Postpartal ovarian activity of healthy cows and those affected by subclinical metabolic disorders

The postpartal ovarian activity was studied in healthy cows and in those affected by subclinical fatty liver disease and subclinical primary ketosis. Twenty-six dairy cows, 4–7 years old, with not less than 5000 l milk yield in the preceding lactation, were monitored for the following blood parameters: aspartate-amino-transferase (AST)/glutamicoxalacetic transaminase (GOT), albumin, total protein, total lipid, cholesterol, non-esterified fatty acid (NEFA) and triglyceride levels, glucose and ketones. The ratios albumin : total protein and NEFA : triglyceride were also calculated. Blood samples were collected once a week between days 255 and 260 of gestation and days 1–15 postpartum, and subsequently on the 30th, 45th and 60th days postpartum. Based on the biochemical parameters, the cows were assigned in retrospect to three groups: (1) healthy (n = 5), (2) subclinical fatty liver disease (n = 6), and (3) subclinical primary ketosis (n = 10). The postpartal ovarian activity was assessed by rectal palpation and by milk progesterone levels, determined by radioimmunoassay in samples collected 2–3 times weekly between days 5 and 70 postpartum.As early as 10–18 days postpartum, ovarian activity was evident in healthy cows. This was confirmed by the progesterone profiles indicating three regular ovarian cycles with normal hormone levels within 70 days postpartum. All five healthy cows were detected in estrus and only one failed to conceive after insemination.The six cows with subclinical fatty liver disease had no regular cycles and only a few small follicles were palpable on their ovaries. The milk progesterone fluctuated around the basal level (? 1.6 nmol/l). The first Graafian follicles appeared as late as days 30–35 postpartum and the luteal function commenced only between days 38 and 60, as indicated by palpation of corpora lutea and a concomitant rise in milk progesterone. The progesterone peaks were at the lower physiological limits characteristic of luteal function.The ovarian activity of the ten cows with subclinical primary ketosis was intermediate between that of healthy and fatty liver cows. The first follicles and corpora lutea were somewhat delayed, and the progesterone levels fluctuated considerably during the luteal phase. One cow in this group resumed ovarian inactivity during the 70-day postpartal observation period.     

Correlation between composition of the bacterial community and concentration of volatile fatty acids in the rumen during the transition period and ketosis in dairy cows

The transition period is a severe challenge to dairy cows. Glucose supply cannot meet demand and body fat is mobilized, potentially leading to negative energy balance (NEB), ketosis, or fatty liver. Propionate produces glucose by gluconeogenesis, which depends heavily on the number and species of microbes. In the present study, we analyzed the rumen microbiome composition of cows in the transition period, cows with ketosis, and nonperinatal cows by terminal restriction fragment length polymorphism (TRFLP) analysis of 16S rRNA genes and quantitative PCR. TRFLP analysis indicated that the quantity of Veillonellaceae organisms was reduced and that of Streptococcaceae organisms was increased in rumen samples from the transition period and ketosis groups, with the number of Lactobacillaceae organisms increased after calving. Quantitative PCR data suggested that the numbers of the main propionate-producing microbes, Megasphaera elsdenii and Selenomonas ruminantium, were decreased, while numbers of the main lactate-producing bacterium, Streptococcus bovis, were increased in the rumen of cows from the transition period and ketosis groups, with the number of Lactobacillus sp. organisms increased after calving. Volatile fatty acid (VFA) and glucose concentrations were decreased, but the lactic acid concentration was increased, in rumen samples from the transition period and ketosis groups. Our results indicate that the VFA concentration is significantly related to the numbers of Selenomonas ruminantium and Megasphaera elsdenii organisms in the rumen.     

Effects of a plant product consisting of green tea and curcuma extract on milk production and the expression of hepatic genes involved in endoplasmic stress response and inflammation in dairy cows

