Plant polyphenol extract supplementation affects performance, welfare, and the Nrf2-oxidative stress response in adipose tissue of heat-stressed dairy cows.

We examined the effects of a supplement of plant polyphenols extracts of green tea, capsicum, and fenugreek, and electrolytes ([Na+, K+]; AXT, Axion ThermoPlus, CCPA, France] during summer heat load on production, welfare, and oxidative stress proteins in adipose tissue (AT) of dairy cows. A total of 42 multiparous mid-lactation cows were divided into 3 groups during summer, and were fed for 2 wk either a standard milking cow diet (CTL, n = 14) or diets supplemented with 100 g/d of AXT (100AXT, n = 14), or 150 g/d of AXT (150AXT, n = 14), while being cooled 5 times a day. Then, half of the cows from each dietary treatment were cooled (CL) or not cooled (NCL) for 2 wk, after which the cooled and uncooled groups were switched for additional 2 wk. Cows were milked 3 times a day, and milk composition was analyzed at the end of each 2-wk period. Vaginal temperature (VT) was measured for 3 consecutive days in each period. Biopsies of subcutaneous AT were taken from 10 NCL cows (5 each of CTL and 150AXT) at the end of the period and examined by liquid chromatography-tandem mass spectrometry proteomics analysis. Data were analyzed with PROC MIXED of SAS (version 9.2, SAS Institute Inc.). The model included the effects of dietary treatment, cooling regimen, period, and their interactions. Protein and mRNA abundances and proteomic data (P ≤ 0.05 and fold change [FC] ± 1.5) were analyzed by t-test. Milk yields and 4% fat-corrected milk (FCM) were higher in 100AXT than in CTL; milk components were not different. Dry matter intake (DMI) was higher in 100AXT than in CTL. The effect of cooling and the interactions of period × cooling were significant for DMI, 4% FCM, energy-corrected milk, and milk/DMI. The proportion of time that VT was >39°C was lower in 100AXT and in 150AXT than in CTL. Daily rumination time was greater in 150AXT than in CTL, and lying time was greater in 100AXT and 150AXT than in CTL. Proteomics of AT demonstrated that 150AXT had increased abundances of peroxidasin (FC = 1.6), microsomal glutathione S-transferase 2 (FC = 2.5), and heme oxygenase 1 (FC = 3.6) compared with CTL. Top enriched canonical pathways included acute phase response signaling, Nrf2-mediated oxidative stress response, and lipopolysaccharide (LPS)/IL-1-mediated inhibition of RXR function. Immunoblots of AT showed a higher abundance of the transient receptor potential vanilloid 1 and of LPS binding protein in AT of 150AXT compared with CTL. Supplementation of AXT increased DMI, milk, and 4% FCM, lowered VT, improved welfare indices, and enriched the AT with Nrf2-oxidative stress response and acute phase response proteins in heat-stressed dairy cows.

Saturated fat supplemented in the form of triglycerides decreased digestibility and reduced performance of dairy cows as compared to calcium salt of fatty acids.

