Concentrations of retinol and tocopherols in the milk of cows supplemented with conjugated linoleic acid

This study was performed to investigate the hypothesis that supplementation of conjugated linoleic acid (CLA) changes the concentrations of retinol and tocopherols in the milk of cows. To investigate this hypothesis, Holstein cows received daily from 3 weeks ante‐partum to 14 weeks post‐partum either 172 g of a CLA‐free rumen‐protected control fat (control group, n = 20) or the same amount of a rumen‐protected CLA fat, supplying 4.3 g of cis‐9, trans‐11 CLA and 3.8 g of trans‐10, cis‐12 CLA per d (CLA group, n = 20). Milk samples (collected at weeks 1, 3, 5, 8 and 11 of lactation) were analysed for retinol, α‐ and γ‐tocopherol concentrations. Milk of cows supplemented with CLA had higher concentrations of retinol (+34%), α‐tocopherol (+44%) and γ‐tocopherol (+21%) than milk of control cows (p < 0.05). The daily output of these vitamins via milk was also greater in cows of the CLA group than in cows of the control group (+36, 50 and 24% for retinol, α‐tocopherol and γ‐tocopherol, respectively, p < 0.05). In agreement with higher concentrations of tocopherols, concentrations of thiobarbituric acid‐reactive substances, determined in milk of week 5, were lower in cows of the CLA group than in control cows, indicative of a lower susceptibility of milk lipids to peroxidation. Plasma concentrations of retinol and α‐tocopherol, determined at 1 and 5 weeks post‐partum, were not different between the two groups of cows. In conclusion, this study shows that supplementing dairy cows with a moderate amount of CLA causes an increase of the concentrations of vitamins A and E in the milk and results in an increased output of those vitamins via milk. These effects might be beneficial with respect to the nutritional value of dairy products and the susceptibility of milk fat to oxidative deterioration.     

Inflammatory and metabolic responses to an intramammary lipopolysaccharide challenge in early lactating cows supplemented with conjugated linoleic acid

Supplementation of dairy cows with trans‐10, cis‐12 conjugated linoleic acid (CLA) allows nutrient repartitioning despite an energy deficiency in early lactation, which might be a benefit for the immune system, too. In this study, we investigated potential nutrient sparing effects of CLA in early lactating cows with low plasma glucose concentrations exposed to an intramammary lipopolysaccharide (LPS) challenge. Fifteen multiparous Holstein cows were exposed to an intramammary LPS challenge in week 4 p.p. Eight cows (CLA) were supplemented daily with 70 g of lipid‐encapsulated CLA (6.8 g trans‐10, cis‐12 and 6.6 g of the cis‐9, trans‐11 CLA isomer; CLA) and seven cows with 56 g of control fat (CON). Blood samples were obtained every 30 min along with rectal temperature, heart and respiratory rate, and milk samples were taken hourly until 10 hr after the LPS application. Plasma was analysed for concentrations of glucose, free fatty acids, beta‐hydroxybutyrate (BHB), cortisol, insulin and glucagon. In milk, somatic cell count and activity of lactate dehydrogenase were determined. Initial plasma glucose concentration was lower in CLA than in CON. During the immunostimulation, CLA had higher glucose concentrations than CON, and BHB decreased distinctly in CLA, whereas CON cows maintained BHB concentration at a lower level. Body temperature in CLA increased earlier, the difference between peak and basal temperature was higher, and the decline thereafter occurred earlier. In conclusion, CLA supplementation of early lactating cows exposed to an intramammary LPS challenge affected local and systemic immune responses. We assume that CLA supplementation triggered glycogen storage. Cows supplemented with CLA provided more glucose and preferentially used BHB as an energy source during the immune response. The more intense metabolic and more concentrated endocrine responses support an immunomodulatory effect of CLA supplementation.     

