Holstein Cows In Late Lactation Research Articles

Impact of dairy cow personality traits and concentrate allowance on their response to training and adaptation to an automated milking system

The objectives of this study were to determine: 1) if dairy cow personality traits and concentrate allowance are associated with the behavior and performance of cows during training to use an automated milking system (AMS); and 2) if these factors were associated with the behavior and performance of cows after AMS training. Twenty-nine mid- to late-lactation Holstein cows (218 ± 49 DIM), who were milking on a rotary parlor and had never previously been milked in an AMS, were enrolled in this study. Cows were assigned to 1 of 2 dietary treatments, consisting of a basal partial mixed ration (PMR) common to both treatment groups, with a concentrate allowance (on dry matter basis) of: 1) 2.0 kg/d in the AMS (L-Tx); or 2) 6.0 kg/d in the AMS (H-Tx). Cows were trained to use the free-traffic AMS, with supervised milkings, over 72 h and were milked in this system for 63 d after training was complete. Variables relating to feeding behavior, milking activity, and production were measured from the start of AMS training until the end of the study. Between 42 and 63 d after AMS introduction, each cow was assessed for personality traits using a combined arena test consisting of exposure to a novel environment, novel object, and novel human. Principal components analysis of behaviors observed during the personality assessment revealed 2 factors (interpreted as boldness and activeness traits) that together explained 85% of the variance; each cow received a score for each trait. Associations between dietary treatment and personality traits with feeding behavior, milking activity, and production were analyzed using mixed-effect linear and logistic regression models. Cows with greater scores for the active trait produced less milk during the 3 d of AMS training compared with cows with lower scores. Within the H-Tx, more active cows had a 3.92 times greater risk of kicking off teat cups during AMS training than less active cows. However, during the 8 wk after training, more active cows had a 1.37 times lesser risk of teat cup kickoffs than those that were less active. Cows on the H-Tx produced 4.4 kg/d more energy-corrected milk compared with cows on the L-Tx in the 8 wk after training. During the 8 wk after AMS training the cows on the H-Tx consumed an average of 21.4 kg/d of PMR and were delivered 4.6 kg/d of AMS concentrate, while the L-Tx cows consumed 23.4 kg/d PMR and were delivered 2.0 kg/d of AMS concentrate. The results indicate that both dairy cow personality traits and AMS concentrate allocation influence their response to AMS training and subsequent feeding and milking behavior and production.

Effects of rumen-protected niacin on inflammatory response to repeated intramammary lipopolysaccharide challenges

Nutritional strategies that improve an animal's resilience to various challenges may improve animal health and welfare. One such nutrient is niacin which has reduced inflammation in mice, humans, and swine; however, niacin's anti-inflammatory effects have not been investigated in cattle. Our objective was to determine whether rumen-protected niacin (RPN) alters lactating dairy cows' inflammatory response to intramammary lipopolysaccharide (LPS) challenges, whether RPN resulted in any carry-over effects, and whether repeated LPS challenges result in signs of immune tolerance or innate immune training. Twenty healthy, late-lactation Holstein cows (232 ± 65 d in milk; 39 ± 5.8 kg/d of milk) were enrolled in a randomized complete block experiment which lasted 70 d. Cows received 26 g/d of RPN or no top-dress (CON) for the first 42 d of the experiment. During the final milking of d 27 and 55, cows were challenged in their rear-right mammary gland (RR) with 100 µg of LPS suspended in 5 mL of phosphate buffered saline. Milk yield, milk conductivity, and feed intake were measured daily. Milk composition was measured on d 14, 23, 24, 30, 37, 45, and 52. Blood samples were collected at 0, 8, 12, 24, 48, 72, 96, and 120 h after each LPS challenge, whereas RR quarter milk samples were collected at 0, 8, 16, 24, 48, 72, 96, 120, 144, and 168 h after each LPS challenge. Body temperature was measured continuously during each challenge with an intravaginal thermometer. Linear mixed models with repeated measures were used to analyze the results. Before LPS challenge, RPN did not affect feed intake or milk production, but it reduced SCS (1.24 ± 0.41 vs. 0.05 ± 0.45). After challenge, RPN did not affect feed intake, milk production, milk composition, SCS, body temperature, plasma glucose, or plasma insulin concentrations. Our results suggest RPN reduced peak plasma haptoglobin and lipopolysaccharide binding protein (LBP) during the 1st LPS challenge. Plasma haptoglobin tended to be less after the 2nd challenge for cows previously supplemented RPN while LBP was similar for each treatment group after the 2nd challenge. The 2nd LPS challenge resulted in decreased plasma haptoglobin compared with the 1st LPS challenge, suggestive of tolerance but it also induced a greater peak SCS than the 1st LPS challenge. Our results suggest that repeated LPS challenges promote a systemic tolerance but heightened local response to LPS-induced mastitis. Feeding RPN reduced SCS before challenge and reduced plasma acute phase proteins after challenge suggesting that RPN may reduce systemic inflammation without altering the local inflammatory responses.

