Augusto Schneider

Associations between resumption of postpartum ovarian activity, uterine health and concentrations of metabolites and acute phase proteins during the transition period in Holstein cows

The resumption of ovarian activity, uterine health, severity of the negative energy balance and the synthesis of inflammatory mediators during the transition period in dairy cows are interrelated. Therefore, the aim of this study was to evaluate the association between the resumption of postpartum ovarian activity and the percentage of polymorphonuclear (PMN) cells in endometrial cytology, lipid mobilization and the secretion of acute phase proteins. For this study, 20 multiparous Holstein cows were used. Blood samples that were collected from 21d before calving to 44d in milk (DIM) were analyzed for serum glucose, non-esterified fatty acids (NEFA), insulin, haptoglobin, albumin, paraoxonase and progesterone. Endometrial cytology was performed at 37±2DIM to evaluate the percentage of PMN cells in the uterine flushing. Cows were divided into two groups: (1) ovulatory cows (n=12), which returned to ovarian activity by 44±2DIM; and (2) anovulatory cows (n=8), which did not resume ovarian activity during this period. Ovulatory cows had a lower (P=0.05) percentage of PMN cells in endometrial cytology than anovulatory cows (26.3±8.3% vs. 53.4±16.9%, respectively). Ovulatory cows had higher serum albumin during the pre- (P=0.03) and postpartum periods (P=0.01), and tended to have lower haptoglobin concentrations in the prepartum period (P=0.07) and higher paraoxonase activity in the postpartum period (P=0.09). In conclusion, cows that resumed ovarian activity early in the postpartum period had higher albumin concentrations in the peripartum period, which were associated with a lower percentage of uterine PMN cells.

The role of transplanted visceral fat from the long-lived growth hormone receptor knockout mice on insulin signaling

Growth hormone receptor knockout mice (GHRKO) are characterized by high insulin sensitivity and extended lifespan. Interestingly, the secretory activity of visceral fat in GHRKO mice is altered, stimulating whole body insulin sensitivity. In this study, we transplanted normal (N) mice with visceral fat pads from GHRKO or N mice to determine the role of visceral fat on the insulin signaling. We found that the transplant of visceral fat from GHRKO mice to N mice (N-GHRKO) improved whole body insulin sensitivity when comparing with sham-operated mice (N-S) and with mice that received visceral fat from N mice (N-N). This was associated with increased hepatic insulin sensitivity as observed by the increased phosphorylated insulin receptor and increased hepatic expression of Pparα and Pparγ. In conclusion, we demonstrated that visceral fat transplant from GHRKO mice into normal mice enhanced insulin sensitivity and glucose tolerance. These results further confirm the differential physiological role played by visceral adipose tissue from GH receptor deficient mice, indicating that the increase of this fat depot can be associated with beneficial effects on insulin signaling and longevity.

Effect of the ovulatory follicle diameter and progesterone concentration on the pregnancy rate of fixed-time inseminated lactating beef cows

The objective of this study was to evaluate the influence of the ovulatory follicle diameter on the reproductive performance of lactating beef cows subjected to low progesterone fixed-time artificial insemination (FTAI) protocols. Ninety-three lactating beef cows (60-80 days postpartum) at random stages of estrous cycle were given a luteolytic dose of prostaglandin F2α (500 μg cloprostenol; PGF) twice, 11 d apart. Ten days after the second PGF treatment, cows were given 1.5 mg of estradiol benzoate im and a progesterone-releasing intravaginal device (Cue-Mate) with a single pod containing 0.78 g progesterone (Day 0). Cows received another luteolytic dose of PGF on Day 0. On Day 8, the Cue-Mate was removed. Fifty-four to 56 hours later, cows received 12.5 mg of porcine LH (pLH) i.m. and were concurrently artificially inseminated. Ultrasound examinations of the ovaries were performed on Days 10 and 17 to evaluate the diameter of ovulatory follicle and corpus luteum, respectively. Cows which presented ovulatory follicle > 19 mm resulted in larger corpus luteum than cows that had ovulatory follicle <15 mm in diameter. However, cows with ovulatory follicle between 13-15 mm had higher pregnancy rate than other categories of ovulatory follicle. Although larger ovulatory follicles result in larger corpus luteum and consequently higher progesterone production, the optimal size of ovulatory follicles (13-15 mm) may result in positive benefits on pregnancy rate for cows subjected to FTAI with low progesterone concentration protocols.

Effect of recombinant bovine somatotropin on plasma concentrations of insulin-like growth factor I, insulin and membrane integrity of bull spermatozoa.

