Prediction of pregnancy viability in bovine in vitro-produced embryos and recipient plasma with Fourier transform infrared spectroscopy

We analyzed embryo culture medium (CM) and recipient blood plasma using Fourier transform infrared (FTIR) metabolomics to predict pregnancy outcome. Individually cultured, in vitro-produced (IVP) blastocysts were transferred to recipients as fresh and vitrified-warmed. Spent CM and plasma samples were evaluated using FTIR. The discrimination capability of the classifiers was assessed for accuracy, sensitivity (pregnancy), specificity (nonpregnancy), and area under the receiver operator characteristic curve (AUC). Within all IVP fresh embryos (birth rate = 52%), high AUC were obtained at birth, especially with expanded blastocysts (CM: 0.80 ± 0.053; plasma: 0.89 ± 0.034). The AUC of vitrified IVP embryos (birth rate = 31%) were 0.607 ± 0.038 (CM, expanded blastocysts) and 0.672 ± 0.023 (plasma, all stages). Recipient plasma generally predicted pregnancy outcome better than did embryo CM. Embryos and recipients with improved pregnancy viability were identified, which could increase the economic benefit to the breeding industry.     

Economic and genetic performance of various combinations of in vitro-produced embryo transfers and artificial insemination in a dairy herd

The objective of this study was to find the optimal proportions of pregnancies from an in vitro-produced embryo transfer (IVP-ET) system and artificial insemination (AI) so that profitability is maximized over a range of prices for embryos and surplus dairy heifer calves. An existing stochastic, dynamic dairy model with genetic merits of 12 traits was adapted for scenarios where 0 to 100% of the eligible females in the herd were impregnated, in increments of 10%, using IVP-ET (ET0 to ET100, 11 scenarios). Oocytes were collected from the top donors selected for the trait lifetime net merit (NM$) and fertilized with sexed semen to produce IVP embryos. Due to their greater conception rates, first ranked were eligible heifer recipients based on lowest number of unsuccessful inseminations or embryo transfers, and then on age. Next, eligible cow recipients were ranked based on the greatest average estimated breeding values (EBV) of the traits cow conception rate and daughter pregnancy rate. Animals that were not recipients of IVP embryos received conventional semen through AI, except that the top 50% of heifers ranked for EBV of NM$ were inseminated with sexed semen for the first 2 AI. The economically optimal proportions of IVP-ET were determined using sensitivity analysis performed for 24 price sets involving 6 different selling prices of surplus dairy heifer calves at approximately 105 d of age and 4 different prices of IVP embryos. The model was run for 15 yr after the start of the IVP-ET program for each scenario. The mean ± standard error of true breeding values of NM$ of all cows in the herd in yr 15 was greater by $603 ± 2 per cow per year for ET100 when compared with ET0. The optimal proportion of IVP-ET ranged from ET100 (for surplus dairy heifer calves sold for ≥$300 along with an additional premium based on their EBV of NM$ and a ≤$100 embryo price) to as low as ET0 (surplus dairy heifer calves sold at $300 with a $200 embryo price). For the default assumptions, the profit/cow in yr 15 was greater by $337, $215, $116, and $69 compared with ET0 when embryo prices were $50, $100, $150, and $200. The optimal use of IVP-ET was 100, 100, 62, and 36% of all breedings for these embryo prices, respectively. At the input price of $165 for an IVP embryo, the difference in the net present value of yr 15 profit between ET40 (optimal scenario) and ET0 was $33 per cow. In conclusion, some use of IVP-ET was profitable for a wide range of IVP-ET prices and values of surplus dairy heifer calves.     

