The role of insulin in milk protein synthesis is unresolved in the bovine mammary gland. This study examined the potential role of insulin in the presence of two lactogenic hormones, hydrocortisone and prolactin, in milk protein synthesis. Insulin was shown to stimulate milk protein gene expression, casein synthesis and (14)C-lysine uptake in mammary explants from late pregnant cows. A global assessment of changes in gene expression in mammary explants in response to insulin was undertaken using Affymetrix microarray. The resulting data provided insight into the molecular mechanisms stimulated by insulin and showed that the hormone stimulated the expression of 28 genes directly involved in protein synthesis. These genes included the milk protein transcription factor, ELF5, translation factors, the folate metabolism genes, FOLR1 and MTHFR, as well as several genes encoding enzymes involved in catabolism of essential amino acids and biosynthesis of non-essential amino acids. These data show that insulin is not only essential for milk protein gene expression, but stimulates milk protein synthesis at multiple levels within bovine mammary epithelial cells.
The aim of this experiment was to quantify the milk production capacity of cows undergoing extended lactations while fed a pasture-based diet typical of those used in the seasonal-calving dairying systems of Victoria, Australia. One hundred twenty-five Holstein cows were randomly assigned to 1 of 5 groups. Breeding was progressively delayed after calving to enable management of the groups for lactation lengths of 10, 13, 16, 19, and 22 mo (equivalent to calving intervals of 12 to 24 mo). Cows were provided with a daily energy intake of at least 180 MJ of metabolizable energy/cow. This was supplied primarily by grazed pasture with supplementary cereal grain, pasture silage, and hay. Cows were dried off when milk volume fell below 30 kg/wk or when they reached 56 d before their expected calving date. Most cows (>96%) could lactate above this threshold for 16 mo, >80% for 19 mo, and >40% for 22 mo. There were negative relationships between lactation length and annual production of milk and milk solids (milk fat + protein), but losses were small until 16 mo. Annualized yields of milk solids were 497, 498, 495, 474, and 463 kg/cow for the 10, 13, 16, 19, and 22 mo groups, respectively. This reduction in annual production of milk solids with increasing lactation length was relatively less than for milk volume because during extended lactation, cows produced milk with higher concentrations of protein. Cows undergoing extended lactations also finished their lactations having gained more body weight and body condition than cows lactating for only 10 mo. The data showed that many cows on pasture-based diets were capable of lactating longer than the 10 mo that is standard for Victorian herds with seasonally concentrated calving patterns. Further, such extended lactations could be achieved with little penalty in terms of annual milk solids production.
Reduced oocyte quality has been associated with poor fertility of high-performance dairy cows during peak lactation, due to negative energy balance. We examined the role of nonesterified fatty acids (NEFAs), known to accumulate within follicular fluid during under- and overnutrition scenarios, in causing endoplasmic reticulum (ER) stress of in vitro maturated cattle cumulus-oocyte complexes (COCs). NEFA concentrations were: palmitic acid (150 μM), oleic acid (200 μM), and steric acid (75 μM). Abattoir-derived COCs were randomly matured for 24 h in the presence of NEFAs and/or an ER stress inhibitor, salubrinal. Total and hatched blastocyst yields were negatively impacted by NEFA treatment compared with controls, but this was reversed by salubrinal. ER stress markers, activating transcription factor 4 (Atf4) and heat shock protein 5 (Hspa5), but not Atf6, were significantly up-regulated by NEFA treatment within whole COCs but reversed by coincubation with salubrinal. Likewise, glucose uptake and lactate production, measured in spent medium samples, showed a similar pattern, suggesting that cumulus cell metabolism is sensitive to NEFAs via an ER stress-mediated process. In contrast, while mitochondrial DNA copy number was recovered in NEFA-treated oocytes, oocyte autofluorescence of the respiratory chain cofactor, FAD, was lower following NEFA treatment of COCs, and this was not reversed by salubrinal, suggesting the negative impact was via reduced mitochondrial function. These results reveal the significance of NEFA-induced ER stress on bovine COC developmental competence, revealing a potential therapeutic target for improving oocyte quality during peak lactation.
