Fixed-time Artificial Insemination Protocol Research Articles

Use of long-acting injectable progesterone in fixed-time artificial insemination protocols

Knowledge of the physiology of the estrous cycle in bovine females has been extremely important in monitoring and improving strategies in the use of drugs to control their estrous cycle and follicular development, providing better recruitment, selection, growth, and maturation until ovulation, with strategies to further optimize estrus and ovulation synchronization protocols. The objective of this work is to carry out a bibliographical analysis comparing the use of slow-release progesterone devices in relation to the use of long-acting injectable progesterone, taking into account their advantages, disadvantages and characteristics within a fixed-time artificial insemination program.

Combining Embryo Transfer and Artificial Insemination to Achieve Twinning in Beef Cattle, and Effects of Different Twin Calf-Raising Methods on Neonatal Behavior and Growth

As the beef industry moves towards efficient animal production to improve sustainability in agriculture, new production and management approaches are emerging. Among the many facets of the beef industry, cow–calf operations have the most opportunity for efficiency improvement, including improvements in fertility. This project accounts for measures and methods of (1) twinning reproductive technologies and (2) twin calf perinatal care and pre-weaning rearing. The overall objective was to produce twin calves using two reproductive technologies—embryo transfer and artificial insemination. The subobjectives were to determine accuracy of twin pregnancies embryo/fetal losses using ultrasonography, evaluate parturition and dystocia, and determine the effects of different twin-raising methods on neonatal behavior and growth. A fixed-time artificial insemination (FTAI) protocol was applied to 77 multiparous Angus-cross cows from a commercial beef herd in northcentral South Dakota during the summer of 2019. Cows were assigned to two different treatments groups: only artificially inseminated (AI) or received an embryo transfer following artificial insemination (ET + AI). They were estrous-synchronized, artificially inseminated (AI) with black Angus semen at day 0, and received and embryo transfer (ET) at day 7. Ultrasound examination detected 56% pregnancy risk for both groups, with sensitivity, specificity, and accuracy of 75%, 100%, and 90.5%, respectively, for bilateral twin detection. Calves were born during spring 2020. Twin calves (n = 34) and singleton calves (n = 11) were assigned to one of three raising methods: (1) twin born and twin raised (TT; n = 16), (2) twin born and single raised (TS; n = 18), and (3) single born and single raised (S; n = 11). Neonatal nursing behavior and birth weights were recorded, and adjusted day 200 and day 280 were calculated measures of vitality and growth. Blood samples were collected at age 24 h for colostrum intake measures (total serum protein, IgG1, and IgM). Twin calves were born 20% (p < 0.05) lighter in body weight than singletons; however, weights did not differ at day 280 between TT and S calves. TS calves had the shortest average latency to stand, but immunoglobulin concentrations did not differ among treatments. At weaning, cows that had birthed and raised twins produced more kilograms of live weight per pregnancy than cows birthing and raising singletons. Using ET + AI proved to increase twinning rate, and growth was maintained when raising both twins with their dam.

130 Effect of an appeasing substance application on growth, stress, handling quality and reproduction of Bos indicus beef heifers synchronized to fixed-time AI

Abstract This experiment evaluated the effects of an analogue of bovine appeasing substance (BAS) on growth, stress, handling quality, and reproduction of Nellore heifers. Twenty-nine heifers [body condition score (BCS) = 4.84 ± 0.13; body weight (BW) = 371 ± 0.77 kg; 23 ± 2 months of age)] were assigned to 1 of 2 treatments: 1) Saline solution (0.9 NaCl; n = 14) or BAS (Secure Cattle, IRSEA Group, Quartier Salignan, France; n = 15). Both solutions were applied topically in the nuchal skin area (5 mL/heifers) at the beginning (d -9) of the fixed-time artificial insemination protocol (FTAI). Heifers were allocated in two different pastures according to the treatment (one pasture per treatment) for 15 d (time of action of BAS) and then combined in a single group (both treatments in the same pasture). Heifers were inseminated on d 0, and BW and BCS were collected on d -9 and 40. The temperament in the squeeze chute (entry and exit scores; scale 1 – 3; chute score; scale 1 – 5; greater values mean more reactive heifers) and blood samples were collected on d -9, 0, 7, 14 and 30. The percentual of aggression, accident, and dirtiness in the perineal region were evaluated on d 0 along with time to inseminate, estrus expression scores (scale 1 – 3; 1 = no expression, 2 = low expression, 3 = greater expression), and dominant follicle size. The corpus luteum size was measured on d 7 and 14, and the pregnancy and embryo size on d 40. The BAS vs. Saline-treated heifers tended to have greater BW change (23.6 vs. 20.8 ± 1.03 kg; P = 0.09) and had greater BCS change (0.05 vs. -0.21 ± 0.05; P < 0.01). The BAS vs. Saline-treated heifers tended to have lesser entry scores (1.42 vs. 1.67 ± 0.09; P = 0.09) and percentual of dirtiness in the perineal region (48.3 vs. 71.1 ± 8.67 %; P = 0.06), and no effects of treatment were detected (P ≥ 0.39) for chute score, exit score, aggression, accident and time to inseminate. The BAS vs. Saline-treated heifers had greater estrus scores (2.00 vs. 1.27 ± 0.21; P = 0.03), and no effects were detected (P ≥ 0.11) for ovarian variables, pregnancy rate, and conceptus size. Furthermore, no treatment effects were detected (P ≥ 0.25) for serum concentration of cortisol and plasma concentration of haptoglobin, ceruloplasmin, and progesterone. Therefore, BAS application at the beginning of the FTAI protocol could decrease the temperament and dirtiness in the perineal region and increase the growth and estrus expression of beef heifers.