Effect of pre‐ and post‐partum supplementation with lipid‐encapsulated conjugated linoleic acid on milk yield and metabolic status in multiparous high‐producing dairy cows

We evaluated the lactation performance, liver lipid content and plasma metabolites indicating the energy balance of dairy cows supplemented with conjugated linoleic acid (CLA) pre‐ and post‐partum (PP) vs. only PP. A total of 60 cows were divided into three groups (n = 20). Daily diet of cows was supplemented with 14 g of CLA (7 g cis‐9, trans‐11 and 7 g trans‐10, cis‐12 isomers) from week 3 before the expected date of calving (group CLA1), or from the day of calving (group CLA2) until 77–91 days PP. Control cows were fed an isocaloric, isonitrogenous and isolipidic diet without CLA. Between week 3 and week 6 PP, the milk yield of cows in both CLA‐treated groups was approximately 4.5 kg higher (p < 0.05) than in control. Milk fat concentrations decreased from week 3 and were lower in both CLA groups than in control (p < 0.01). Body condition score loss was lower (p < 0.05) in the CLA1 than in the control group on week 5 PP. By week 11 PP, the body condition of both CLA1 and CLA2 groups exceeded that of control. Plasma non‐esterified fatty acid was lower in CLA1 compared to CLA2 and control during the early PP period (p < 0.05), while this difference faded away by the late PP period. Beta‐hydroxybutyrate (BHBA) increased rapidly in all groups following calving. In CLA1 group, it began to decrease sooner than in CLA2 and control. The prevalence of subclinical ketosis (BHBA > 1.2 mm ) was lower in CLA1 group than in CLA2 and control (p < 0.05). Liver biopsy analyses showed that CLA1 treatment decreased (p < 0.05) the total lipid content of liver compared to control at week 5 after calving. Our results show that CLA supplementation is more efficient in alleviating body mass mobilization and decreasing the incidence of subclinical ketosis when applied as early as 3 weeks before calving than started feeding after calving.     

Altered Expression of 3β‐HSD,CYP17 and 17β‐HSD in the Foetal Porcine Gonads in Response to Anti‐androgen Flutamide Exposure

We investigated whether the limited access to androgens during late prenatal period alters expression of steroidogenic enzymes involved in androgen production: 3β‐hydroxysteroid dehydrogenase/Δ5‐Δ4 isomerase (3β‐HSD), cytochrome P450 17α‐hydroxylase/17,20‐lyase (CYP17) and 17β‐hydroxysteroid dehydrogenase type 1 (17β‐HSD1) or type 3 (17β‐HSD3) in the foetal porcine gonads. Pregnant gilts were injected with anti‐androgen flutamide (for seven days, 50 mg/day/kg bw) or corn oil (control) starting at 83 (GD90) or 101 (GD108) gestational day. To assess 3β‐HSD, CYP17 and 17β‐HSD1 or 17β‐HSD3 expression, real‐time PCR and immunohistochemistry were performed. In testes from flutamide‐treated foetuses, increased 3β‐HSD and CYP17 mRNA expression was observed in the GD90 group, while decreased 3β‐HSD and 17β‐HSD3 mRNA expression and increased CYP17 mRNA expression were found in the GD108 group. CYP17 and 17β‐HSD3 were localized in Leydig cells. Following flutamide administration, the intensity of CYP17 immunostaining was higher in both treated groups, while 17β‐HSD3 intensity was lower in the GD108 group. In ovaries from flutamide‐treated foetuses in the GD90 group, mRNA level for 3β‐HSD was elevated, but it was diminished for CYP17 and 17β‐HSD1. In the GD108 group, flutamide treatment led to lower mRNA level for 3β‐HSD but higher for CYP17. 3β‐HSD was found in granulosa cells, while CYP17 was localized within egg nests and oocytes of forming follicles. Following flutamide treatment, the intensity of 3β‐HSD and CYP17 immunostaining was higher in the GD90 and GD108 groups, respectively. Immunohistochemical staining for 3β‐HSD was restricted to the ovary. Concluding, diminished androgen action in the porcine foetal gonads during late gestation induces changes in steroidogenic enzymes expression, which may led to changes in gonadal function. However, it seems that androgens exert diverse biological effects depending on the gestational period.     

