Reproductive Performance and Culling Rate of Purebred Holstein Cows and Their Crosses With Fleckvieh and Brown Swiss Cows Under Subtropical Conditions

This study aimed to elucidate the reproductive performance of purebred Holstein (HO) cows with their crosses with Fleckvieh (FV) and Brown Swiss (BS) cows under subtropical conditions. A total of 677 cows [487 HO, 104 HO × FV (HFV); 50% FV and 50% HO and 86 HO × BS (HB); 50% BS and 50% HO] were enrolled in this study. Pure HO cows had significantly greater service per conception (S/C; 3.69), days open (147.9 days), and calving interval (449.6 days), than the HFV (2.89, 116.7, and 407.4 days, respectively) and HB (3.07, 134.3, and 434.2 days, respectively) crossbred cows. At day 28, the conception percentage was significantly greater among HFV crossbred cows vs. pure HO cows [crude odds ratios (COR) = 2.16], but embryonic loss, abortion percentage, calving difficulty, and retained placenta percentage were similar (p > 0.05) among pure HO cows and their crosses. HFV crossbreds had significantly lower incidence of endometritis (COR = 0.70, p = 0.035), mastitis (COR = 0.69, p = 0.015), and ketosis (COR = 0.53, p = 0.004) vs. other cows. HB and pure HO cows had a similar incidence of mastitis, lameness, and ketosis (COR = 0.76, 0.75, and 0.81; p = 0.223, 0.468, and 0.492, respectively). HFV crossbred cows had a lower risk of culling rate than HB crossbred cows. In summary, HFV cows demonstrated the best reproductive performance in terms of S/C, days open, calving interval, conception at 28 days, mastitis percentage, ketosis percentage, and endometritis.

Identification of novel candidate factor SNX5 specific for pregnancy failure in Holstein heifers

Pregnancy loss predominantly occurs during periods between blastocyst hatching and conceptus (embryo plus extraembryonic membranes) implantation to the endometrium in cattle. Insufficient biochemical communication between conceptus and endometrium has been suspected as the primary cause for early embryonic losses. If molecules regulating this communication were identified, molecular mechanisms associated with early pregnancy success or loss could be better understood. To identify novel factors as detection markers of non-pregnant or females undergoing embryonic loss, blood sera from embryo-transferred heifers on day 7 (day 0 = day of estrus) were collected on day 17, 20, or 22, which were subjected to metabolome and global proteome iTRAQ analyses. On each sample, the metabolome analysis partly divided serum components into pregnant or not. In the iTRAQ analysis, heatmap analysis with 25 unique proteins separated into pregnant or not on day 20 or 22. Furthermore, receiver operating characteristic curve (ROC) analysis identified five candidate proteins detecting non-pregnant heifers, of which SNX5 in day 22 sera had the highest area under the curve (AUC), 0.983. We also detected SNX5 in day 22 sera from non-pregnant heifers using western blotting. These results suggest that high SNX5 in day 22 sera could predict early pregnancy loss in heifers.

Effects of parity and body condition score (BCS) on oestrus expressions and post breeding reproductive performance of Katjang crossbred goat

