Characterization of conception rate after embryo transfer in comparison with that after artificial insemination in dairy cattle.

Artificial insemination (AI) and embryo transfer (ET) are important in the reproduction of dairy cows. The conception rate after AI or ET is an essential indicator when selecting appropriate breeding methods. However, information on the environmental factors affecting ET conception rate when compared with AI is limited. We aimed to investigate environmental factors affecting ET conception rate and characterize the differences in environmental factors between AI and ET. Records of the first AI (n = 1,870,143) and ET (n = 29,922) from Holstein nulliparous, primiparous, and multiparous cows in Hokkaido, Japan, were analyzed using separate multivariable logistic regression models. For each breeding method, we grouped primiparous and multiparous cows according to milk yield at peak lactation (PY; < 25, 25-30, 30-35, ≥ 35 kg in primiparous, < 40, 40-45, 45-50, ≥ 50 kg in multiparous) and the interval from calving to first AI or ET (CFI/CFT; < 60, 60-79, 80-99, ≥ 100 d) to evaluate the effects of PY and CFI/CFT on conception rate. AI conception rate decreased with increasing PY in primiparous and multiparous cows, whereas ET conception rate did not decrease significantly. Additionally, the ET conception rate did not decrease even in primiparous and multiparous cows slightly earlier than 60 d in CFI/CFT when compared with those in CFI/CFT after 60 d, which differed from the AI conception rate. Collectively, breeding by ET leads to the avoidance of negative effects of high milk yield and calving on the conception rate, indicating that cows are fertile by ET within 60 d after calving.

Estimation of direct and maternal covariance along with genetic and phenotypic trends of reproduction traits in Murrah buffalo.

The research utilized data from 662 Murrah buffaloes meticulously recorded over 24 years (1996-2019) from historical pedigree sheets maintained at the buffalo farm of the Department of Livestock Production and Management (LPM) at Lala Lajpat Rai University of Veterinary and Animal Sciences (LUVAS), Hisar. A series of six univariate animal models were employed to calculate estimates of (co)variance components and heritability for first lactation reproduction traits. Among these models, Model 2 was identified as the best fit for age at first calving (AFC) and calving interval (CI), while Model 1 proved optimal for service period (SP) and conception rate (CR). The heritability estimates for AFC, SP, CR and CI across the models were ranged between 0.11 and 0.32, 0.01 and 0.03, 0.05 and 0.06, and 0.01 and 0.06, respectively. Maternal effects (m2 ) were observed in AFC and CI, ranging from 0.10 to 0.20 and 0.01 to 0.03, respectively. Across all three traits, there was a consistent negative genetic correlation (-0.75 to -0.92) between direct additive and maternal effects. The breeding values for AFC, SP, CR and CI varied within specific ranges from -32.85 to 44.33 days, -15.61 to 28.42 days, -7.41 to 6.48% and -20.64 to 35.79 days, respectively. Significantly, Spearman's rank correlation analysis revealed highly significant relationships (p < .01) between the breeding values of different models, indicating strong and consistent associations within these traits. These findings underscore the stable and reliable connections observed within the breeding values for these essential reproductive traits across the various models used in the study. The majority of reproductive traits showed favourable negative trends, indicating a positive outcome. A decrease in AFC, SP and CI suggests an extended economic life for the animals. Additionally, the upward trends in CR reflect positive indications of effective management practices and skilled operational procedures.

Ovarian response and conception rate in Boran and Boran*Holstein cows treated by Gonadotrophin-realizing hormone and ProstaglandinF2α with and without exogenous progesterone.

