Addition Of eCG Research Articles

Addition of equine chorionic gonadotropin in controlled internal drug-release-based synchronization improves reproductive performance of Nili-Ravi buffalo in the sub-tropics

This study evaluated the effect of equine chorionic gonadotropin (eCG) and factors such as body condition score (BCS), cyclicity, and breeding season in a controlled internal drug release (CIDR)-based oestrus synchronization protocol on ovarian follicle dynamics, oestrus expression, ovulation rate, and pregnancy per artificial insemination (AI) (P/AI) in Nili-Ravi buffalo. Adult buffaloes (n = 88), 6.69 ± 1.17 years, parity 3.36 ± 0.97, and BCS 2.76 ± 0.45, were randomly administered eCG or saline, concurrent with prostaglandin F2α (PGF2α) treatment on Day 6 in a CIDR synchronization protocol during the breeding season and low breeding season. Follicular dynamics and oestrus expression were recorded from eCG administration until ovulation. Fixed-time artificial insemination (FTAI) was performed at 48- and 60-hour intervals after CIDR withdrawal. On Day 35 post-AI, pregnancy was diagnosed with ultrasonography. There was no significant difference in the size of the ovulatory follicle between eCG-treated buffaloes (ETB) and saline-treated buffaloes (STB). The growth rate of the dominant follicle, oestrus response and intensity, ovulation, and P/AI were higher (P <0.05) in eCG-treated buffaloes. It was concluded that eCG improved the growth rate of the dominant follicle, oestrus expression, and P/AI in a CIDR-based FTAI programme in Nili-Ravi buffalo. The deleterious effects of poor BCS, acyclicity, and low breeding season could be reduced with a progesterone-based fixed time AI protocol with the addition of eCG.

Effect of FSH and eCG on Alpaca (Vicugna pacos) Oocyte Maturation in vitro

This study evaluated alpaca oocytes that developed to metaphase II (MII), using different concentrations of follicle stimulating hormone (FSH) with and without equine chorionic gonadotropin (eCG) during in vitro maturation. Oocytes were obtained from ovaries of slaughtered alpacas. Oocytes were matured in vitro for 36h in TCM-99 and supplemented in groups with different doses of FSH: 0.5, 0.25, 0μg.mL-1 and eCG: 15, 5, 0IU.mL-1. Oocytes were stained with 2% lacmoid, and examined for their nuclear status. The parameter for comparisons between groups was the percentage of oocytes in MII. There was an interaction between FSH and eCG (P<0.05). A higher percentage of oocytes in MII were obtained when using 0.5µg.mL-1 of FSH with any concentration of 15, 5 or 0IU.mL-1 of eCG [58.4±1.94% (n=78); 59.5±1.94% (n=85); 54.3±0.56% (n=81); P<0.05]. A similar percentage was also found using 0.25µg.mL-1 of FSH plus 15IU.mL-1 of eCG [52.3±1.66% (n=86); P>0.05] followed by 0.25µg.mL-1 of FSH with 5 or 0IU.mL-1 of eCG [47.3±1.94% (n=82); 45.3±1.94% (n=86); P<0.05]. The lowest maturation percentages were found using any concentration of eCG without FSH [8.2±0.73% (n=84); 8.0±0.66% (n=74); 11.4±1.23% (n=71); P<0.05]. In conclusion, the addition of eCG to the maturation medium would reduce the amount of FSH required in the in vitro maturation of alpaca oocytes.

114 Short-term hormonal protocol efficiency either with or without equine chorionic gonadotrophin to promote oestrous synchronization in cyclic dairy goats

