E. Block

Invited review: Recommendations for reporting intervention studies on reproductive performance in dairy cattle: Improving design, analysis, and interpretation of research on reproduction

Abundant evidence from the medical, veterinary, and animal science literature demonstrates that there is substantial room for improvement of the clarity, completeness, and accuracy of reporting of intervention studies. More rigorous reporting guidelines are needed to improve the quality of data available for use in comparisons of outcomes (or meta-analyses) of multiple studies. Because of the diversity of factors that affect reproduction and the complexity of interactions between these, a systematic approach is required to design, conduct, and analyze basic and applied studies of dairy cattle reproduction. Greater consistency, clarity, completeness, and correctness of design and reporting will improve the value of each report and allow for greater depth of evaluation in meta-analyses. Each of these benefits will improve understanding and application of current knowledge and better identify questions that require additional modeling or primary research. The proposed guidelines and checklist will aid in the design, conduct, analysis, and reporting of intervention studies. We propose an adaptation of the REFLECT (Reporting Guidelines for Randomized Controlled Trials for Livestock and Food Safety) statement to provide guidelines and a checklist specific to reporting intervention studies in dairy cattle reproduction. Furthermore, we provide recommendations that will assist investigators to produce studies with greater internal and external validity that can more often be included in systematic reviews and global meta-analyses. Such studies will also assist the development of models to describe the physiology of reproduction.

Effect of fat additions to diets of dairy cattle on milk production and components: A meta-analysis and meta-regression

The objectives of this study were to critically review randomized controlled trials, and quantify, using meta-analysis and meta-regression, the effects of supplementation with fats on milk production and components by dairy cows. We reviewed 59 papers, of which 38 (containing 86 comparisons) met eligibility criteria. Five groups of fats were evaluated: tallows, calcium salts of palm fat (Megalac, Church and Dwight Co. Inc., Princeton, NJ), oilseeds, prilled fat, and other calcium salts. Milk production responses to fats were significant, and the estimated mean difference was 1.05 kg/cow per day, but results were heterogeneous. Milk yield increased with increased difference in dry matter intake (DMI) between treatment and control groups, decreased with predicted metabolizable energy (ME) balance between these groups, and decreased with increased difference in soluble protein percentage of the diet between groups. Decreases in DMI were significant for Megalac, oilseeds, and other Ca salts, and approached significance for tallow. Feeding fat for a longer period increased DMI, as did greater differences in the amount of soluble protein percentage of the diet between control and treatment diets. Tallow, oilseeds, and other Ca salts reduced, whereas Megalac increased, milk fat percentage. Milk fat percentage effects were heterogeneous for fat source. Differences between treatment and control groups in duodenal concentrations of C18:2 and C 18:0 fatty acids and Mg percentage reduced the milk fat percentage standardized mean difference. Milk fat yield responses to fat treatments were very variable. The other Ca salts substantially decrease, and the Megalac and oilseeds increased, fat yield. Fat yield increased with increased DMI difference between groups and was lower with an increased estimated ME balance between treatment and control groups, indicating increased partitioning of fat to body tissue reserves. Feeding fats decreased milk protein percentage, but results were heterogeneous. An increased number of milkings increased the milk protein percentage, whereas the difference between the treatment and control groups in duodenal concentrations of 18:2 fatty acids and dietary Mg concentration reduced the milk protein percentage. None of the fat treatments influenced milk protein production. The range of responses to different fats fed approached or exceeded 5 standard deviations from the mean and differed in point direction for all variables studied, indicating the varied and profound biological effects of fats. Responses to fat feeding were highly heterogeneous for all variables studied and heterogeneity was present within responses to individual fat groups. The lower DMI combined with higher milk and milk fat production showed that fats could improve the efficiency of milk production. More studies are required to more completely characterize sources of variation in responses to fats.

Short Communication: Characterization of Madin-Darby Bovine Kidney Cell Line for Peroxisome Proliferator-Activated Receptors: Temporal Response and Sensitivity to Fatty Acids