Using rumen probes to examine effects of conjugated linoleic acids and dietary concentrate proportion on rumen pH and rumen temperature of periparturient dairy cows

The study aimed to examine the influence of supplemented conjugated linoleic acids (CLA) to periparturient cows receiving different concentrate proportions antepartum on rumen pH (RpH) and rumen temperature (RT). Twenty pregnant German Holstein cows were equipped with rumen probes for continuous RpH and RT measurement in a frequency of 15 min to investigate effects of dietary concentrate and CLA around parturition and the impact of parturition itself on RpH and RT. Cows had ad libitum access to partial mixed rations, 3 weeks prior to calving until day 7 post‐partum. Antepartum, cows received 100 g/day control fat (CON) or CLA supplement, either in low (20%; CON‐20, CLA‐20) or high concentrate diet (60%; CON‐60, CLA‐60). Post‐partum, concentrate proportion was adjusted to 50% while fat supplementation continued. Compared with adapted feeding, high concentrate proportions antepartum tended to increase DMI and reduced RpH. Groups CON‐60 and CLA‐60 spent more than 4 h per day below RpH 5.6 during late pregnancy, indicating the presence of subacute rumen acidosis (SARA). The RT remained unaffected antepartum. Before calving, cows spent less time below RpH 5.6 and SARA could be detected in each group post‐partum. Mean RpH increased slightly antepartum, whereas few hours before parturition a sharp decrease in RpH could be observed, accompanied with increased RT. Overall, it seems that CLA supplementation influences RpH and RT. Bearing in mind that rumen parameters fluctuate during day and herd level must be known, rumen probes for continuous RpH and RT measurement could be a useful management tool for animal health surveillance and may also help to predict parturition.     

Conjugated linoleic acids (CLA) moderate negative responses of heat-stressed cows

The objective of this study is to examine whether CLA would help to improve negative responses in lactating cows during heat stress. During a 2-week adjustment period, all cows were fed a control diet supplemented with 400 g of calcium salts of palm oil fatty acids. After the adjustment period, cows (n = 9 per group) were randomly assigned by block to one of four treatments: control or three diets containing 100, 200, or 400 g of CLA (70% of which were mixed isomers of CLA). Milk fat percentage and yield decreased by supplementation of CLA (P < 0.01), but milk yield and other nutrient components did not vary (P > 0.05). The concentrations of short and medium-chain fatty acids (C4–C14) decreased and long-chain fatty acids (C18) increased in milk fat for cows fed CLA. Moreover, the contents of cis9, trans11 and trans10, cis12-CLA in milk fat increased markedly (P < 0.01). Respiration rate was lower (P < 0.05) for cows fed 200 g of CLA and rectal temperature decreased (P < 0.01) in all CLA groups during day 15 to day 28. Contents of aspartate aminotransferase, creatine kinase, thyroxin, potassium, sodium, calcium and chlorine were altered by CLA supplements, whereas other plasma parameters were not affected (P > 0.05). Lower concentrations of aspartate aminotransferase and creatine kinase in blood for cows fed 200 g or 400 g of CLA suggested that CLA protected cows from muscle damage. In addition, higher concentrations of electrolytes and thyroxin in blood in these groups suggested that CLA could improve heat stress situations in cows. Overall, the results showed that supplementations with 200 or 400 g/day of CLA moderated the negative responses in heat-stressed cows.     

Effect of Oestrus Synchronization with PGF2α/eCG/hCG on Luteal P4 Synthesis in Early Pregnant Gilts

Administration of hormones to synchronize oestrus is a useful tool in animal breeding. However, exogenous ovarian stimulation may be detrimental to reproductive function. This study was aimed to examine whether an oestrus synchronization with PGF2α/eCG/hCG could affect luteal P4 synthesis in early pregnant gilts. Corpora lutea (CLs) were collected on days 9, 12 and 16 of pregnancy from gilts with natural (n = 16) and synchronized (n = 18) oestrus and analysed for (i) the expre‐ssion of steroidogenic acute regulatory protein (StAR), cytochrome P450 family 11 subfamily A polypeptide (CYP11A1), and 3β‐hydroxysteroid dehydrogenase (3βHSD); (ii) the concentration of P4 in the luteal tissue and blood; and (iii) the expression of luteinizing hormone receptors (LHR) and oestrogen receptors (ERα and ERβ). Additionally, the effect of LH on P4 secretion from CL slices collected from synchronized and naturally ovulated animals has been studied in vitro. PGF_2α/eCG/hCG administration increased mRNA expression of StAR, CYP11A1, 3βHSD, and LHR on day 9 and CYP11A1 and LHR on day 12 of pregnancy compared with the control group (p < 0.05). CYP11A1, 3βHSD, LHR, ERα and ERβ proteins were not affected by synchronization; only StAR protein increased in hormonally treated animals (p = 0.017). The concentration of P4 in luteal tissue was greater on day 9 (p < 0.01), but lower on day 16 (p < 0.05) in gilts with hormonally induced oestrus compared with control animals. Blood serum levels of P4 were lower in synchronized than control gilts (p < 0.001). Synchronization did not affect LH‐stimulated P4 secretion from luteal slices; however, greater basal concentration of P4 in incubation medium was detected for CLs collected from synchronized than control gilts (p < 0.05). In conclusion, synchronization of oestrus with PGF2α/eCG/hCG protocol in gilts did not impair the expression of luteal P4 synthesis system, although decreased P4 concentration in the blood.     