This experiment goal was to identify the effect of parity and body condition score (BCS) on oestrusexpressions (OE), post breeding reproductive performance (PBRP) of Katjang crossbred andchanges of progesterone (P4) concentration. Twenty-four does were treated with 14 dayssynchronisation followed by visual observation of oestrus expressions (standing heat (SH),mounted by other buck but not standing (MBNS), vocalization (VC), tail swaging (TW), andsniffing of the genital of other buck (SGB)) and blood sampling twice daily up to 72 hours followingControl Internal Drug Release (CIDR) removal. PBRP data was recorded for evaluation whichincluded pregnancy rate (PR), abortion rate (AR), mortality rate (MR), litter size (LS), litter birthweight (LBW) and gestation length (GL). Result revealed that there was no significant effect(P>0.05) of parity and BCS on OE, PBRP and progesterone concentration except for birth weightwhich had been affected by parity. Based on observation of OE of SH, it was found that accuracyof pregnancy prediction was up to 50%. Analysis of OE established reliability of OE in wassequence from SH>SGB>TW>MBNS>VC and onset of oestrus began approximately 33h post toCIDR removal. Analysis of PBRP shown parity?2 had heavier birth weight in single litter size thanparity?3 which was associated with no MR. Progesterone concentration indicated all of the doeswere on oestrus and highly susceptible as early as 48 h post to CIDR removal. In conclusion, thisstudy demonstrated that both non-genetic factors; parity and BCS are very poor in determining theeffect of OE, PBRP and P4 concentration due to limitation of sample size and greater influences ofexternal factors such as weather, extreme environmental condition and farm management. It isrecommended to improve program in future by focusing on enhancing the fertilization rate andreducing embryonic loss for optimum conception rate.

Truncation of IFT80 causes early embryonic loss in cattle

Recessive alleles represent a substantial risk in populations that have undergone significant bottleneck events. Analyzing millions of genotypes and hundreds of sequenced bulls, we identified one pernicious recessive variant that achieved 1.66% frequency in the US Holstein population and confirmed loss of embryonic development via CRISPR-Cas9 editing. In the process, we identified the developmental significance of two protein domains of the IFT80 gene in the embryonic development of cattle.

Peripheral leucocyte molecular indicators of inflammation and oxidative stress are altered in dairy cows with embryonic loss

Objective of experiment was to determine whether oxidative stress (OS) and inflammation altered embryonic loss in dairy cows. Blood samples were collected at days 0, 16, 32 and 60 after timed (AI) from 200 Holstein cows to determine embryonic loss based on interferon-stimulated gene-15 (ISG15) mRNA expression (day 16) and ultrasound at day 32 and day 60. Leucocyte expressions of mRNA TLR2, TLR4, TNF-α, IL1B, IL10, STAT3 (inflammation), PTGS2, PTGES (prostaglandin synthesis), and PLA2G4A and ALOX5AP (eicosanoid metabolism) at days 0 and 16 were determined. Plasma redox status for antioxidant enzymatic activities of glutathione peroxidase (GPX), superoxide dismutase (SOD), total antioxidant capacity (TAC), and concentrations of malondialdehyde (MDA) were determined at days 0, 16, 32 and 60. All antioxidant-redox responses were beneficially significant in pregnant cows diagnosed pregnant at day16 and sustained pregnancy to day 60 compared to non-pregnant cows at day16 or pregnant at day16 and lost embryos by days 32 or 60. The leucocyte mRNA expressions of TLR2, TLR4, STAT 3, IL1B, PTGS2, PLA2G4A and ALOX5AP were greater and PTGES was lower at day16 in pregnant cows that lost embryos early (P < 0.05). In conclusion peripheral leucocyte molecular indicators of inflammation and plasma indicators of OS were altered in pregnant cows undergoing embryonic losses compared to cows with a sustained pregnancy.

Impact of trace mineral source on beef replacement heifer growth, reproductive development, and biomarkers of maternal recognition of pregnancy and embryo survival

