Timed Artificial Insemination Research Articles

49 Prediction of Beef Heifer Reproductive Potential Using Molecular Markers

Abstract Heifer infertility has a negative impact on beef cattle production and profit on the livestock industry. The ability to identify heifers early based on reproductive potential would help improve the sustainability of beef production. The omics technologies have provided opportunities to improve reproductive efficiency. Previously, we have explored the ability of plasma metabolic profiles and mRNA profiles in the peripheral white blood cells (PWBCs) to serve as molecular markers of reproductive potential, at the time of artificial insemination (AI). Recently, we have integrated these data to identify the potential genetic pathways involved. We used RNA-Seq paired data and untargeted metabolomics from six AI-pregnant (AI-P) and six nonpregnant (NP) Angus-Simmental crossbred heifers at AI. Based on network co-expression analysis, we identified 17 and 37 hub genes in the AI-P and NP groups, respectively. We also identified TGM2, TMEM51, TAC3, NDRG4, and PDGFB as more connected in the NP heifers’ network. Metabolomic analysis identified 18 and 15 hub metabolites in the AI-P and NP networks. Tryptophan in the NP network, and allantoic acid in the AI-P network, exhibited a connectivity gain. Gene-metabolite integration identified tocopherol-a as positively correlated with ENSBTAG00000009943 in the AI-P group. Conversely, tocopherol-a was negatively correlated in the NP group with EXOSC2, TRNAUIAP, and SNX12. In the NP group, α-ketoglutarate-SMG8 and putrescine-HSD17B13 were positively correlated, whereas a-ketoglutarate-ALAS2 and tryptophan-MTMR1 were negatively correlated. These multiple interactions identified novel targets and pathways underlying fertility in bovines. Next, we hypothesized that gene expression from PWBCs at weaning could potentially predict the future fertility potential of beef heifers. We used the RNA-Seq approach to gain insights into the gene expression profile of PWBC at weaning from Angus-Simmental crossbred heifers. The heifers were retrospectively classified as pregnant through artificial insemination (AI-P, n = 8) and non-pregnant (NP, n = 7). We identified 92 differentially expressed genes between the groups, including MORN4, TFF2, MXD1, PHF8, KAT2B and CLEC4D. Based on a network co-expression analysis, we identified 14 and 52 hub targets, out of which ENSBTAG00000052659, OLR1, TFF2 and NAIP were exclusive to the AI-P group while 42 hubs were exclusive to the NP group. The differential connectivity between the networks of each group revealed a gain in connectivity due to the rewiring of major regulators in the NP group. The rewiring of major regulators unveiled in this study likely modulates the expression of gene targets as a response to infertility. The exclusive hub targets from AI-P are over-represented for the CXCR chemokine receptor pathway and inflammasome complex. In contrast, the 42 hub targets are over-represented for immune response and cytokine production pathways. These multiple interactions identified novel targets and pathways predicting fertility potential at an early stage of heifer development.

31 Correlation of the Reproductive Microbiome and Cytokine Profiles with Plasma Progesterone Concentrations in Postpartum Beef Cows Prior to Timed Artificial Insemination