Human Chorionic Gonadotrophin (hCG) induces changes in IFN-pathway and Interferon-Stimulated Genes (ISGs) on the bovine endometrium at Day 18 of pregnancy.

We hypothesized that the hCG modulates the expression of IFNT-pathway and ISGs in bovine endometrium during early pregnancy. The aim of the current study is to evaluate the effect of hCG on IFNT-pathway signals and ISGs expression in endometrial cells. For this, 29 non-lactating cross-bread cows were used in the study and submitted to a 9-day fixed-time artificial insemination (FTAI) protocol. The day of the AI was considered Day 0 (D0), and five days (D5) after the FTAI, the cows were allocated into two groups: Control and hCG group, when a hCG group received a single dose of 2.500UI of hCG. On day 18 after FTAI (D18) cows were slaughtered and endometrial tissue samples were collected. There was no difference between the embryo recovery rate of the cows in C compared to the hCG. The hCG group increased the accessory corpus luteum formation rate. The hCG resulted in greater serum progesterone concentration in the hCG group compared to the C on Day 14. Only the expression of IFNAR2 and STAT1 were upregulated on pregnant cows of the hCG group compared to the C group. The pathway genes (JAK1, STAT2, and IRF9) were not regulated. The mRNA abundance of ISG15, MX1, MX2, and OAS1 was upregulated in pregnant cows for hCG group, compared to C group. The results show that the administration of hCG, 5 days after AI, in addition to increasing the serum progesterone, modulates the expression of IFNT-pathway and ISGs on bovine endometrium on Day 18 of pregnancy.

Characterizing pregnancy losses in Bos indicus beef females receiving a fixed-timed artificial insemination protocol

This experiment evaluated pregnancy losses from day 30 of gestation to calving in Bos indicus females, and evaluated if serum haptoglobin concentrations during early gestation impacts subsequent pregnancy losses. A total of 4926 Nelore females were used, being 1802 nulliparous heifers inseminated as yearlings (precocious heifers), 1356 nulliparous heifers inseminated at 24 months of age (conventional heifers), 887 suckling primiparous cows, and 881 suckling multiparous cows. Cows were assigned to an ovulation synchronization + fixed-time artificial insemination (FTAI) protocol from day −11 to 0 of the experiment. Pregnancy status was verified using transrectal ultrasonography on days 30 and 60 after FTAI, via transrectal palpation on day 150 after FTAI, and according to calf birth. Blood samples were collected from all animals diagnosed pregnant on day 30 after FTAI, and analyzed for serum concentrations of haptoglobin and pregnancy associated glycoproteins (PAG). Pregnancy loss was greater (P < 0.01) from day 60–150 of gestation (10.2 %) compared with day 30–60 of gestation (6.0 %) and with day 150 of gestation to calving (7.4 %), and differed (P = 0.04) between these latter two periods. Pregnancy loss from day 30–60 of gestation did not differ (P = 0.26) among parities, whereas total pregnancy losses (day 30 to calving) were greater (P < 0.01) in precocious (28.4 %) and conventional (27.1 %) heifers compared with primiparous (16.4 %) and multiparous (13.0 %) cows. Serum PAG concentrations on day 30 after FTAI were less (P ≤ 0.03) in cows that lost the pregnancy (5.63 ng/mL) from day 30–60 of gestation, as well as those that lost the pregnancy from day 30 to calving (8.59 ng/mL) compared with cohorts that maintained the pregnancy (9.39 and 9.32 ng/mL, respectively). No differences in serum PAG concentrations on day 30 after FTAI were noted (P ≥ 0.23) according to pregnancy losses from day 60 to calving. Serum haptoglobin concentration on day 30 after FTAI also did not differ (P ≥ 0.48) between cows that maintained or lost the pregnancy. This experiment provides novel information about pregnancy losses after day 30 of gestation in B. indicus cattle, with most losses occurring as fetal mortality and not affected by systemic inflammation during early gestation. Pregnancy losses were nearly doubled in precocious and conventional heifers compared with parous cows, demonstrating the relevance of this reproductive failure to B. indicus replacement heifers.