Effect of nicotinic acid supplementation on digestion, metabolism, microbiome, and production in late-lactation Holstein cows

The purpose of this experiment was to determine if nicotinic acid (NA) effects on dairy cows and rumen microbial characteristics are forage type dependent (corn silage, CS; grass silage, GS). Four late lactation (days in milk = 225 +/- 12 d) Holstein cows were used in a 4 × 4 Latin square design with a 2 × 2 factorial arrangement of treatments. The main effects were a CS (66.10% CS) based diet or a GS (79.59%) based diet with or without 12 g/d NA. Each experimental period lasted for 28 d. Milk production and milk components, blood metabolites, apparent total-tract nutrient digestibilities, minutes rumen pH were below 5.8 as an indicator of ruminal acidosis, and body temperature changes were analyzed as indicators of heat stress. Nicotinic acid supplementation did not improve apparent total-tract nutrient digestibility. Feeding a GS-based diet improved NDF and hemicellulose digestibility. Feeding a CS-based diet increased the apparent total-tract digestibility of fat, and minutes rumen pH below 5.8 for a greater proportion of the time. The CS-based diet also improved milk yield, milk fat and protein yields, and energy-corrected milk yield; however, somatic cell count and BHB were also increased. Supplementing NA tended to decrease nonesterified fatty acids, especially when combined with GS where DMI was low. There was a trend for the total protozoa population to increase when GS and NA were fed but decreased when CS and NA were fed. Nicotinic acid tended to decrease rumen protozoal populations of Dasytricha, but increased populations of Ophryoscolex and Diplodiniinae with GS diets and decreased with CS diets. Entodiniinae were increased with CS but NA had no effect. Body temperature was increased when a CS-based diet was fed when compared with a GS-based diet. More research is needed to determine how NA can affect rumen microbial protein synthesis and what kind of diets will provide the optimum effect.

Ruminal inocula with distinct fermentation profiles differentially affect the in vitro fermentation pattern of a commercial algal blend.

The in vitro rumen batch technique is widely used for screening novel feed sources; however, it remains unclear to what extent the in vitro fermentability of non-conventional feed sources is affected by non-adapted ruminal inocula. Thus, in this study, we evaluated the effects of distinct ruminal inocula on the in vitro fermentation parameters of a sustainable non-conventional feed, a commercially available algal blend composed of microalgae (Chlorella vulgaris and Nannochloropsis oceanica) and seaweeds (Ulva sp. and Gracilaria gracilis). First, four late-lactation Holstein cows were fed four forage-based diets varying only in the proportions of basal forage (100% corn silage, 70% corn silage and 30% haylage, 30% corn silage and 70% haylage, and 100% haylage) in a 4 × 4 Latin square design with the last square omitted. After 3 weeks of adaptation, haylage-based diets resulted in ruminal fermentation parameters distinct from those promoted by corn silage-based diets, as reflected in increased pH, ammonia-N contents, and acetate proportions. Individual ruminal fluids derived from each of the four diets were further used as inocula in in vitro incubations. Here, a 1:1 mixture of corn silage and haylage was supplemented with 0, 5, 10, or 15% algal blend and incubated with each inoculum for 24 h in a 4 × 4 factorial design. Total gas and methane production decreased with inocula from cows fed haylage-based diets and with increasing algal blend supplementation levels. The fermentation pH increased and the ammonia-N contents decreased with inocula from cows fed haylage-based diets; however, these parameters were not affected by algal blend inclusion levels. The interaction between the ruminal inoculum source and the algal blend supplementation level affected the total volatile fatty acids (VFA) and the proportions of most individual VFA. Total VFA production decreased with increasing algal supplementation levels, particularly with inocula from cows fed 30% corn silage and 70% haylage; the acetate, propionate, and valerate proportions were only affected by algal blend levels under incubation with 100% corn silage inocula. Overall, our findings highlight the importance of the ruminal inoculum source when assessing the fermentability of non-conventional feed as well as the potential of the algal blend as a natural modulator of ruminal fermentation.