Milk fatty acid profile from cows fed with mixed rations and different access time to pastureland during early lactation

Milk fatty acid (FA) profiles were determined in Holstein cows (n = 27) fed total mixed rations (TMR) ad libitum (G0) or diet composed by TMR (50% dry matter [DM] offered) plus grazing of pasture with 6 hr of access time to paddock in one session (G1) or 9 hr in two sessions (G2) at 45 days in milk (DIM). Moreover, milk FA was determined at 65 DIM when G0 cows turned out to G1 diet without adaptation period (Post‐G0), G1 remained as controls. Milk FA was quantified using gas chromatography and mass spectrometry. Preformed FA at 45 DIM was greater (+27%) for G2 than G0 cows (p < .05). Stearic acid (C18:0) was 30% greater for G2 cows (p < .05). De novo FA was lowest for G2 cows (p < .05). Conjugated linoleic acid (CLA) did not differ (p < .12), while vaccenic acid (C18:1trans) was twofold greater for grazing treatments (p < .01). Linolenic acid [C18:3(n‐3)] was greatest for G2 and lowest for G0 cows (p < .01). Omega 6 FA was greater for G0 than grazing cows, mainly due to linoleic acid [18:2cis(n‐6); p < .05]. These results determined that n‐6/n‐3 ratio was almost threefold greater for G0 than grazing cows (p < .001). When diet of G0 cows changed to include pasture (Post‐G0), preformed FA increased (p < .05), explained mainly by the increase (p < .05) of stearic (C18:0) and C18:1trans, while de novo FA tended to decrease (p < .1). Moreover, the amount of CLA and C18:3(n‐3) tended to increase (p < .1) in Post‐G0 cows. Offering 50% of dietary DM from pasture modified milk FA profile in early lactation potentially beneficial for human health. When TMR‐fed cows were turned out to 50% pasture, milk FA profile reflected dietary change without need of an adaptation period.     

Ovarian localization of 11β-hydroxysteroid dehydrogenase (11βHSD): effects of ACTH stimulation and its relationship with bovine cystic ovarian disease

Cystic ovarian disease (COD) is an important cause of infertility in cattle, and ACTH has been involved in regulatory mechanisms related to ovarian function associated with ovulation, steroidogenesis, and luteal function. Here, we examined the localization of 11β-hydroxysteroid dehydrogenase type 1 (11βHSD1) and 11βHSD2 proteins in the ovary of healthy cows and animals with spontaneous and ACTH-induced COD and the in vitro response of the follicular wall exposed to ACTH. After stimulation by ACTH, we documented changes in 11βHSD expression and cortisol secretion by the follicular wall of large antral and follicular cysts. Follicular cysts showed a higher constitutive expression of both enzymes, whereas ACTH induced an increase in 11βHSD1 in tertiary follicles and follicular cysts and a decrease in 11βHSD2 in follicular cysts. Moderate expression of 11βHSD1 was observed by immunohistochemistry in granulosa of control animals, with an increase (P < 0.05) from primary to secondary, tertiary, and atretic follicles. The level of immunostaining in theca interna was lower than that in granulosa. The expression of 11βHSD2 was lower in the granulosa of primary follicles than in that of secondary, tertiary, and atretic follicles and was lower in the theca interna than in the granulosa. In ACTH-induced and spontaneously occurring follicular cysts, differences from controls were observed only in the expression of 11βHSD1 in the granulosa, being higher (P < 0.05) than in tertiary follicles. These findings indicate that follicular cysts may be exposed to high local concentrations of active glucocorticoids and indicate a local role for cortisol in COD pathogenesis and in regulatory mechanisms of ovarian function.     