BACKGROUND: Difference in breed, nutrition status and climate in which animals are managed result in differences in response to reproductive hormones. Fertility rate to artificial insemination is very low in Ethiopian Boran and Boran*Holstein crosses. This partly maybe due to adopting estrus and/or ovulation synchronization developed for temperate taurine cattle. Experimental study was conducted to evaluate ovarian response to combinations of Gonadotrophin-Realizing Hormone agonist (gonadorelin) and ProstaglandinF2α (PGF2α) with or without progesterone (Controlled Internal Drug Release/CIDR), and conception rate to timed AI. Postpartum native Ethiopian Boran (n = 60) and Boran*Holstein cross (n = 66) cows were randomly assigned to four treatment groups as Ovsynch (gonadorelin on day of start, PGF2α seven days later, 2nd gonadorelin at 48 h of PGF2α and insemination at 19 h of the 2nd gonadorelin); CIDR + Ovsynch (same as Ovsynch but CIDR device was inserted into vagina for 7 days); Cosynch (same as Ovsynch but insemination was made at the 2nd gonadorelin) and CIDR + Cosynch (same as Cosynch but CIDR was inserted for 7 days). RESULT: There was no difference (P > 0.05) in ovulation rate to day 9 gonadorelin (88.33% in Boran; 78.79% in Boran*Holstein) and interval from day 9 gonadorelin to ovulation (36.5 ± 1.13 h in Boran and 36.057 ± 1.11 h in Boran*Holstein). Dominant follicle immediate to ovulation (14.95 ± 0.19 mm Vs 19.12 ± 0.49 mm) and corpus luteum size (16.31 ± 0.33 mm Vs 20.28 ± 0.43 mm ) respectively were smaller (P < 0.05) in Boran than Boran*Holstein. Plasma progesterone concentration at PGF2α was higher (P < 0.05) in Boran (11.91 ± 0.74ng/mL) than Boran*Holstein (6.13 ± 0.27ng/mL) but luteolysis rate was lower (P < 0.05) in Boran (87.9%) than Boran-Holstein (96.9%). Cows with CIDR had higher conception rate than cows without CIDR (72.00% Vs 39.02% in Boran*Holstein and 74.07%, Vs 51.52% in Boran respectively). Insemination at 19 h of gonadorelin administration resulted in higher conception rate (78.6% for Boran; 71.43% for Boran*Holstein) than insemination at gonadorelin (69.29% for Boran; 66.67% for Boran*Holstein). CONCLUSION: Boran cows have smaller preovulatory follicles, smaller corpus luteum, large amount of progesterone and lower rate of luteolysis to PGF2α compared to Boran*Holstein. The CL of Boran cattle seems les reactive to PGF2α than Boran*Holstein CL. CIDR significantly improved conception rate in Boran and Boran*Holstein cows.

Effects of the temperature-humidity index on conception rates in Holstein heifers and cows receiving in vitro-produced Japanese Black cattle embryos.

We investigated the effect of the temperature-humidity index (THI) on the conception rate (CR) in Holstein heifers and cows receiving in vitro-produced (IVP) Japanese Black cattle fresh embryos. IVP embryos were transferred to Holstein heifers (n = 1,407) and cows (n = 3,189) on 245 commercial farms. The monthly average ambient temperature (AT) and THI ranged from 4.7 to 29°C and 41 to 81, respectively; both were the highest in August. The monthly CR ranged from 16.3% to 46.7% in cows and 23.8% to 74.1% in heifers. The CR of heifers was unaffected by THI, AT, or the month of embryo transfer. However, these parameters affected the CR of cows. The CR at THI values of 61-65 and 71-75 was greater than that at THI > 75, whereas other THI values had no effect. The CR at temperatures > 25°C was lower (P = 0.008) than that at temperatures of 15-20°C and 20-25°C. Moreover, the CR was lowest (P = 0.003) in July. THI and parity (P = 0.057 and P = 0.001, respectively) and AT and parity (P = 0.019 and P = 0.001, respectively) showed significant effects on CR; however, there was no interaction between these two factors. In conclusion, AT > 25°C and THI > 75 adversely affect the CR outcome in cows but not in heifers.

Differentially expressed microRNAs in biochemically characterized FrieswalTM crossbred bull semen.

