Jin G. Gong

Impact of Dietary Fatty Acids on Oocyte Quality and Development in Lactating Dairy Cows

Abstract The purpose of this study was to examine the effects of level of rumen inert fatty acids on developmental competence of oocytes in lactating dairy cows. Estrous cycles were synchronized in 22 cows on a silage-based diet supplemented with either low (200 g/day) or high (800 g/day) fat. A total of 1051 oocytes were collected by ultrasound-guided ovum pickup (OPU) in seven sessions/cow at 3–4 day intervals. Oocytes were matured, fertilized, and cultured to the blastocyst stage in vitro. Embryo quality was assessed by differential staining of Day 8 blastocysts. The high-fat diet reduced numbers of small and medium follicles. There was no effect on the quality of oocytes (grades 1–4) or cleavage rate. However, high fat significantly improved blastocyst production from matured (P < 0.005) and cleaved (P < 0.05) oocytes. Blastocysts from the high-fat group had significantly more total, inner cell mass and trophectoderm cells than the low-fat group (P < 0.05). Regression analysis showed negative effects of milk yield (P < 0.001), dry matter intake (P < 0.001), metabolizable energy intake (P < 0.005), and starch intake (P < 0.001) on blastocyst production in the low-fat group but not in the high-fat group. Within the low-fat group, blastocyst production was negatively related to growth hormone (P < 0.05) and positively related to leptin (P < 0.05). The low-fat group had higher nonesterified fatty acids than the high-fat group (P < 0.05). In conclusion, higher milk yields were associated with reduced developmental potential of oocytes in cows given a low-fat diet. Provision of a high-fat diet buffered oocytes against these effects, resulting in significantly improved developmental potential.

Oocyte quality in lactating dairy cows fed on high levels of n-3 and n-6 fatty acids.

Different fatty acid (FA) sources are known to influence reproductive hormones in cattle, yet there is little information on how dietary FAs affect oocyte quality. Effects of three dietary sources of FAs (supplying predominantly palmitic and oleic, linoleic (n-6) or linolenic (n-3) acids) on developmental potential of oocytes were studied in lactating dairy cows. A total of 12 Holstein cows received three diets containing rumen inert fat (RIF), soyabean or linseed as the main FA source for three periods of 25 days in a Latin-square design. Within each period, oocytes were collected in four ovum pick-up sessions at 3-4 day intervals. FA profiles in plasma and milk reflected profiles of dietary FA sources, but major FAs in granulosa cells were not affected. Dietary FA source did not affect plasma concentrations of leptin, insulin, IGF1, GH, or amino acids. RIF led to a higher proportion of cleaved embryos than soya or linseed, but blastocyst yield and embryo quality were not affected. It is concluded that the ovary buffers oocytes against the effects of fluctuations in plasma n-3 and n-6 FAs, resulting in only modest effects on their developmental potential.

The effect of increased dietary intake on superovulatory response to FSH in heifers

We have previously shown that the number of ovarian follicles <4 mm in diameter can be increased by enhanced dietary intake in heifers. This study investigated the effect of the same dietary treatment on superovulatory response. The estrous cycles of 24 mature Hereford x Friesian heifers were synchronized by a standard progesterone plus prostaglandin protocol. The animals were fed with either 100% (group M, n = 12) or 200% (group 2M, n = 12) maintenance requirements for a 3-week period. Starting from day 4 of the synchronized estrous cycle, all the animals were superovulated using a standard 4-day FSH regime followed by an injection of GnRH analogue (GnRHa) to induce ovulation. Rectal ultrasound scanning was carried out to assess ovarian follicular populations at the start of FSH treatment and on the day of GnRHa injection, and to determine the number of corpora lutea 5 days after GnRHa injection. The body weight (BW) and body condition score (BCS) were recorded weekly and plasma samples were collected throughout the experimental period. There were no differences in either BW or BCS between two groups at the start of the experiment. The BW and BCS were maintained during the experiment in the group M, whilst animals in the group 2M showed a non-significant (P > 0.05) increase in BW and BCS. Circulating concentrations of insulin were significantly (P < 0.01) higher in heifers from the group 2M throughout the controlled feeding period. The group 2M had significantly (P < 0.05) more follicles 2-4 mm in diameter at the start of FSH treatment and more (P < 0.01) follicles >9 mm in diameter on the day of GnRHa injection, when compared with the group M. Similarly, 5 days after GnRHa injection there were significantly (P < 0.01) more corpora lutea in the group 2M (18.1+/-2.2) than in the group M (10.6+/-3.0). In addition, plasma progesterone concentrations following GnRHa injection were significantly (P < 0.01) higher in heifers from the group 2M. In conclusion, these results confirm that increased dietary intake can enhance the recruitment of ovarian follicles in heifers. This treatment may provide a valuable approach to improving superovulatory response in cattle.