The aim of this study was to measure the effect of type of diet and level of energy intake on the performance of cows undergoing extended lactations. Ninety-six Holstein-Friesian cows that calved in July and August 2004 were assigned randomly to 1 of 8 groups each of 12 cows (including 4 primiparous cows). Two of the 8 groups were assigned to each of 4 treatments that varied in lactation length (300 or 670 d) and diet (3 diets: control, high, or full total mixed ration (TMR). The 4 treatments were 1) control 300: cows were managed for a 300-d lactation and grazed pasture supplemented with grain and forage to provide a minimum daily dietary intake of 160 MJ of ME/cow; 2) control 670: as for control 300 except that cows were managed for a 670-d lactation; 3) high 670: cows were managed for a 670-d lactation and pasture was supplemented with grain and forage to provide a minimum daily dietary intake of 180 MJ of ME/cow; 4) full TMR 670: cows were managed for a TMR system that included a high body condition score at calving with cows offered a TMR during a 670-d lactation. The TMR was initially offered ad libitum indoors until about 440 DIM when the amount of TMR offered was reduced by about 2 kg of DM/d to prevent excessive weight gain. The proportions of cows still milking at the end of a 670-d lactation were similar for the control and high dietary groups. The full TMR group had fewer cows milking at 600 DIM: 17 cows milking compared with 24 cows in the control 670 group and 22 cows in the high 670 group. For the period 1 to 670 DIM, increasing the energy level in the diet (control 670 vs. high 670) resulted in a similar yield of milk and a similar fat concentration in the milk, but greater yields of milk fat and protein and greater milk protein percentage of the milk. The full TMR 670 group produced greater yields of milk and milk components (fat, protein, and lactose) and also protein percentage in the milk than the other groups. The milk solids (fat + protein) ratio for the 3 extended-lactation groups, defined as production achieved during the 24-mo calving interval divided by 2 yr (annualized production) expressed as a ratio of that produced in the normal 12-mo calving interval, was not affected by increasing the level of grain in the pasture-based diets (0.93 vs. 0.90 for control and high diets, respectively), but decreased with the TMR diet (0.79). The control 670 group produced 7.1% less milk, but only 2.4% less milk solids than the control 300 group over the 2-yr period of the study. Combining our data with that from 2 earlier studies of extended lactation demonstrated that Holstein cows with a high proportion of Northern Hemisphere genes offered pasture-based diets could achieve a high milk solids ratio, a greater proportion of cows milking at drying-off, and lower body weight gain over the lactation.
Abstract. Synchronization programs have become standard components in the current breeding management of cows in the dairy herds of most dairy industries. Many are based on protocols that allow timed inseminations (TAI) so as to circumvent the practical difficulties associated with estrus detection. These difficulties are exacerbated in modern herds of high producing cows either because of increasing herd size in which individual animal monitoring is difficult and often subjective, or because small intensively managed herds are milked in robotic systems that minimize animal: staff interactions. Additional reasons arise from high producing cows having less obvious symptoms of estrus, partly because of housing systems combined with intensive feeding and milking, partly because of higher metabolic clearance rates of reproductive hormones like estradiol and partly because of the increasing prevalence of prolonged post-partum anestrus and reproductive tract pathology. The most recently developed programs include protocols for resynchronization following first or subsequent inseminations. These re-synchronization protocols may involve selected forms of hormonal intervention during the diestrous and pro-estrous periods following TAI, or following pregnancy diagnosis by ultrasound from 28 days after TAI. The latter form of re-synchronization has become increasingly important with the recognition that late embryonic/early foetal death has become a major factor compromising the reproductive performance of high producing Holstein cows in many dairy industries. Although cows detected in estrus without any hormonal treatment before insemination have higher conception rates than those inseminated following synchronization and TAI, the low detection rates combined with embryonic death means that intervals from calving to conception (days open) are usually less when synchronization programs have been successfully implemented. One of the significant factors affecting a program's success is the compliance rate that may sometimes be less than 70%. Almost all programs involve strategically timed injections of prostaglandin F2α (PGF) and gonadotropin releasing hormone (GnRH). Injections of an estradiol ester and progesterone supplementation per vaginum may be included in some programs. The basic program is the "Ovsynch" regimen. Numerous variations have been tested and developed. Many involve increasingly complex protocols that increase the risk of non-compliance, none has consistently achieved conception rates that exceed 40% and few have reduced the incidence of embryonic death. These synchronization programs are the best that are currently available. They have not been able to overcome the consequences of lowered fertility associated with high levels of milk yield, forms of nutrition and environmental factors like heat stress that have profound effects on the physiology and metabolism of the high producing dairy cow.
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