During the periparturient phase, cows are typically in an inflammation-like condition, and it has been proposed that inflammation associated with the induction of stress of the endoplasmic reticulum (ER) in the liver contributes to the development of fatty liver syndrome and ketosis. In the present study, the hypothesis that supplementation of dairy cows with a plant product consisting of green tea (95%) and curcuma extract (5%) rich in polyphenols attenuates inflammation and ER stress in the liver during early lactation was investigated. Twenty-seven cows were assigned to two groups, either a control group (n = 14) or a treatment group (n = 13). Both groups of cows received a total mixed ration, and the ration of the treatment group was supplemented with 0.175 g of the plant product per kg dry matter from week 3 prepartum to week 9 postpartum. Dry matter intake and energy balance during week 2 to week 9 postpartum were not different between the two groups. However, cows supplemented with the plant product had a greater amount of energy-corrected milk during week 2 to week 9 postpartum and lower concentrations of triacylglycerols and cholesterol in the liver in week 1 and week 3 postpartum than cows of the control group (p < 0.05). Cows supplemented with the plant product showed a trend towards a reduced mRNA concentration of haptoglobin (p < 0.10), while relative mRNA concentrations of eight genes of the unfolded protein response considered in the liver were not different between the two groups of cows. Relative hepatic mRNA concentration of fibroblast growth factor, a stress hormone induced by various stress conditions, was reduced at week 1 and week 3 postpartum in cows supplemented with the plant product (p < 0.05). Overall, the data of this study suggest that – although there were only minor effects on the occurrence of ER stress and inflammation – a supplementation of polyphenols might be useful to improve milk yield and prevent fatty liver syndrome in dairy cows.     

Effects of feed restriction and supplementary folic acid and vitamin B12 on immune cell functions and blood cell populations in dairy cows

Cows undergoing a negative energy balance (NEB) often experience a state of immunosuppression and are at greater risk of infectious diseases. The present study aimed to evaluate the impact of a folic acid and vitamin B₁₂ supplement and feed restriction on several immune parameters. Sixteen cows at 45 ± 3 days in milk were assigned to 8 blocks of 2 cows each according to each cow’s milk production in the previous week, and within each block, the cows randomly received weekly intramuscular injections of either saline or 320 mg of folic acid and 10 mg of vitamin B₁₂ for 5 weeks. During week 5, the cows were fed 75% of their ad libitum intake for 4 days. Blood samples were taken before the beginning of the experiment, just before feed restriction and after 3 days of feed restriction, in order to evaluate blood cell populations, the phagocytosis capacity and oxidative burst of polymorphonuclear leukocytes (PMNs), the proliferation of peripheral blood mononuclear cells (PBMCs) and concentrations of non-esterified fatty acids (NEFAs) and β-hydroxybutyrate. The vitamin supplement did not affect any of the tested variables except milk fat and lactose content. Feed restriction reduced milk production and increased the concentration of NEFAs. Feed restriction did not affect blood cell populations but did reduce the percentage of PMN positive for oxidative burst after stimulation with phorbol 12-myristate 13-acetate. The proliferation of PBMCs was reduced when the cell culture medium was supplemented with sera collected during the feed restriction. In conclusion, feed restriction affected the functions of PMN and PBMC and this effect was not prevented by the folic acid and vitamin B₁₂ supplement. These results support the hypothesis that the greater risk of infectious diseases in cows experiencing a NEB is related to impaired immune cell functions by high circulating concentration of NEFAs.     

Concentration of the mammalian lignans enterolactone and enterodiol in milk of cows fed diets containing different concentrations of whole flaxseed

A total of 32 lactating Holstein cows with mean body weight of 622 kg (s.e. = 24) were allotted, at week 25 of lactation, to eight groups of four cows blocked for similar days in milk. The objective of the experiment was to determine the effect of feeding four dietary concentrations (0, 50, 100 or 150 g/kg of dry matter) of whole flaxseed, which contains the plant lignan precursor secoisolariciresinol diglucoside (SDG), on concentrations of two mammalian lignans (enterodiol and enterolactone) in milk. The effects of the four diets on feed intake, milk production, milk composition and digestion were also studied. Cows within each block were assigned to one of the four isonitrogenous and isoenergetic total mixed diets and the experiment was carried out from week 25 to 29 of lactation. Diets were fed for ad libitum intake. Enterolactone was the mammalian lignan, of the two metabolites studied, detected in the milk of cows and its concentration in milk tended (P = 0.08) to increase linearly with higher intake of SDG in the diet. Feed intake, milk yield and milk composition were similar among diets. Milk fatty acid profile was slightly improved by feeding flaxseed, as shown by higher concentrations of fatty acids (e.g. n-3) recognized as being beneficial for human health. Those results suggest that feeding of whole flaxseed may result in changes in milk fatty acid composition and enterolactone content, which offer benefits for consumers.     