Influence of conjugated linoleic acids and vitamin E on milk fatty acid composition and concentrations of vitamin A and α‐tocopherol in blood and milk of dairy cows

The objective of this trial was to investigate the influences of conjugated linoleic acid (CLA ) and vitamin E (Vit. E) and their interactions on fatty acid composition and vitamins in milk (α‐tocopherol, retinol and β‐carotene) as well as on α‐tocopherol in blood of pluriparous cows from week 6 ante partum until week 10 post‐partum (p.p.). We assigned 59 pluriparous German Holstein cows to four treatment groups with the treatment factors CLA and Vit. E at two levels in a 2 × 2 factorial design. Milk fatty acid composition and milk vitamins were analysed on lactation days 7 and 28. α‐tocopherol in blood serum was analysed on days ?42, ?7, 1, 7, 14, 28 and 70 relative to parturition. Milk concentration of α‐tocopherol was influenced by Vit. E (p  < .001) and CLA (p  = .034). Percentage of cis ‐9, trans ‐11 CLA in total milk fat was influenced by treatment with CLA (p  < .001), while for percentage of trans‐ 10, cis ‐12 CLA an interaction between treatment and day (p  = .019), driven by an increase in both CLA groups from day 7 to day 28, was found. Serum ratios of α‐tocopherol to cholesterol were influenced by Vit. E (p  < .001). Results suggest that treatment with CLA during late pregnancy and early lactation is suitable to enhance the proportion of trans‐ 10, cis ‐12 CLA in milk and thereby influencing nutritional properties. As treatment with Vit. E did not have an impact on milk fatty acid composition, it might be possible to increase the antioxidative capacity of the dairy cow without affecting milk properties. Consequently, combined treatment with CLA and Vit. E might elicit synergistic effects on the cow and milk quality by increasing the proportion of CLA in milk fat as well as the excretion of Vit. E and the Vit. E levels in serum.     

Expression of 11β‐Hydroxysteroid Dehydrogenase Type 1 and Glucocorticoid Receptors in Reproductive Tissue of Male Horses at Different Stages of Sexual Maturity

Glucocorticoids (GCs) as mediators of the stress response may affect Leydig cell function by inhibiting either luteinizing hormone receptor expression or testosterone biosynthesis. The isozymes 11β‐hydroxysteroid dehydrogenase (11βHSD) 1 and 11βHSD2 control the intracellular cortisol levels. Little is known about the effects of stress on fertility in the equine. The objective of the present study was to determine the presence and cellular localization of glucocorticoid receptors (GCR) and glucocorticoid‐metabolizing enzymes (11βHSD1 and 11βHSD2) in equine epididymal and testicular tissue with special regard to sexual maturation. Testicular and epididymal tissue was collected from 21 healthy stallions, and four age groups were designed: pre‐pubertal, young, mature and older horses. Immunohistochemistry (IHC) analysis and quantitative real‐time PCR (qRT‐PCR) were used. Pre‐pubertal horses showed higher testicular gene expression of 11βHSD1, 11βHSD2 and GCR than horses of all other groups (p < 0.05). A positive intranuclear immunoreaction for GCR was seen in epithelial cells of caput, corpus and cauda epididymidis and in Leydig cells. Significant differences (p < 0.05) between age groups occurred. The number of Leydig cells staining positive for GCR was highest in immature stallions (p < 0.05). The enzyme 11βHSD1 was localized in epithelial cells of the caput and corpus epididymidis and in Leydig cells. As determined by enzyme assay, nicotinamide adenine dinucleotide (NAD)‐dependant dehydrogenase (oxidation) activity was not detected in testicular tissue from immature stallions but in all other age groups (n = 3 per group). Results of this study suggest a contribution of GCs to maturation of male reproductive tissue in horses. In mature stallions, expression of 11βHSD enzymes and the oxidative 11βHSD activity in Leydig cells and epididymal basal and principal cells suggest a protective role on these tissues contributing to physiological intracellular glucocorticoid concentrations.     