This study aimed to evaluate the effect of the exogenous recombinant bovine somatotropin (rbST) on plasma concentrations of insulin-like growth factor I (IGF-I), insulin and semen quality of bulls. Twenty bulls (Aberdeen Angus and Brangus) were divided by breed into two groups. Placebo group was injected with NaCl 0.9% (s.c.) and treatment group with rbST (s.c., 500 mg) at days 0 and 14 of the experiment. Immediately after semen collection, blood samples were taken on days 0, 14, 28, 42 and 56 of the experiment. Semen was also collected on day 70 of the experiment. Evaluation of sperm motility was performed at pre-freezing and post-thawing stage, whereas assessment of sperm membrane integrity was performed after freezing and thawing. Analysis of data revealed that the effect of treatment and treatment-by-collection day on plasma concentrations of IGF-I and insulin was not significant. However, mean plasma concentrations of IGF-I and insulin were affected (p < 0.0001) by days of blood sampling. Effect of treatment and treatment-by-collection day on motility of spermatozoa was similar (p > 0.05) at pre-freezing and post-thawing stage. Intactness of plasmalemma and tail membrane of spermatozoa at post-thawing stage was higher (p < 0.05) in rbST-treated group than in control. In conclusion, rbST did not affect plasma concentrations of IGF-I and insulin, however, it did improve post-thaw sperm membrane integrity.

Insulin-like growth factor and growth hormone receptor in postpartum lactating beef cows

The objective of this study was to evaluate the plasma concentrations of insulin-like growth factor-I (IGF-I), and the mRNA hepatic expression of IGF-I and of the growth hormone receptors GHR and GHR 1A, in postpartum beef cows. Four Angus and four crossbred (Angus x Nelore) postpartum suckled beef cows were used. Liver and blood samples were collected every 10 days, from calving to 40 days postpartum, for gene expression and for β‑hydroxybutyrate and IGF‑I assays, respectively. Samples for progesterone assay were collected every other day, from day 10 to 40 postpartum. Three cows ovulated before 40 days postpartum. IGF-I concentration was higher in Angus x Nelore than in Angus cows. There was no difference in the expression of GHR, GHR 1A and IGF-I according to breed or ovulatory status. IGF-I concentrations were higher in crossbred cows, but have not changed according to postpartum ovulatory status. Moreover, changes in postpartum IGF-I concentrations are not associated with changes in liver GHR, GHR 1A and IGF-I mRNA expression in either breed.

Efeito da duração do tratamento com progestágeno e da maturidade sexual na taxa de prenhez em novilhas de corte: avaliação econômica e biológica

The aim of this experiment was to evaluate the effect of the duration of progestagen (P4) exposure and sexual maturity of prepubertal and pubertal heifers submitted to timed artificial insemination (TAI). A hundred and fourteen 16- month old Bradford heifers were classified, according treatment and sexual maturity, in 4 groups: PPLPG (prepubertal long exposure to P4, n=21), PPSPG (prepubertal short exposure to P4, n=19), PLPG (pubertal long exposure to P4, n=34) and PSPG (pubertal short exposure to P4, n=40). On day -8, the heifers from PPLPG (prepubertal long exposure to progestagen) and PLPG (pubertal long exposure to progestagen) received an intravaginal sponge impregnated with medroxiprogesterone acetate (MAP). On day 0 the sponge was removed and a new sponge was inserted, together with an injection of 2.5 mg estradiol benzoate (EB). On day 8 the sponge was removed and the heifers received an injection of 0.5 mg EB and 250 ig sodium cloprostenol. The TAI was performed 54 hours after sponge withdrawal. The heifers from PPSPG and PSPG received the same synchronization treatment, except for the additional MAP from day -8 to day 0. The pregnancy rate was higher in pubertal (60.3%) than in pre-pubertal (35.0%) heifers. However, there was no effect of the duration of treatment in the pregnancy rate between pubertal and prepubertal heifers. The second part of this study aimed to evaluate the economical return of the protocols and compare them with the traditional breeding system at 26 months of age. A better economic response with the use of protocols associating P4 in heifers at 16 months was observed compared to the traditional system at 26 months of age. The prolonged exposure to P4 did not improved pregnancy rate in heifers submitted to TAI.