Abstract Female sex steroid hormones have been shown to interact with cytokines within the immune system to help influence fertility. Previous research has indicated that shifts in commensal bacteria are associated with compromised reproductive performance during the postpartum period, yet evaluation of how the female sex hormone progesterone (P4) influences these microbial shifts is not fully understood. The objectives of this study were to determine 1) the relationship between P4 concentrations and cytokine concentrations, and 2) the relationship between P4 concentrations and the reproductive microbiome. Twenty Bos indicus-influenced beef cows (~60 days postpartum and free of health issues) were weighed, body condition scored and then subjected to the Bos indicus prostaglandin 5 day (d) + CIDR protocol beginning on d -8 and timed artificially inseminated on d 0. Blood samples were collected on d-3, -1, 0, and 28. Cytokines and reproductive microbiome were evaluated using vaginal and uterine flushes collected on d -3 and d-1. Experimental groups were assigned by d 28 pregnancy status, determined by transrectal ultrasonography, resulting in Open (n = 13) and Pregnant (n = 7) females. Bacterial community analyses were conducted targeting the V4 hypervariable region of the 16S rRNA gene. Plasma P4 concentrations on d -3 and d -1 were quantified per manufacturers’ instructions using a double-antibody RIA kit. Cytokine analyses of plasma and flushes were conducted using the RayBiotech Quantibody Bovine Cytokine Array Q1 kit per the manufacturer’s instructions. Statistical analyses were conducted using the GLM procedure and CORR procedure in SAS. Plasma P4 concentrations did not differ by pregnancy status (P > 0.05) but did decrease from d -3 to d -1 (0.89 vs. 0.08 ± 0.23 ng/mL, respectively; P = 0.02). There were no significant correlations between plasma P4 and plasma cytokines (P > 0.05). Interferon-alpha (IFNα) and P4 concentrations were positively correlated in Open and Pregnant uterine flushes (r = 0.41; P = 0.01). Concentrations of P4 interleukin (IL)-1F5 were negatively correlated in Open and Pregnant vaginal flushes (r = -0.32; P = 0.04). The relative abundance of genus Methanobrevibacter in uterine flushes was positively correlated with P4 concentrations in both experimental groups (r = 0.35; P = 0.02). Within vaginal flushes, there was a tendency for a negative correlation between the relative abundance of genus Bacteroides and P4 concentrations (r = -0.31; P = 0.05). Alternatively, there is a positive correlation between P4 concentration and relative abundance of genus Streptococcus (r = 0.47; P = 0.01) and genus Methanobrevibacter (r = 0.37; P = 0.02) in vaginal flushes. These results suggest there may be relationships between several cytokines and circulating P4 concentrations; therefore, potentially suggesting immunomodulatory activities of P4. Overall, this research alludes to P4 having a vital role in reproductive microbial shifts and associated immune responses. Further investigation on the local effects of P4 within the reproductive tract on immune responses and bacterial shifts could provide greater insight into mechanisms to improve fertility in beef cattle.

132 Weight and Reproduction Response to a Peanut Skins Plus Corn Supplement Offered to Heifers Grazing Spring Wheat Pasture During Artificial Insemination Breeding

Abstract The objective was to assess heifer weight and reproduction response to a peanut skins plus corn supplement during spring wheat grazing. In spring of 2021 and 2022, 10 and 8 pastures (1.62 ha/pasture) of fall overseeded wheat were randomly assigned to supplement (SUP) or control (CONT) constrained to the same number of replications per treatment. Pastures were stocked with 6 heifers/pasture each year. The initial goal was to use peanut skins as a single ingredient; however, testing palatability in year 1, before spring grazing, demonstrated a need to create a mixed supplement for improved acceptance. Over 5 weeks from initiation of grazing to breeding, heifers were transitioned from a 33:67 to a 50:50 peanut skins:corn mix. Supplement was offered at 0.5% BW, as-fed, at the start of spring wheat grazing in early March. An additional 2.4% and 8.3% inclusion of dried molasses was incorporated to further enhance acceptance in year 1 and 2, respectively. Estrus synchronization (ES) began March 31 and April 5 for each year. Data were analyzed across year with year considered as a random effect and pasture the experimental unit. Initial weight was 12 ± 5.8kg greater (P = 0.05) for SUP. Weight at ES tended (P = 0.11) to be 9.4 ± 5.55kg greater for SUP. Reproduction tract score was similar (P = 0.55), averaging 4.2 ± 0.13, and heifers were of similar (P = 0.33) age (405 ± 4.1 days) at time of insemination. Average daily gain from initial wheat grazing until removal for natural service breeding did not differ between treatments (P = 0.92) and averaged 1.0 ± 0.21 kg. Body condition score was similar (P = 0.43) at ES and averaged 5.7 ± 0.7. The heavier initial BW of SUP heifers was sustained through wheat grazing (P = 0.05). Ultrasound determined pregnancies averaged 66.7 and 50.0 ± 6.6% for SUP and CONT, respectively, for an odds ratio of 2.1 ± 0.79 (P = 0.08) at 30 days after insemination. Overall pregnancy rates following natural service averaged 88.9 and 79.6 ± 4.88% for SUP and CONT, respectively with an odds ratio of 2.1±1.12 (P=0.19). High dietary protein has been associated with reduced reproductive performance. Using the peanut skins plus corn supplement was hypothesized to improve reproduction on a high protein diet due to the possible effect of peanut skins tannins on dietary protein utilization and corn altering the rumen degradable protein to organic matter ratio. While the fixed time artificial insemination pregnancy rates for CONT would not be considered poor, there was a tendency for SUP to secure more pregnancies among heifers of similar age, reproductive tract score, and body condition.

