IMM versus CON (P < 0.05) at D60 and D150. Sperm motility in the IMM group was ∼50 and 10% of that in the CON group on D60 and D150, respectively. For the IMM group, serum testosterone concentrations were similar on D0 and D15 (but ∼50% of CHE or SUR on D0, attributed to the first treatment on D-30), and had decreased ∼70% on D60 and D150, whereas in the CHE and SUR groups, there were ∼80-90% decreases in testosterone after D0. In conclusion, immunological castration was a viable alternative to surgical castration, as it supressed testosterone production and spermatogenesis, with the benefits of being much less invasive, with better animal welfare and less stress.

Eficacy of ivermectin, moxidectin and febendazole in equine in Brazil.

The objectives of the present study were to evaluate the efficacy of three anthelmintic drugs, i.e. fenbendazole, ivermectin and moxidectin; to determine the genera and species of the most abundant strongyles; and to investigate parasite control measures used in herds of horses in the western region of the state of São Paulo, Brazil. This study was onducted between February and December 2013 on 10 farms in this region. Coprological evaluations were conducted for counting the numbers of eggs per gram of feces (EPG) and coprocultures were made in order to identify third-stage larvae (L3) of strongyles. Parasite control measures were investigated by surveying each farm's management and characteristics. A fecal egg count reduction test (FECRT) was used to determine the anthelmintic efficacy based on EPG count results before and after treatment. The FECRT showed that anthelmintic resistance to fenbendazole was present and that ivermectin and moxidectin had high effectiveness in the western region of the state of São Paulo. Identification of L3 revealed that there was high prevalence of cyathostomins among strongyle larvae. The highest prevalence of Strongylus vulgaris occurred on farms where the intervals between deworming were long. The questionnaire applied showed that ivermectin was the drug most used among these farmers.

Cross comparison of seminal plasma proteins from cattle and buffalo (Bubalus bubalis).

The objective of this study was to evaluate seminal plasma proteins from cattle and buffalo (Bubalus bubalis), to identify differences between related species. Sixteen buffaloes and 16 cattle between 30 and 60 months of age were used. Semen collection was performed by electroejaculation, followed by macroscopic and microscopic subjective analyses. After analysis, the samples were centrifuged at 800 g for 10 min, and the supernatant (seminal plasma) was recentrifuged at 10,000 g for 30 min at 4°C. The total protein concentration was determined by the Bradford method, and the proteins were digested in solution for mass spectrometry (nLC-MS/MS). Multivariate statistical analysis was used to evaluate the proteomics results by non-hierarchical clustering the considering exponentially modified protein abundance index (emPAI). Principal component analysis (PCA) and partial least squares discriminant analysis (PLS-DA) were used for clustering. Proteomics identified 78 proteins, and multivariate analysis showed 4 that were over-expressed in buffaloes (cystatin C, prosaposin, peptide YY and keratin type II cytoskeletal 5) and 9 in cattle (spermadhesin-1, seminal plasma protein PDC-109, ribonuclease 4, metalloproteinase inhibitor 2, acrosin inhibitor 1, seminal ribonuclease, C-type natriuretic peptide, angiogenin-1 and osteopontin). Among the proteins identified in seminal plasma, the C-type natriuretic peptide and metalloproteinase inhibitors were described for the first time in buffaloes. Some protease inhibitors were found over-expressed in buffaloes, and important proteins in seminal plasma of cattle were not identified or were found at lower expression levels in buffaloes, which can contribute to reproductive performance in this species.

Proteomic analysis of amniotic and allantoic fluid from buffaloes during foetal development.

The objective of this study was to describe the dynamic changes in protein composition and protein abundance in amniotic and allantoic fluids from buffaloes during gestation. Amniotic and allantoic fluids were collected during the first, second and third trimesters of gestation. The foetuses were measured and weighed. Fluid samples were centrifuged at 800 g for 10 min and then at 10,000 g for 60 min at 4°C. The supernatant was collected to determine the total protein concentration. Based on total protein concentration, an aliquot (50 µg) was used for in-solution tryptic digestion, and mass spectrometry analysis (nano-LC-MS/MS) was performed. A multivariate statistical analysis of the proteomic data was conducted. Across the different stages of buffalo gestation, fifty-one proteins were found in the amniotic fluid, and twenty-one were found in the allantoic fluid. A total of twelve proteins were common among the stages, and four presented significant differences (VIP score α > 1). Fibronectin and alpha-1-antiproteinase were more abundant in the amniotic fluid than in the allantoic fluid. Alpha-2-macroglobulin and alpha-2-HS-glycoprotein were more abundant in the allantoic fluid than in the amniotic fluid. Alpha-2-macroglobulin participates in remodelling and growth of the uterus at beginning of the gestation (first trimester), and these findings indicate that can serve as a potential tool for the early diagnosis of pregnancy in buffaloes.

Effect of estrous cycle phase on vulvar, orbital area and muzzle surface temperatures as determined using digital infrared thermography in buffalo.