Recombinant bovine interferon-τ enhances in vitro development of bovine embryos by upregulating expression of connexin 43 and E-cadherin

Interferon-τ (IFNT), produced in ruminants by embryonic trophoblastic cells before implantation, is involved in the maternal recognition of pregnancy. It is a pleiotropic molecule that alters the synthesis of endometrial proteins and inhibits the proliferation of some cells. The present study investigated the effects of recombinant bovine IFNT on the development of early-stage bovine embryos and the molecular mechanism underlying this effect. This study demonstrated that expression of mRNA encoding type I IFN receptor subunits was detectable from d 4 to 8 in in vitro fertilized (IVF) bovine embryos. A considerable number of IVF (n = 1,941) and parthenogenetic activated (n = 1,552) bovine embryos demonstrated that supplementing the culture medium with IFNT (100 ng/mL) produced a greater percentage of blastocysts, and the total cell number within the resulting blastocysts was higher. In addition, IFNT upregulated the expression levels of both mRNA and protein for connexin 43 (GJA1) and E-cadherin (CDH1) and expression levels for granulocyte-macrophage colony-stimulating factor and insulin-like growth factor 2 mRNA but not for their proteins in d 8 embryos. However, IFNT inhibited mRNA expression for leukemia inhibitory factor (LIF), LIF receptor α, and the sodium/potassium-transporting ATPase subunit β-1. We concluded that IFNT promoted the development of bovine embryos by upregulating the expression of GJA1 and CDH1. Thus, supplementing embryo cultures or transfer medium with IFNT may stimulate embryo development and improve embryo transfer efficiency.     

Pregnancy and bovine somatotropin in nonlactating dairy cows: II. Endometrial gene expression related to maintenance of pregnancy

The objective was to evaluate the effects of pregnancy and bovine somatotropin (bST) on endometrial gene and protein expression related to maintenance of pregnancy in nonlactating dairy cows at d 17. In endometrial tissues, treatment with bST increased the steady state concentration of oxytocin receptor (OTR) mRNA; bST-treated cyclic (bST-C) cows had greater OTR mRNA than bST-treated pregnant (bST-P) cows. Estradiol receptor alpha (ERalpha) mRNA was reduced in bST-P cows compared with control P and C (no bST) cows. Western blotting revealed that pregnancy decreased the abundance of ERalpha protein, and bST stimulated an increase in ERalpha protein in C and P cows. Treatment with bST increased steady state concentrations of progesterone receptor (PR) mRNA. No differences were detected in steady state mRNA concentrations of prostaglandin H synthase-2 (PGHS-2), prostaglandin E synthase, and prostaglandin F synthase due to pregnancy or bST treatment. However, PGHS-2 protein was increased in response to pregnancy and bST treatment. Immunostaining indicated that P decreased ERalpha protein in luminal epithelium and increased PR protein in epithelial cells of the uterine glands. The PR protein response in the glands was less in bST-P cows than in P cows. In the stromal layer of the endometrium, bST decreased PR protein abundance in C and P cows. The PGHS-2 protein was localized exclusively in the luminal epithelium cells of endometrium and was increased in P cows. In conclusion, distinctly different mRNA and protein responses were detected between C and P cows related to prostaglandin biosynthesis, and bST-induced changes may potentially impact mechanisms associated with maintenance of pregnancy in nonlactating cows.     

Disruption of bovine oocytes and preimplantation embryos by urea and acidic pH

Feeding cattle diets high in degradable crude protein (CP) or in excess of requirements can reduce fertility and lower uterine pH. Objectives were to determine direct effects of urea and acidic pH during oocyte maturation and embryonic development. For experiment 1, oocytes were matured in medium containing 0, 5, 7.5, or 10 mM urea (0, 14, 21, or 28 mg/dl urea nitrogen, respectively). Cleavage rate was not reduced by any concentration of urea. However, the proportion of oocytes developing to the blastocyst stage at d 8 after insemination was reduced by 7.5 mM urea. In addition, the proportion of cleaved oocytes becoming blastocysts was decreased by 5 and 7.5 mM urea. For experiment 2, putative zygotes were collected -9 h after insemination and cultured in modified Potassium Simplex Optimized Medium (KSOM). Urea did not reduce the proportion of oocytes developing to the blastocyst stage, although 10 mM urea reduced cleavage rate slightly. For experiment 3, dimethadione (DMD), a weak nonmetabolizable acid, was used to decrease culture medium pH. Putative zygotes were cultured in modified KSOM containing 0, 10, 15, or 20 mM DMD for 8 d. DMD reduced cleavage rate at 15 and 20 mM and development to the blastocyst stage at all concentrations. Results support the idea that feeding diets rich in highly degradable CP compromises fertility through direct actions of urea on the oocyte and through diet-induced alterations in uterine pH.     

