Andrew Ponter

Does supplementing dairy cows with β-carotene during the dry period affect postpartum ovarian activity, progesterone, and cervical and uterine involution?

β-carotene is the main natural precursor of vitamin A and plays an important role in reproductive efficiency and immune function in dairy cows. The objective of this study was to investigate whether a supplement of β-carotene given during the dry period is able to 1) increase blood concentrations of β-carotene postpartum, 2) improve ovarian function and progesterone production, and 3) enhance uterine involution and uterine health. This study was conducted using 40 Holstein cows. On the day of drying-off, cows were allocated to one of two dietary treatments: control diet (C, n = 20) or control diet plus 1g/d β-carotene (BC, n = 20). The β-carotene supplement was given individually to the cows until calving. Blood samples were obtained regularly before and after calving from the cows to measure the concentrations of β-carotene. The diameters of the cervix and uterine horns were measured regularly using ultrasonography. Endometrial cytology samples were acquired from the cervix and uterus to determine uterine health. Milk samples were obtained three times per week for progesterone assay. Additional blood samples were taken on the day of calving, 7 and 21 days postpartum to determine the plasma concentrations of amino acids. Blood concentrations of β-carotene were not different before the start of the experiment (C, 3.03 ± 0.22 mg/L vs BC, 3.12 ± 0.22 mg/L, P > 0.05). Blood concentrations of β-carotene in the BC group peaked (7.45 ± 0.24 mg/L) 1 month after drying-off while the concentrations in the C group remained constant. β-carotene concentrations then decreased in both groups. The difference in blood concentrations of β-carotene between groups became significant 2 weeks after the start of the supplement until 2 weeks postpartum. There was no significant difference in the interval from calving to ovulation between groups (C, 27.8 ± 3.46 d vs BC, 35.8 ± 3.55 d, P > 0.05). The dietary supplement of β-carotene during the dry period had no effect on ovarian activity, progesterone production, cervix and uterine horn diameters. Plasma concentrations of hydroxyproline in the BC group were higher than in the C group on day 21 postpartum (BC, 20.8 ± 1.33 μmol/L vs C, 15.0 ± 1.33 μmol/L; P < 0.01). On day 28 postpartum the percentage of neutrophils in the BC group was lower than in the C group (cervical smear; C, 21.0 ± 3.22% vs BC, 9.7 ± 3.14%, P < 0.05 and uterine smear; C, 32.0 ± 3.86% vs BC, 20.9 ± 3.76%, P < 0.05). In the present experiment a dietary supplement of β-carotene had no effect on ovarian activity. However, due to effects of β-carotene on hydroxyproline profiles and their potential relationship with uterine function we speculate that uterine involution may have been more complete and that uterine inflammation may have been reduced in cows which received the β-carotene compared to controls.

Short-term effects of maternal feed restriction during pregnancy on goat kid morphology, metabolism, and behavior

Morphometric, metabolic, and behavioral modifications were studied in goat kids after maternal feed restriction during the last one-third of pregnancy. At birth, only kids from twin and triplet litters were studied [n=40 kids born to control dams (CONT) and n=38 born to restricted dams (REST)] and only males thereafter (n=13 CONT and 15 REST kids) until slaughter at 6 wk of age. Kids born to restricted goats had a smaller abdominal girth at birth (P<0.01) and tended to have a smaller body mass index (P=0.10) and a smaller density index (P=0.09) than kids born to CONT goats. Male REST kids had a lighter birth weight (P=0.03) than male CONT kids, but no differences (all P>0.10) were found for BW and morphometric measurements thereafter. Decreased NEFA concentrations suggested that male REST kids mobilized their body reserves less than CONT kids at birth (P<0.01). No modifications in drinking tests at 3 and 5 wk of age were observed, or in feeding behavior and emotional reactivity at 5 wk of age (all P>0.10). In conclusion, maternal feed restriction in the last one-third of pregnancy resulted in a decrease in birth size, but male kids rapidly caught up, and there were no changes in behavior, morphology, or metabolism during rearing.