Uterine microbial profiles in healthy postpartum dairy cows do not vary with sampling techniques or phases of estrous cycle

In this study, we aimed to compare uterine microbial profiles in postpartum dairy cows, determined by bacteriological culture and next-generation sequencing, using three uterine sampling techniques (swab, cytobrush, and lavage) and induced phases of the estrous cycle (estrus and diestrus). Fifteen healthy postpartum dairy cows at 53 ± 5 days postpartum were enrolled in the study. Uterine samples were collected during a fixed-time artificial insemination protocol. Viable bacteria were aerobically cultured from part of each sample, and bacterial isolates were identified through Sanger sequencing of the 16S rRNA gene. Total genomic DNA was extracted from the remainder of undiluted samples to quantify bacterial load using 16S rRNA qPCR and characterize the microbiome by metagenomic sequencing of the V1–V3 region of the 16S rRNA gene. Microbial profiles and composition were analyzed using the Shannon-Weaver diversity index and principal component analysis, respectively. Out of 87 samples, 88 % (77/87) were culture positive. The proportion of culture-positive uterine samples did not differ between sampling techniques (P = 0.39) or estrous cycle phases (P = 0.99). However, swab, cytobrush, and lavage techniques yielded 1.5, 9 and 9 times greater bacterial loads (P < 0.01), respectively, during diestrus than estrus phase. Moreover, during diestrus phase, the cytobrush method yielded 3 and 6 times more bacteria (P < 0.01) than both the lavage and swab methods. The most abundant bacterial genera identified from both bacteriological culture and metagenomic sequencing were Bacillus and Enterococcus, regardless of sampling technique or phases of the estrous cycle. Bacterial genera in moderate to low abundance through metagenomic sequencing included Streptococcus, Oscillospiraceae, and Lachnospiraceae. Notably, the uterine microbial profiles and composition, determined by metagenomic sequencing, did not differ by sampling techniques (P = 0.55 and P = 0.60, respectively) or estrous cycle phases (P = 0.34 and P = 0.17, respectively). In conclusion, our results suggest that any of the sampling techniques can be reliably used to study the uterine microbiome of healthy cows at random phases of the estrous cycle. However, it is important to consider potential differences in bacterial yield as a confounding factor.

171 Supplementing Ca Salts of Soybean Oil Via Low-Moisture Molasses-Based Blocks to Improve Reproductive Performance and Overall Productivity of Beef Cows

Abstract This experiment evaluated reproductive and productive responses of beef cows receiving self-fed low-moisture blocks (LMB) enriched or not with Ca salts of soybean oil (CSSO) throughout the breeding season. Non-pregnant, suckling multiparous Angus-influenced cows (n = 554) were assigned to a fixed-time artificial insemination (AI) protocol (day -10 to 0) followed by natural service (day 15 to 70). Cows were managed in 12 groups (46 ± 4 cows/group) maintained in individual pastures, and groups received LMB enriched with 25% (as-fed basis) of CSSO or ground corn (CON) from day -10 to 100. Both treatments were designed to yield a daily LMB intake of 0.454 kg/cow (as-fed basis). Groups were enrolled in an estrus-synchronization + fixed-time AI protocol from day -10 to 0 (AI occurred on day 0) of the experiment and beginning 15 days after AI cows were exposed to mature Angus bulls for 55 d. Pregnancy status was verified on day 55 and 100 of the experiment, and cows that gave birth during the initial two weeks were considered pregnant to fixed-time AI. Calf birthdate and weight was recorded during the subsequent breeding season. All calves were weaned at the same time, and a calf BW and age were recorded. Blood samples were also collected from a subsample of cows on days -10, 0 (fixed-time AI), and 55 (pregnancy diagnosis). All data were analyzed using group as the experimental unit and Satterthwaite approximation to determine the denominator df for the tests of fixed effects. Quantitative data were analyzed with the MIXED procedure of SAS, and binary data analyzed with the GLIMMIX procedure of SAS with a binomial distribution and logit link function. Data were analyzed with group (treatment × location), cow(group), and location as random variables. Significance was set at P ≤ 0.05 and tendencies were determined if P &amp;gt; 0.05 and ≤ 0.10. Cows receiving CSSO had greater (P &amp;lt; 0.01) mean concentrations of ω-6 fatty acids in plasma samples collected on days 0 and 55. Cows receiving CSSO had greater (P = 0.05) pregnancy rate to fixed-time AI (67.2 vs. 59.3%), whereas final pregnancy rate did not differ (P = 0.92) between treatments. Pregnancy loss was less (P = 0.03) in CSSO cows (4.50 vs. 9.04%), which also calved earlier during the calving season (treatment × week; P = 0.04). Weaning rate tended to be greater (P = 0.09) in CSSO (84.8 vs. 79.4%), although calf weaning age and weight did not differ (P ≥ 0.72) between treatments. Kilos of calf weaned/cow exposed was greater (P = 0.04) in CSSO cows (234 vs. 215 kg). Therefore, supplementing CSSO to beef cows via LMB during the breeding season improved their reproductive and overall productivity during a cow-calf cycle.