In addition to the transmission of paternal genome, spermatozoa also carry coding as well as noncoding microRNAs (miRNAs) into the female oocyte during the process of biological fertilization. Based on RNA deep sequencing, a total 28 number of differentially expressed miRNAs were cataloged in categorized FrieswalTM crossbred (Holstein Friesian X Sahiwal) bull semen on the basis of conception rate (CR) in field progeny testing program. Validation of selected miRNAs viz. bta-mir-182, bta-let-7b, bta-mir-34c and bta-mir-20a revealed that, superior bull semen having comparatively (p < .05) lower level of all the miRNAs in contrast to inferior bull semen. Additionally, it was illustrated that, bta-mir-20a and bta-mir-34c miRNAs are negatively (p < .01) correlated with seminal plasma catalase (CAT) activity and glutathione peroxidase (GPx) level. Interactome studies identified that bta-mir-140, bta-mir-342, bta-mir-1306 and bta-mir-217 can target few of the important solute carrier (SLC) proteins viz. SLC30A3, SLC39A9, SLC31A1 and SLC38A2, respectively. Interestingly, it was noticed that all the SLCs were significantly (p < .05) expressed at higher level in superior quality bull semen and they are negatively correlated (p < .01) with their corresponding miRNAs as mentioned. This study may reflect the role of miRNAs in regulating few of the candidate genes and thus may influence the bull semen quality traits.

Intensity of estrus expression - valuable obvious determinant of fertility in Bos indicus cows.

This study aimed to characterize estrus response and to establish relationships between intensity of estrus, preovulatory follicle (POF) size and estradiol (E2) concentrations on day of AI, luteal profiles and pregnancy outcome in lactating Hariana breed of cows. 200 cyclic cows were subjected to Ovsynch (n = 54) and Pre-OV treatment (n = 146). Ovsynch: Buserelin acetate (BA; 10 µg), Cloprostenol (500 µg) and BA (10 µg) were injected i.m. on day 0, 7 and 9, respectively, irrespective of treatment. Pre-OV: BA (10 µg) and Cloprostenol (500 µg) was also injected i.m. simultaneously 7 days prior to initiate Ovsynch. On the basis of estrus behavior, the cows were classified into three groups: weak, moderate and intense. Artificial insemination performed at 18-24 hours after 2nd BA of Ovsynch in both treatments. The average duration of estrus did not differ (p > 0.05) between Ovsynch and Pre-OV treatment. A positive correlation was observed between estrus response and POF size, concentration of E2 on day of AI and luteal profiles on day 12 post-AI. First service conception rate was higher in cows exhibited intense (45.46%) and moderate (42.56%) estrus response than weak (28.57%) estrus response. In conclusion, intensity of estrus expression could be considered as important determinant for deciding pregnancy outcomes in Bos indicus cows.

Seasonal influence on expression of heat shock proteins (HSP70 and HSP90) vis-à-vis functional competence of Gir bull semen.

The success of assisted reproduction relies on functional competence of frozen-thawed semen. Heat stress affects protein folding leading to aggregation of mis-folded proteins. Hence, a total of 384 (32 ejaculates/bull/season) ejaculates from six matured Gir bulls were used to evaluate physico-morphological parameters, the expression of HSPs (70 and 90) and fertility of frozen-thawed semen. The mean percent individual motility, viability and membrane integrity were significantly (p < 0.01) higher in winter compared to summer. Out of 1200 Gir cows inseminated, 626 confirmed pregnant and the mean conception rate of winter (55.04 ± 0.35) was significantly (p < 0.001) higher than summer (49.33 ± 0.32). A significant (p < 0.01) difference in concentration of HSP70 (ng/mg of protein) but not HSP90was observed between the two seasons. The HSP70 expression in pre-freeze semen of Gir bulls had significant positive correlation with motility (p < 0.01, r = 0.463), viability (p < 0.01, r = 0.565), acrosome integrity (p < 0.05, r = 0.330) and conception rate (p < 0.01, r = 0.431). In conclusion, the season influences physico-morphological parameters and expression of HSP70 but not HSP90 in Gir bull semen. The HSP70 expression is positively correlated with motility, viability, acrosome integrity and fertility of semen. The semen expression of HSP70 may be utilized as biomarker for thermo-tolerance, semen quality and fertilizing capacity of Gir bull semen.

Improving national fertility evaluations by accounting for the rapid rise of embryo transfer in US dairy cattle.