The use of equine chorionic gonadotrophin (eCG) in oestrous induction protocols is associated with antibody production against eCG. Moreover, there are important equine welfare issues involved in its production and the development of viable protocols without eCG in dairy goats is essential. The aim of this study was to assess oestrus and ovulatory responses of short-term oestrous synchronization protocol with or without eCG in nulliparous (NUL) and multiparous (MUL) dairy goats during the breeding season. On a random day of oestrous cycle (Day 0), Alpine (n=10 NUL and 10 MUL), Nubian (n=10 NUL and 10 MUL), and Saanen (n=10 NUL and 6 MUL) goats received 30μg of d-cloprostenol intramuscularly (IM) and an intravaginal sponge containing 60mg of medroxyprogesterone acetate, which was maintained for 6 days (Day 6). At 24h before sponge removal (Day 5), goats received IM 200IU of Novormon 5000 (G-eCG) or saline solution (G-Control). NUL and MUL goats of each breed were assigned equally into one of the two treatments. Transrectal ultrasonography was used to assess ovarian parameters, and teaser goats detected oestrus every 12h from sponge removal to ovulation or until 96h after sponge removal. The results are described as mean±s.e.m., and categorical results are presented as percentages. The normal distribution of the residues of all variables was determined by the Shapiro-Wilk test and for homoscedastic by Levene test. To evaluate interaction between treatments and parity order, the general linear model (GLM) was applied. Parametric variables were subjected to Student’s t-test. Nonparametric variables were analysed by Mann–Whitney test, Chi-squared test, and Fisher’s exact test. Values of P<0.05 were considered significant. According to the breed and parity order, Nubian and Saanen goats showed no difference (P>0.05) in reproductive parameters. Alpine goats had a longer (P<0.05) interval from sponge removal to onset of oestrus (SROE) in MUL (61±14.0h) than NUL (38.5±12.7h), longer interval from onset of oestrus to ovulation (IEOV) in NUL (33.5±11.9h) than MUL (19.5±13.4h), and the number of ovulations was greater (P<0.05) in MUL (2.2±0.9) than in NUL (1.3±0.5). In goats either receiving eCG or not, Alpine and Nubian goats had no difference (P>0.05) in SROE, IEOV, interval from sponge removal to ovulation (SROV), percentage of animals that ovulated, diameter of the ovulatory follicle, and number of ovulations. Saanen goats had a higher (P<0.05) percentage of animals that ovulated in G-eCG (87.5%) than G-control (25.0%). Regardless of parity order and breed, SROV was longer (P<0.05) in G-control (73.2±9.8h) than in G-eCG (65.1±10.2h), and the percentage of animals that ovulated was greater (P<0.05) in G-eCG (96.4%) than in G-control (67.9%). In summary, in short-term protocols for oestrous synchronization in cyclic dairy goats, the addition of eCG led to a shorter interval from sponge removal to ovulation and increased number of animals that ovulated. This research was supported by CNPq (310166/2012-8) and Embrapa (20.19.01.004.00.03.001).

Screening of additives to reduce grain damage risk on unhairing by proteinase K

Enzymatic unhairing is a cleaner strategy for leather-making. It is a potential alternative to the traditional hair-burning process. However, several shortcomings, such as uncontrolled enzymatic reaction, and risk of grain looseness and damage have restricted the broad application of enzymatic unhairing. In this work, metal ions and organic additives were screened for lessening the hydrolytic activity of proteinase K to collagen fiber. Then, the selected additives were applied to the enzymatic unhairing process for bovine hide. The results showed that a suitable concentration of metal ions (Cu (II), Fe (III) and Al (III)) and organic additives (salicylate, laurate, adipate, gallate and epicatechin (ECG)) could diminish approximately 35% of the hydrolytic activity of proteinase K to collagen fibers. Then, the additives were applied for the bovine hide enzymatic unhairing process. Hydroxyproline determination in the unhairing float shows that applying additives could reduce collagen hydrolysis. The morphology results showed that the grain damage could be significantly reduced with the addition of the screened additives in the proteinase K enzymatic unhairing system, whereas the addition of ECG and gallate significantly slowed down the unhairing speed. This outcome provides new potential to reduce the risk of grain damage in enzymatic unhairing process.Graphical abstract

Removal of ECG artifacts from EMG signals with different artifact magnitudes by template subtraction

AbstractThis work investigates the performance of eventsynchronous noise cancelling algorithms to separate ECG artifacts from thoracic EMG recordings. In focus are the precise detection of heart beats, the exact time alignment of the QRS segments and the construction of a template QRS signal. Among the utilized methods is a modified structural intensity (SI) approach based on the observation of extrema in low pass filtered versions of the second derivative as a detector. An adoption of this approach is also used to obtain the exact time alignment of QRS segments. Artificial test signals are constructed based on addition of ECG and sEMG with different magnitudes. We show that approaches based on the observation of extrema in different scales yield superior results.

Equine chorionic gonadotropin (eCG) enhances reproductive responses in CIDR-EB treated lactating anovular Nili-Ravi buffalo during the breeding season

The objective of this study was to evaluate the effect of equine chorionic gonadotropin (eCG) on the estrous response, ovulatory follicle development, ovulation rate, corpus luteum (CL) diameter, concentrations of progesterone (P4) and pregnancy per AI (P/AI) in CIDR-EB treated lactating anovular Nili-Ravi buffalo during the breeding season. Buffalo (n = 87), 60 to 200 Days postpartum were treated with CIDR at a random stage of the estrous cycle (Day 0) and assigned to two groups; with eCG (+eCG; n = 44), or without eCG (-eCG; n = 43). In both groups, each animal received 2 and 1 mg estradiol benzoate (EB) on Day 0 and then on Day 8, respectively. On Day 6, eCG and PGF2α were administered. On Day 7, CIDR were removed and FTAI was performed at 48 and 60 h later and pregnancy was diagnosed 35 Days post AI. The results revealed that eCG treatment increased (P < 0.05) the CL diameter on 15, 18, and 21 after the ovulation that resulted from imposing the estrous synchronization protocol than when eCG was not administered. Similarly, treatment with eCG resulted in an increased (P < 0.05) mean concentration of P4 as compared with animals not treated with eCG on Days 18 and 21 after the ovulation that occurred subsequent to imposing the estrous synchronization protocol. The ovulation rate was greater in eCG-treated buffalo as compared with those not treated with eCG (93% compared with 74%, respectively; P < 0.05). The P/AI was greater in buffalo treated with eCG than those not treated with eCG (56% compared with 31%, respectively; P < 0.05). In conclusion, the addition of eCG along with EB and P4 based treatments enhanced the estrous response, follicular and luteal function, ovulation rate, concentrations of P4, and P/AI in lactating anovular Nili-Ravi buffalo.