Yeast estrogen screen assay applied in the assessment of estrogenic activity removal from dairy cattle wastewater treated by anaerobic digestion

The presence of estrogen-like endocrine disrupting chemicals (EEDC) in surface waters, soils, and groundwater has been frequently reported in the literature. Since these compounds can be excreted by humans and animals, the release of domestic sewage and animal wastewaters may be an important source of EEDC in the environment. In this context, there is a growing concern regarding the presence of these substances in the environment and their potential adverse effects on human health. However, most studies have been focused on the presence of EEDC in domestic wastewaters and the available information regarding EEDC in livestock wastewater is still limited. Therefore, this work aimed to quantify the estrogenic activity (EA) in the liquid phase of dairy cattle wastewaters and evaluate the removal of EA during anaerobic digestion, applying an adapted methodology based on the yeast estrogen screen (YES) assay, which has been widely used to analyze EA in domestic sewage. Influent and effluent dairy cattle wastewater samples from a full-scale anaerobic digester placed in an experimental farm were collected during seven months. EA concentrations in the liquid phase of raw wastewater ranged from 228 to 2182 ng.L−1 E2-eq. Anaerobic digestion was not efficient in EA removal, achieving effluent concentrations ranging from 109 to 946 ng.L−1 E2-eq. Results also indicate that the application of an estradiol-based fixed time artificial insemination protocol did not statistically contribute to the increase of EA concentrations in dairy cattle wastewater which reuse in cleaning of animal feeding operation facilities and fertigation are beneficial and economically advantageous to dairy production systems, reducing water demand in parallel to promoting organic matter and nutrient recycling. However, the potential environmental and health risks associated with the presence of EEDC in dairy cattle wastewaters should be better elucidated and evaluated.

PSVI-21 Effects of Calcium Salts of Soybean Oil Supplementation to Nellore Heifers During Second and Third Trimesters of Gestation and Offspring Sex on Milk Production and Performance of the Offspring

Abstract The study aimed to evaluate the effects of supplementing calcium salts of soybean oil to Nellore heifers during the second and third trimesters of gestation and offspring sex on cow productivity and their male and female offspring growth. Forty-eight pregnant Nellore heifers [body weight (BW) = 387 ± 38.4 kg] were assigned to this experiment at day 123 ± 13 of gestation. All heifers were conceived during the same fixed-time artificial insemination protocol at 14 and 24 months of age. Heifers were randomly used in a completely randomized block design with a 2 × 2 factorial treatment arrangement, consisting of supplementing or not (CON) calcium salts of soybean oil (CSSO; 3 g/kg of BW; Table 1) and the sex of the offspring (male or female), totaling four treatments and twelve reps per treatment. Throughout the experimental period, heifers were maintained in a pasture composed of Brachiaria brizantha cv.Marandu. After birth, all animals remained under the same pasture conditions and received the same supplementation until weaning. Data were analyzed using the MIXED procedure of SAS and heifer as the experimental unit. Treatment × offspring sex interaction was not detected (P &amp;gt; 0.36) for any of the variables evaluated. Calves born from heifers supplemented with CCSO were 2 kg heavier (P = 0.03) at birth and 5 kg heavier (P = 0.03) at 120 days of age compared with calves born from CON heifers. At weaning (210 days), no dietary treatment effects were observed (P = 0.17) in offspring BW. Male offspring were 3.5 kg, 9 kg, and 12 kg heavier (P ≤ 0.01) at birth, day 120, and at weaning, respectively, compared with female offspring (Table 2). Throughout the experiment, male offspring gained 0.040 kg more daily (P = 0.01) than female offspring. The female offspring had 0.1 thicker subcutaneous fat per 100 kg of BW (P = 0.02) than the male offspring. Heifer milk production was not impacted (P &amp;gt; 0.10) by the dietary treatments or sex of the offspring in any of the periods evaluated (days 30 and 210; Table 3). Hence, supplementing CSSO to Nellore heifers during the second and third trimester of gestation altered offspring BW up to 120 days after birth, suggesting programming effects on postnatal offspring growth resultant from the CSSO treatment. Male offspring were heavier from birth to weaning compared with female offspring. These outcomes indicate that supplementing CSSO to Nellore heifers during pregnancy might be a feasible alternative to optimize offspring productivity, and carcass merit in grazing systems.