Reducing dietary crude protein: Effects on digestibility, nitrogen balance, and blood metabolites in late-lactation Holstein cows

Our objectives were to determine the effects of reducing dietary crude protein (CP) concentration on nutrient digestibility, rumen function, N balance, and serum AA concentration for dairy cows in late lactation. At the initiation of the experimental period, we stratified Holstein cows (n = 128; mean ± standard deviation 224 ± 54 d in milk) by parity and days pregnant (86 ± 25 d) and assigned them to 1 of 16 pens. For 3 wk, all cows received a covariate diet containing 16.9% CP [dry matter (DM) basis]. For the subsequent 12 wk, we assigned pens to 1 of 4 treatments containing 16.2, 14.4, 13.4, or 11.9% CP (DM basis) in a randomized complete block design. Diets were fed as a total mixed ration once daily. To reduce dietary CP, we replaced soybean meal with soybean hulls in the concentrate mix (DM basis). Diet evaluations suggested that several EAA, especially His, limited productivity as dietary CP declined. Digestibility of DM and CP decreased linearly with dietary CP reduction. Digestibility of neutral detergent fiber and potentially digestible neutral detergent fiber tended to respond in a quadratic pattern with the greatest digestibility at intermediate treatments. The reduction in dietary CP did not affect ruminal pH, but ruminal ammonia-N and branched-chain VFA concentrations declined linearly. The concentration of milk urea-N and plasma urea-N, secretion of milk N, and excretions of fecal N, urinary N, urinary urea-N, and unaccounted N decreased linearly with the reduction in dietary CP concentration. Urinary N expressed as a percentage of N intake was unaffected by dietary CP. Serum concentrations of total essential AA and non-essential AA were unaffected by dietary CP concentration. However, the ratio of essential to non-essential AA decreased with decreasing dietary CP. Serum 3-methylhistidine concentration increased linearly with decreasing dietary CP concentration, indicating greater skeletal muscle breakdown. Although our trial confirmed that reducing dietary CP decreased absolute excretion of urinary N, diet evaluations suggested that milk protein production decreased as certain essential AA became increasingly limited. Thus, reduced-CP diets have the potential to lessen reactive-N outputs of late lactation cows, but more research is needed to design diets that minimize deleterious effects on productivity.

Effects of substitution of millet straw for corn silage and alfalfa hay on lactation performance, ruminal fermentation, and blood metabolites in late-lactation Holstein dairy cows

The study was conducted to evaluate the effect of replacing dietary corn silage (CS) and alfalfa hay (AH) with millet straw (MS) on the production performance and the ruminal fermentation patterns of Holstein cows in late lactation. For this purpose, forty-eight multiparous Holstein dairy cows blocked by milk yield (32 ± 3.2 kg/d) and days in milk (179 ± 19 d) were randomly allocated to one of four treatments. The diets were formulated with a forage-to-concentrate ratio of 48:52 [dry matter (DM) basis] and contained the same concentrate mixtures, with different forage sources (on a DM basis): (1) Control (60% CS, 40% AH, and 0% MS; on 100% roughage); (2) LMS (54% CS, 36% AH, and 10% MS), (3) MMS (48% CS, 32% AH, and 20% MS), and (4) HMS (42% CS, 28% AH, and 30% MS). The dry matter intake and milk yield were lower in the HMS diet than those in the Control, LMS, and MMS diets (P < 0.01), whereas there was no difference in the Control, LMS, and MMS diets. Compared with the Control diet, 4% FCM, milk protein yield, and N conversion ratio decreased in the MMS and HMS diets (P < 0.01), whereas no difference between the Control and LMS diets was found. Besides, cows fed the HMS diet had lower milk fat yield, lactose percentage, and feed efficiency than those in the Control and LMS diets (P < 0.05), with no difference between the Control, LMS, and MMS diets. The percentage of milk fat and protein was not affected by the diets. Compared with the Control diet, the total VFA concentration was decreased, whereas the ammonia-N concentration was increased in the HMS diet (P < 0.05), with no difference between the Control, LMS, and MMS diets. The molar proportion of acetate and the ratio of acetate to propionate were lower (P < 0.01), while the molar proportion of propionate was higher in the Control diet than that in the MMS and HMS diets (P < 0.01), with no difference between the LMS and MMS diets. Compared with the Control, the apparent digestibility of DM, CP, NDF, and ADF was decreased, while the blood urea nitrogen concentration was increased in the HMS diet (P < 0.05), with no differences in the Control, LMS, and MMS diets. Thus, it was concluded that it is feasible to use MS as a fiber source for late-lactation dairy cows. However, considering the milk production performance and the feed digestibility, the replacement of CS and AH with MS should not exceed 20% of the roughage.

Changes in plasma and milk choline metabolite concentrations in response to the provision of various rumen-protected choline prototypes in lactating dairy cows.