Pregnancy, bovine somatotropin, and dietary n-3 fatty acids in lactating dairy cows: II. Endometrial gene expression related to maintenance of pregnancy

The objectives were to examine the effects of bovine somatotropin (bST), pregnancy, and dietary fatty acids on expression of key endometrial genes and proteins regulating prostaglandin synthesis in lactating dairy cows. Two diets were fed, at about 17 d in milk (DIM), in which oil of whole cottonseed (control diet) was compared with calcium salts of fish oil-enriched lipid (FO). Ovulation was synchronized in cows with a presynchronization plus Ovsynch protocol and cows were inseminated artificially or not inseminated on d 0 (d 0 = time of synchronized ovulation; 77 ± 12 DIM). On d 0 and 11, cows received bST (500 mg) or no bST, and were slaughtered on d 17 to recover uterine secretions and endometrial tissue. Number of cows in the control diet: 5 bST-treated cyclic (bST-C), 5 non-bST-treated cyclic (no bST-C), 4 bST-treated pregnant (bST-P), and 5 non-bST-treated pregnant (no bST-P) cows and in the FO diet: 4 bST-treated FO-cyclic (bST-FO-C) and 5 non-bST-treated cyclic (no bST-FO-C) cows. The FO diet increased progesterone receptor (PR) mRNA, and treatment with bST increased PR mRNA concentration in endometrium of no bST-C, but not in no bST-FO-C or no bST-P cows. Concentrations of estrogen receptor-α (ERα) mRNA and protein, and oxytocin receptor (OTR) mRNA were decreased in no bST-P cows compared with no bST-C cows. Treatment with bST tended to increase OTR and ERα mRNA concentrations in cyclic cows fed control or FO diets. Immunohistochemistry demonstrated effects of bST, FO, and pregnancy on distributions of ERα and PR proteins in endometrium. Pregnancy and FO feeding decreased ERα abundance in luminal epithelium. Prostaglandin H synthase-2 (PGHS-2) protein was elevated in pregnant cows and localized to the luminal epithelium. Both FO and bST treatments reduced staining intensity of PGHS-2 protein. Concentrations of prostaglandin E synthase mRNA were elevated in either cyclic or pregnant cows in response to bST, whereas bST decreased prostaglandin F synthase mRNA in pregnant cows. Uterine lumen fluids had more PGF_2α and prostaglandin E₂ in pregnant than cyclic cows. Uterine lumen fluids of bST-P cows contained more prostaglandin E₂ than those from no bST-P cows. In summary, both pregnancy and bST altered endometrial gene expression, and cyclic cows responded differently to bST than pregnant cows. Feeding FO modulated PR, ERα, and PGHS-2 expression and distribution among endometrial cell types in a manner that may favor establishment and maintenance of pregnancy.     

Pregnancy, bovine somatotropin, and dietary n-3 fatty acids in lactating dairy cows: I. Ovarian, conceptus, and growth hormone-insulin-like growth factor system responses