The peroxisome proliferator-activated receptors (PPAR) are critical for lipid metabolism, and many fatty acids are PPAR agonists. Madin-Darby bovine kidney (MDBK) cells were tested as an in vitro bovine model for PPAR activation, and preliminary evaluation of the effect of fatty acids on bovine PPAR was performed. Cells were treated with Wy-14643 (WY, specific PPARalpha agonist) and rosiglitazone (ROSI, specific PPARgamma agonist). The gene expression of specific PPARalpha-responsive genes such as carnitine palmitoyl transferase-1 (CPT1A) and acetyl coenzyme A oxidase (ACOX1) and of PPARgamma-responsive gene lipoprotein lipase (LPL) were analyzed using real-time reverse transcription PCR. It was found that CPT1A exhibited a significant increase in cells treated with WY, whereas the ACOX1 gene expression was not altered. The LPL gene expression showed an increase in response to ROSI. Interestingly, LPL was almost undetectable in MDBK cells not treated with ROSI. The potency of different fatty acids in activating PPARalpha as assessed by CPT1A mRNA abundance in MDBK cells was also tested. The mRNA of CPT1A (2.5- to 1.4-fold) was significantly increased by fatty acids in the order of palmitate > linolenate > linoleate > conjugated linoleate, and oleate. The results demonstrated MDBK cells to be responsive to PPAR agonists and thus a promising model to evaluate the role of PPAR in bovine cells. In addition, fatty acids were proven to have a different potency in modulating expression of CPT1A through PPARalpha.

Effect of prepartum anionic supplementation on periparturient feed intake, health, and milk production

Our objectives were to determine if dietary cation-anion difference (DCAD) and source of anions influence periparturient feed intake and milk production of dairy cattle during the transition period. Diets differed in DCAD (cationic or anionic) and anionic supplement. The 4 diets used prepartum were (1) control [DCAD +20 mEq/100g of dry matter (DM)], (2) Bio-Chlor (DCAD -12 mEq/100g of DM; Church & Dwight Co. Inc., Princeton, NJ), (3) Fermenten (DCAD -10 mEq/100g of DM; Church & Dwight Co. Inc.), and (4) salts (DCAD -10 mEq/100g of DM). Urine pH was lower for cows that consumed an anionic diet prepartum compared with control. Prepartum diet had no effect on prepartum dry matter intake (DMI) of multiparous or primiparous cows. Postpartum DMI and milk yield for multiparous cows fed anionic diets prepartum were greater compared with those fed the control diet. Postpartum DMI and milk yield of primiparous cows were similar for prepartum diets. Feeding prepartum anionic diets did not affect plasma Ca at or near calving. However, cows fed anionic diets began their decline in plasma Ca later than control cows. Postpartum β-hydroxybutyrate and nonesterified fatty acids were lower for primiparous cows fed prepartum anionic diets compared with those fed the control diet. Prepartum and postpartum plasma glucose concentrations were not affected by prepartum diet for all cows. Liver triglyceride differed for parity by day. Parities were similar at 21 d prepartum, but at 0 d and 21 d postpartum, levels were greater for multiparous cows. Results indicate that decreasing the DCAD of the diet during the prepartum period can increase postpartum DMI and milk production of multiparous cows without negatively affecting performance of primiparous cows.

Calcium salts of polyunsaturated fatty acids deliver more essential fatty acids to the lactating dairy cow.

Recent research has focused on the importance of supplying essential fatty acids to the lactating dairy cow. The addition of essential fatty acids, specifically linoleic and linolenic acid, to dairy cow diets has been investigated as a method to increase reproductive efficiency. Rumen bacteria, however, biohydrogenate polyunsaturated fatty acids (PUFA) to saturated fatty acids. This is an important issue because it can also lead to milk fat depression when unsaturated fatty acids are fed. The formation of Ca salts has previously been shown to partially protect unsaturated fatty acids from rumen biohydrogenation. The objective of this experiment was to evaluate feed intake, milk production, and milk composition of cows fed Ca salts of palm fatty acids (CS) compared with those fed Ca salts of palm fatty acids with an increased content of PUFA (CS+PUFA). Nineteen lactating Holstein cows were used in a switchback experiment to determine any differences between CS and CS+PUFA on milk production and composition. This experiment consisted of 3 consecutive periods of 14 d. Treatments were formulated to provide 450 g/d (dry matter basis) of the Ca salt supplement and were mixed with the same basal ration. Milk weights and feed intakes were recorded daily for each cow. Milk samples were collected the last 2 d of each period and analyzed for milk composition and fatty acids. Dry matter intake [28.0 vs. 27.0 kg/d; standard error of the mean (SEM) = 0.4] and milk production (44.4 vs. 44.0 kg/d; SEM = 0.7) were not different between treatments for CS and CS+PUFA, respectively. Milk fat percentage (3.34 vs. 3.22%; SEM = 0.07) and milk protein percentage (2.78 vs. 2.80%; SEM = 0.01) were not different for CS- and CS+PUFA-fed cows. Feeding CS+PUFA reduced the concentration of palmitic acid in milk fat (28.3 vs. 26.8 wt%; SEM = 0.3). Supplementation of CS+PUFA increased the linoleic acid concentration (3.96 vs. 4.61 wt%; SEM = 0.1) of milk fat, indicating that linoleic acid was partially protected from rumen biohydrogenation. Concentrations of conjugated linoleic acid were also increased (0.44 vs. 0.52 wt%; SEM = 0.02) when cows consumed CS+PUFA, indicating that some biohydrogenation did occur. Supplementing CS+PUFA did not alter milk production, milk fat percentage, or dry matter intake when compared with CS. The CS+PUFA supplement supplied more linoleic acid to the small intestine for milk fat synthesis.