Ultra-structural characteristics of bovine granulosa cells associated with maintenance of oestradiol production in vitro

We investigated whether the maintenance of oestradiol production by bovine granulosa cells (GC) in vitro was related to GC ultra-structure, and studied the effects of inclusion of serum as a cell attachment factor on oestradiol secretion, cell morphology and ultra-structure. Bovine granulosa cells from medium-sized follicles (4-8 mm diameter), in a serum-free (SF) culture system, maintained oestradiol production for 6 days, whereas oestradiol secretion by cells cultured in serum-coated (SC) wells declined rapidly with time, in culture. SF cells formed clumps consisting of two types of cells. Cells within clumps presented a phenotype similar to GC in vivo, being spherical, tightly joined by extensive gap junctions and interdigitated pseudopodia/microvilli, had abundant rough and smooth endoplasmic reticulum (ER) and mitochondria with trabecular cristae. In contrast, cells cultured in either SC wells or in the flattened base of cell clumps from SF cultures were enlarged, containing less rough ER, had fewer mitochondria (which tended to be round) and contained endosome-like structures, morphological characteristics suggestive of early luteinisation.

Nutrition, Metabolism, and Fertility in Dairy Cows : 3. Amino Acids and Ovarian Function

Plasma insulin concentrations influence resumption of ovarian activity in postpartum dairy cows, and plasma insulin can be manipulated by changing dietary starch and fat supply. The objective of this experiment was to investigate the role of dietary amino acids in altering peripheral metabolic hormones and ovarian function. Thirty-two cows were fed a standard diet from calving until 40 d in milk (DIM), and then 8 cows were transferred to each of 4 dietary treatments until 70 DIM. The 4 diets were designed to supply either low (diets 1 and 2) or high (diets 3 and 4) levels of metabolizable protein (MP), containing either low (diet 1 and 3) or high (diets 2 and 4) proportions of Leu. Leucine was manipulated with heat-treated lupins and corn gluten meal. Estrus was synchronized at 60 DIM. Between 60 and 70 DIM, energy intake and energy balance were similar among diet groups, although cows receiving high MP containing high Leu had a greater milk yield than other groups (means: 37.8, 37.1, 37.4, 39.4 +/- standard error 0.85 kg/d for diets 1 to 4, respectively). Interactions between MP and Leu were found for insulin, glucagon, and the ratio between them. Insulin was not affected by Leu in diets with low MP but was decreased by greater Leu in diets with high MP (means: 0.37, 0.32, 0.46, 0.39 +/- SE 0.031 ng/mL for diets 1 to 4, respectively). Glucagon was not affected by MP in diets with low Leu but was increased by greater MP in diets with high Leu (means: 92, 81, 88, 95 +/- SE 6.0 pg/mL for diets 1 to 4, respectively). For the low-MP treatments, the insulin-to-glucagon ratio was greater with high Leu; for the high-MP treatments, the insulin-to-glucagon ratio was greater with low Leu (means: 4.28, 5.42, 5.16, 4.22 +/- SE 0.456 for diets 1 to 4, respectively). There was no effect of MP or Leu on ovarian follicle numbers or reproductive hormones. Based on hormonal and ovarian responses, we conclude that altering metabolic hormones through manipulation of amino acid supply and balance is unlikely to have a significant effect on ovarian function in dairy cows.