PSIX-20 Differential Gene Expression in Peripheral White Blood Cells of Fertile and Sub-Fertile Beef Heifers

Abstract Infertility in beef heifers remains a major economic loss to the cow-calf production industry. Identifying beef heifers with superior genetic potential for improved fertility would increase profitability. Therefore, the study aimed to identify differences in transcriptome profile from peripheral white blood cells (pWBCs) of beef heifers with varying reproductive potential. To accomplish this, Angus-Simmental crossbred heifers were subjected to an estrus synchronization and fixed-time artificial insemination (AI) protocol (7-Day CO-Synch + CIDR). Fourteen days after AI, all heifers were exposed to fertile bulls for a 60-day natural breeding season. Depending on the presence or absence of conceptus 120 days post-AI, heifers were classified as fertile (pregnant heifers) or sub-fertile (non-pregnant by AI or bull-breeding). Pregnancies were terminated, and all animals in both groups were cycling when pWBCs were isolated. Total RNA was extracted from the PWBCs and subjected to the library preparation and sequencing on the Nova-Seq platform. The read counts were obtained after data filtering and quality control using FastQC v0.11.9 and MultiQC v1.12 and alignment to the Ensemble’s ARS UCD1.2 Bos taurus genome reference using STAR aligner v2.7.5 for fertile (n = 8) and sub-fertile (n = 5) beef heifers. The filtered data were subjected to differential expression analysis using DESeq2. We identified 300 significantly differentially expressed genes (DEGs) with P-value ≤ 0.05 and absolute (log2 fold change) ≥ 0.5. Among them, 34 genes including FKBP5, ADCY3, REPS1, LAMP1, CTSH, ADGRE5 were identified with padj &amp;lt; 0.05. DEGs were over-represented for pathways such as response to thyroid stimulus, immune response, response to lipid, neutrophil degranulation, and calcium ion binding. Some of these genes were reported as fertility-related, while the other genes are novel and have the potential to predict heifers with varying reproductive potential. A detailed understanding of the underlying biological mechanisms of the top genes and a follow-up study with a higher sample size at different time points could validate the potential candidates identified in this study.

180 Differences in Gene Expression in Granulosa Cells Harvested from Fertile and Sub-Fertile Beef Heifers

Abstract Infertility in beef replacement heifers is one of the leading contributors to economic losses in the cow-calf sector of the beef production industry. Successful reproduction is complex, involving multiple tissues and biological systems. This complexity makes identifying the leading causes of subfertility difficult. One potential source of fertility issues is poor oocyte health. As developing oocytes are dependent on granulosa cells (GCs) and physically coupled with GCs, we hypothesized that the gene expression profile would be different between the GCs from fertile heifers and those from sub-fertile heifers. The objective of this study was to identify differences in gene expression in GCs collected from beef heifers of differing fertility status, either fertile or sub-fertile. To gather this information, Angus-Simmental crossbred heifers underwent an estrus synchronization and fixed-time artificial insemination (AI) protocol (7-Day CO-synch + CIDR). At 41 days post-AI, heifers were pregnancy checked through ultrasound and palpation. Heifers pregnant to AI were classified as fertile (n = 8). Any heifers that remained non-pregnant following AI were exposed to bulls of proven fertility for 61 days. Bull-exposed heifers were evaluated for pregnancy 34 days after the bulls were removed. The non-pregnant heifers following both AI and natural service were classified as sub-fertile (n = 5). Pregnancies were terminated in fertile heifers, and heifers from both groups were allowed to resume normal cyclicity before GCs being harvested. RNA was extracted from GCs and quality was checked for each sample. Total RNA from GCs of 13 heifers was subjected to the library preparation and sequencing on the Nova-Seq platform. Through the application of DESeq2, 79 genes were identified as significantly differentially expressed genes (DEGs) with P-value &amp;lt; 0.05 and absolute (log2 fold change) &amp;gt; 0.5 including DNER, HOXD10, CD52, CXCL12, and HOXD11. DEGs, along with co-expressed genes, were over-represented in pathways associated with the regulation of MAP kinase activity, regulation of serine/threonine kinase activity, MHC protein binding, and plasma membrane signaling receptor complex. Through the pursuit of knowledge regarding the function of top DEGs, increased sample size, and more diverse sample collection time points, we may improve our understanding of the causation of heifer fertility issues. Improving our knowledge of heifer infertility has the potential to develop therapeutic options, and in turn, increase the efficiency and sustainability of cow-calf production systems.

Upgrading the fixed-time artificial insemination (FTAI) protocol in Romanian buffaloes.