Dairy producers have improved fertility of their herds by selecting bulls with higher conception rate evaluations. This research was motivated by the rapid increase in embryo transfer (ET) use to 11% of recent births and >1 million total births, with >5 times as many ET calves born in the United States in 2021 compared with just 5 yr earlier. Historical data used in genetic evaluations are stored in the National Cooperator Database. Recent records in the national pedigree database revealed that only 1% of ET calves have corresponding ET records in the breeding event database, 2% are incorrectly reported as artificial inseminations, and 97% have no associated breeding event. Embryo donation events are also rarely reported. Herd years reporting >10% of calves born by ET but less than half of the expected number of ET breeding events were removed to avoid potential biases. Heifer, cow, and sire conception rate evaluations were recalculated with this new data set according to the methods used for the official national evaluations. The edits removed about 1% of fertility records in the most recent 4 yr. Subsequent analysis showed that censoring herd years with inconsistent ET reporting had little effect on most bulls except for the highest ranking, younger bulls popular for ET use, and with largest effects on genomic selection. Improved ET reporting will be critical for providing accurate fertility evaluations, especially as the popularity of these advanced reproductive technologies continues to rise.

A Million-Cow Genome-Wide Association Study of Three Fertility Traits in U.S. Holstein Cows.

A genome-wide association study (GWAS) of the daughter pregnancy rate (DPR), cow conception rate (CCR), and heifer conception rate (HCR) using 1,001,374-1,194,736 first-lactation Holstein cows and 75,140-75,295 SNPs identified 7567, 3798, and 726 additive effects, as well as 22, 27, and 25 dominance effects for DPR, CCR, and HCR, respectively, with log10(1/p) > 8. Most of these effects were new effects, and some new effects were in or near genes known to affect reproduction including GNRHR, SHBG, and ESR1, and a gene cluster of pregnancy-associated glycoproteins. The confirmed effects included those in or near the SLC4A4-GC-NPFFR2 and AFF1 regions of Chr06 and the KALRN region of Chr01. Eleven SNPs in the CEBPG-PEPD-CHST8 region of Chr18, the AFF1-KLHL8 region of Chr06, and the CCDC14-KALRN region of Chr01 with sharply negative allelic effects and dominance values for the recessive homozygous genotypes were recommended for heifer culling. Two SNPs in and near the AGMO region of Chr04 that were sharply negative for HCR and age at first calving, but slightly positive for the yield traits could also be considered for heifer culling. The results from this study provided new evidence and understanding about the genetic variants and genome regions affecting the three fertility traits in U.S. Holstein cows.

Ovarian activity and reproductive responses of lactating Angus cows due to a mineral supplementation throughout a timed AI protocol.

Two experiments were conducted to evaluate the effect of intramuscular administration of minerals during a TAI program on the reproductive responses of lactating Angus cows. All cows (n=353) were subjected to a 9-day TAI program based on CIDR insertion plus injections of estradiol, cloprostenol, and eCG, and then TAI 48 h later. In experiment 1, two groups were randomly created, one control with a placebo injection (CON, n=109), and the second received 10 mL of Fosfosan® (MIN, n=172) on day 0 of the synchronization. Conception rate (66.9 vs. 55%) and estrus percentage (55.8 vs. 44%) were higher (P≤0.05) in MIN than in CON cows. Given these results, a second experiment was conducted randomly assigning the cows to two treatments (n=36 each): a single injection of 10 mL of Fosfosan® (MIN-O) on day 0 or two injections of 10 mL of Fosfosan® (MIN-T) on synchronization days 0 and 7. Four cows of each treatment were randomly selected to be scanned with transrectal ultrasound before and during the synchronization protocol to assess ovarian structures and cyclicity, and at day 39 post-TAI for pregnancy diagnosis. Also, blood samples were obtained for the determination of serum minerals and progesterone (P4) concentrations. The number of mineral injections did not affect conception rate (P≥0.1229) conception rate, serum mineral and P4 concentrations, number, and size of emerging follicles, or follicle size according to 1 to 4 classifications. The MIN-T promoted (P<0.05) earlier follicular wave emergence than MIN-O. However, MIN-O cows had a dominant follicle of 15.12 mm, which is more significant (P<0.05) than that in MIN-T cows (13.5 mm). In conclusion, providing a single mineral injection of Fosfosan® at the start of a TAI program is an excellent reproductive strategy in lactating Angus cows to improve the dominant follicle growth, estrus response, and conception rate.

Reproductive performance of Horro and Horro-crossbred dairy cows in Ethiopia's subhumid tropical environments.