Effects of intramammary infusion of Bifidobacterium breve on mastitis pathogens and somatic cell response in quarters from dairy cows with chronic subclinical mastitis

The present study assessed the effects of intramammary infusion of Bifidobacterium breve (B. breve) on mastitis‐causing pathogens and on the somatic cell counts (SCC) in lactating cows with chronic subclinical mastitis. The bacteriological cure rates of 42 quarters from 42 cows infected with Staphylococcus aureus, Corynebacterium bovis, coagulase‐negative staphylococci, and environmental streptococci were 18.2% (2/11), 14.3% (1/7), 58.8% (10/17), and 28.6% (2/7), respectively, on day 14 after B. breve infusion. In a second trial, B. breve was infused into 18 quarters from 18 cows with chronic subclinical mastitis from which pathogens had not been isolated; the rates of quarters showing SCC > 50 × 10⁴ cells/ml prior to B. breve infusion that decreased to < 30 × 10⁴ cells/ml after infusion were significantly (p < .01) increased to 61.1% (11/18) on day 14 compared to that prior to infusion (0/18). The intramammary infusion of B. breve appears to be a non‐antibiotic approach for elimination of minor pathogens and decreasing SCC in quarters with chronic subclinical mastitis in dairy cows.     

Hyperglycaemia in transition dairy cows: Effects of lactational stage and conjugated linoleic acid supplementation on glucose metabolism and turnover

Supplementing conjugated linoleic acid ( CLA ) is supposed to spare glucose due to the milk fat‐depressing effect of the trans ‐10, cis ‐12 CLA isomer, and allows repartitioning nutrients despite an energy deficiency in early lactation. However, there is still a lack of knowledge in terms of the dynamic pattern of the glucose turnover in transition dairy cows. We hypothesized that dairy cows supplemented with CLA have an altered rate of glucose turnover and insulin sensitivity during early lactation. We conducted three consecutive hyperglycaemic clamps (HGC ) in weeks ?2, +2 and +4 relative to parturition in Holstein cows supplemented daily either with 70 g of lipid‐encapsulated CLA (6.8 g trans ‐10, cis ‐12 and 6.6 g of the cis ‐9, trans ‐11 CLA isomer; CLA ; n  = 11) or with 56 g of control fat ( CON ; n  = 11). From week ?3 up to week +4 relative to parturition, milk yield and dry matter intake (DMI ) were recorded daily, while body weight (BW ) and milk composition were obtained once weekly. Blood samples were taken once weekly and every 30 min during the HGC . Plasma was analysed for concentrations of glucose, fatty acids (FFA ), beta‐hydroxybutyrate (BHB ), insulin, triglycerides and cholesterol. The CLA supplementation did not affect performance and metabolic parameters except for BHB and cholesterol. Furthermore, insulin concentrations and insulin sensitivity were affected by treatment. During the HGC in early lactation, insulin response was lower and decrease in FFA and BHB greater compared with the HGC in week ?2 although glucose target concentration achieved during the steady‐state period was similar for all three HGC . Our findings in terms of insulin and cholesterol suggest that body reserves are preserved through CLA feeding without restraining animal's performance. Furthermore, CLA effects on cholesterol and triglyceride concentrations indicated beneficial effects on hepatic lipid export contributing to an improved efficiency of prevailing metabolites in circulation.     

Diagnostic opportunities for evaluation of the exposure of dairy cows to the mycotoxins deoxynivalenol (DON) and zearalenone (ZEN): reliability of blood plasma,bile and follicular fluid as indicators

To investigate the usefulness of follicular fluid (FF) in relation to blood plasma and bile as indicators of exposure of dairy cows to ZEN, DON and their metabolites, a dose–response study was performed with 30 dairy cows. The cows, 10 in each group (named CON; FUS‐50, FUS‐100), received a diet with three different concentrations of Fusarium toxin‐contaminated maize. Thereby, the following dietary concentration were reached: CON (0.02 mg ZEN and 0.07 mg DON, per kg dry matter, DM), FUS‐50 (0.33 mg ZEN and 2.62 mg DON, per kg DM) and FUS‐100 (0.66 mg ZEN and 5.24 mg DON, per kg DM). ZEN, DON and de‐epoxy‐DON (de‐DON) were detected in FF. Based on the linear regression between toxin concentration in plasma and FF, it seems that about 50 % (m = 0.5) of ZEN present in plasma is present in FF while an increase of 1 ng/ml DON or de‐DON in plasma is paralleled by an increase of 1.5 ng/ml DON or 1.1 ng/ml de‐DON in FF. ZEN, DON and their metabolites, except zearalenone (ZAN), were also detected in bile. Contrary to DON and de‐DON, ZEN and its metabolites were accumulated in bile so that the concentration of ZEN and metabolites was much higher than for DON and de‐DON. The main compound was β‐zearalenol (β‐ZEL). The biliary ZEN, α‐zearalenol (α‐ZEL) and β‐ZEL concentration correlated linearly with each other with an uncertainty of <15 % (r² ≥ 0.86), whereas the ratio between ZEN: α‐ZEL: β‐ZEL was about 1.5:1:11. With the help of established linear relationship between toxin intake and toxin concentration, bile could be used as diagnostic indicator to assess the exposure of cows.     