The objective was to examine effects of bovine somatotropin (bST), pregnancy, and dietary fatty acids on reproductive responses in lactating dairy cows. Beginning at approximately 17 d in milk (DIM), a comparison was made of isoenergetic diets comprising supplementary lipids of whole cottonseed vs. calcium salts of fish oil enriched lipid (FO). Ovulation was synchronized in cows with a presynchronization plus Ovsynch protocol, and cows were inseminated artificially by appointment or not inseminated (d 0 = time of synchronized ovulation; 77 ± 12 DIM). On d 0 and 11, cows received bST (500 mg) or no bST. All cows were slaughtered on d 17. Number of cows in each group was as follows: control diet had 5 bST-treated cyclic (bST-C), 5 non-bST-treated cyclic (no bST-C), 4 bST-treated pregnant (bST-P), and 5 non-bST-treated pregnant (no bST-P) cows; and cyclic cows fed FO diet had 4 bST-treated (bST-FO) and 5 non-bST-treated cyclic (no bST-FO-C) cows. Feeding FO increased milk production, number of class 1 follicles (2 to 5 mm), and decreased insulin during the period before d 0 compared with control-fed cows. The bST increased milk production, pregnancy rate [83% (5/6) vs. 40% (4/10)], conceptus length (45 vs. 34 cm), and interferon-τ in the uterine luminal flushings (9.4 vs. 5.3 μg) with no effect on interferon-τ mRNA concentration in the conceptus. Treatment with bST increased plasma growth hormone (GH) and insulin-like growth factor (IGF)-I. Among control-fed cows (cyclic and pregnant), bST decreased progesterone concentration in plasma. Cows fed FO had less plasma insulin than control-fed cyclic cows, and FO altered the plasma GH (bST-FO > bST-C) and IGF-I (bST-C > bST-FO-C) responses to bST injections. Endometrial IGF-I mRNA was reduced in pregnant cows and tended to decrease in those fed FO. The IGF-II mRNA was increased in the endometrium of pregnant and bST-treated cows fed the control diet. Cows fed FO had increased concentrations of IGF-II mRNA, when bST was not injected. The insulin-like growth factor binding protein-2 (IGFBP-2) mRNA was increased in bST-P cows, whereas bST decreased the IGFBP-2 mRNA in all cyclic cows. In summary, bST and FO seemed to modulate reproductive responses that may be beneficial to the developing conceptus and pregnancy rate.     

The effects of conjugated linoleic acid isomers cis-9,trans-11 and trans-10,cis-12 on in vitro bovine embryo production and cryopreservation

Conjugated linoleic acid (CLA) isomers can affect the lipid profile and signaling of cells and thereby alter their function. A total of 5,700 bovine oocytes were used in a structured series of experiments to test the effects of CLA cis-9,trans-11 and CLA trans-10,cis-12 in vitro. In experiment 1, high doses of each CLA isomer during in vitro maturation (IVM) were compared with high or low doses during the entire in vitro culture (IVC) of parthenogenetic embryos. High doses of the CLA isomers ranged from 50 to 200 μM and low doses were 15 and 25 μM. In experiment 2, the low doses of each CLA isomer were tested during IVM/IVC on embryos produced by in vitro fertilization (IVF). Experiment 3 compared the effects of 15 μM doses of each CLA isomer during IVM or IVC of IVF embryos. In experiment 4, post-rewarming survival rates and blastomere counts were assessed for embryos supplemented with each CLA isomer during IVM or for 36 h before vitrification. In experiment 1, when either CLA isomer was provided only during IVM, we observed no effects on overall rates of maturation, cleavage, or blastocysts (92.2 ± 1.6%, 78.3 ± 4.1%, and 28.9 ± 5.1%, respectively). However, high doses of each CLA isomer, but not low doses, during the entire embryo culture period decreased blastocyst rates (5–20%) in a dose-dependent manner. Cleavage rates improved with 15 or 50 μM CLA trans-10,cis-12. Progesterone concentrations in maturation media were significantly increased by high doses of each CLA isomer compared with control, but low doses of CLA isomers had no effect. In experiment 2 with IVF embryos, low doses of each CLA isomer did not alter cleavage rates (average 84.9 ± 1.9%) and only 25 μM CLA trans-10,cis-12 during IVC reduced blastocyst rates below those of controls (25.5 ± 2.1 vs. 38.2 ± 2.3%). The lipid content of embryos was increased and relative expression of the BIRC5 (baculoviral IAP repeat containing 5) gene was depressed by CLA trans-10,cis-12. In experiment 3, low doses (15 μM) of each CLA isomer during IVC significantly reduced blastocyst rates (20.6 ± 2.4% and 27.7 ± 1.2% vs. 34.18 ± 1.2% for CLA trans-10,cis-12 and CLA cis-9,trans-11 compared with control, respectively) with less effect of each CLA during IVM. In experiment 4, adding 100 μM CLA cis-9,trans-11 during the final 36 h of culture resulted in a high survival rate after rewarming and culture, and the higher embryo blastomere count was comparable to that of control embryos not undergoing vitrification. In conclusion, supplementation with either CLA isomer did not improve embryo production, but inclusion of CLA cis-9,trans-11 before vitrification improved the quality of bovine IVF embryos after rewarming and culture.     