Timed artificial insemination in blocks: A new alternative to improve fertility in lactating beef cows

The aim of this study was to evaluate whether changing the interval from CIDR removal to timed artificial insemination (TAI) according to the diameter of the preovulatory follicle (POF) would improve pregnancy per AI in cows. In Study 1, a retrospective analysis of TAI experiments (n=96 cows) was performed to characterize the time of ovulation according to the diameter of the dominant follicle. It was observed that cows with a larger POF had ovulations earlier than cows with smaller POF, according to the equation: y=0.72x(2)-26.74x+264.54 (R(2)=0.63; P<0.001). In Study 2, lactating Nelore cows (n=412) were subjected to an EB-CIDR based TAI protocol. On the morning of Day 10 (time of TAI), cows were randomized into Control (n=209) and Block (n=203) groups; (1) Cows in the Control Group were TAI 48 h after CIDR removal (08:00 am on Day 10), and; (2) Cows in the block group were inseminated once at one of the following time points, according to the diameter of the POF on Day 10: B0 (POF≥15mm, TAI 0 h after convetional TAI), B1 (POF 13-14.9 mm, TAI 6h later), B2 (POF 10.1-12.9 mm, TAI 24h later) and B3 (POF≤10mm, TAI 30 h later). The cows of the Block Group had greater pregnancy rates per AI than the Control Group (129/203, 63.5% when compared with 102/209, 48.8%, respectively; P<0.01). In conclusion, results of the present study demonstrate that adjusting the timing of TAI according to the diameter of the POF can be an effective practice for improving fertility of cows in TAI protocols.

The effect of the paraoxonase 1 (PON1) T(−107)C polymorphism on serum PON1 activity in women is dependent on fatty acid intake

Paraoxonase 1 (PON1) is an enzyme that prevents the peroxidation of lipoprotein and cell membranes. Our hypothesis is that the effect of the PON1 T(-107)C polymorphism on serum PON1 activity in healthy adult women is dependent on their fatty acid intake profile. This study included women (n = 39) who completed a food frequency questionnaire. Fatty acid intake was estimated based on the interview and a nutrient reference table. Blood samples were collected for genotyping and to measure serum PON1 activity. Serum PON1 activity was different among genotypes and was higher for women of the CC genotype (P < .001). Women in the study were categorized in 2 groups according to the median nutrient intake. Overall, there was a difference (P < .05) in serum PON1 activity between the CC and TT genotypes in women ingesting either above or below the median total fat, saturated fatty acids, monounsaturated fatty acids, polyunsaturated fatty acids, omega 3 (n-3) and omega 6 (n-6; P < .05). However, genotype effects on serum PON1 activity were not observed in women ingesting below the median (15:1) ratio of n-6/n-3 (P > .05) but were observed in women ingesting above the ratio of n-6/n-3 (P < .05). This is partly because women of the CC genotype had decreased PON1 activity when ingesting a lower ratio of n-6/n-3 diet (P < .05), while women of the TT genotype had increased PON1 activity (P < .05). In conclusion, the overall presence of the C allele was associated with increased serum PON1 activity, although a diet with high saturated fatty acid or a low ratio of n-6/n-3 reduced PON1 activity in women with the CC genotype.

Characterization of single nucleotide polymorphisms in the promoter region of the bovine paraoxonase 1 (PON1) gene affecting serum enzyme activity in dairy cows

The aim of the present study was to characterize single nucleotide polymorphisms (SNPs) in the promoter region of the bovine paraoxonase 1 (PON1) gene and determine their relationship with PON1 serum activity. Blood samples were collected from 47 Holstein cows during the periparturient period. Serum PON1 was measured and DNA extracted for PCR and sequencing of the promoter region of the PON1 gene. Seven novel SNPs were identified and of these, five SNPs, at positions -105, -221, -392, -611 and -674, upstream of the start of the published mRNA sequence (NM_001046269.2), were found to be associated with variability in serum PON1 activity (P < 0.05). The SNPs at positions -221 and -611 were located in regions predicted to bind to transcription factors linked to the acute phase response.

Simple Method to Genotype the ACTN3 r577x Polymorphism

The alpha-actinin-3 r577x polymorphism (rs1815739) is one of the most important polymorphisms associated with athletic performance. This single-nucleotide mutation leads to a premature stop codon, resulting in a nonfunctional protein product. The presence of the dominant R allele is associated with full power skeletal muscle contraction. Homozygosity for the X allele is correlated with more efficient energy disposure. Restriction fragment length polymorphism and real-time polymerase chain reaction (PCR) are the standard methods used to genotype this polymorphism, but they are expensive and require special equipments. Here, we present a simple and cost-efficient method to genotype the ACTN3 r577x polymorphism by a single PCR. External primers yield a 690-bp product that indicates the template quality. Internal primers produce a 413-bp product if the R allele is present and a 318-bp product if the X allele is present. Our four-primer genotyping PCR was validated by the standard real-time PCR, generally used to genotype this single-nucleotide polymorphism, demonstrating the accuracy of this method. This protocol is perfect for small- or large-scale cohort genotyping of the ACTN3 r577x polymorphism.