Trace minerals are known to play important roles in early embryo development. The study objective was to determine effects of trace mineral source on heifer reproductive performance. Beef heifers (n = 129) were randomly assigned to one of two treatments. From weaning through breeding, all heifers were individually fed a basal diet supplemented with Co, Cu, Mn, and Zn either from organic sources (COMP; Cu, Mn, and Zn amino acid complexes and Co glucoheptonate; Availa-4, Zinpro Corporation, Eden Prairie, MN) or inorganic sources (INORG; Cu, Mn, and Zn hydroxychlorides; Intellibond C, M, and Z, Micronutrients, Indianapolis, IN) and Co as CoSO4. Blood samples and a reproductive tract score (RTS) were collected to determine pubertal status. All animals were synchronized and artificially inseminated (AI). Pregnancy status was determined by lymphocyte gene expression, circulating concentrations of pregnancy associated glycoproteins (PAG), and by transrectal ultrasonography after AI. Embryonic loss was defined as when a previously pregnant animal was subsequently diagnosed not pregnant. Data were analyzed using the MIXED procedure in SAS. Puberty (P = 0.44), pelvic area (P = 0.74), RTS (P = 0.49), and estrus expression (P = 0.82) were not influenced by treatment. There was no effect of treatment (P = 0.37) or treatment by time (P = 0.19) on pregnancy, but there was a tendency (P = 0.13) for decreased embryonic loss among COMP heifers (27 ± 6%) compared to INORG heifers (38 ± 6%). There was a treatment by pregnancy status by time interaction (P &lt; 0.01) on circulating PAG concentrations with PAG concentrations tending (P = 0.08) to be greater on d 25 among heifers in the COMP treatment compared to heifers in the INORG group. In summary, source of trace mineral did not affect puberty, RTS, pelvic area, or overall pregnancy success, but feeding complexed trace minerals tended to increase circulating PAG concentrations and embryo survival.

Pre-implantation exogenous progesterone and pregnancy in sheep: I. polyamines, nutrient transport, and progestamedins

Background Administration of exogenous progesterone (P4) to ewes during the pre-implantation period advances conceptus development and implantation. This study determined effects of exogenous P4 on transport of select nutrients and pathways that enhance conceptus development. Pregnant ewes (n = 38) were treated with either 25 mg P4 in 1 mL corn oil (P4, n = 18) or 1 mL corn oil alone (CO, n = 20) from day 1.5 through day 8 of pregnancy and hysterectomized on either day 9 or day 12 of pregnancy. Endometrial expression of genes encoding enzymes for synthesis of polyamines, transporters of glucose, arginine, and glycine, as well as progestamedins was determined by RT-qPCR. Results On day 12 of pregnancy, conceptuses from P4-treated ewes had elongated while those from CO-treated ewes were spherical. The mRNA expression of AZIN2, an arginine decarboxylase, was lower in endometria of P4-treated than CO-treated ewes on day 9 of pregnancy. Expression of FGF10, a progestamedin, was greater in endometria of CO and P4-treated ewes on day 12 of gestation in addition to P4-treated ewes necropsied on day 9 of gestation. Treatment with P4 down-regulated endometrial expression of amino acid transporter SLC1A4 on day 12 of pregnancy. Conclusions Results indicated that administration of exogenous P4 during the pre-implantation period advanced the expression of FGF10, which may accelerate proliferation of trophectoderm cells, but also was correlated with decreased expression of glycine and serine transporters and polyamine synthesis enzyme AZIN2. Further research with increased sample sizes may determine how differential expression affects endometrial functions and potentially embryonic loss.

The Effect of Gonadotropin Releasing Hormone Administration on Fertility and Embryonic Loss in Goats during the Anoestrus Period