Effects of intravenous triacylglycerol emulsions on hepatic metabolism and blood metabolites in fasted dairy cows

The objective was to determine the effects of intravenous infusion of triacylglycerol (TAG) emulsions derived from different lipid sources on energy metabolism during a 4-d fast. Six nonpregnant, nonlactating multiparous Holstein cows were randomly assigned to treatments in a replicated 3 x 3 Latin Square design. Treatments included intravenous infusion of tallow, linseed oil, or fish oil emulsions at a rate of 0.54 g of TAG/kg of body weight per day; infusions were concurrent with a 4-d fast. The emulsions were administered for 20 to 30 min every 4 h throughout the 4-d fast. Cows were fed ad libitum for 24 d between the fast/infusion periods. Infusion of tallow, linseed oil, or fish oil emulsions increased plasma concentrations of palmitic acid, linolenic acid, and eicosapentaenoic and docosahexaenoic acids, respectively. Infusion of linseed oil emulsion decreased plasma TAG concentrations compared with tallow and fish oil treatments, which were similar. Infusion of the tallow emulsion resulted in the highest concentrations of plasma nonesterified fatty acid (NEFA), insulin, and glucose, whereas the infusion derived from linseed oil had the lowest NEFA and beta-hydroxybutyric acid concentrations. The different TAG emulsions had no effect on total or peroxisomal oxidation of [1-(14C)]oleic acid in liver homogenates. Liver TAG content increased 12.0, 7.8, and 14.1 microg/microg of DNA during the fast for tallow, linseed oil, and fish oil treatments, respectively; linseed oil was different from fish oil and tended to be different from tallow.     

Pregnancy and bovine somatotropin in nonlactating dairy cows: I. Ovarian, conceptus, and insulin-like growth factor system responses

Nonlactating dairy cows were used to examine effects of bovine somatotropin (bST) on components of the insulin-like growth factor (IGF) system. Estrus was synchronized in cows with a Presynch + Ovsynch protocol and timed AI (TAI; n = 55) or not TAI (cycling, C; n = 23) on d 0 (time of synchronized ovulation). On d 0 and 11, cows received bST (500 mg) or no bST, and were sacrificed on d 17. Pregnancy rates were less in bST cows (27.2%, 9 of 33) than in controls (63.6%; 14 of 22). In contrast, conceptuses were larger in bST-treated cows (39.2 +/- 4.8 cm) than in controls (20 +/- 4.3 cm). Total interferon-tau in uterine luminal flushings (ULF) was greater in bST-treated cows (7.15 > 2.36 microg). Number of class 2 follicles (6 to 9 mm) was less in bST-C cows on d 7 and 16. On d 17, corpus luteum (CL) weight tended to be greater in bST-treated cows. Concentrations of progesterone were greater after d 10 in C than in pregnant (P) cows. In the ULF, IGF-binding protein-3 was greater in bST-P cows than in pregnant cows. A tendency for an increase in IGF-I hormone concentrations in the ULF was detected on d 17 in bST-treated and cyclic cows. Endometrial mRNA for IGF-I, IGF-II, IGFBP-2, and IGFBP-3 increased in bST-C, but not in bST-P cows. Treatment with bST increased plasma concentrations of insulin, IGF-I, and growth hormone (GH). In conclusion, bST may have hyperstimulated plasma IGF-I and insulin to cause asynchrony between conceptus and uterus that was detrimental to pregnancy.     