The two most popular rumen-protected fatty acid supplements in dairy cow rations are calcium salts of palm oil fatty acid calcium salts of palm oil fatty acid (CSFA) and prilled saturated fatty acids (SFAs). The objectives of this study were to determine the effects of supplementing SFA in the form of triglycerides (TSFA), as compared to CSFA, on yields, efficiency and diet digestibility in high-yielding dairy cows. Twenty-eight (14 cows in each group) multiparous cows were fed a basal diet supplemented (on DM basis) with either 12 g/kg TSFA (~350 g/cow per day - contained 980 g/kg fat; 882.3 g/kg SFAs) or 14 g/kg CSFA (~440 g/cow per day - contained 800 g/kg fat; 566.4 g/kg SFAs). The supplement amounts in the diet were balanced according to fat content. Rumen samples were taken for measurements of ammonia and volatile fatty acids concentrations, and fecal samples were taken for digestibility measurements. The CSFA cows produced 3% higher milk yields (47.6 v. 46.2 kg/day; P < 0.0001) and 4.7% higher 4% fat-corrected milk (FCM; 44.7 v. 42.7 kg/day; P = 0.02) than the TSFA cows. No difference in milk-fat content was observed, but milk-protein content was higher in the TSFA than CSFA cows. Yields of fat and protein were similar, but lactose yields were higher in TSFA cows. There were no differences in dry matter intake or efficiency calculations between groups. The ruminal ammonia concentrations were similar between groups, whereas acetate concentrations and acetate : propionate ratio were greater for CSFA than TSFA cows. The apparent total-tract digestibility of dry (P < 0.0007) and organic matters (P < 0.0003), fat (P < 0.0001), NDF and ADF (P = 0.02) were lower in the TSFA v. CSFA cows. In conclusion, the CSFA-supplemented cows produced 3% higher milk and 4.7% higher 4% FCM than the TSFA cows. However, TSFA supplementation did not depress milk-protein content. The apparent total-tract digestibility was lower for all dietary components in the TSFA cows, which was probably due to the effects of both degree of saturation and triglyceride form of the TSFA supplement. Considering that diets were balanced according to the fat content of the supplements, the lower yields of milk and FCM observed in the TSFA than CSFA cows were likely due to the lower digestibility of the fat and other nutrients in the TSFA cows, which might have negatively influenced the dietary energy content.

Reducing milking frequency from thrice to twice daily in early lactation improves the metabolic status of high-yielding dairy cows with only minor effects on yields.

Reducing milk production during early lactation might be of interest to improve the energy balance (EB) of high-yielding dairy cows. Therefore, the objective of this study was to determine how reducing the milking frequency (MF) of high-yielding dairy cows from thrice to twice a day during the first 30 d in milk (DIM) affects yields, intake, efficiency, metabolic status, and carryover effects. To this end, 42 multiparous cows were divided into 2 groups according to their previous lactation performance, parity, and body weight. The control cows were milked 3 times a day (3ML) and the treated cows were milked twice a day (2ML) until 30 DIM and then both groups were milked 3 times a day. Milk samples were taken twice a week from 2 or 3 consecutive milkings until 45 DIM for analysis of milk solids, and both groups were followed until 100 DIM to determine the carryover effects of MF until 30 DIM. Individual dry matter intake (DMI), milk yield, and body weight were recorded daily. Blood samples were taken 3 times weekly from 14 d prepartum until 45 DIM. Milk yield during the first 30 DIM was 8.6% higher (49.3 and 45.4 kg/d, respectively), milk fat percentage was lower (3.96 and 4.27%, respectively), and the yields of all milk solids were higher in the 3ML cows than in the 2ML cows. Dry matter intake and 4% fat-corrected milk were similar between groups. The EB during the first 30 DIM was lower in the 3ML cows than in the 2ML cows, and milk yield, but not 4% fat-corrected milk yield, per unit of DMI was higher in the 3ML cows. No differences were observed between groups from 31 to 100 DIM in milk yield (∼56.3 kg/d for both groups), milk solids yield, DMI, or milk/DMI; however, fat percentage was lower and EB was higher in the 3ML cows. Blood glucose concentrations between 0 and 30 DIM were lower and ß-hydroxybutyrate concentrations were higher in the 3ML cows than in the 2ML cows, but nonesterified fatty acids concentrations were lower, which may be attributed to the lower clearance frequency of nonesterified fatty acids from the blood stream in the 2ML cows. A lower proportion of the 3ML cows (10%) ovulated ≤15 DIM compared with the 2ML cows (40%), with no beneficial effects on preovulatory follicle characteristics. Reducing the MF from thrice to twice a day during the first 30 DIM improved EB and metabolic status, with only minor effects on production.

Proteome dataset of subcutaneous adipose tissue obtained from late pregnant dairy cows during summer heat stress and winter seasons.