Effect of supplementing fat to pregnant nonlactating cows on colostral fatty acid profile and passive immunity of the newborn calf.

The objectives were to evaluate the effect of supplementing saturated or unsaturated long-chain fatty acids (FA) to nulliparous and parous Holstein animals (n=78) during late gestation on FA profile of colostrum and plasma of newborn calves and on production and absorption of IgG. The saturated FA supplement (SAT) was enriched in C18:0 and the unsaturated FA supplement (ESS) was enriched in the essential FA C18:2n-6. Fatty acids were supplemented at 1.7% of dietary dry matter to low-FA diets (1.9% of dietary dry matter) during the last 8 wk of gestation. Calves were fed 4 L of colostrum within 2h of birth from their own dam or from a dam fed the same treatment. Feeding fat did not affect prepartum dry matter intake, body weight change, or gestation length. Parous but not nulliparous dams tended to give birth to heavier calves if fed fat prepartum. Parous dams were less able to synthesize essential FA derivatives, as evidenced by lower desaturase indices, compared with nulliparous dams, suggesting a greater need for essential FA supplementation. The FA profile of colostrum was modified to a greater degree by prepartum fat feeding than was that of neonatal calf plasma. The placental transfer and synthesis of elongated n-3 FA (C20:5, C22:5, and C22:6) were reduced, whereas the n-6 FA (C18:2, C18:3, and C20:3) were increased in plasma of calves born from dams fed ESS rather than SAT. Supplementing unsaturated FA prepartum resulted in elevated concentrations of trans isomers of unsaturated monoene and diene FA, as well as C18:2n-6 in colostrum. Serum concentrations of IgG tended to be increased in calves born from dams fed fat compared with those not fed fat, and prepartum feeding of SAT tended to improve circulating concentrations of IgG in newborn calves above the feeding of ESS. Apparent efficiency of absorption of IgG was improved in calves born from dams fed fat, and SAT supplementation appeared more effective than supplementation with ESS. Feeding SAT prepartum may be of greater benefit based upon greater circulating IgG concentrations of calves after colostrum feeding. Feeding moderate amounts of saturated or unsaturated long-chain FA during the last 8 wk of gestation changed the FA profile of colostrum and plasma of neonates to reflect that of the supplements.

Effectiveness of potassium carbonate sesquihydrate to increase dietary cation-anion difference in early lactation cows

The effect of additional dietary potassium in early lactation dairy cows was evaluated with the addition of potassium carbonate sesquihydrate, which increased dietary K from 1.3 to 2.1% of dry matter (DM) from wk 3 to 12 of lactation. Cows fed potassium carbonate sesquihydrate in the form of DCAD Plus (Church & Dwight Co. Inc., Princeton, NJ) had increased DM intake, milk fat percentage and yield, energy-corrected milk, and efficiency of milk production per unit of DM intake. Milk fat of cows fed higher dietary K had a lower concentration of trans fatty acids, suggesting a role for potassium carbonate sesquihydrate in the rumen in the biohydrogenation processes converting linoleic to stearic acid. Cows fed the diet with 2.1% K had greater apparent balance of K, and no effects were noted on the concentration of blood Mg or amount of fecal Mg. The data support the feeding of greater amounts of K in the early lactation cow.

Effect of feeding calcium salts of long-chain fatty acids, from palm fatty acid distillate or soybean oil, to high producing dairy cows on milk yield and composition, and on selected blood and reproductive parameters