AMSSM Position Statement on Cardiovascular Preparticipation Screening in Athletes: Current evidence, knowledge gaps, recommendations and future directions

Cardiovascular screening in young athletes is widely recommended and routinely performed prior to participation in competitive sports. While there is general agreement that early detection of cardiac conditions at risk...

The utility of presentation and 4-hour high sensitivity troponin I to rule-out acute myocardial infarction in the emergency department

ObjectivesInternational guidance recommends that early serial sampling of high sensitivity troponin be used to accurately identify acute myocardial infarction (AMI) in chest pain patients. The background evidence for this approach is limited. We evaluated whether on presentation and 4-hour high-sensitivity troponin I (hs-cTnI) could be used to accurately rule-out AMI. Design and methodshs-cTnI was measured on presentation and at 4-hours in adult patients attending an emergency department with possible acute coronary syndrome. We determined the sensitivity for AMI for at least one hs-cTnI above the 99th percentile for a healthy population or alone or in combination with new ischemic ECG changes. Both overall and sex-specific 99th percentiles were assessed. Patients with negative tests were designated low-risk. Results63 (17.1%) of 368 patients had AMI. The median (interquartile range) time from symptom onset to first blood sampling was 4.8h (2.8–8.6). The sensitivity of the presentation and 4h hs-cTnI using the overall 99th percentile was 92.1% (95% CI 82.4% to 97.4%) and negative predictive value 95.4% (92.3% to 97.4%) with 78.3% low-risk. Applying the sex-specific 99th percentile did not change the sensitivity. The addition of ECG did not change the sensitivity. ConclusionHs-cTnI >99th percentile thresholds measured on presentation and at 4-hours was not a safe strategy to rule-out AMI in this clinical setting irrespective of whether sex-specific 99th percentiles were used, or whether hs-cTnI was combined with ECG results.

170 INFLUENCE OF OOCYTE QUALITY AND DIFFERENT MEDIA SUPPLEMENT ON IN VITRO MATURATION, CLEAVAGE, AND EMBRYO DEVELOPMENT OF BUFFALO (BUBALUS BUBALIS) OOCYTES

A series of 4 factorial-arranged experiments were conducted to study the effect of oocyte quality and different in vitro maturation (IVM) media supplements on IVM, cleavage, and embryo development of buffalo (Bubalus bubalis) oocytes. Buffalo ovaries were collected at a local abattoir in a warm (32–35°C) saline (0.9% NaCl), and oocytes were aspirated using an 18-gauge needle. In experiment 1, oocytes (n = 320) were classified according to the number of cumulus cell layers and morphology of ooplasm as excellent, good, or fair. Oocytes were cultured for IVM, fertilization, and embryo culture (IVMFC) in TCM-199 + 10% FCS. In experiment 2, excellent quality oocytes (n = 237) were subjected to IVM in TCM-199 enriched with either 10% FCS or oestrous buffalo serum (EBS; 20–40 pg mL–1) and then fertilized using frozen–thawed buffalo semen capacitated in Bracket and Oliphant's (BO) medium containing heparin (0.02 mg mL–1) and sodium caffeine benzoate (3.89 mg mL–1). In experiment 3, oocytes (n = 290) were classified into 2 groups; Group 1, without gonadotropins, served as a control; Group 2, in which IVM medium was supplemented with 20 IU mL–1 equine chorionic gonadotropins (eCG). Experiment 4 was carried out to examine the suitable capacitating agent added to BO medium, either heparin or caffeine or both (n = 210 fertilized oocytes). In all experiments (multiple replicates), oocytes (2–6 mm in diameter) were kept at 39°C under 5% CO2 for IVMFC and examined several times for cleavage and embryo development (morula and blastocyst). Statistical analysis was carried out using Chi-squared test. Excellent and good quality oocytes produced a higher (P &lt; 0.05) maturation, cleavage, and morula development rates than poor quality oocytes (70% and 65% v. 33.3%), (50% and 46.2% v. 25%), and (42.9% and 33.3% v. 10%), respectively. Blastocyst production rate was also higher (P &lt; 0.05) for excellent compared with good quality oocytes (28.6% v. 16.7%, respectively). In experiment 2, the IVM and cleavage rates were significantly higher (P &lt; 0.05) in IVM medium plus 10% EBS than those cultured in 10% FCS (73% v. 45% and 50% v. 33.3%, respectively). In experiment 3, the addition of eCG to maturation medium increased (P &lt; 0.05) developmental competence of buffalo oocytes (IVMFC) compared with control medium (16% v. 4%). In experiment 4, the addition of heparin together with caffeine to BO medium produced significantly higher (P &lt; 0.05) cleavage and embryo developmental rates compared with heparin or caffeine alone (56.3% v. 33.3% and 35.7%, respectively; 22.2% v. 10% and 8%, respectively). In conclusion, excellent quality oocytes cultured in IVM medium supplemented with either EBS or eCG and fertilized with capacitated buffalo spermatozoa in BO medium enriched with heparin and caffeine progressively enhanced developmental competence of buffalo oocytes.