Estrus detection, timing of insemination, feeding, and health care practices are all factors that can significantly affect cattle breeding efficiency and lifetime production. In Ethiopia's subhumid environment, the age at first calving, number of days open, duration from calving to first heat, and number of services per conception are economic traits that affect reproductive performance of Horro and Horro-crossbred dairy cows. Data collected at Ethiopia's Bako Agricultural Research Center between 1980-2019 was used to study the reproductive performance of Horro and Horro-crosses with Holstein Friesian and Jersey dairy cows. We identified overall mean ± standard error values of factors that affect reproductive performance, including age at first service (AFS) (29.2 ± 0.2 months), age at first calving (AFC) (39.8 ± 0.2 months), number of services per conception (NSC) (1.76 ± 0.4 months), calving interval (CI) (13.2 ± 0.3 months), days open (DO) (94.3 ± 4.3 days), conception rate (CR) (75.0 ± 1.3%), and replacement rate (RR) (28.4 ± 0.3%). The odds ratios of the 60-day and 90-day nonreturn rates (NRR) were 0.22 and 0.96, respectively. Sire and dam breed and birth period had significant (P < 0.001) effects on AFS and AFC, while season and dam parity had significant effects on CI and DO. Inconsistencies regarding feeding, heat detection, inseminator skills, insemination timing, animal health, and other husbandry practices may extend periods of AFS, AFC, CI, and DO. To optimize reproductive performance in Horro cows and Horro crossbreeds in Ethiopia's subhumid environments, improving management practices should be a priority.

No adverse effect of confirmation of ovulation by rectal palpation and ultrasonography after artificial insemination on formation, development, and function of the corpus luteum and conception rate in cows.

The effect of confirmation of ovulation by rectal palpation and ultrasonography after artificial insemination (AI) on the development of the corpus luteum (CL) and conception rate was investigated in cows. A total of 90 clinically healthy Holstein-Friesian dairy cows were examined in this study. After AI, the cows were divided into three groups (30 cows per group). In Group I, ovulation was confirmed by rectal palpation at 24 h after AI. In Group II, ovulation was confirmed using transrectal ultrasonography 24 h after AI. In Group III, ovulation was not confirmed after AI. Day 0 was defined as the day when ovulation was confirmed in Groups I and II, and as the day after AI was performed in group III. Transrectal ultrasonography was performed on days 3, 5, 7, and 14 to measure the CL diameter, tissue area, and CL blood flow area, and the ratio of CL blood flow area to CL tissue area was calculated. On the day of CL measurement, blood samples were collected to determine the plasma concentrations of progesterone (P4) and estradiol-17ß (E2). Pregnancy was diagnosed at 28 and 60 days after AI. A high conception rate of approximately 80% was achieved in Groups I and II, in which confirmation of ovulation was conducted. There were no differences in the diameter, tissue area, or blood flow area of the CL between the three groups. These results indicate that the confirmation of ovulation by rectal palpation and transrectal ultrasonography did not affect the formation and function of the CL or conception rate.

Environmental factors affecting the conception rates of nulliparous and primiparous dairy cattle.

Dairy cattle must allocate energy to milk production and reproduction. Therefore, understanding the environmental factors that affect conception rates in nulliparous and primiparous cows is helpful in appropriate feeding management strategies before and after calving. Accordingly, the aim of this study was to investigate the influence of environmental factors before and after the first calving on the conception rate, representing the starting point of milk production. The records of the first artificial insemination (AI) from Holstein nulliparous cows (n = 533,672) and primiparous cows (n = 516,710) in Hokkaido, Japan, were analyzed using separate multivariable logistic regression models. The mean conception rates for nulliparous and primiparous cows from 2012 to 2018 were 55.2 and 39.2%, respectively. In both nulliparous and primiparous cows, the conception rate of crossbreeding using Japanese Black (JB) semen was significantly higher than that for purebred Holstein breeding. The conception rate using sexed semen decreased in the warmer months only in nulliparous cows. Moreover, we grouped primiparous cows according to milk yield during peak lactation (PY; < 25, 25-30, 30-35, ≥35 kg) and the interval from calving to first insemination (CFI; < 60, 60-79, 80-99, ≥100 d), and evaluated their combined effect on the conception rate. Both PY and CFI strongly affected the conception rate in primiparous cows, which decreased with an increase in PY, even for the group with CFI ≥100 d; however, the conception rate increased for a CFI ≥60 d regardless of PY. Taken together, this study demonstrates the long-term effect of PY and an independent effect of CFI on the conception rate of cows. These results provide guidance for management to execute appropriate AI implementation strategies before and after lactation.