Dietary supplementation with safflower seeds differing in fatty acid composition differentially influences serum concentrations of prostaglandin F metabolite in postpartum beef cows

Synthesis and secretion of prostaglandin F2alpha (PGF2alpha) is elevated following parturition and exerts divergent effects on the re-establishment of fertile estrous cycles in cows. The objective of these experiments was to determine if oil seed supplements differing in fatty acid composition differentially influence serum concentrations of the specific PGF2alpha metabolite, PGFM. Safflower seed supplements were formulated to provide 5% of dry-matter intake as fat. In Trial 1, 24 multiparous beef cows were individually fed control (beet pulp-soybean meal) or cracked high-linoleate safflower seed (78% 18:2n-6) supplements for 80 d postpartum. Linoleate supplemented cows had greater (P < 0.001) serum concentrations of PGFM than control cows. In Trial 2, primiparous beef cows (n = 36) were individually fed control (cracked corn-soybean meal), cracked high-linoleate (76% 18:2n-6) or -oleate (72% 18:1n-9) safflower seed supplements for 92 d postpartum. As in Trial 1, serum concentrations of PGFM were greater (P < or = 0.04) in linoleate than control or oleate supplemented cows. Serum concentrations of PGFM, however, did not differ (P = 0.40) among oleate and control supplemented cows. Although potential impacts on reproductive performance remain to be proven, dietary oil supplements high in linoleate, but not oleate, increased serum concentrations of PGFM compared to control supplements.     

High- and low-purity glycerine supplementation to dairy cows in early lactation: effects on silage intake,milk production and metabolism

This study evaluated the effects of supplemental low- and high-purity glycerine on silage intake, milk yield and composition, plasma metabolites and body condition score (BCS) in dairy cows. A total of 42 cows of the Swedish Red Breed, housed in individual tie stalls, were fed 0.25 kg of low- or high-purity glycerine on top of concentrate, twice daily, during the first 4 weeks of lactation. One-third of the cows acted as controls, receiving no glycerine. Silage was fed for ad libitum intake and concentrate was fed at restricted level of intake, about 6 kg/day for primiparous cows and 7 kg/day for multiparous cows. Feed refusals were weighed daily. Cows were milked twice daily, milk yield was recorded on four occasions per week and milk samples were collected simultaneously. Blood samples were drawn from the coccygeal vessel once a week. Low- and high-purity glycerine had no effect on silage or total dry matter intake (P = 0.38 and P = 0.75, respectively) or on BCS (P = 0.45). Cows fed high-purity glycerine tended to have higher milk yield than control cows (P = 0.06). Milk composition tended to differ among treatments. No main effects of treatment on concentration of glycerine (P = 0.44), glucose (P = 0.78), insulin (P = 0.33), non-esterified fatty acids (P = 0.33) and β-hydroxybutyrate (P = 0.15) in plasma. These data indicate that high-purity glycerine has the potential to increase milk yield, as well as enhance the milk protein concentration and milk fat + protein yield.     

Effects of feeding dry propylene glycol to early postpartum Holstein dairy cows on production and blood parameters

In all, 18 multiparous and 19 primiparous Holstein dairy cows were used in a completely randomized design with restrictions to evaluate the effects of feeding propylene glycol (PG) as a dry product, via two delivery methods, on production and blood parameters. PG treatments were administered from parturition through 21 days postpartum. Treatments were: (i) control, no PG; (ii) top dress, 162.5 g PG/day by top dressing onto the total mixed ration (TMR) and; (iii) mixing, 162.5 g PG/day as a part of the TMR by incorporating it into the TMR. PG used was a dry product which contained 65% pure PG and 35% silicon dioxide as the dry carrier. Coccygeal blood was sampled on 4, 7, 14 and 21 days in milk (±1.50 pooled s.d.). Supplementation of dry PG by top dressing onto, or incorporating into, the TMR had no effects on average dry matter intake, milk yield and composition, serum insulin, serum and plasma metabolites and milk ketones. Concentrations of urine ketones tended (P = 0.10) to be reduced by PG supplementation from 41.5 to 15.2 mg/dl. Supplementation of PG tended (P = 0.07) to decrease the incidence for subclinical ketosis from 39% to 24% and 13% for cows fed a TMR supplemented with no dry PG, with dry PG as a top dress and dry PG as a part of the TMR, respectively. It is concluded that supplementing PG as a dry product via incorporating into the TMR is as effective as when used as a top dress, based on the efficacies of both delivery methods to numerically reduce urine ketones concentrations and, therefore, the incidence for subclinical ketosis during the first 21 days of lactation. However, it should be noted that the number of cows used in the current study was minimal, and more cows are needed to confirm the efficacy of supplementing PG as a dry product on reducing the prevalence of subclinical ketosis in dairy cows during the first month of lactation.     