Oocyte and embryo production and quality after OPU-IVF in dairy heifers given diets varying in their n-6/n-3 fatty acid ratio

Dietary fat supplementation can improve oocyte quality in ruminants. The influence of the type of dietary fat on the number and quality of oocytes collected by ovum pick-up and on the production of embryos in vitro was investigated in Holstein heifers. Heifers were given hay plus one of two dietary supplements for 42 days. The supplements were linseed (L, rich in linolenic acid, C18:3n-3, n = 9) or soya bean (S, rich in linoleic acid, C18:2n-6, n = 9). Oocytes were collected by ovum pick-up (OPU) for 6 wks (2 sessions/wk) and morphologic quality assessed. Half the oocytes were frozen and the other half was used to produce embryos. Blood samples were analyzed for: insulin, insulin-like growth factor-1, glucose, non-esterified fatty acids, β-hydroxy butyrate and urea and the proportions of fatty acids. Neither growth rate nor plasma hormone and metabolite concentrations were affected by dietary supplement. However, L significantly increased the proportion of plasma C18:3n-3 while S significantly increased the proportion of C18:2n-6(P < 0.001). Neither oocyte characteristics (number, their quality and number fertilized and cleaved per heifer per session) nor embryo characteristics (number and quality per heifer per session) and embryo development stages were affected by dietary treatment. Real-time RT-PCR was performed on immature and mature cumulus-oocyte complexes (COC). Prostaglandin E synthase-1 expression increased in L compared to S heifers. In conclusion, the type of fatty acid did not modify the numbers of oocytes and embryos produced by OPU-IVF and their quality in dairy heifers. Upregulation of prostaglandin E synthase-1 may ensure sufficient PGE(2) production for oocyte maturation even when its precursor is low.

Postpartum Variations of Plasma IGF and IGFBPs, Oocyte Production and Quality in Dairy Cows: Relationships With Parity and Subsequent Fertility

The aim of this study was to determine whether postpartum variations of plasma IGF-1 and IGFBP concentrations, oocyte production and quality were related to parity and subsequent conception rate in Holstein dairy cows. Holstein dairy cows [10 primiparous (PP) and 22 multiparous (MP)] were allotted in six batches and sampled once weekly between calving and oestrous synchronization treatment started at 71.2 ± 2.0 days postpartum. During the 3 weeks before treatment, ovum pick-up (OPU) was performed twice weekly. Oocytes were scored on a 4-point scale, and oocytes from OPU1, 3 and 5 were fertilized in vitro. Seventeen cows became pregnant after first and second AI and were considered as fertile (F), while the others were considered to be subfertile (SF). Logistic regression was carried out to investigate the relationships between repeated measurements and fertility including parity and batch effects in the models. Likelihood of fertility significantly increased when plasma urea and IGFBP-3 concentrations decreased and was higher in PP compared with MP cows. There was a trend for fertility to increase when plasma IGF-1 concentrations increased (p = 0.07). In vitro cleavage and development rates were similar between SF and F cows (46.4% and 28.3% in SF vs 55.0% and 22.1% in F). Parity had an effect on plasma IGF-1 concentrations (PP: 61.65 ± 2.67 vs MP: 41.63 ± 5.81 ng/ml, p < 0.001), mean number of follicles aspirated per session (PP: 5.7 ± 1.3 vs MP: 9.5 ± 0.8, p < 0.05) and fertility (PP: 8/10 = 80% vs MP: 9/22 = 41%, p < 0.05) but not on the number of oocytes recovered per session nor their quality. In conclusion, postpartum plasma urea and IGFBP-3 concentrations, but not oocyte production and quality before breeding, were related to subsequent conception rate in our experimental design. Parity had a significant effect on energy status, follicular growth and fertility and needs to be considered when investigating relationships between nutrition and reproduction.

Long-term consequences of feed restriction during late pregnancy in goats on feeding behavior and emotional reactivity of female offspring.

Feed restriction during pregnancy can have detrimental effects on offspring development both during the juvenile period and during adult life. Long-term effects of maternal feed restriction during the last third of pregnancy on growth, metabolism and behavior of female kids, with a focus on feeding behavior and emotional reactivity, were studied in goats. Female kids born to control (CONT, n=17) or born to feed restricted goats (REST, n=15) were artificially reared and monitored from birth to 24 months of age. Maternal feeding restriction globally reduced live weight (P<0.001) and body condition score (P=0.02) of REST compared to CONT offspring. Females from the REST group had a higher daily feed intake (P=0.05) and tended to eat more rapidly (P=0.09) than females from the CONT group at 12 months of age. One year later, REST goats still ate more than CONT goats (P=0.05). Glucose metabolism did not appear to be modified as no differences were observed in glucose or insulin responses to an intravenous glucose tolerance test. No differences in time budget were found at 12 months of age. However, the HPA axis response to an ACTH injection was greater in REST than in CONT goats: higher peak cortisol concentration (P=0.02) and a greater area under the curve were found (P=0.01) at 14 months of age. In conclusion, maternal feed restriction during late pregnancy modified both feeding behavior and the stress physiology of female offspring for up to 2 years of age. However, the changes observed were small.