144 The effect of exogenous GnRH at the time of artificial insemination on luteinizing hormone in lactating Holstein cows

144 The effect of exogenous GnRH at the time of artificial insemination on luteinizing hormone in lactating Holstein cows

Review: Current status of corpus luteum assessment by Doppler ultrasonography to diagnose non-pregnancy and select embryo recipients in cattle.

A number of potentials uses of Doppler ultrasonography have been explored in the last decades, both as research tools in reproductive physiology investigations and for the reproductive management of farm animals. The objective of this review was to address some of the recent strategies developed in fixed-time reproductive programs and resynchronization of ovulation in cattle, based on the evaluation of corpus luteum function by color-Doppler ultrasound imaging. Recent studies in dairy and beef cattle pointed out to a high accuracy when Doppler ultrasonography is used to assess the functionality of the corpus luteum and identify non-pregnant females at 20-24days after breeding. Therefore, super-early resynchronization programs starting in the second week after timed-artificial insemination or embryo transfer have been developed and are being implemented in commercial assisted reproduction programs; thus, anticipating conception with proven semen or genetically superior embryos. In addition, assessment of corpus luteum blood perfusion can be used for identifying high fertility embryo recipients in fixed-time embryo transfer programs.

Review: Manipulation of follicle development to improve fertility of cattle in timed-artificial insemination programs.

The development of an ovulatory follicle is a fundamental premise for any reproductive management program that aims to optimize fertility in cattle. Controlling follicular development comprises the synchronized emergence of a new follicular wave, selection and growth of the dominant follicle, and synchronized ovulation of a high-quality oocyte. All these follicular events, primarily driven by gonadotropin secretion, occur under a very dynamic hormonal environment. In this sense, controlling follicular development demands essentially a precise manipulation of the hormonal environment to modulate gonadotropin secretion. Furthermore, the effectiveness of hormonal manipulation strategies in the management of follicular development depends on specific particularities of each situation, which can vary widely according to genetic groups (Bos taurus vs Bos indicus), nutritional, metabolic, and reproductive status. In this regard, the constant search for the refined synchrony between the hormonal treatments and reproductive events, considering these distinctions and particularities, have provided valuable information that contributed to the development of efficient reproductive programs. This manuscript discusses the physiological bases behind the development of fine-tuned timed-artificial insemination protocols for beef and dairy cattle that resulted in great improvements in reproductive efficiency of beef and dairy herds.

117 Effect of delayed timing of artificial insemination with sex-sorted semen on pregnancy per artificial insemination in synchronized dairy heifers managed in a seasonal-calving pasture-based system

117 Effect of delayed timing of artificial insemination with sex-sorted semen on pregnancy per artificial insemination in synchronized dairy heifers managed in a seasonal-calving pasture-based system

The parity affects the relationship between the insemination-ovulation interval and the conception rate in lactating dairy cows.

The aim of the present study was to investigate the relationship between parity and the optimal insemination-ovulation interval (IOI) in lactating dairy cows, as well as the relationship between the optimal timing of artificial insemination (AI) based on the onset of estrus and parity. A total of 183 postpartum lactating Holstein cows participated in the study and underwent 522 AI procedures using unsorted frozen semen between 2011 and 2018. Transrectal ultrasound was used to detect ovulation times in 326 AI procedures, and a pedometer was used to detect estrus onset times in 470 AI procedures. An analysis was also performed on data from 265 AI procedures in which all the times of estrus onset, estrus end, and ovulation were detected. The results showed that ovulation occurring 6-12 hr after AI was associated with the numerical highest conception rate in multiparous cows. Moreover, the conception rate was higher when ovulation occurred between 6 and 12 hr after AI than when it occurred between 0 and 6 hr or 18 and 24 hr after AI. In contrast, the effect of IOI on the conception rate in primiparous cows was insignificant. There was no effect of AI timing based on the estrus onset on the conception rate in either primiparous or multiparous cows. Furthermore, the interval from estrus onset to ovulation was similar in primiparous and multiparous cows. The difference in optimal IOI between primiparous and multiparous cows may be due to the larger uterus size in multiparous cows. These findings suggest that the optimal timing for AI in multiparous cows is more restricted than in primiparous cows. Therefore, it is essential to carefully determine the timing of AI when it is performed in multiparous cows.