The present study describes the challenges of assisted reproduction in Romanian buffaloes while increasing the efficacy of artificial insemination by choosing the most suitable method. The modified fixed-time artificial insemination (FTAI) protocol with sexed semen was used to increase the conception rate. This study included a total of 80 buffalo heifers that received ovarian stimulation using the OvSynch protocol. Two groups (n = 40), namely, a control group, in which the classic FTAI method was performed, and an experimental group, in which deep intrauterine AI was performed in cows that had developed a dominant follicle (US+UcFTAI), were randomly selected. The conception rate (CR) was 63.6% in the experimental group, which was statistically higher (P < 0.05) than the control group (30%). The ultrasound examination indicated that, using the OvSynch protocol, 82.5% (33 out of 40) of buffaloes developed a dominant follicle (DF) while the distribution between the warm and cold seasons was 75 and 90%, respectively. The CR was 60% during the hot season and 66.6% during the cold season. At calving, 92.5% female fetuses were born. The improved FTAI method in this study enhanced the results by reducing the waste of sexed semen and maximizing the response to OvSynch, making it a recommendation for practitioners. This study presents preliminary results and highlights that genetic progress is difficult to achieve. A systematic approach is needed in order to choose the most suitable biotechnological method for each farm.

Evaluation of two beef cow fixed-time AI protocols that utilize presynchronization

Presynchronization was evaluated as a method to improve estrus response before fixed-time AI (FTAI). The objective was to compare FTAI results in beef cows from two different presynchronization approaches. Blood samples were collected on Day −14 (Day 0 = CIDR removal) to determine progesterone concentration (≥1 ng/mL = high, <1 ng/mL = low). In a subset (n = 1289), an additional blood sample was collected between Day −21 and −29 to determine cyclicity (if both the Day −14 and Day −21 to −29 samples were classified as low progesterone cows were classified as noncycling). Cows (n = 1388) from 30 herds were grouped by days postpartum (DPP) and age, and randomly assigned to either of two protocols. Cows assigned to the PG 6-day CIDR & FTAI protocol (PG6d) received prostaglandin F2α (PG) on Day −9, CIDR insertion and GnRH on Day −6, and CIDR removal and PG on Day 0. Cows assigned to the 7&7 Synch protocol (7&7) were administered PG and CIDR insertion on Day −14, GnRH on Day −7, and CIDR removal and PG on Day 0. For both protocols, FTAI occurred concurrently with GnRH 66 h after second PG. Pregnancy was determined by transrectal ultrasonography 30–40 d after FTAI. The GLIMMIX procedure of SAS was used to detect differences in estrus response and pregnancy success with herd as a random variable. Estrus response (0–66 h) was analyzed with two models, one included cyclicity and another replaced cyclicity with progesterone concentration at Day −14. In both models, cows assigned to the 7&7 protocol had greater (P < 0.01) estrus response than cows assigned to the PG6d protocol. The model including cyclicity, estrus response was impacted by the cyclicity by DPP interaction (P = 0.03), cyclicity by protocol interaction (P = 0.04), and the tendency of BCS by protocol interaction (P = 0.08). In the estrus response model that included progesterone concentration at Day −14, significant variables included the protocol by progesterone concentration at Day −14 (P = 0.01), and BCS (P < 0.01), while DPP (P = 0.08) and progesterone concentration at Day −14 (P = 0.07) were tendencies. Pregnancy success was influenced by estrual status (P < 0.01), body condition score (P = 0.04), and cycling status (P = 0.02), but was not influenced by protocol (P = 0.75; PG6d = 38 ± 5% and 7&7 = 37 ± 5%). In conclusion, effectiveness of presynchronization method depended on a cows’ physiological status, and the 7&7 protocol increased estrus response compared with PG6d, but there was no difference in pregnancy success.

Environmental and behavioral effects on pregnancy rate of Nellore cows synchronized to fixed-time artificial insemination protocol: multivariate analysis

The aim of the study was to evaluate the influence of behavioral reactivity, body condition score (BCS), corral temperature (CT), difficulty and time of insemination on pregnancy rate of Nellore females raised in the Pantanal area and submitted to a fixed-time artificial insemination (FTAI) protocol. Thus, 167 females, kept in an extensive grazing system were evaluated. The variables recorded were BCS, escape velocity (EV), CT, insemination difficulty (ID), chute behavior (CB), insemination time (IT) and pregnancy rate. Initially, Fisher's Exact Test was performed to verify differences between expected and observed for BCS, CB, EV, ID, IT and CT, in which the influence of each variable on pregnancy rate was individually analyzed and it was not significant influence of these variables was observed (P&lt;0.05). Subsequently, the result of the pregnancy diagnosis (success = 1 and failure = 0) was used to study the probability of pregnancy. Multivariate analysis was used to assess the influence of several characteristics together on the conception rate, creating different evaluation scenarios. In none of the scenarios extracted from the cluster analysis was there a statistically relevant influence on the number of pregnancies. Under the conditions in which the study was conducted, there was no influence of corral temperature, reactivity, time and difficulty of insemination, body condition score and animal category on pregnancy rate of bovine females submitted to a FTAI protocol.

Comparison of Gonadotropin-Releasing Hormone versus Estrogen-Based Fixed-Time Artificial Insemination Protocols in Grazing Bos taurus Suckled Beef Cows.