Pregnancy, bovine somatotropin, and dietary n-3 fatty acids in lactating dairy cows: III. Fatty acid distribution

Our objective was to examine effects of exogenous bovine somatotropin (bST), pregnancy, and dietary fatty acids on fatty acid distribution in various tissues of lactating dairy cows. Two diets were fed, starting about 17 d in milk (DIM), in which oil of whole cottonseed (control diet) was compared with a calcium salt of fish oil-enriched lipid (FO; 1.9% of dietary DM). Starting at 44 ± 5 DIM, ovulation was synchronized with a presynchronization plus Ovsynch protocol (d 0 = time of synchronized ovulation). Some cows were inseminated (77 ± 12 DIM) to create a pregnant group. On d 0 and 11, cows received bST (500 mg) or no bST, and were killed on d 17 (94 ± 12 DIM). Number of cows in control group was 5 bST-treated cyclic (bST-C), 5 non-bST-treated cyclic (no bST-C), 4 bST-treated pregnant (bST-P), and 5 non-bST-treated pregnant (no bST-P) cows; and for the FO diet: 4 bST-treated (bST-FO-C) and 5 non-bST-treated cyclic (no bST-FO-C) cows. At slaughter, samples of endometrium, liver, muscle, s.c. adipose, internal adipose, and mammary gland were collected. Milk was collected at 75 ± 5 DIM. Gas chromatography was used to determine fatty acid percentages in tissues and milk fat. Endometrium from the cows fed FO had increased proportions of C20:5 and C22:6, whereas C20:4 was decreased. Injections of bST reduced both C18:2 and the n-6:n-3 ratio, but increased C22:6 in endometrium of cyclic control-fed, but not pregnant cows. In addition, FO decreased the n-6:n-3 ratio in all tissues and milk fat except for s.c. and internal adipose tissue. Cows fed FO also had increased C18:3, C20:5, and C22:6 in the liver and mammary tissue, and C18:3 and C22:6 were increased in the milk fat. The FO diet decreased the Δ⁹-desaturase index [(product of Δ⁹-desaturase]/(product of Δ⁹-desaturase + substrate of Δ⁹-desaturase]; DIX) in muscle and s.c. tissues, accompanied by an increase in saturated fatty acid (SFA) percentage. In addition, FO diet decreased DIX in the endometrium. In mammary and internal adipose tissues, bST increased DIX in cyclic control-fed cows, whereas bST decreased DIX in FO-fed cows, with no difference in the concentration of SFA and UNSFA. Cis-9, trans-11 conjugated linoleic acid was increased in milk fat, but decreased in the muscle and s.c. adipose tissue of FO-fed cows. The FO-enriched lipid, bST treatment, and early pregnancy can alter fatty acid percentages and distributions that may alter tissue functionality and functional nutrients of consumer products.     

Milking frequency, estradiol cypionate, and somatotropin influence lactation and reproduction in dairy cows