Mineral supplementation (injectable) improved reproductive performance in Holstein cows managed in a warm summer environment.

Sustainability of dairy production depends largely on reproductive efficiency that is affected by heat stress due to high ambient temperature and humidity during summer. Supplementation of minerals has been proposed as a management strategy to minimize adverse impact of heat stress on fertility. The objective of this study was to determine the effects of an injectable mineral supplement (Fosfosan) containing selenium (Se), copper (Cu), potassium (K), magnesium (Mg) and phosphorus (P) on the ovarian structures, reproductive hormones and conception rate of heat-stressed Holstein cows. Sixteen cows were assigned during summer to one of two treatments, a control group (CON; n = 8) and a mineral-supplemented group (SUP; n = 8). Ambient temperature and relative humidity records were collected and processed to estimate the temperature-humidity index (THI), which confirmed a heat-stressed environment during the study (avg. THI = 79.4). Cows were subjected to a fixed-time artificial insemination (FTAI) program using the CIDR-Synch synchronization protocol. Traits indicative of ovarian activity were recorded during and after this protocol, as well as serum concentrations of reproductive hormones. Pregnancy diagnosis was made 28 and 35 d after FTAI. A completely randomized block design with repeated measures over time was performed to study ovarian functional structures and its hormonal profiles. Correlations and regressions were estimated to study relationships between ovarian structures and related hormones. Mineral supplementation did not increase follicular diameter or follicular populations (p > .05), yet tended to increase corpus luteum diameter (p < .10), and it enhanced (p < .01) oestrogen and progesterone serum concentrations and improved (p < .05) cow's conception rate. Diameter of dominant follicles and corpus luteum was correlated (p < .05) with oestrogen and progesterone levels, respectively, but only in mineral-treated cows. Two additional dairy herds were evaluated to confirm that mineral supplementation improved conception rate during the heat stress period (objective 2). Cows from dairy 1 received FTAI during winter (n = 401) and summer (n = 240), whereas cows from dairy 2 were bred after natural detected oestrus during winter (n = 558) and summer (n = 314). Conception rates were higher (p < .05) in winter than summer and they improved (p < .05) with mineral supplementation, but only in cows managed during summer. In conclusion, supplementation of minerals enhanced hormonal secretion from ovarian structures and improved conception rate in Holstein cows exposed to summer heat stress.

Presynchronization with simultaneous administration of GnRH and prostaglandin F2α (PGF2α) 7 days prior to Ovsynch improves reproductive profile in Hariana zebu cow.

This study is aimed at assessing the impact of simultaneous administration of GnRH and prostaglandin F2α (PGF2α) 7 days prior to Ovsynch in Hariana cow. Two hundred cyclic cows (> 4 months postpartum) were assigned to control (n = 54) and pre-OV (n = 146). As per Ovsynch protocol, buserelin acetate (10 µg), cloprostenol (500 µg), and buserelin acetate (10 µg) were injected i.m. on days 0, 7, and 9, respectively, in cows irrespective of treatment. But in pre-OV cows, buserelin acetate (10 µg) and cloprostenol (500 µg) were also injected i.m. simultaneously 7 days prior to initiate the Ovsynch protocol. Artificial insemination was performed between 18 and 24 h after the 2nd GnRH of Ovsynch in both treatments. Ultrasonography and blood sampling for hormonal analysis were done on each day of treatment, on day of AI, and 12 days post-AI. Pre-OV treatment resulted to increased (45.20% vs 29.62%; P < 0.05) pregnancy outcomes and higher (P < 0.01) ovulation rate to first GnRH of Ovsynch than control. Cows showing complete luteolysis in response to PGF2α of Ovsynch were also higher (P < 0.05) in pre-OV than control. Greater (P < 0.05) synchronization rate was recorded in pre-OV than control (86.76% and 68.75%). The circulating concentrations of estradiol on day of AI and progesterone on day 12 post-AI were higher (P < 0.01) in cows diagnosed pregnant than non-pregnant in both control and pre-OV treatment. In conclusion, simultaneous administration of GnRH and PGF2α 7 days before Ovsynch improved the synchronization rate and luteal profile in terms of CL area and hence resulted in higher conception rate in Hariana zebu cow.