Background: Following the induction of oestrus out of season in small ruminants, low fertility and variations in fertility rates are associated with embryonic losses. One of the main causes of embryonic loss is luteal dysfunction. Gonadotropin Releasing Hormone (GnRH) supports the luteal structure, and increasing progesterone levels may be beneficial in terms of promoting embryonic life. The main objective of the present study was to evaluate the efficacy of GnRH administration following an oestrus induction protocol in the anoestrus season for preventing embryonic loss in goats having failure to conceive during the season. Materials, Methods & Results: In the study, 106 Damascus goats aged 3-5 years and weighing 45-60 kg were used. The oestrus of 106 goats in the anoestrous group was stimulated with progesterone and pregnant mare serum gonadotropin (PMSG) treatment. Out of breeding season, goats were divided into the 4 following groups: GnRH0 (n = 27), GnRH7 (n = 26), GnRH0+7 (n = 27) and control (n = 26). In each goat, an intravaginal sponge (IS) containing 20 mg of fluorogestone acetate (FGA) was placed into the vagina and left for 9 days. With the withdrawal of the sponge, 550IU PMSG and 125 μg of d-cloprostenol were injected intramuscularly. Oestrus detection was made via teaser bucks for 3 days starting 24 h after withdrawal of the IS. Eighteen bucks known to be fertile were used for breeding. Goats in the oestrus period were mated via natural breeding. The GnRH analogue lecirelin was injected intramuscularly at breeding in the GnRH0 group, on day 7 post-breeding in the GnRH7 group, and both at breeding and on day 7 post-breeding in the GnRH0+7 group. No injections were given to the control group. Blood samples for progesterone measurement were taken by jugular vena puncturing on days 3, 6, 7, 10, 13, 16, and 19 after breeding from 10 randomly chosen goats in all groups. The goats with a level of > 3.5 ng/mL of progesterone on day 21 post-breeding were evaluated as pregnant. Pregnancy was also viewed on day 50 after breeding by real-time ultrasonography (USG) with a 5-7.5 MHz convex probe. The oestrus rate was 96.23% (102/106) in the goats. The rates of onset of oestrus between 36-48 h, 48-60 h and 60 h and beyond were 38.7% (41/106), 21.7% (23/106) and 35.8% (38/106), respectively. The total pregnancy rate was 35.8% (38/106). There were no statistically significant differences (P > 0.05) found for the pregnancy rate, embryonic death rate or progesterone concentration of the groups. However, serum progesterone levels were statistically different in the GnRH7 group compared with the control group (P < 0.05).Discussion: After synchronisation, various anti-luteolytic strategies can be used to support corpus luteum development and elevate progesterone concentration in the luteal phase to decrease embryonic loss and increase reproductive performance. Therefore, application of GnRH to support the luteal structure and to increase progesterone levels may be beneficial in terms of supporting embryonic life. The results showed that GnRH treatment on the day 7 post-breeding following oestrus induction, including FGA and PMSG, can increase serum progesterone levels in Damascus goats in the anoestrus period. However, following oestrus induction in the anoestrus period, it was seen that GnRH treatment at breeding or on day 7 after breeding did not have any positive effect on embryonic loss or reproductive performance. In conclusion, it was considered that this protocol could be implemented successfully, yielding a 35% pregnancy rate in Damascus goats in the anoestrus period, but embryonic loss must be deeply studied in detail.

374 Early embryonic loss is an important mechanism for maximizing litter size in the pig

Litter size of commercial dam lines of pigs has seen a significant increase over the last 20 years. Although the current large litter size of gilts and sows can be attributed to improved genetic selection and nutrition, much of the increase in total number born has occurred through the continued increase in ovulation rate over the last four decades. Ovulation rate has increased from 14 in the 80’s to today’s average of 20–25 in commercial dam lines. However, high ovulation rate not only causes increased competition between embryos (uterine crowding), but the number of potential piglets born can out strip the number available teats. In addition, fetal crowding can reduce fetal growth and comprise health and survivability of lighter weight piglets at birth. Since there is usually minimal embryo loss observed up to blastocyst formation and hatching, the majority (20 to 30%) of embryonic loss occurs during Days 10 to 30 of gestation, which encompasses a critical period of conceptus development and differentiation, pregnancy recognition signaling, and onset of implantation. This period of early embryo loss is actually essential for establishing adequate uterine space to support fetal survival for a large litter. The period of rapid conceptus elongation on Day 12 of pregnancy provides a key “selection point” for regulating potential litter size in the pig. Establishing the fundamental roles of conceptus- and uterine-secreted factors is an essential step to develop strategies to increase placental and fetal growth to improve overall health and survivability of piglets before and after birth. During the peri-implantation period of pregnancy, porcine conceptuses produce interleukin-1B2, estrogens, prostaglandins and interferons. CRISPR/Cas9 genomic engineering technology has provided a direct method to evaluate the role of key pig conceptus genes. Loss-of-function studies have increased our understanding of the multiplicity of uterine/conceptus factors that are involved with maintenance of pregnancy.