Our objectives were to determine lactational and reproductive outcomes in response to increased milking frequency (MF), injection of estradiol cypionate (ECP), and treatment with bovine somatotropin (bST). Lactating dairy cows (n = 144) were blocked by lactation number (1 vs. 2+) and assigned randomly to a 2 × 2 × 2 factorial experiment consisting of 8 treatment combinations: 1) MF consisting of 4× daily milking (4×) for the first 30 d in milk (DIM) vs. 2× daily milking (2×), with all cows milked 2× after 30 DIM; 2) 10 mg of ECP given postpartum at 8 ± 3 DIM versus controls that received ECP diluent (oil); and 3) biweekly bovine somatotropin (bST), starting sometime after 60 DIM, versus no bST. Ovulation before the first artificial insemination was synchronized by using Heatsynch (GnRH injection 7 d before PGF_2α followed in 24 h by ECP), and cows were artificially inseminated after detected estrus or at 48 h after ECP, whichever came first. Pregnancy was assessed by transrectal ultrasonography 28 to 30 d after artificial insemination. Daily yield and weekly components of milk were measured during the first 90 DIM. Intervals to first and second postpartum ovulation were unaffected by treatment, but cows were in estrus earlier after 2× (24 ± 4 d) than 4× (41 ± 4 d) daily MF, and sooner after ECP (25 ± 3 d) than after oil (39 ± 4 d) treatment. Pregnancy rates among 4× cows increased for ECP versus oil (52.8 vs. 27.8%) more than for cows with 2× MF treated with ECP versus oil (50.0 vs. 39.4%). Increased MF increased daily milk yields and energy-corrected milk yields during the first 30 DIM. Although milk yields were increased acutely by ECP during the 10 d after its injection, subsequent milk yields were decreased for ECP-treated cows previously milked 4× daily. Treatment with bST increased overall daily milk yields most in cows previously milked 2× daily and treated with oil and those milked 4× daily and treated with ECP. We concluded that early postpartum ECP injection increased pregnancy rates, but generally had detrimental effects on milk yields after 30 DIM for ECP-treated cows previously milked 4× daily, unless those cows also were treated with bST.     

Symposium review: Predicting pregnancy loss in dairy cattle

Several tools exist to diagnose pregnancy in dairy cattle. However, substantial pregnancy loss occurs within the first 60 d of gestation in cattle, and these losses have a profound adverse economic impact on the dairy and beef cattle industries. Detecting these impending pregnancy losses could offer producers an opportunity to reduce costs associated with this source of reproductive inefficiency. Several of the pregnancy diagnostic tools currently available and new technologies are being examined for their ability to predict pregnancies at risk for failing in early pregnancy. This review provides a synopsis of work undertaken recently to predict pregnancy losses in cattle. Currently, opportunities to predict pregnancy loss include (1) using transrectal ultrasonography to detect loss of the fetal heartbeat, floating debris within the placental fluids, and reductions in fetal size; (2) observing reductions in circulating progesterone concentrations; (3) detecting reductions in concentrations of circulating placental products; namely, pregnancy-associated glycoproteins and microRNAs; and (4) detecting reductions in the early pregnancy-dependent increase in interferon-stimulatory gene expression in peripheral blood leukocytes. An achievable goal may be to identify markers of embryo mortality so that researchers and clinicians can focus their efforts on developing intervention strategies for cows identified to be at risk for pregnancy failure.     

Epidemiologic and economic analyses of pregnancy loss attributable to mastitis in primiparous Holstein cows

The main objective of the study reported here was to examine the association between pregnancy loss (PL) and previous exposure to clinical or subclinical mastitis before breeding or during gestation in primiparous Holstein cows. A secondary objective was to estimate the cost of clinical mastitis during gestation, including that of PL attributable to mastitis in study cows. A total of 687 primiparous Holstein cows from 1 dairy farm were included in a matched case-control study. Study cows were declared pregnant via ultrasound on d 33 after timed artificial insemination (TAI). Case cows (n = 78) were those diagnosed as nonpregnant by rectal palpation on d 47 or 75 after TAI. Control cows were those confirmed as pregnant by rectal palpation on d 47 and 75 after TAI. Case cows were matched with eligible controls according to year of calving and calving-to-conception interval ±3 d. Cows were assigned to 1 of 3 groups: (1) cows not affected with clinical or subclinical mastitis; (2) cows affected with subclinical mastitis (Dairy Herd Improvement Association somatic cell score >4.5); and (3) cows affected with clinical mastitis during 2 exposure periods, 1 to 42 d before breeding or during gestation (1 to PL diagnosis day for case cows, and 1 to 75 d for control cows). Conditional logistic regression was used to model the odds of PL as a function of previous exposure to mastitis in study cows. Mastitis before breeding was not associated with PL. The odds of PL were 2.21 times greater in cows affected with clinical mastitis during gestation (95% confidence interval = 1.01, 4.83), compared with cows without mastitis, after controlling for breeding type and lameness. The cost of clinical mastitis during gestation was $149, which includes the cost ($27) of PL attributable to mastitis. In conclusion, this study provides evidence that clinical mastitis during gestation can cause PL in primiparous dairy cows leading to economic losses.     