Adipose tissue has a central role in the regulation of metabolism in dairy cows, and many proteins expressed in this tissue are involved in metabolic responses to stress (Peinado et al., 2012) [1]. Environmental heat stress is one of the main stressors limiting production in dairy cattle (Fuquay, 1981; West, 2003) [2], [3], and there is a complex interaction between heat stress and the transition period from late pregnancy to onset of lactation, which is manifested in heat-stressed late-gestation cows (Tao and Dahl, 2013) [4]. We recently defined the proteome of adipose tissue in peripartum dairy cows, identifying 586 proteins of which 18.9% were differentially abundant in insulin-resistant compared to insulin-sensitive adipose tissue (Zachut, 2015) [5]. That study showed that proteomic techniques constitute a valuable tool for identifying novel biomarkers in adipose tissue that are related to metabolic adaptation to stress in dairy cows. The objective of the present work was to examine the adipose tissue proteome under thermo-neutral or seasonal heat stress conditions in late pregnant dairy cows. We have collected subcutaneous adipose tissue biopsies from 10 late pregnant dairy cows during summer heat stress and from 8 late pregnant dairy cows during winter season, and identified and quantified 1495 proteins in the adipose tissues. This dataset of adipose tissue proteome from dairy cows adds novel information on the variety of proteins that are abundant in this tissue during late pregnancy under thermo-neutral as well as heat stress conditions. Differential abundance of 107 (7.1%) proteins was found between summer and winter adipose. These results are discussed in our recent research article (Zachut et al., 2017) [6].

Seasonal heat stress affects adipose tissue proteome toward enrichment of the Nrf2-mediated oxidative stress response in late-pregnant dairy cows.

Environmental heat stress and metabolic stress during transition from late gestation to lactation are main factors limiting production in dairy cattle, and there is a complex interaction between them. Many proteins expressed in adipose tissue are involved in metabolic responses to stress. We aimed to investigate the effects of seasonal heat stress on adipose proteome in late-pregnant cows, and to identify biomarkers of heat stress. Late pregnant cows during summer heat stress (S, n=18), or during the winter season (W, n=12) were used. Subcutaneous adipose tissue biopsies sampled 14days prepartum from S (n=10) and W (n=8) were analyzed by intensity-based, label-free, quantitative shotgun proteomics (nano-LC-MS/MS). Plasma concentrations of malondialdehyde and cortisol were higher in S than in W cows. Proteomic analysis revealed that 107/1495 proteins were differentially abundant in S compared to W (P<0.05 and fold change of at least ±1.5). Top canonical pathways in S vs. W adipose were Nrf2-mediated oxidative stress response, acute-phase response, and FXR/RXR and LXR/RXR activation. Novel biomarkers of heat stress in adipose tissue were found. These findings indicate that seasonal heat stress has a unique effect on adipose tissue in late-pregnant cows. SIGNIFICANCE: This work shows that seasonal heat stress increases plasma concentrations of the oxidative stress marker malondialdehyde and cortisol in transition dairy cows. As many proteins expressed in the adipose tissue are involved in metabolic responses to stress, we investigated the effects of heat stress on the proteome of adipose tissue from late-pregnant cows during summer or winter seasons. We demonstrated that heat stress enriches several stress-related pathways, such as the Nrf2-mediated oxidative stress response and the acute-phase response in adipose tissues. Thus, environmental heat stress has a unique effect on adipose tissue in late-pregnant cows, as part of the regulatory adaptations to chronic heat load during the summer season. In addition, this study presents the widest available dataset of adipose tissue proteome in dairy cows, and revealed several novel biomarkers of heat stress in adipose tissue of dairy cows, the use of which awaits further validation.

Dietary α-linolenic acid from flaxseed oil improved folliculogenesis and IVF performance in dairy cows, similar to eicosapentaenoic and docosahexaenoic acids from fish oil.