Changes of leukocyte counts and expression of pro‐ and anti‐inflammatory cytokines in peripheral leukocytes in periparturient dairy cows with retained fetal membranes

In dairy cows, retained fetal membranes (RFM) affect reproductive performance. The aim of this study was to examine the leukocyte counts and the gene expression of tumour necrosis factor α (TNFα), interleukin 1β (IL‐1β), IL‐8, and IL‐10 in polymorphonuclear leukocytes (PMNs) and peripheral blood mononuclear cells (PBMCs) in cows with (n = 5) or without (n = 5) RFM during the peripartum period. The lymphocyte counts in RFM cows were higher than those in control cows throughout the experiment (p < .05). The expression of IL‐8 in PMNs of control cows was higher (p < .05) compared with that of RFM cows postpartum. In cows with RFM, IL‐1β expression was higher (p < .05) in PMNs at 6 weeks postpartum whereas the expression of IL‐1β was lower (p < .05) in PBMCs at 4 weeks postpartum. The expression of IL‐10 in PBMCs of control cows was higher (p < .05) than that of RFM cows at 2 weeks prepartum and 4 weeks postpartum. Taken together, our data indicate that changes of gene expression of pro‐ and anti‐inflammatory cytokines in RFM cows might be associated with the delayed placental separation and development of uterine inflammation in RFM cows.     

Effects of Feeding Dry Glycerol to Primiparous Holstein Dairy Cows on Follicular Development,Reproductive Performance and Metabolic Parameters Related to Fertility during the Early Post‐partum Period

This study examined the effects of dry glycerol supplementation on follicular growth, post‐partum interval to first ovulation, concentration of serum metabolites and hormones related to fertility, body condition score (BCS) and body weight (BW) in primiparous Holstein dairy cows. Sixty primiparous Holstein dairy cows were randomly assigned to two groups (control: n = 30 and glycerol supplemented: n = 30). Dry glycerol (250 g/day/cow) was fed as a top dressing to the common lactating total mixed ration (TMR) from parturition to 21 days post‐partum. Ovaries were examined four times using ultrasonography on days 13, 19, 25 and 36 post‐partum to determine ovarian follicular growth. Concentration of serum metabolites and hormones was determined weekly. Body condition score was evaluated weekly from weeks 1 to 5 after parturition, and BWs were recorded three times on days 1, 11 and 21 during the experimental period. The cows fed dry glycerol had more large follicles (p < 0.0001) and corpora lutea (CL) (p = 0.02) compared with the control cows. Days to the first ovulation (p = 0.06), days to first oestrus (p = 0.05), services per conception (p = 0.06) and days open (p = 0.004) were positively affected by dry glycerol supplementation. Serum concentration of glucose and insulin was higher in dry glycerol‐supplemented cows (p = 0.1; p = 0.06, respectively). Feeding glycerol had no effect on mean serum concentrations of β‐hydroxybutyrate, non‐esterified fatty acids and IGF‐1 during the experimental period. However, significant differences were observed at concentration of BHBA and IGF‐1 (p = 0.02 and p = 0.04, respectively) between two groups on day 21 after calving. The cows in the glycerol‐fed group had higher serum progesterone concentrations on days 33 (p = 0.007) and 36 (p = 0.004) after calving. Supplemented cows had lower body condition loss during weeks 1–5 after calving compared with the control cows (0.34 vs 0.41 BCS). In week 13 post‐partum, the proportion of cycling cows was 83.3 and 69.9% for those which received supplemented or non‐supplemented diet, respectively. These results demonstrated that feeding dry glycerol as a glucogenic supply may be useful to improve negative energy balance and reproductive efficiency in young cows which calve with high requirement of energy.     