Follicular dynamics during the pre-ovulatory and subsequent first follicular wave stages affect the pregnancy outcome in Japanese Black cows.

The diameters of the pre-ovulatory follicles (PF) and the largest follicle during the subsequent first follicular wave (W1LF), and plasma estradiol-17ß (E2) concentrations were monitored on Days 0, 1, 3, 5, and 7 (ovulation = Day 1). Pregnancy was diagnosed on Day 30. Cows were classified into two groups according to the location of the dominant follicle ipsilateral (IG) or contralateral (CG) to the corpus luteum on Day 7. From Days 3 to 7, some follicles that had been determined as the subordinate in the previous examination exceeded the W1LF located in the opposite ovary in terms of the diameter. These were defined as switching (SW), whereas others were defined as non-switching (NSW). The diameter of PF was significantly smaller in pregnant (P) animals than in non-pregnant (NP) animals. The plasma E2 concentration on Day 0 was significantly higher in P animals than in NP animals and tended to be higher in NSW than in SW. In addition, plasma E2 concentrations around Days 3 to 7 tended to be higher in P animals of NSW than in NP animals of SW. The conception rates did not differ between IG and CG but were significantly higher in NSW than in SW. In the IG group, the conception rate tended to be higher in NSW than in SW.

Targeted sequencing reveals candidate causal variants for dairy bull subfertility.

Bull fertility is a key factor for successful reproductive performance in dairy cattle. Since the semen from a single bull can be used to inseminate hundreds of cows, one subfertile bull could have a major impact on herd reproductive efficiency. We have previously identified five genomic regions, located on BTA8 (72.2 Mb), BTA9 (43.7 Mb), BTA13 (60.2 Mb), BTA17 (63.3 Mb), and BTA27 (34.7 Mb), that show large dominance effects on bull fertility. Each of these regions explained about 5-8% of the observed differences in sire conception rate between Holstein bulls. Here, we aimed to identify candidate causal variants responsible for this variation using targeted sequencing (10 Mb per region). For each genomic region, two DNA pools were constructed from n ≈ 20 high-fertility and n ≈ 20 low-fertility Holstein bulls. The DNA-sequencing analysis included reads quality control (using FastQC), genome alignment (using BWA and ARS-UCD1.2), variant calling (using GATK) and variant annotation (using Ensembl). The sequencing depth per pool varied from 39× to 51×. We identified a set of nonsense mutations, missense mutations, and frameshift variants carried by low-fertility bulls. Notably, some of these variants were classified as strong candidate causal variants, i.e., mutations with deleterious effects located on genes exclusively/highly expressed in testis. Genes affected by these candidate causal variants include AK9, TTLL9, TCHP, and FOXN4. These results could aid in the development of novel genomic tools that allow early detection and culling of subfertile bull calves.

Evaluation of bull fertility in Italian Brown Swiss dairy cattle using cow field data.

Dairy bull fertility is traditionally evaluated using semen production and quality traits; however, these attributes explain only part of the differences observed in fertility among bulls. Alternatively, bull fertility can be directly evaluated using cow field data. The main objective of this study was to investigate bull fertility in the Italian Brown Swiss dairy cattle population using confirmed pregnancy records. The data set included a total of 397,926 breeding records from 1,228 bulls and 129,858 lactating cows between first and fifth lactation from 2000 to 2019. We first evaluated cow pregnancy success, including factors related to the bull under evaluation, such as bull age, bull inbreeding, and AI organization, and factors associated with the cow that receives the dose of semen, including herd-year-season, cow age, parity, and milk yield. We then estimated sire conception rate using only factors related to the bull. Model predictive ability was evaluated using 10-fold cross-validation with 10 replicates. Interestingly, our analyses revealed that there is a substantial variation in conception rate among Brown Swiss bulls, with more than 20% conception rate difference between high-fertility and low-fertility bulls. We also showed that the prediction of bull fertility is feasible as our cross-validation analyses achieved predictive correlations equal to 0.30 for sire conception rate. Improving reproduction performance is one of the major challenges of the dairy industry worldwide, and for this, it is essential to have accurate predictions of service sire fertility. This study represents the foundation for the development of novel tools that will allow dairy producers, breeders, and artificial insemination companies to make enhanced management and selection decisions on Brown Swiss male fertility.