Multimodal information improves the rapid detection of mental fatigue

One of the major challenge in the detection of mental states is improving the accuracy of brain activity-based detectors with additional information from extracranial signals. We assessed the suitability, for real-time mental fatigue detection, of EEG, EOG and ECG measurements, taken separately or together. Thirteen subjects performed six blocks of switching tasks. For each participant, the block with the lowest error rate from the first two blocks and the block with the highest error rate from the last three blocks were discriminated with a machine learning algorithm (support vector machine). The classification scores obtained with ECG or EOG were greater than would be expected by chance (>50%) for time windows of at least 8s. EEG was the best single mode of detection, with classification scores ranging from 80±3% with a 4s time window to 94±2% with a 30s time window. The addition of ECG and EOG features to EEG features significantly increased classification scores for short time windows (e.g., to 86±3% with a 4s time window, p<0.001). For short time windows (up to 12s), ECG significantly increased the discriminatory power of EEG, whereas EOG did not. These results demonstrate that mental state detection on the basis of extracerebral measurements is feasible and that a combination of EEG and ECG is particularly appropriate for the rapid detection of mental fatigue.

310 DOES PROGESTERONE OR EQUINE CHORIONIC GONADOTROPIN IMPROVE THE SUPEROVULATORY RESPONSE AND EMBRYO QUALITY IN WOOD BISON DURING THE ANOVULATORY SEASON?

Superovulation protocols are being developed in wood bison (Bison bison athabascae), a threatened Canadian species. In initial studies, 2 doses of FSH diluted in hyaluronan given 48 h apart were successful for inducing ovarian superstimulation in wood bison, and ovulation rate was improved by final treatment with hCG instead of LH (Palomino et al. 2012 Reprod. Fertil. Dev. 24, 226). In that study, exogenous progesterone had no effect on the number of ovulations, but embryo quality could not be evaluated because of the low number of embryos collected. In beef cattle, replacement of the final doses of FSH with eCG has resulted in the recovery of a greater number of ova/embryos. The objective of the present study was to evaluate the effect of exogenous progesterone (PRID) on embryo quality and to determine if the addition of eCG increases the ovulation rate in superstimulated wood bison. Follicular ablation was done to synchronize follicular wave emergence in 26 wood bison cows during the anovulatory season (May). Cows were assigned randomly to 3 groups: PRID/no eCG (n = 8), PRID/eCG (n = 9), and no PRID/eCG (n = 9). A PRID was inserted on the day of follicular ablation (Day –1) in respective groups. In all bison, FSH diluted in hyaluronan (5 mg mL–1, MAP-5, Bioniche Animal Health Inc., Belleville, Ontario, Canada) was given intramuscularly on Day 0 (300 mg) and Day 2 (100 mg). On Day 3, a single dose of 450 IU of eCG (Pregnecol, Bioniche Animal Health Inc.) was given intramuscularly and the PRID were removed in the corresponding groups. On Day 5, all bison were given 2500 IU of hCG (Chorulon, Merck, Whitehouse Station, NJ, USA) intramuscularly to induce ovulation. Bison were inseminated with chilled semen 12 and 24 h later. Nonsurgical embryo collection was performed on Day 14. Transrectal ultrasonography was done to record the ovarian response, ovulation rate, and number of corpora lutea (CL). Results were compared by ANOVA and Chi-squared test (Table 1). The number of ovulatory-sized follicles (≥9 mm) on Day 5 did not differ among groups. Ovulation rate was lowest in bison treated with both a PRID and eCG (P &lt; 0.05). There were no differences among groups in the number of CL on Day 14, the number of ova/embryos collected, or the number of transferable embryos. The superovulatory response and embryo collection rate in the present study were higher than in any previous reports in bison. The ovulation rate was not improved by the addition of eCG treatment, and exogenous progesterone had no effect on embryo quality. Table 1.Response of superstimulated wood bison (mean ± SEM) to treatment with exogenous progesterone (PRID) and/or eCG