Supplementation of dairy cows with a fish oil containing supplement and sunflower oil to increase the CLA content of milk produced at pasture

It has been shown that the cis 9, trans 11 isomer of conjugated linoleic acid (CLA) can be increased in milk by supplementation with fish oil and vegetable oils. Feeding a high level of oil, however, can impact negatively on gross milk composition. The principal aim of this study was to determine if relatively low levels of fish oil or sunflower oil, either alone or in combination, offered to dairy cows on pasture would increase the C18:2 cis 9, trans 11 CLA concentration in milk. Forty autumn-calved cows on a diet of grazed grass were assigned to 4 supplementation treatments: (i) No supplement (P), (ii) 255 g/day of sunflower oil (SO), (iii) 255 g/day of sunflower oil + 52.5 g/day of fish oil (SOFO), and (iv) 105 g/day of fish oil (FO). The fish oil was supplied in a proprietary product called Omega-3 Supplement which is a mixture of marine oils and an extracted oilseed meal and contains 500 g/kg of oil. The oils were fed in a concentrate mixture, which was offered at a rate of 3.0 kg/cow per day. The production of the cows was measured for 54 days and the milk fatty acid composition was determined on day 0 (immediately before the supplements were introduced) and on days 14, 28 and 42 after the treatments were imposed. Supplementation increased the yield of milk (P < 0.01), protein (P < 0.05) and lactose (P < 0.001), decreased milk fat (P < 0.05) and protein (P < 0.01) concentrations and increased (P < 0.01) lactose concentration. Type of oil did not significantly affect any production variable. The concentration of C18:1 trans 9 + C18:1 trans 11 (mainly C18:1 trans 11) (P < 0.001) and C18:2 cis 9, trans 11 CLA (P < 0.01) were greater on supplemented treatments than on P and the concentration of both were greater (P < 0.05) on FO than on SO. The results confirm that the concentration of C18:2 cis 9, trans 11 CLA can be increased further, from an already relatively high concentration in milk from pasture, by offering supplements containing a low level of fish oil either alone or in combination with sunflower oil.     

Partially replacing sorghum silage with cactus (Opuntia stricta) cladodes in a soybean oil-supplemented diet markedly increases trans-11 18:1, cis-9, trans-11 CLA and 18:2 n-6 contents in cow milk

Based on low 18:0 contents observed in milk fat of cows fed cactus cladodes (CC), we hypothesized that including Opuntia stricta cladodes in a soybean oil (SO)-supplemented diet would promote incomplete rumen biohydrogenation of supplemental PUFA, leading to increased trans-11 18:1 and cis-9, trans-11 CLA contents in milk. Twelve Holstein cows were used in a two-period study: (a) Baseline: all cows received a total mixed ration (TMR) composed of sorghum silage (SS) and a concentrate containing no SO for 14 days; (b) Treatment: cows received one of the following SO-supplemented diets for 21 days: (1) SS-TMR: a TMR composed of SS and a SO-enriched concentrate, (2) CC-TMR: a TMR containing CC as a partial substitute for SS plus the SO-enriched concentrate, and (3) CC-PMR: same diet as in treatment 2, but CC were mixed with the SO-enriched concentrate and fed as a partial mixed ration (PMR). Both CC diets increased relative abundances of trans-11 18:1, cis-9, trans-11 CLA, and 18:2 n-6 in milk fat, whereas opposite effects were observed on 18:0 and cis-9 18:1. Proportion of 18:2 n-6 increased, and cis-9, trans-11 CLA tended to increase with CC-PMR as compared to CC-TMR, whereas 18:3 n-3 was higher with CC-PMR than with SS-TMR. Proportions of several odd- and branched-chain fatty acids, certain 18:1 isomers, and trans-9, cis-11 CLA changed with CC diets, notably with CC-PMR. Milk yield and intake of most nutrients (except fibre) increased or tended to increase with the CC diets, whereas gross milk composition was unaltered. Stearoyl-CoA desaturase-1 index for C18 (SCD₁₈) was higher with CC-PMR than with SS-TMR, and milk n-6:n-3 FA ratio and apparent transfer of 18:2 n-6 to milk increased with CC diets. These results indicate that Opuntia stricta cladodes can be a valuable feed ingredient for improving the nutraceutical value of milk fat.