The objectives of this study were to determine the differential incorporation of various omega-3 (n-3) fatty acids (FAs) supplemented to dairy cows into ovarian compartments and assess the effects on IVF. Forty-two 256-day pregnant cows were supplemented with encapsulated fats, in treatments designated as i) SFA - saturated fat at 240 and 560 g/day per cow, prepartum and post partum (PP) respectively; ii) FLX - flaxseed oil at 300 and 700 g/day per cow prepartum and PP respectively; and iii) FO - fish oil at 300 and 700 g/day per cow prepartum and PP respectively. Commencing at 60 days in lactation, ovum pickup (OPU) was performed twice weekly (20 sessions; five cows per group) and in vitro maturation and IVF were conducted. The proportion of α-linolenic acid (ALA) was greater in follicular fluid (FF), granulosa cells, and cumulus-oocyte complexes (COCs) of FLX cows than in other groups (P<0.001). The proportion of docosahexaenoic acid (DHA) was 6.7 times as great in FF of FO as in other groups (P<0.001); docosapentaenoic acid n-3 and DHA were detected in COCs of FO but not in others. The follicle number during OPU was higher in FLX and FO than in SFA (P<0.05), and the oocyte cleavage rate was higher in FLX and FO than in SFA (P<0.01). Also, the percentage of oocytes that developed to blastocysts tended to be higher in both n-3 groups than in SFA (P<0.1). In conclusion, both dietary n-3 FAs similarly improved folliculogenesis and IVF performance; therefore, ALA-rich botanical n-3 seems to be a satisfactory approach to improve oocyte quality.

Performance and welfare of high-yielding dairy cows subjected to 5 or 8 cooling sessions daily under hot and humid climate.

The objectives were to determine the effects of cooling of high-yielding dairy cows under a hot and humid climate on intake, milk yield, rumination time, and welfare parameters. Forty-two multiparous Israeli Holstein dairy cows were divided into 2 treatment groups and were housed in an open barn divided into 2 pens. The groups were subjected to different cooling schedules, in a crossover design as follows: cows were exposed to 5 or 8 cooling sessions per day (designated 5CS and 8CS, respectively) in the holding area of the milking parlor. Each period lasted 4 wk, and then treatments were switched for another 4-wk period. Each cooling session lasted 45 min, comprising cycles of 30s of showering and 4.5 min of ventilation without showering. Respiration rate and rectal temperature were recorded twice per week (Monday and Thursday) at 0630 and 1600 h. Rumination and lying times were recorded automatically. Rectal temperatures were 0.16 and 1.08°C lower in 8CS than in 5CS cows in the morning and afternoon, respectively. Respiration rate was lower in 8CS than in 5CS cows in the morning (49.1 and 54.6 breaths/min, respectively), and more so in the afternoon (50.0 and 83.0 breaths/min, respectively). Dry matter intake and milk yields were 9.3 and 9.6% higher in the 8CS than in the 5CS cows (27.0 vs. 24.7 and 40.1 vs. 36.6 kg/d, respectively), with no differences in milk fat and protein contents. Daily rumination time was 7.4% longer in the 8CS than in the 5CS (440.1 and 409.6 min/d, respectively); however, rumination time per unit of dry matter or neutral detergent fiber consumed was higher in the 5CS than in the 8CS cows. Although the 8CS cows moved 3 times more to the milking area for extra cooling sessions than the 5CS ones, they spent 9.9 min/d more than the 5CS ones in lying down (484.4 and 474.5 min/d, respectively), and used more of their free time (excluding milking and feeding time) in resting than the 5CS cows: 52.0 and 43.9%, respectively. It appears that increasing the cooling frequency from 5 to 8 times per day improved their feeling of welfare, so they could spend more time lying and ruminating. In conclusion, increasing the cooling frequency of high-yielding dairy cows under hot and humid conditions from 5 to 8 times a day increased their intake and milk yield, and lowered their respiration rate and rectal temperature. Moreover, the 8CS cows spent more time resting than 5CS cows, an indication that increasing cooling frequency improved animal welfare.