Progesterone-based strategies to induce ovulation in prepubertal Nellore heifers

Four experiments were conducted to evaluate hormonal strategies to induce ovulation in Nellore heifers. In experiment 1, heifers (N = 1039) received a controlled internal drug release (CIDR) of fourth use (CIDR-4) on Day −12 or no CIDR (CIDR-0). The CIDR was removed on Day 0 in the CIDR-4 treatment, and estrus detection and AI were performed from Days 1 to 7. On Day 8, heifers not detected in estrus were evaluated for CL presence and received the same treatment again, followed by estrus detection and AI from Days 21 to 27. All heifers in experiments 2 (N = 896), 3 (N = 839), and 4 (N = 948) received the CIDR-4 treatment on Day −12. In experiment 2, heifers were randomly assigned to a control group (no additional treatment) or to receive equine chorionic gonadotropin (eCG; 200 IU eCG im) on Day 0. In experiment 3, heifers received the same treatments as in experiment 2, or a treatment that included eCG and estradiol cypionate (ECP) (eCG+ECP; 200 IU im eCG plus 0.5 mg ECP im) on Day 0. In experiment 4, heifers received the treatments described in experiment 3 or only ECP (0.5 mg) on Day 0. In experiments 2 and 3, estrus detection and AI was performed from Days 1 to 7 and on Day 8, heifers not detected in estrus were evaluated for CL presence. In experiment 4, heifers were evaluated for presence of a CL between Days 10 and 14. In experiment 1 heifers treated with CIDR-4 had greater estrus detection, ovulation induction, and pregnancy rates than in the CIDR-0 group. In experiment 2, heifers treated with eCG had greater estrus detection, ovulation induction, and pregnancy rates in 7 days than heifers in the control group. In experiment 3, heifers treated with eCG+ECP had greater estrus detection, ovulation induction, and pregnancy rates than the control and eCG treatments. In experiment 4, ovulation induction was greater for heifers treated with eCG and eCG+ECP relative to control, but did not differ from the ECP treatment. In conclusion, the use of a CIDR of fourth use for 12 days and the addition of eCG and/or ECP at CIDR removal efficiently induced ovulation and increased pregnancy rates in prepubertal Nellore heifers.

225 ADDITION OF EQUINE CHORIONIC GONADOTROPIN TO A TRADITIONAL FOLLICLE STIMULATING HORMONE PROTOCOL FOR SUPEROVULATION OF BOS TAURUS BEEF COWS

This study examined the superovulatory responses of Bos taurus beef cows maintained in a commercial embryo transfer facility. Donors were superovulated 1 to 3 times each with either a traditional 8 injection FSH protocol (controls, n = 126) or 6 injections of FSH with the seventh or eighth FSH treatments replaced by 2 injections of eCG (treatment, n = 134). During the 5-month study, 132 donors were alternatively assigned to a control or treatment group for a single superovulation and an additional 62 animals were superovulated 2 (n = 58) or 3 times (n = 4) in a crossover design. Although 14 beef breeds were represented in the study, 87% of the cows were Angus, Red Angus, Polled Hereford, or Charolais. All donors were synchronized on Day 0 with a CIDR, 5 mg of oestradiol-17β and 100 mg of progesterone. Starting on Day 4 (p.m.), controls were injected twice daily for 4 days with descending doses of porcine FSH (Folltropin-V®, Bioniche Animal Health, Belleville, Ontario, Canada). Cows received 750 μg of cloprostenol (Estrumate®, Intervet Schering-Plough, Summit, NJ, USA) at the seventh FSH injection and the CIDR was removed at the eighth FSH injection. Based on previous experience with specific, individual animals, total FSH dose per donor ranged from 240 to 400 mg. However, 74% of treatments involved 380 mg for controls and 310 mg for treated donors that received eCG. In addition, donors that were superovulated more than once received the same FSH dose in the crossover, treatment-control design. In the treatment group, 200 IU of eCG (Pregnecol™ 6000, Bioniche Animal Health) was substituted for the seventh and eighth FSH injections. Inseminations were conducted on a timed AI basis, with one unit of semen 32 h and a second 48 h following CIDR removal. Results were analysed by ANOVA as shown in Table 1. Although more ova/embryos and unfertilized ova (UFO) were recovered in the control group, the control and treatment groups did not differ in the number of grade 1, 2, or 3 embryos or in the number of degenerate embryos. Previous superovulation studies in South America using eCG to replace the last 2 injections of FSH resulted in more total ova/embryos in Nelore cows but not heifers and in more embryos in Brangus and Sindhi cows. The mean embryo production for the control cows in this study was high and the addition of eCG in the protocol did not improve embryo production. Failure of eCG to increase the number of embryos for Bos taurus cows in this study compared with previous studies may be due to differences with Bos taurus versus Bos indicus breeds or differences in management factors between Canada and South America. The high response rate in the controls may also have contributed to the failure of any advantage of adding eCG to the treatment protocol. Table 1.Mean numbers (± SEM) of ova and embryos recovered from Bos taurus females superovulated with 2 different protocols