Economical implications and the impact of gonadotropin-releasing hormone administration at the time of artificial insemination in cows raised in the extensive system in North Romania.

Artificial insemination (AI) is the first and the most used biotechnologies in reproduction all over the world. Many studies reported the beneficial role of gonadotropin-releasing hormone (GnRH) administrated some hours before the AI or at the artificial insemination time. This study aimed to assess the effect of GnRH analogs given at the time of insemination on the first, second, and third AIs and to assess the economical implications of GnRH administration. We hypothesized that administration of GnRH at the time of insemination would increase ovulation and pregnancy rate. The study was conducted on small farms in northwestern Romania and included animals of the Romanian Brown and Romanian Spotted breeds. Animals in estrous at the first, second, and third inseminations were randomly divided into groups that received GnRH at insemination and groups that did not. A comparison between the groups was performed and the cost of GnRH administration to obtain one gestation was calculated. The GnRh administration increased the pregnancy rate at the first and the second inseminations by 12 and 18%, respectively. For one pregnancy, the cost of GnRH administration was approximately 49 euros for the first insemination group and around 33 euros for the second insemination group. No improvement of the pregnancy rate was observed after the GnRH administration for the cows at the third insemination, so, for this group, no economic statistics were performed.

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.

Timed artificial insemination in Serrana nanny-goats: the effects of vaginal specula and artificial insemination technicians

The success of artificial insemination (AI) depends on the equipment used, especially regarding the facilitation of artificial insemination technicians’ work and the positioning of insemination guns in the genital tract of animals. This study aimed to evaluate how vaginal specula and artificial insemination technicians affected the fertility rates of timed artificially inseminated Serrana Transmontano goats (which are indigenous to Portugal). For this, 58 adult nanny-goats aged between three and nine years were used. They were reproductively controlled by a short progestogen treatment (FGA) (seven days) with equine chorionic gonadotropin (eCG). Timed cervical AI (43 hours after the end of the hormonal treatment) was performed with chilled semen. Nanny-goats were inseminated by two artificial insemination technicians (A vs. B) alternating two vaginal specula (Minitub vs. “Reyes”). Pregnancy was diagnosed by ultrasound 41 days later. About 98% of nanny-goats responded to our treatment with FGA and eCG. In total, 82.8% of nanny-goats were pregnant 41 days after AI. Neither vaginal specula (Minitub: 81.2% vs. “Reyes”: 84.6%) nor artificial insemination technicians (A: 82.8% vs. B: 82.8%) affected fertility rates. Thus, we conclude that neither the vaginal specula used in this research nor the artificial insemination technicians affected Serrana goat pregnancy rates.

The transcriptomic response of bovine uterine tissue is altered in response to sperm from high- and low-fertility bulls†.

Despite stringent quality control checks, some bulls with apparently normal semen quality yield lower than expected pregnancy rates. This study profiled the transcriptome and performed histological analysis of the bovine uterus in response to sperm from high-fertility (HF) and low-fertility (LF) bulls. Postmortem uterine biopsies and uterine explants were collected from heifers 12h after a fixed time artificial insemination (AI) to a synchronized estrus with frozen-thawed semen from five HF (fertility rate 4.01% ± 0.25) and five LF (fertility rate - 11.29% ± 1.11; mean ± SEM) bulls. Uterine biopsies were also collected from control (CTRL) heifers, which were not inseminated. RNA-sequencing and histological analysis were performed for differential gene expression and neutrophil quantification. In the HF treatment relative to CTRL heifers, there were 376 genes significantly differentially expressed in the endometrium with just one gene differentially expressed in the LF treatment relative to CTRL heifers. Comparing the HF and LF treatments directly, there were 40 significantly differentially expressed genes (P < 0.05). Transcriptomic analysis shows a predominant role for the inflammatory marker Interleukin-1 alpha, which was further confirmed by immunohistochemistry. Quantification of neutrophils in the endometrium showed a significant effect of sperm; however, there was no difference in neutrophil numbers between HF and LF groups. In conclusion, this novel study clearly shows a distinct inflammatory response to sperm in the endometrium and a divergent transcriptomic response to semen from HF and LF bulls.