Transition Diseases in Grazing Dairy Cows Are Related to Serum Cholesterol and Other Analytes

The objectives of this study were to describe the incidence of postpartum disease and to evaluate the association with serum cholesterol concentrations during the first 3 weeks after calving in grazing dairy cows. The association between non-esterified fatty acids (NEFA), β-hydroxybutyrate (BHBA), calcium and postpartum diseases was also evaluated. A total of 307 Holstein dairy cows from 6 commercial grazing herds in Osorno, Chile, were monitored from calving until 21 days in milk. Cases of retained placenta, clinical hypocalcemia and clinical mastitis were recorded by the farmer using established definitions. Twice weekly, cows were evaluated for metritis by the same veterinarian based on vaginal discharge and body temperature. Postpartum blood samples were collected weekly and analyzed for serum concentrations of cholesterol, NEFA, BHBA and calcium. Cows were considered as having subclinical ketosis if BHBA >1.2 mmol/L, and subclinical hypocalcemia if calcium <2.0 mmol/L in any of the 3 weekly samples. Overall, 56% of the cows studied developed at least one clinical or subclinical disease after calving. Incidence of individual diseases was 8.8% for retained placenta, 4.2% for clinical hypocalcemia, 11.7% for clinical mastitis, 41.1% for metritis, 19.9% for subclinical hypocalcemia and 16.6% for subclinical ketosis. Lower postpartum cholesterol in cows was associated with developing severe metritis or having more than one clinical disease after calving. For every 0.4 mmol/L decrease in serum cholesterol cows were nearly twice as likely to be diagnosed with multiple clinical diseases after calving. Higher BHBA concentrations and lower calcium concentrations during week 1 were associated with severe cases of metritis. Low serum calcium concentration during week 1 was also associated with developing more than one clinical disorder after calving. In conclusion, the incidence of postpartum diseases can be high even in grazing herds and lower serum cholesterol concentrations were associated with occurrence of clinical postpatum disorders.     

White adipose tissue genome wide-expression profiling and adipocyte metabolic functions after soy protein consumption in rats

Obesity is associated with an increase in adipose tissue mass due to an imbalance between high dietary energy intake and low physical activity; however, the type of dietary protein may contribute to its development. The aim of the present work was to study the effect of soy protein versus casein on white adipose tissue genome profiling, and the metabolic functions of adipocytes in rats with diet-induced obesity. The results showed that rats fed a Soy Protein High-Fat (Soy HF) diet gained less weight and had lower serum leptin concentration than rats fed a Casein High-Fat (Cas HF) diet, despite similar energy intake. Histological studies indicated that rats fed the Soy HF diet had significantly smaller adipocytes than those fed the Cas HF diet, and this was associated with a lower triglyceride/DNA content. Fatty acid synthesis in isolated adipocytes was reduced by the amount of fat consumed but not by the type of protein ingested. Expression of genes of fatty acid oxidation increased in adipose tissue of rats fed Soy diets; microarray analysis revealed that Soy protein consumption modified the expression of 90 genes involved in metabolic functions and inflammatory response in adipose tissue. Network analysis showed that the expression of leptin was regulated by the type of dietary protein and it was identified as a central regulator of the expression of lipid metabolism genes in adipose tissue. Thus, soy maintains the size and metabolic functions of adipose tissue through biochemical adaptations, adipokine secretion, and global changes in gene expression.     