Fixed-timed artificial insemination (FTAI) protocols for beef cattle in South America are primarily based on estradiol esters and intravaginal progesterone-releasing devices (IVPD). The objective of this study was to determine the optimal gonadotropin-releasing hormone (GnRH)-based protocol as an alternative to the use of estrogen-based protocols in grazing Bos taurus suckling beef cows. All cows received an IVPD on the day of protocol initiation and prostaglandin F2α (PG) plus equine chorionic gonadotropin (eCG) treatments at the time of IVPD removal. In Experiment 1, cows (n = 235) were randomly assigned to one of four treatments: (i) 7-day estradiol = 2 mg of estradiol benzoate (EB) at IVPD insertion on Day 9 and 1 mg of estradiol cypionate (ECP) at IVPD removal on Day 2; (ii) 7-day GnRH = 10 µg of GnRH at IVPD insertion on Day 10, IVPD removal on Day 3 and GnRH at FTAI; (iii) 7 & 7 estradiol = PG at IVPD insertion on Day 16, EB on Day 9 and ECP at IVPD removal on Day 2; (iv) 7 & 7 GnRH = PG at IVPD insertion on Day 17, GnRH on Day 10, IVPD removal on Day 3 and GnRH at FTAI. In Experiment 2, cows (n = 462) were randomly assigned to one of four treatments: (i) 6-day estradiol = EB at IVPD insertion on Day 9, IVPD removal on Day 3 and GnRH at FTAI; (ii) 7-day estradiol; (iii) 7-day GnRH; (iv) 7 & 7 GnRH. In Experiment 1, plasma progesterone concentrations and percentage of cows with a corpus luteum (CL) at IVPD removal, and pregnancy per AI (P/AI) were greater for cows subjected to GnRH-based protocols compared with cows subjected to estrogen-based protocols (p < 0.01). In Experiment 2, cows subjected to the 7 & 7 GnRH protocol had the greatest P/AI (p < 0.01). In summary, GnRH-based FTAI protocols resulted in similar or greater P/AI compared to estrogen-based FTAI protocols in grazing postpartum Bos taurus suckled beef cows. The greatest P/AI was attained with the 7 & 7 GnRH protocol.

Increased accuracy of estrus prediction using ruminoreticular biocapsule sensors in Hanwoo (Bos taurus coreanae) cows

Visual estrus observation can only be confirmed at a rate of 50%–60%, which is lower than that obtained using a biosensor. Thus, the use of biosensors provides more opportunities for artificial insemination because it is easier to confirm estrus than by visual observation. This study determines the accuracy of estrus prediction using a ruminoreticular biosensor by analyzing ruminoreticular temperature during the estrus cycle and measuring changes in body activity. One hundred and twenty-five Hanwoo cows (64 with a ruminal biosensor in the test group and 61 without biosensors in the control group) were studied. Ruminoreticular temperatures and body activities were measured every 10 min. The first service of artificial insemination used gonadotropin-releasing hormone (GnRH)-based fixed-time artificial insemination protocol in the control and test groups. The test group received artificial insemination based on the estrus prediction made by the biosensor, and the control group received artificial insemination according to visual estrus observation. Before artificial insemination, the ruminoreticular temperature was maintained at an average of 38.95 ± 0.05°C for 13 h (−21 to −9 h), 0.73°C higher than the average temperature observed at −48 h (38.22 ± 0.06°C). The body activity, measured using an indwelling 3-axis accelerometer, averaged 1502.57 ± 27.35 for approximately 21 h from −4 to −24 h before artificial insemination, showing 203 indexes higher body activity than −48 hours (1299 ± 9.72). Therefore, using an information and communication techonology (ICT)-based biosensor is highly effective because it can reduce the reproductive cost of a farm by accurately detecting estrus and increasing the rate of estrus confirmation in cattle.

Effect of melatonin implantation prior to ovsynch plus cidr protocol on subsequent ovarian activity and pregnancy rate in summer anestrous Murrah buffaloes

For assessing the impact of melatonin pre-treatment in summer anestrous Murrah buffaloes on subsequent reproductive activity of a fixed time artificial insemination (FTAI) protocol, 83 anestrous Murrah buffaloes were subcutaneously inserted 2x4 mm size absorbable melatonin (MT) implants (18 mg/50 kg b wt) at the base of left ear and 38 Murrah buffaloes were used as non-implanted controls. On day 35 post-MT, the treatment and control group Murrah buffaloes were subjected to FTAI protocol (Ovsynch plus CIDR). Ovarian ultrasonography and jugular vein blood sampling was carried out on day 7 and 10 of protocol and on day 7-post AI. Overall pregnancy rate on day 90 post-AI in MT group was higher compared to control group (55.4 vs 31.6%, P&lt;0.05), along with the higher (P&gt;0.05) plasma progesterone on day 7 of FTAI protocol in Murrah buffaloes of the former group which conceived subsequently compared to their non-pregnant counterparts. Moreover, MT group had higher (P&gt;0.05) increase in dominant follicle diameter between day 7 and 10 of protocol compared to control group. On day 7 post-AI, the luteal profile in pregnant murrah buffaloes of MT group was better (P&lt;0.05) compared to their counterparts in control group (CL: 14.80±0.70 vs 10.82±0.46 mm, P4: 1.54±0.07 vs 1.04±0.08 ng/ml). In conclusion, melatonin pre-treatment in summer anestrous Murrah buffaloes could be associated with better luteal activity and higher pregnancy rate in the Ovsynch+CIDR protocol.