Effect of Timing of Artificial Insemination on Fertility of Cows and Calf Sex Ratio in Arsi Cows

An on-station study was conducted to investigate the effect of the interval between artificial insemination (AI) and the beginning of estrus on the fertility and sex ratio of offspring in Arsi cows. Standing to be mounted was considered the beginning of estrus, and artificial inseminations were made 6 h (Group 1), 12 h (Group 2), 18 h (Group 3) and 24 h (Group 4) after the onset of estrus in the cows. Overall, 50 cows with parity between 2 and 4, body weight ranging from 250 to 300 kg were used for the purpose of this study. Ovulation was synchronized by administration of PGF2α hormone. Pregnancy diagnosis was performed by rectal palpation at 90 to 120 days post AI. Pregnancy rates were 60%, in each of the Groups 1, 2, 3 and 50% in group 4. The highest female to male ratio (P=0.041) of calves was obtained in Group 1. Whereas the highest male to female ratio was recorded in group 4. The number of artificial inseminations per pregnancy was within the economical limits in all groups. In conclusion, the results of this study indicated that the variation in time interval between the onset of estrus and artificial insemination alters the sex ratio of offspring in Arsi cows without affecting fertility parameters.

Influence of Ovarian Status and Steroid Hormone Concentration on Day of Timed Artificial Insemination (TAI) on the Reproductive Performance of Dairy Cows Inseminated with Sexed Semen.

This study aimed to evaluate the effect of the ovarian status and steroid hormone concentration on the day of TAI on the reproductive performance of dairy cows subjected to estrus synchronization treatment and timed artificial insemination with sexed semen. Seventy-eight cyclic Holstein cows pre-treated with PGF2α-GnRH were divided in two groups-I (Preselect-OvSynch, n = 38) and II (OvSynch+PRID-7-day+eCG, n = 40)-and inseminated with sexed semen. The presence of preovulatory follicle (PF) with or without corpus luteum (CL), the PF diameter, the estradiol (E2) and progesterone (P4) concentrations on the day of TAI, the pregnancy rate (PR) and embryo loss were determined. On the day of TAI, 78.4% of all the pregnant cows presented a PF (mean size 1.80 ± 0.12 cm) without CL, low P4 (0.59 ± 0.28 ng/mL) and high E2 (12.35 ± 2.62 pg/mg) concentrations. The positive correlation between the size of the PF and the level of E2 in the pregnant cows from group II was stronger than that of group I (R = 0.82 vs. R = 0.52, p < 0.05). The pregnancy rate on day 30 (57.5% vs. 36.8%) and day 60 (50% vs. 26.3%; p < 0.05) and the embryo losses (13% vs. 28.5%) showed better effects of treatment in group II. In conclusion, the ovarian status and the steroid hormone concentration on the day of TAI influence the pregnancy rates of dairy cows subjected to estrus synchronization and timed artificial insemination with sexed semen.

Assisted reproductive technologies (ARTs) in Mithun (Bos frontalis): What progress has been made so far? An overview.

Mithun, a unique bovine species, endemic to parts of North East India and plays an important role in the socioeconomic, cultural and religious fabrics of the local tribal population. To date, Mithuns are reared in a traditional free-range system by communities and increased deforestation, agricultural commercialization, disease outbreaks and indiscriminate slaughtering of elite Mithun for table purposes have significantly decreased its habitat and the elite Mithun population. Greater genetic gain is achieved with the implementation and effective use of assisted reproductive technologies (ARTs); however, presently it is limited to organized Mithun farms. At a slow pace, Mithun farmers are adopting semi-intensive rearing systems and interest in the use of ARTs is gradually escalating in Mithun husbandry. This article reviews the current status of ARTs such as semen collection and cryopreservation, estrus synchronization and timed artificial insemination (TAI), multiple ovulation and embryo transfer and in vitro embryo production and future perspectives in Mithun. Mithun semen collection and cryopreservation have been standardized, and estrus synchronization and TAI are suitable technologies that can be easily implemented under field conditions in near future. The establishment of an open nucleus-breeding system under community participatory mode along with the introduction of the ARTs is an alternative to the traditional breeding system for rapid genetic improvement of Mithun. Finally, the review considers the potential benefits of ARTs in Mithun and future research should include the use of these ARTs which will provide additional opportunities for improved breeding regimens in Mithun.