Medium-term effects of repeated exposure to stray voltage on activity, stress physiology, and milk production and composition in dairy cows

The medium-term effects of permanent or random exposure to stray voltage applied to the water trough were evaluated on milk production and stress physiology in lactating dairy cows. Seventy-four Holstein cows were assigned during two 8-wk experimental periods to 1 of 3 treatments. The treatments were permanent exposure to voltage (PERM, 1.8 V, n=23) applied to the water trough, random exposure to voltage (RAND, 1.8 V, 36 h/wk, n=25), and no exposure to voltage (control, n=26). On the first day of voltage exposure, PERM cows had higher activity levels than control cows (9.8+/-2.70 vs. -2.3+/-2.74 14-s periods of movement/h). During the eighth week of exposure, RAND cows had higher activity levels than control cows (4.2+/-3.64 vs. -7.7+/-3.54 14-s periods of movement/h) and higher milk cortisol concentration than PERM cows (0.21+/-0.024 vs. 0.14+/-0.020 ng/mL). No differences were observed between treatments for cortisol response after an ACTH challenge during the seventh week of exposure. No effects of voltage exposure were observed on production traits and daily water intake. There was a transient decrease in milk yield on the second day of exposure in PERM cows (-1.4+/-0.74 kg) and on the third day of exposure in RAND cows (-3.5+/-1.03 kg) compared with control cows. In dairy cows, permanent or random exposure to stray voltage (1.8 V; 3.6 mA) could induce a transient stress response. Moreover, unpredictable voltage exposure could be considered a mild stressor, with slight modifications in stress physiology and activity but no impairment in production in the medium term.

Dietary propylene glycol and in vitro embryo production after ovum pick-up in heifers with different anti-Müllerian hormone profiles.

Rapid genetic improvement in cattle requires the production of high numbers of embryos of excellent quality. Increasing circulating insulin and/or glucose concentrations improves ovarian follicular growth, which may improve the response to superovulation. The measurement of anti-Müllerian hormone (AMH) can help predict an animals response to superovulation treatment. The aim of the present study was to investigate whether increasing circulating insulin concentrations, through propylene glycol (PG) drenches, could improve in vitro embryo production in oestrus-synchronised superovulated heifers with different AMH profiles. Holstein heifers were grouped according to pre-experimental AMH concentrations as low (L) or high (H). The PG drench increased circulating insulin and glucose concentrations and reduced β-hydroxybutyrate and urea concentrations compared with the control group. AMH was a good predictor of follicle and oocyte numbers at ovum pick-up (OPU), and of oocyte and embryo quality (AMH H>AMH L). PG in the AMH H group increased the number of follicles and blastocyst quality above that in the control group, but did not improve these parameters in the AMH L group. These results indicate that short-term oral PG supplementation modifies an animals metabolic milieu and is effective in improving in vitro embryo production, after superovulation-OPU, more markedly in heifers with high rather than low AMH concentrations.

Oral propylene glycol modifies follicular fluid and gene expression profiles in cumulus–oocyte complexes and embryos in feed-restricted heifers

Dietary supplementation with propylene glycol (PG) increases in vitro production of high-quality embryos in feed-restricted heifers. The aim of the present study was to evaluate the effects of PG in feed-restricted heifers on follicular fluid insulin and insulin-like growth factor (IGF) 1 concentrations, expression of IGF system genes in oocytes and cumulus cells and the expression of selected genes in blastocysts. Feed-restricted (R) heifers were drenched with water or PG during induced oestrous cycles (400mL of PG or water/drench, daily drenching at 1600 hours for the first 9 days of the oestrous cycle). Ovum pick-up (OPU) was performed after superovulation to produce in vitro embryos and without superovulation to recover oocytes, cumulus cells and follicular fluid. OPU was also performed in a control group (not feed restricted and no drenching). Follicular fluid IGF1 concentrations were reduced by R, and PG restored IGF1 concentrations to those seen in the control group. In cumulus cells, expression of IGF1, IGF1 receptor (IGF1R) and IGF binding protein 4 (IGFBP4) was decreased in the R group, and fully (IGF1 and IGF1R) or partially (IGFBP4) restored to control levels by PG. Blastocyst perilipin 2 (PLIN2; also known as adipophilin), Bcl-2-associated X protein (BAX), SCL2A1 (facilitated glucose/fructose transporter GLUT1), aquaporin 3 (AQP3), DNA (cytosine-5)-methyltransferase 3A (DNMT3A) and heat shock 70-kDa protein 9 (HSPA9B) expression were decreased in R heifers; PG restored the expression of the last four genes to control levels. In conclusion, these results suggest that, during follicular growth, PG exerts epigenetic regulatory effects on gene expression in blastocyst stage embryos.