Metabolomic differences of seminal plasma between boars with high and low average conception rates after artificial insemination.

Seminal plasma is a complex biological fluid containing many metabolites including amino acids, fructose, carbohydrates and lipids Metabolites play important roles in multiple biological processes, but details and significance of the seminal plasma metabolome related to boar fertility are unknown. The aim of the present study was to compare the comprehensive metabolome of seminal plasma from boars with different conception rate after artificial insemination and to identify the potential biomarkers. Semen samples were collected from boars which divided into two groups according to the conception rates in the offspring. Seminal plasma metabolites were isolated, purified, and then subjected to Ultra-high Performance Liquid Chromatography-Quadrupole Time-of-Flight Mass Spectrometry (UHPLC-qTOF-MS) procession. A total of 576 (Positive ion mode) and 377 (Negative ion mode) metabolites were identified in seminal plasma. Metabolites were identified and categorized according to their major chemical classes, including carboxylic acids and derivatives, organooxygen compounds, amino acids, peptides, and alogues, fatty amides, fatty acyls, benzene and substituted derivatives, purine nucleotides, pyrimidine nucleotides, glycosyl compounds, fatty acids and conjugates. The results showed that 4-Aminobenzoate, Pro-Asn, Ile-Tyr, Homoveratric acid and D-Biotin were higher in semen of boar with higher conception rate (HG) versus lower conception rate (LG) (p < .05), whereas L-Serine, Butoxyacetic acid, S-Methyl-5'-thioadenosine, Capsaicin and 1-O-(cis-9-Octadecenyl)-2-O-acetyl-sn-glycero-3-phosphocholine (PAF) were lower in HG than in LG (p < .05). These metabolites may be considered as candidate biomarkers for different fertility in boars.

Effects of age and season on conception rate of Mediterranean Italian Dairy Buffalo (Bubalus bubalis) following oestrus synchronization and fixed-time artificial insemination.

The objective of this project was to report the effects of season and age on conception rate amongst Italian Mediterranean Buffalo subjected to an Ovsynch/Resynch (O/R) reproductive management protocol. The study utilized nulliparous (heifers), primiparous buffalo cows (PBC) and multiparous buffalo cows (MBC). The primiparous and multiparous groups were subjected to the synchronization protocol throughout the entire year, but heifers were synchronized and inseminated only during the spring/summer seasons. The conception rate obtained following the OvSynch oestrus synchronization protocol, applied during spring, was 68.4% for heifers, 83.3% for PBC and 67.7% for MBC. The overall total conception rates following the complete O/R protocol were 84.27%, 94.4% and 79%. Conception rates achieved during summer were heifers 52%, PBC 47.2% and MBC 49%, whilst overall conception rates following the full O/R protocol were 72%, 69.8% and 58.2% respectively. In the autumn seasons, PBC conceived 58.9% and MBC 52.1% following initial Ovsynch, which improved to total overall conception rates of 87.5% and 78.7% following the full O/R protocol. Similarly, in the winter season, PBC experienced a conception rate of 47.5% following Ovsynch and 72.5% after a follow-up Resynch. MBC experienced 60.0% and 74.4% conception following Ovsynch and full O/R, respectively, during winter. Total conception rates during all seasons were quite acceptable following the O/R protocol. There was a significant decrease from spring to summer in conception rate for all parity groups, but heifers were not a severely affected as older buffalo cows. This finding agrees with that of other investigators indicating that heifer fertility is not as negatively impacted by long photoperiod and higher ambient temperature as that of older animals. The O/R protocol as utilized in this study is an effective means of reproductive management for dairy buffalo cows and is effective for improving fertility during out-of-season breeding.