GnRH dose at initiation of a 5-day CO-Synch + P4 for fixed time artificial insemination in suckled beef cows

The objective of the present study was to evaluate the effect of GnRH dose administered at initiation (GnRH-1) of a 5-day CO-Synch + P4 protocol on ovulatory response, expression of estrus, and fertility in suckled beef cows. Suckled beef cows (n = 1101) at four locations were randomized to receive either 100 or 200 µg of gonadorelin acetate at initiation (D-8) of a 5-day CO-Synch + P4 protocol concurrently with insertion of an intravaginal progesterone (P4) device. On D-3 the P4 device was removed, two doses of prostaglandin F2α were administered concurrently and a patch was applied to evaluate expression of estrus. Artificial insemination was performed 72 h after P4 device removal (D0) simultaneously with the administration of 100 µg of gonadorelin acetate (GnRH-2). Increasing GnRH dose at initiation of a 5-day CO-Synch + P4 did not enhance ovulatory response (P = 0.57) to GnRH-1, expression of estrus (P = 0.79), nor pregnancies per AI (P/AI; P = 0.91). Both follicle size (quadratic) and circulating P4 (linear) affected (P < 0.01) ovulatory response to GnRH-1 independent of dose. Cows that had ovulation to GnRH-1 had smaller (P < 0.001) follicle size on D-3 and reduced (P = 0.05) expression of estrus compared to cows that did not have ovulation to GnRH-1, however, P/AI did not differ (P = 0.75). In conclusion, increasing the dose of GnRH-1 in the 5-day CO-Synch + P4 protocol did not enhance ovulatory response, expression of estrus, or P/AI in suckled beef cows.

Effect of a progesterone-releasing intravaginal device (PRID) for 8 days during a modified Ovsynch protocol on pregnancy outcomes in lactating Holstein cows

Our objective was to evaluate the effect of a progesterone-releasing intravaginal device (PRID) in a 7-d Ovsynch protocol on pregnancy per artificial insemination (P/AI) and pregnancy loss, compared with a standard 7-d Ovsynch protocol without progesterone supplementation. We hypothesized that progesterone supplementation during an Ovsynch protocol would increase P/AI and decrease pregnancy loss. Data were collected on lactating Holstein cows (n = 716) that either received a 7-d Ovsynch protocol (control: d 0, 100 µg of GnRH; d 7, 500 µg of cloprostenol; d 9, µg of GnRH; n = 360) or a modified Ovsynch protocol with addition of a PRID (PRIDsynch; d 0, 100 µg of GnRH + PRID; d 7, 25 mg of dinoprost; d 8, PRID removal; d 9, 100 µg of GnRH; n = 356). All cows received timed artificial insemination (TAI) approximately 16 h after the second GnRH treatment. Pregnancy diagnosis was performed via ultrasonography on d 38 ± 3 after TAI and rechecked on d 80 ± 7 after TAI. Reproductive performance differed between treatments, with PRIDsynch cows having greater (38.9%) P/AI compared with control cows (31.7%) at d 38 ± 3 and also at d 80 ± 7 (34.6% vs. 28.9%, for PRIDsynch and control cows, respectively). Pregnancy loss did not differ among treatments.

Artificial intelligence as object of intellectual property in Indonesian law

AbstractArtificial intelligence (AI) has an important role in digital transformation worldwide, including in Indonesia. AI itself is a simulation of human intelligence that is modeled in machines and programmed to think like humans. At the time AI and the Internet of Things are connected, it can, in principle, act on data without the need for human intervention. AI was originally created by humans as a consequence of the massive digital revolution. Recently, AI has also developed rapidly to perform functions like humans and even produce works of intellectual property like humans, such as creating songs, making new inventions, making industrial designs, and so on. Thus, it is a question whether AI can be qualified as a legal subject of creator, inventor or designer and then register as intellectual property. This research answers this question from the perspective of Indonesian law based on Copyright Law, Patent Law, Industrial Design Law, as well as Trademark Law, and Geographical Indications as the existing Indonesian laws.