Effects of body condition score on direct and indirect measurements of insulin sensitivity in periparturient dairy cows

Reductions in insulin sensitivity in periparturient dairy cows develop as a means to support lactation; however, excessive mobilization of fatty acids (FA) increases the risk for peripartal metabolic disorders. Our objectives were to investigate the effect of prepartum body condition score (BCS) on systemic glucose and insulin tolerance, and to compare direct and indirect measurements of insulin sensitivity in peripartal lean and overweight dairy cows. Fourteen multiparous Holstein cows were allocated into two groups according to their BCS at day −28 prepartum: lean (n = 7; BCS ≤ 3.0) or overweight; (n = 7; BCS ≥ 4.0). Liver biopsies were performed on day −27, −14 and 4, relative to expected parturition. Intravenous insulin or glucose tolerances tests were performed following each liver biopsy. Relative to lean cows, overweight cows exhibited lower dry matter intake, lost more BCS and displayed increased plasma FA and β-hydroxybutyrate concentrations and elevated liver lipid content during peripartum. Glucose clearance rate was lower for all cows postpartum. Prepartum BCS had minimal effects on insulin and glucose tolerance; however, the ability of the cow to restore blood glucose levels following an insulin challenge was suppressed by increased BCS. Glucose-dependent parameters of insulin and glucose tolerance were not correlated with surrogate indices of insulin sensitivity. We conclude that prepartum BCS had minimal effect on systemic insulin sensitivity following parturition. The observed inconsistency between surrogate indices of insulin sensitivity and direct measurements of insulin and glucose tolerance adds support to growing concerns regarding their usefulness as tools to estimate systemic insulin action in periparturient cows.     

Diets enriched in unsaturated fatty acids enhance early embryonic development in lactating Holstein cows

We hypothesized that a diet enriched in alpha-linolenic acid would enhance embryonic development relative to diets enriched in linoleic or saturated fatty acids. Twenty-four lactating Holstein cows (86+/-22 d postpartum) were assigned to one of three diets containing saturated fatty acids (SAT; high in palmitic and stearic acids), whole flaxseed (FLX; high in alpha-linolenic acid) or sunflower seed (SUN; high in linoleic acid). Rations were formulated to provide 750 g supplemental fat/cow/d in all dietary groups. Ovulation (Day 0) was synchronized approximately 20 d after diets began. Ultrasound-guided follicular ablation of all follicles >8 mm was performed 5 d after ovulation; super stimulatory treatments began 2 d after follicular ablation, and embryos were collected non-surgically 7 d after AI. Fertilization rate, numbers of follicles and ovulations, and total and transferable embryos did not differ (P>0.05) among dietary groups. Sixty-one transferable embryos were stained and total blastomere number determined. Blastomere number was affected by diet (P<0.01); without regard to stage of development, embryos collected from cows fed SAT had lower (P<0.01) blastomere numbers (mean+/-S.E.M.; 77.1+/-3.9) than those from cows fed FLX (93.4+/-3.3) or SUN (97.2+/-3.5). Differences were most evident in the expanded blastocyst stage; at this stage, embryos of cows fed FLX and SUN diets had more blastomeres (P<0.02) than those of cows fed SAT (115.4+/-6.3, 132.3+/-8.3, and 89.3+/-9.6 cells, respectively). Although our hypothesis was only partially supported, embryonic development was enhanced in Holstein cows fed unsaturated fatty acids compared to those fed saturated fatty acids.     

Effects of dietary energy source on energy balance, metabolites and reproduction variables in dairy cows in early lactation

This paper summarizes three recent studies by the same authors with the objective to study the effect of dietary energy source on the energy balance (EB) and risk for metabolic and reproductive disorders in dairy cows in early lactation. The first study, a literature survey, illustrated that feeding extra glucogenic nutrients relative to lipogenic nutrients, decreased milk fat and seems to decrease plasma non-esterified fatty acid (NEFA) and beta-hydroxybutyrate (BHBA) concentration. However studies are scarce and mostly confound the effect of energy source with level of energy intake, compromising eventual effects on EB and fertility. Therefore, in the second study, 16 dairy cows were either fed a glucogenic or a lipogenic diet (isocaloric) and EB was determined in climate-controlled respiration chambers from week 2 until 9 of lactation. Glucogenic diet decreased milk fat yield and milk energy and tended to decrease body fat mobilization compared with lipogenic diet. The objective of the third study was to study the effect of dietary energy source on EB, metabolites and reproduction variables. Dairy cows (n=111) were fed glucogenic, lipogenic or mixed diet from week 3 until week 9 relative to calving. Multiparous cows fed glucogenic diet had lower milk fat yield, higher calculated EB, and lower plasma NEFA, BHBA and liver triacylglyceride concentration and tended to have fewer days to first postpartum ovulation. In conclusion, increasing the glucogenic nutrient availability improved the EB and had potential to reduce the risk for metabolic disorders and to improve reproductive performance in dairy cows.