43 COSTS RELATED TO ESTRUS SYNCHRONIZATION AND AI PROTOCOLS IN CROSSBRED HOLSTEIN-ZEBU DAIRY COWS

The objective of this paper was to compare 3 bovine AI protocols, evaluating the cost of each treatment and the pregnancy rates obtained, to demonstrate the importance of routine utilization of estrus synchronization techniques. The experiment was conducted in the Farm School of Cachoeiras de Macacu of the Federal Fluminense University, in the state of Rio de Janeiro, during the period from 2006 to 2007 (August to June). One hundred sixteen crossbred female Holstein-Zebu dairy cows in cycle, with body condition between 2.5 to 4 (on a scale from 1 to 5), were selected and divided randomly in 3 groups (G). Animals in G1 [n = 60; 21 heifers (H), 13 primiparous (P), and 26 multiparous (M)], with palpable corpus luteum, received 25 mg of prostaglandin F2 intravulvar, and AI was done 12 h after estrus detection. For animals in G2 (n = 30; 11 H, 6 P, and 13 M) an intravaginal progesterone device (CIDR®, Pfizer Animal Health, New York, NY, USA) was inserted and 2 mg of estradiol benzoate (EB) was administered i.m. on Day 0. On Day 8 the device was removed and 25 mg of prostaglandin was administered i.m. on Day 9, 2 mg of EB was administered, and 30 h later, timed AI of all animals was performed. For G3 (n = 26; 9 H, 5 P, and 12 M), the protocol used was similar to that for G2, except that on Day 8 400 IU of eCG was administered (in addition to the removal of the device) and on Day 9 the dose of EB was reduced to 1 mg and timed AI was done 27 h later. AI was done with semen from 2 sires with proven fertility, by the same technician. The pregnancy diagnoses were done on Day 45 by palpation per rectum and transrectal ultrasonography. The cost-benefit relation was calculated according to the cost of pharmaceuticals multiplied by the number of animals in each experimental group divided by the number of pregnancies obtained in each one, without considering the manual labor costs in all the groups and considering 3 uses of the CIDR. The statistical analyses were done using chi-square test. The pregnancy rate obtained in G2 (60%, 18/30) was superior to that in G1 (30%, 18/60) (P &lt; 0.05), demonstrating the efficiency of the treatment that utilized progesterone. G3 (53.85%, 14/26) was similar (P &gt; 0.05) to G2, despite the fact that the eCG was added in this group. In reference to cost-benefit, considering only the pharmaceuticals costs and the value for pregnancy obtained in each group, the costs found, respectively, were G1 U$7.02 and U$23.66; G2 U$28.57 and U$47.59; and G3 U$47.96 and U$89.05. The lowest cost in the estrus synchronization was based on the use of prostaglandins alone; the use of progesterone increased the synchronization cost, but also increased the pregnancy rate and eliminated the need for estrus observation. The addition of eCG in cows that are cycling does not necessarily demand a increase in pregnancy rate. Failure in estrus detection, using prostaglandin alone, increased the interval from calving, causing a daily loss in milk production, which justified the investment in protocols of estrus and ovulation synchronization, contributing to increase the reproductive efficiency of a dairy cow herd.

Taxa de prenhez de vacas Nelore lactantes tratadas com progesterona associada à remoção temporária de bezerros ou aplicação de gonadotrofina coriônica eqüina

Avaliaram-se os efeitos da remoção temporária de bezerros (RTB) ou da aplicação de gonadotrofina coriônica eqüina (eCG) na taxa de prenhez (TP) de vacas Nelore lactantes tratadas com um dispositivo intravaginal liberador de progesterona (DILP). No experimento 1, 83 vacas Nelore e 102 mestiças (Nelore × Red Angus) foram distribuídas em três grupos. Em dia aleatório do ciclo estral (D0), os animais do grupo 1 foram tratados com benzoato de estradiol (BE; 2,5 mg, IM, Estrogin®) e um DILP (1 g de progesterona, DIB®), removido no D9, quando também se administrou d-cloprostenol (150 µg, IM, Prolise®) e aproximadamente 12 horas após identificação do estro realizou-se a inseminação artificial. No grupo 2 (IATF), o tratamento foi semelhante ao aplicado no grupo 1, porém, administrou-se uma segunda dose de benzoato de estradiol (1 mg) no D10 e 30-36 horas mais tarde realizou-se a inseminação artificial em tempo fixo (IATF). No grupo 3 (IATF/RTB), os bezerros foram removidos a partir do D9 até a inseminação artificial com tempo fixo (IATF). As taxas de prenhez para as vacas Nelore e mestiças foram, respectivamente, de 7,69 e 41% (Controle); 23,52 e 59,57% (IATF); 69,44 e 55,81% (IATF/RTB). No experimento 2, 255 vacas Nelore lactantes foram distribuídas em três grupos. Os animais do grupo 4 (IATF) foram tratados com benzoato de estradiol (2 mg) e um DILP (1,9 g de progesterona, CIDR-B®) no D0 e no D8 sofreram remoção do DILP no D8 e administração de 25 mg de dinoprost (IM, Lutalyse®). No D9, foi aplicado benzoato de estradiol (1 mg) realizando-se a IATF 30-36 horas mais tarde. No grupo 5 (IATF/RTB), os bezerros foram removidos no D9 até a IATF. O grupo 6 (eCG) foi semelhante ao IATF, exceto pela aplicação de eCG no D9 (400 UI, IM, Novormon®). As taxas de prenhez foram de 50,57 (IATF), 53,57 (IATF/RTB) e 54,76% (eCG). A associação de RTB ao tratamento hormonal com DILP aumenta as taxas de prenhez, enquanto a adição de eCG ao tratamento não melhora as taxa de prenhez de vacas Nelore lactantes ciclando e em boa condição corporal.