Application of Exogenous GnRH in Food Animal Production.

Over several decades, exogenous GnRH and agonists have been employed for controlling reproductive cascades in animals, and treating some reproductive morbidities. The administration of GnRH is used in animals to counter ovarian dysfunction, induce ovulation, and to increase conception and pregnancy rates. GnRH and its agonists are used in the treatment of cystic ovarian degeneration and repeat breeder syndrome. The development of protocols for GnRH administration by intramuscular injection, intramuscular or subcutaneous implants, and intravaginal deposition has empowered their clinical use worldwide. Currently, exogenous GnRH products are a central part of several pre- and post-breeding programs for the enhancement of fertility, including the control of estrous cycles and timing of ovulation, development of fixed-time artificial insemination protocols, improved embryo survival, and the treatment of reproductive morbidity. The aim of the present review is to summarize the application of exogenous GnRH agonists in food animal production.

Effect of Different Protocols of Fixed-Time Artificial Insemination on Mucus, Ovarian Size, and Pregnancy of Mixed-Breed Cows in the Humid Tropics of Peru.

The objective of the research was to evaluate three synchronization protocols for fixed-time artificial insemination (FTAI) in Bos indicus with Bos taurus crossbred cows. Forty-five cows of 5 ± 1.6 years were selected. The Jsynch treatment consisted of the application of an intravaginal device plus benzoate; on day 8, the device was removed and PGF2α, estradiol cypionate plus eCG was applied, and the FTAI was performed at 48-52 hours after. Conventional treatment consisted of the application of an intravaginal device plus benzoate; on day 8, the device was removed and PGF2α plus eCG was injected, and on day 9, estradiol benzoate was administered, and FTAI was performed at 30 hours. The Ovsynch treatment consisted of the application of GnRH, and on day 8, PGF2α was applied, on day 9, GnRH was applied again, and on day 10, FTAI plus GnRH was applied. The diagnosis of pregnancy was determined by transrectal ultrasound 45 days after the FTAI. The Jsynch (39.1%) and conventional (39.1%) treatment showed higher pregnancy compared to the Ovsynch treatment (21.7%) of pregnancy. The presence of crystalline mucus at the time of the FTAI was significantly associated with pregnancy. The results of this investigation indicate that it is possible to obtain acceptable pregnancy rates with the FTAI in B. indicus with B. taurus crossbred milk-producing cows, and treatments with progesterone-releasing devices plus eCG can improve the reproductive performance of cows.

Supplementing Ca salts of soybean oil via low-moisture molasses-based blocks to improve reproductive performance and overall productivity of beef cows

This experiment evaluated reproductive and productive responses of beef cows receiving self-fed low-moisture blocks (LMB) enriched or not with Ca salts of soybean oil (CSSO) throughout the breeding season. Non-pregnant, suckled multiparous Angus-influenced cows were assigned to a fixed-time artificial insemination (AI) protocol (day −10 to 0) followed by natural service (day 15–70). Cows were managed in 12 groups (46 ± 4 cows/group) maintained in individual pastures, and groups received LMB enriched with 25 % (as-fed basis) of CSSO or ground corn (CON) from day − 10 to 100. Both treatments were designed to yield a daily LMB intake of 0.454 kg/cow (as-fed basis). Cows receiving CSSO had greater (P < 0.01) mean concentrations of ω-6 fatty acids in plasma samples collected on days 0 and 55. Cows receiving CSSO had greater (P = 0.05) pregnancy rate to fixed-time AI (67.2 vs. 59.3 %), whereas final pregnancy rate did not differ (P = 0.92) between treatments. Pregnancy loss was less (P = 0.03) in CSSO cows (4.50 vs. 9.04 %), which also calved earlier during the calving season (treatment × week; P = 0.04). Weaning rate tended to be greater (P = 0.09) in CSSO (84.8 vs. 79.4 %), although calf weaning age and weight did not differ (P ≥ 0.72) between treatments. Kilos of calf weaned/cow exposed was greater (P = 0.04) in CSSO cows (234 vs. 215 kg). Therefore, supplementing CSSO to beef cows via LMB during the breeding season improved their reproductive and overall productivity during a cow-calf cycle.