The objective of this study was to evaluate variations in the orbital area, muzzle and vulva surface temperatures and progesterone (P4) concentrations during follicular and luteal phases in Murrah buffalo and whether these temperatures are influenced by the weather patterns. Forty cows were submitted to P4-based hormonal protocol. After P4 device withdrawal transrectal ultrasonography and infrared digital thermography were performed daily until day 16 and on days 20, 24, 28 and 32 to follow the ovulation as well as the vulva, orbital area and muzzle temperatures. In addition, the weather variables were evaluated, as well as rectal temperature (RT) and P4 and cortisol concentration. Vulva, muzzle and orbital area temperatures correlated positively with RT and with weather data. Greater temperatures of the vulva, orbital area and muzzle were detected during the period of estrus. The vulvar surface temperature (VST) was not influenced to a great extent by weather factors during the morning, so this period was chosen to evaluate the influence of the phase of the estrous cycle on VST. The VST was less during days 16, 20, 24 and 28 (diestrus) and P4 concentration was inversely proportional to the VST. Muzzle, orbital area and RT, however, were not of the same pattern. Negative correlations were observed between VST and P4 concentrations. It is concluded that VST undergoes changes during the reproductive phases, correlating with P4 concentration. The weather factors influence the temperatures of the body surface areas, and the morning is the most desirable time to perform the thermographies.

Functional insights into the role of seminal plasma proteins on sperm motility of buffalo.

The objective of the present study was to describe the proteins from the seminal plasma of buffalo and correlate these proteins with sperm motility. Ejaculates from sixteen Murrah buffalo were used. Semen collection was performed by electroejaculation, and the ejaculate was evaluated by macroscopic (volume) and microscopic analysis (subjective motility and vigor, as well as sperm concentration). After the analysis, the samples were centrifuged (800g for 10 min and 10,000 for 30 min at 4 °C), and the supernatant (seminal plasma) was used to determine total protein concentration by the Bradford method. Based on total protein concentration, an aliquot (50 µg) was taken to conduct protein in-solution digestion for nano-LC-ESI-Q-TOF mass spectrometry analysis. Samples were divided into two groups, minimal (little sperm motility) and greater (typical sperm motility), based on non-hierarchical clustering considering motility and emPAI protein value. The data were analyzed by multivariate statistical analysis using principal component analysis (PCA) and partial analysis of minimum squares discrimination (PLS-DA). Forty-eight proteins were detected in the seminal plasma, and fifteen were common to two groups. There were six proteins that were significantly different between the groups. The main functions of proteins in seminal plasma were catalytic and binding activity. Spermadhesin protein, ribonuclease, 14-3-3 protein zeta/delta and acrosin inhibitor were in greater amounts in seminal plasma from the group with greater sperm motility; prosaposin and peptide YY were in greater amounts in the group with little sperm motility. The proteins detected in the greater motility group were correlated with sperm protection, including protection against oxidative stress, lipid peroxidation, protease inhibition and prevention of premature capacitation and acrosome reaction. In the group with little sperm motility, one of the identified proteins is considered to be an antifertility factor, whereas the function of other identified protein is not definitive. Results from the present study add to the knowledge base about the molecular processes related with sperm motility, and these findings can be used for determining potential markers of semen quality.

Evaluation of buffaloes' follicular dynamics and stress state under different ovulation synchronization protocols.

The aims of the present study were to analyze the effect of different hormonal protocols using an implant containing Norgestomet, at the ovulation synchronization in buffaloes and to verify the effect of the stress caused by manipulation of the herd during the experiment. Twenty-four female Murrah breed buffaloes, lactating, multiparous, aged from three to eight years, with a body condition score of 3.5 or higher (0 to 5) and with more than 45 days post partum, were used. These females were randomly distributed into one of the three groups (group 1, group 2 and group 3) with eight subjects in each. On day 0 (day 0), all animals received an ear progesterone implant with 3.0 mg of Norgestomet and an intramuscular (IM) injection with 2.0 mg of estradiol benzoate (EB). The implants were removed after nine days (day 9) and one single dose of PGF2α (0.15 mg d-cloprostenol, IM) was administered to all animals. On the same day, the group´1 and group 3 buffaloes were treated with 500 UI of eCG (IM). Two days later (day 11), 1000UI of hCG (IM) were administered to the group 1 and group 2 buffaloes. After the implant had been removed, an ultrasound evaluation was performed every 12 h, in order to access the ovarian follicular dynamics, using an Aloka 500 equipment with a 5 MHz transrectal probe. Blood samples were also taken on days 0, 9 and 11 to determine the plasmatic concentrations of cortisol and progesterone. No difference was observed in the ovulation rate (group 1: 62.5%, group 2: 50%, group 3: 75%). However, the size of the preovulatory follicles and the plasmatic concentration of cortisol had (P < 0.05). G3 was the most efficient group in promoting the ovulation synchronization, which suggests that this protocol can be used in Fixed Timed Artificial Insemination programs (FTAI) among postpartum, lactating Murrah breed buffaloes'.