22 PREGNANCY RATES IN DAIRY COWS TREATED WITH PROGESTERONE-RELEASING DEVICES AND DIFFERENT GONADOTROPIN-RELEASING HORMONE OR ESTRADIOL PLUS EQUINE CHORIONIC GONADOTROPIN FIXED-TIME ARTIFICIAL INSEMINATION PROTOCOLS

We have previously shown that the addition of eCG improved pregnancy rates following fixed-time artificial insemination (FTAI) in lactating dairy cows treated with estradiol benzoate (EB) and progesterone releasing devices (Veneranda et al. 2006 Reprod. Fertil. Dev. 18, 118). An experiment was designed to compare pregnancy rates in lactating dairy cows treated with progesterone-releasing devices and EB plus eCG with different gonadotropin-releasing hormone (GnRH)-based protocols. Five-hundred lactating dairy cows between 30 and 51 days postpartum (when prostaglandin F (PGF) treatments were given in the Pre-Synch groups), with a milk yield of 29.5 � 7.0 kg per day (range 13.0 to 46.0 kg) and a body condition score (BCS) between 2.5 to 3.5 out of 5 were used. Cows were blocked by days postpartum and randomly assigned to five treatment groups. Cows in the Pre-Synch group received PGF 28 and 14 days prior to the insertion of a DIB device (1 g progesterone, Syntex SA, Buenos Aires, Argentina) and the administration of 50 µg of Lecirelin (GnRH, Ovusin, Syntex SA) IM (Day 0). On Day 7, DIB were removed and cows received PGF (0.5 mg cloprostenol, Ciclase, Syntex SA). On Day 9, cows received a second GnRH treatment and were FTAI 16 h later (60 h after device removal). Cows in the Modified Pre-Synch group were treated similarly except that they received a DIB for 7 days prior to the second PGF injection and no DIB at the time of the first GnRH. Cows in the P4-Synch group were treated as those in the Pre-synch group but did not receive the two doses of PGF 28 and 14 days earlier. Cows in the P4 + EB + eCG groups received a DIB for 8 days, 2 mg of EB at DIB insertion (Day 0), and PGF and 400 IU eCG (Novormon, Syntex SA) IM at DIB removal (Day 8); they were subdivided to receive 1 mg of EB on Day 9 (P4 + EB + eCG + EB group) or GnRH on Day 10 (P4 + EB + eCG + GnRH group). Cows in these latter two groups were also FTAI 60 h after device removal. Pregnancy was determined by rectal palpation 50 days after FTAI and data were analyzed by logistic regression. Pregnancy rates were not affected by BCS, days postpartum, or milk yield (P &gt; 0.5). However, there was a significant group effect due to a higher (P &lt; 0.05) pregnancy rate in the P4 + EB + eCG + EB (49%) and P4 + EB + eCG + GnRH (44%) treatment groups than in those in the Modified Pre-Synch group (30%). Pregnancy rate in the P4-Synch group (39%) was lower (P &lt; 0.05) than those in the P4 + EB + eCG + EB group but not different from the others. Finally, pregnancy rate in Pre-Synch group (41%) was intermediate and not different from the other treatment groups. Results of these experiments suggest that treatments with progesterone-releasing devices, EB, and eCG result in pregnancy rates that are comparable to or better than those obtained in GnRH-based protocols in lactating dairy cows. Further studies are needed to investigate whether eCG could increase pregnancy rates in cows treated with GnRH and progesterone-releasing devices. The authors thank Syntex SA for providing the hormones used in the study.