孕期传统和新兴真菌毒素暴露的胚胎发育毒性及其生物监测的研究现状

真菌毒素是某些真菌菌种在特定条件下产生的有毒次级代谢产物。研究发现多种真菌毒素的联合暴露可引起胎儿发育迟缓、胚胎停育甚至流产等胚胎发育毒性。目前,尚没有关于孕期真菌毒素暴露对宫内胎儿胚胎发育毒性的影响和生物监测情况的系统报道。因此,为预防和降低孕期真菌毒素暴露带来的危害,本文梳理了孕期传统真菌毒素如黄曲霉毒素、脱氧雪腐镰刀菌烯醇、玉米赤霉烯酮和新兴真菌毒素如交链孢毒素、白僵菌素和恩镰孢菌素等暴露引起的胚胎发育毒性,以及以生物标志物如暴露标志物和效应标志物为基础的生物监测技术的研究现状。     

牛α-乳白蛋白-油酸复合物对肿瘤细胞能量代谢的影响

本研究以人肝癌细胞(HepG2)为对象,分为α-LA-OA处理组、牛-α乳白蛋白(α-LA)处理组和溶剂对照组,以细胞的糖酵解和线粒体有氧呼吸2个过程中胞外酸化率和细胞耗氧率的变化水平为衡量指标,评估α-LA-OA对肿瘤细胞能量代谢的影响.结果表明,α-LA-OA能够显著性降低人肝癌细胞的糖酵解能力以及线粒体有氧呼吸能...     

Presence and metabolism of endogenous androgenic–anabolic steroid hormones in meat-producing animals: a review

The presence and metabolism of endogenous steroid hormones in meat-producing animals has been the subject of much research over the past 40 years. While significant data are available, no comprehensive review has yet been performed. Species considered in this review are bovine, porcine, ovine, equine, caprine and cervine, while steroid hormones include the androgenic–anabolic steroids testosterone, nandrolone and boldenone, as well as their precursors and metabolites. Information on endogenous steroid hormone concentrations is primarily useful in two ways: (1) in relation to pathological versus ‘normal’ physiology and (2) in relation to the detection of the illegal abuse of these hormones in residue surveillance programmes. Since the major focus of this review is on the detection of steroids abuse in animal production, the information gathered to date is used to guide future research. A major deficiency in much of the existing published literature is the lack of standardization and formal validation of experimental approach. Key articles are cited that highlight the huge variation in reported steroid concentrations that can result when samples are analysed by different laboratories under different conditions. These deficiencies are in most cases so fundamental that it is difficult to make reliable comparisons between data sets and hence it is currently impossible to recommend definitive detection strategies. Standardization of the experimental approach would need to involve common experimental protocols and collaboratively validated analytical methods. In particular, standardization would need to cover everything from the demographic of the animal population studied, the method of sample collection and storage (especially the need to sample live versus slaughter sampling since the two methods of surveillance have very different requirements, particularly temporally), sample preparation technique (including mode of extraction, hydrolysis and derivatization), the end-point analytical detection technique, validation protocols, and the statistical methods applied to the resulting data. Although efforts are already underway (at HFL and LABERCA) to produce more definitive data and promote communication among the scientific community on this issue, the convening of a formal European Union working party is recommended.