Abstract Calcium salts of long-chain fatty acids from palm fatty acid distillate (CaPFAD) and soybean oil (CaSOFA) were compared using 14 first lactation and 26 multiparous Holstein cows housed in a free-stall barn. It was hypothesized that if replacing CaPFAD with CaSOFA, a rich source of linoleic acid, could result in increased conjugated linoleic acid (CLA) content in milk without losing the lactational and reproductive benefits of feeding CaPFAD, this would be beneficial to the dairy industry and public health, as well as potentially enhance the consumption of dairy products. Cows were group-fed one of two total mixed rations (TMR) containing 1.7% CaPFAD or CaSOFA (DM basis) for ad libitum intake following recommendations from the manufacturer of the fat supplements. Fatty acid profiles (g/100xa0g of fatty acid) of CaPFAD and CaSOFA, respectively, were 16:0: 48.1, 12.1; 18:0: 4.27, 5.34; 18:1: 35.7, 23.9; and 18:2: 8.9, 51.2. Cows spent about 45 days in the fresh group, after which they were moved to high producing group where they stayed up to 10 weeks post-partum within their treatment assignments. The TMR (DM basis) were 50:50 forage:concentrate for both the fresh group and high producing group cows and contained (DM) 28% NDF and 18% CP. There were no treatment differences in milk yield, milk fat percentage, milk CLA content, milk true protein percentage, milk lactose percentage, milk urea N, linear somatic cell count and change in body condition score. Cows fed CaSOFA produced milk containing a higher content of 18:1(t) ( P P P =0.03). Cows had similar concentrations of serum nonesterified fatty acids and β-hydroxybutyrate, and similar 30 days uterine score, days to first ovulation, days to first service, first service conception rate and progesterone concentration. In conclusion, while cows fed CaSOFA had similar lactational and reproductive performance to cows fed CaPFAD, the anticipated beneficial effect of enhancing content of CLA in milk was not realized from feeding CaSOFA at 1.7% of expected DM intake.

Responses of the estrous cycle in dairy cows exposed to electric and magnetic fields (60 Hz) during 8-h photoperiods.

To study the effects of exposure to extremely low frequency (ELF) electric and magnetic fields (EMF) on the estrous cycle of dairy cows under short-day photoperiod, 16 non-lactating, non-pregnant Holstein cows were exposed to a vertical electric field of 10 kV/m and a horizontal magnetic field of 30 microT for 16 h per day in a cross-over design consisting of two sequences. Each sequence included three periods, and each period corresponded to the duration of one estrous cycle. All animals were maintained under short photoperiod (8 h light/16 h dark) during the trial. Exposure to EMF had an impact on the duration of a complete estrous cycle (P<0.01) and on the duration of the luteal phase (P<0.01). The mean duration of one cycle was 19.5+/-0.4 for the control and 21.3+/-0.4 days for the exposed animals, respectively. The mean duration of the luteal phase was 15.4+/-0.4 days for the control and 17.2+/-0.4 days for the exposed group. The total area under the progesterone (P(4)) curve, the amplitude of the curve or the slope of the P(4) rise at the onset of the luteal phase were not affected by EMF exposure. Results indicate that exposure to EMF may increase the duration of the estrous cycle.

Extended negative dietary cation-anion difference feeding does not negatively affect postpartum performance of multiparous dairy cows

Low postpartum blood calcium remains one of the largest constraints to postpartum feed intake, milk yield, and energy balance in transitioning dairy cows. Supplemental dietary anions decrease the dietary cation-anion difference (DCAD) and reduce the risk for postpartum hypocalcemia. Prepartum management strategies aiming to minimize social stress and diet changes have resulted in a need to explore the effects of extended exposure to a negative DCAD (>21 d) diet. Holstein and Holstein-cross dairy cows (n=60) were assigned to 1 of 3 treatments 42 d before expected calving to evaluate effects of supplying anions for 21 or 42 d during the dry period on energy status, postpartum production, and Ca homeostasis. Treatments included (1) a control diet (CON; DCAD=12 mEq/100 g of DM), (2) a 21-d negative DCAD diet (21-ND; DCAD=12 and -16 mEq/100 g of DM), and (3) a 42-d negative DCAD diet (42-ND; DCAD=-16 mEq/100 g of DM). Cows fed CON were fed positive DCAD prepartum for 42 d. Cows fed 21-ND received the positive DCAD (12 mEq/100 g of DM) diet for the first 21 d of the dry period and the anionic diet (-16 mEq/100 g of DM) from d 22 until calving. Cows fed 42-ND received the anionic diet for the entire dry period. Control and anionic diets were formulated by using 2 isonitrogenous protein mixes: (1) 97.5% soybean meal and (2) 52.8% BioChlor (Church & Dwight Co. Inc.), 45.8% soybean meal. Supplementing anions induced a mild metabolic acidosis, reducing urine pH for 21-ND and 42-ND compared with CON. Prepartum DMI was not different among treatments. Postpartum DMI was higher for 21-ND compared with CON (20.8 vs. 18.1±1.1 kg/d), and 42-ND had similar DMI compared with 21-ND. During the first 56 d of lactation 21-ND had greater average milk production compared with CON (44.8 vs. 39.2±2.1 kg/d). Average milk production by 42-ND was similar to 21-ND. Postpartum total blood Ca concentration was greater for 42-ND. Cows fed anionic diets prepartum tended to have lower lipid accumulation in the liver after calving compared with CON. These data suggest low-DCAD diets fed for 21 or 42 d during the dry period can have positive effects on postpartum DMI, Ca homeostasis, and milk production.