19 PREGNANCY RATES IN DAIRY COWS TREATED WITH INTRAVAGINAL PROGESTERONE DEVICES AND DIFFERENT FIXED-TIME AI PROTOCOLS

Two experiments were designed to evaluate the effect of different treatments for the synchronization of ovulation on pregnancy rates following fixed-time AI (FTAI) of lactating dairy cows. In Experiment 1, 394 Holstein cows that were 61.7 � 13.6 days postpartum (range 35 to 94 days), with a milk yield of 30.7 � 6.8 liters per day (range 12.0 to 52.4 liters) and a body condition score (BCS) between 2.5 to 3.5 out of 5, were used. Cows were blocked by days postpartum and milk yield and randomly assigned to one of four treatment groups. Cows in P4+EB treatment groups received an intravaginal DIB device (1 g P4; Syntex, Argentina) and 2 mg of estradiol benzoate (EB; Syntex) i.m. on Day 0. On Day 8, DIB devices were removed, and cows received PGF (150 �g D+cloprostenol: Ciclase; Syntex), and were subdivided to receive 400 IU eCG (Novormon 5000, Syntex) i.m. or no further treatment at that time. On Day 9, all cows received 1 mg of EB and were FTAI 60 h after DIB removal. Cows in P4-Synch groups received a DIB device and 50 �g of GnRH (Lecirelina, Gonasyn; Syntex) i.m. on Day 0. On Day 7, DIB devices were removed; cows received PGF and were divided to receive 400 IU eCG i.m. or no further treatment. On Day 9, all cows received a second GnRH treatment and were FTAI 60 h after DIB removal. Blood samples were taken on Days -10 and 0 to determine plasma P4 concentrations; 93% of the cows had &gt;1 ng/mL P4 in at least one sample. In Experiment 2, 200 lactating cows from the same farm were treated with the P4+EB+eCG and P4-Synch without eCG treatments with either a DIB or a CIDR-B (1.9 g P4; Pfizer Animal Health, Groton, CT, USA) in a 2 � 2 factorial design. Cows were examined by rectal palpation 50 days after FTAI to determine pregnancy status, and data were analyzed by chi-square Mantel-Haenszel test. There was an EB/GnRH by eCG interaction (P &lt; 0.05) which was attributed to a higher pregnancy rate in the P4+EB+eCG group (44/98; 44.9%) than in the P4+EB without eCG group (30/100; 30.0%) and P4-Synch+eCG group (30/98; 30.6%); the P4-Synch without eCG group had an intermediate pregnancy rate that did not differ from the other treatment groups (37/98; 38.8%). In Experiment 2, no significant differences (P = 0.40) in pregnancy rates were detected between cows treated with DIB (51/100; 51.0%) or CIDR-B (42/100; 42.0%), and there was no difference (P = 0.18) between P4+EB+eCG (52/100; 52.0%) and P4-Synch without eCG (41/100; 41.0%) treated groups. Results suggest that the addition of eCG will improve pregnancy rates following FTAI in lactating dairy cows treated with EB but not in those treated with GnRH at the time of insertion and after removal of a P4 releasing device. Although treatment with P4+EB+eCG resulted in numerically higher pregnancy rates, results were not different from those obtained in the GnRH-based treatment, without the addition of eCG.

Protective Activity of (−)-Epicatechin 3- O -gallate against Peroxynitrite-mediated Renal Damage

The protective effect of ( m )-epicatechin 3- O -gallate (ECg) against peroxynitrite (ONOO m )-mediated damage was examined using an animal model and a cell culture system. In rats subjected to lipopolysaccharide (LPS) administration plus ischemia-reperfusion, the plasma 3-nitrotyrosine level, an indicator of ONOO m production in vivo, was elevated, whereas it declined significantly and dose-dependently after the oral administration of ECg at doses of 10 and 20 w moles/kg body weight/day for 20 days prior to the process. Moreover, oral administration of ECg significantly enhanced the activities of the antioxidant enzymes, superoxide dismutase, catalase and glutathione peroxidase, and the antioxidant glutathione, showing enhancement of the biological defense system against the damage induced by ONOO m . In addition, the significant increase in the renal mitochondrial thiobarbituric acid-reactive substance level of LPS and ischemic-reperfused control rats was attenuated in rats given ECg. Furthermore, the elevations in the plasma urea nitrogen and creatinine (Cr) levels and the urinary methylguanidine/Cr ratio induced by the procedure were attenuated markedly after oral administration of ECg, implying amelioration of renal impairment. The addition of ECg (25 or 125 w M) prior to 3-morpholinosydnonimine (SIN-1, 800 w M) exposure reduced ONOO m formation and increased the viability of cultured renal epithelial (LLC-PK 1 ) cells in a dose-dependent manner. In particular, ECg inhibited ONOO m -mediated apoptotic cell death, which was confirmed by decreases in the DNA fragmentation rate and the presence of apoptotic morphological changes, i.e. small nuclei and nuclear fragmentation. Furthermore, adding ECg before SIN-1 treatment regulated the cell cycle by enhancing G 2 /M phase arrest. This study provides evidence that ECg has protective activity against the renal damage induced by excessive ONOO m in cellular and in vivo systems.