Comparison of the 7-day CO-Synch and 8-day estradiol-based protocols for estrus synchronization and timed artificial insemination in suckled Bos taurus cows

Two experiments were conducted to compare, follicle diameter (FD) on Day −1, corpus luteum (CL) area on Day 7, progesterone (P4) concentration on Day 7 and 18, pregnancy per timed artificial insemination (TAI) on Day 30, and pregnancy loss (PL) between Days 30 and 60 after TAI (TAI, Day 0) using two different synchronization protocols. In Experiment 1, Angus cows (n = 1148) were randomly assigned to either 7-d progesterone CO-Synch (7-d CO-Synch) or 8-d progesterone + estradiol (8-d P + ES) synchronization protocols for TAI. On Day −10, cows in the 7-d CO-Synch treatment group (n = 574) received a progesterone-releasing intravaginal device (PIVD; 0.5 g P4) and GnRH (0.105 mg), on Day −3 the PIVD was removed and cows received cloprostenol (0.150 mg), then, on Day 0 (64 h after PIVD removal), cows received GnRH (0.105 mg) and were TAI. On Day −10, cows in the 8-d P + ES treatment group (n = 574) received a PIVD (0.5 g P4) and estradiol benzoate (2.0 mg), on Day −2 the PIVD was removed, and cows received cloprostenol (0.150 mg) and estradiol cypionate (0.5 mg), then, on Day 0 (48 h after PIVD removal), cows were TAI. Pregnancy per TAI was determined on Days 30 and 60. In a subset of cows (7-d CO-Synch, n = 41; 8-d P + ES, n = 40), serum P4 concentration was evaluated on Day 18. In Experiment 2, anestrus (n = 34) and cyclic (n = 34) suckled beef cows were selected and submitted at random on Day −10, to either 7-d CO-Synch or 8-d P + ES treatment groups. Follicle diameter on Day −1, CL area, and serum P4 concentration on Day 7 were determined. In Experiment 1, pregnancy per TAI on Day 30 did not differ (7-d CO-Synch = 48.9%; 8-d P + ES = 45.6%) between treatments but it was greater for cows with BCS ≥5 (P < 0.01). Pregnancy loss between Days 30 and 60 did not differ between treatment groups but tended to be greater in cows with BCS <5.0 (P < 0.1). In a subset of cows, serum P4 concentration on Day 18 did not differ between treatment groups but tended to be lower (P < 0.1) in cows that had PL between Days 30 and 60 compared to cows that had no PL. In Experiment 2, FD tended to be greater (P < 0.1) and CL area was greater (P = 0.05) in anestrus cows from 7-d CO-Synch treatment. In cyclic cows, the treatment did not affect the FD or CL area. In conclusion, there was no difference in pregnancy per TAI on Day 30 and PL between Days 30 and 60 between cows using 7-d CO-Synch + PIVD or 8-d estradiol-based + PIVD protocols for estrus synchronization and TAI.

Application of gender-ablated semen during timed artificial insemination following oestrous synchronization in dairy and beef cows.

The objectives of this study were to compare oestrous synchronization expression and conception rate following timed artificial insemination (TAI) with frozen-thawed X-sexed or unsexed semen in dairy and beef cows. For this study, 227 cows (dairy, n= 130 and beef, n= 97) were assigned to a 9-day Ovsynch + controlled intravaginal drug release (CIDR) protocol. All cows were TAI using X-sexed or unsexed semen from 8sires. Each semen type was obtained from 4 sires [2 dairy (Holstein Friesian) and 2 beef (Angus)]. Pregnancy detection was performed on Days 35, 65 and 95 following TAI by transrectal ultrasonography and hand palpation. The proportion of oestrus expression was higher in dairy (85.3%) cows compared with beef (65.0%) cows (p< .05). Overall, dairy (X-sexed, 61.9% and unsexed, 62.0%) cows had greater conception rates on Day 35 compared to beef (X-sexed, 56.0% and unsexed, 52.2%) cows (p<.05). Concurrently, on Day 95, overall conception rates in dairy (X-sexed, 41.4% and unsexed, 48.5%) cows were greater than beef (X-sexed, 38.0% and unsexed, 37.0%) cows (p< .05). Pregnancy/embryo losses between Days 35 and 65 in dairy (X-sexed, 33.3% and unsexed, 18.2%) cows and beef (X-sexed, 28.6% and unsexed, 29.2%) cows were recorded (p< .05). Dairy (X-sexed, 7.7% and unsexed, 8.3%) cows had higher incidence of pregnancy losses between Days 66 and 95 when compared to beef (X-sexed, 5.0% and unsexed, 0.0%) cows (p< .05). Oestrous expression and conception rates in dairy and beef cows were satisfactory. Advanced reproductive biotechnologies can successfully utilize gender-ablated semen in organized emerging cattle farming systems.