Choline feeding in the form of rumen-protected choline (RPC) has been shown to increase milk production and improve measures of metabolic health (e.g., liver triglyceride) in dairy cows. The objective was to characterize changes in plasma and milk choline and choline metabolite concentrations, including microbial-derived trimethylamine N-oxide (TMAO), in response to increasing ruminal spot-doses, different types of RPC, and ruminal stability of RPC in lactating cows. For experiment 1, 12 mid-lactation (121 ± 16.3 d in milk) Holstein cows were balanced by total plasma choline concentrations and milk yields. Cows were assigned to 1 of 3 lipid-encapsulated RPC products (main plots): prototypes P1, P2, and P3 (containing 59, 56, and 30% choline chloride, respectively). Within each main plot, cows were assigned to a sequence of doses in a 4 × 4 Latin square design: 0, 18, 36, or 54 g of choline chloride. Treatments were preconditioned with ground corn and administered as a single ruminal bolus once per experimental period 1 h postfeeding of a total mixed ration. For experiment 2, we compared a control (0 g of choline chloride) versus P2, and P4 and P5 (60 and 62% choline chloride, respectively) in a repeated 4 × 4 Latin square design. Experiment 2 followed a similar design as experiment 1 with modifications: 12 late-lactation (228 ± 7.10 d in milk) Holstein cows were used; treatments were administered as part of a premeal; and cows received a daily allowance of a total mixed ration as equal provisions every 4 h within 24 h before and after treatment. For both experiments, plasma and milk samples were collected for choline and choline metabolite quantification. Data were analyzed using a mixed model including fixed effects of treatment, period, and time. Contrast statements were used to test for linearity of dose and differences between prototypes for experiment 1 and 2, respectively. Plasma and milk TMAO concentrations increased with RPC dose (peak by h). Milk choline and betaine yields increased with RPC dose in a quadratic manner; albeit, dependent upon RPC type. Milk phosphocholine (PCho) and glycerophosphorylcholine (GPC) yields changed by select RPC dose (experiment 1), however Met, PCho, GPC, phosphatidylcholine, and total choline concentrations in milk, and plasma Met and sphingomyelin concentrations were not responsive. We conclude that plasma or milk choline, betaine, and TMAO concentrations are responsive to RPC type, dose, and stage of lactation evaluated.

Cows enrolled in a pilot randomized clinical trial evaluating pegbovigrastim administered to dairy cows at dry-off had reduced incidence of intramammary infection.

The objective of this pilot study was to determine if an alternative dosing schedule of pegbovigrastim (PEG; Imrestor; Elanco Animal Health) affects mammary gland health, rear udder width, or milk production of healthy dairy cows. 20 pregnant late-lactation Holstein cows in November 2019 through April 2020. Cows were randomly assigned to receive subcutaneous injections with either 15 mg of PEG (PEG group; n = 10) or a sham injection with saline (0.9% NaCl) solution (control group; 10) administered 7 days before dry-off and at dry-off. Quarter milk samples were collected for bacterial culture and somatic cells before and after dry-off and after calving. Mammary gland width was assessed before and after dry-off. Daily milk yields were evaluated after calving. The incidence of intramammary infection was 5 times greater for quarters of cows in the control group than for quarters of cows that received PEG. The effect of treatment on somatic cell count was not significant, but the effects of period and a treatment-by-period interaction were identified. Treatment did not significantly affect milk production in the subsequent lactation, but the effects of period and an interaction of treatment by period were identified. Rear udder width after dry-off was not significantly affected by treatment, but an effect of period was identified. In this pilot study, cows treated with PEG using an alternative dosing schedule had reduced incidence of intramammary infection and an interaction of treatment by sampling period was observed for milk yield. These results suggest that further studies with larger numbers of cows are warranted.

Effects of acute intravenous lipopolysaccharide administration on the plasma lipidome and metabolome in lactating Holstein cows experiencing hyperlipidemia.

The effects of lipopolysaccharides (i.e., endotoxin; LPS) on metabolism are poorly defined in lactating dairy cattle experiencing hyperlipidemia. Our objective was to explore the effects of acute intravenous LPS administration on metabolism in late-lactation Holstein cows experiencing hyperlipidemia induced by intravenous triglyceride infusion and feed restriction. Ten non-pregnant lactating Holstein cows (273 ± 35 d in milk) were administered a single bolus of saline (3 mL of saline; n [Formula: see text] 5) or LPS (0.375 [Formula: see text]g of LPS/kg of body weight; n [Formula: see text] 5). Simultaneously, cows were intravenously infused a triglyceride emulsion and feed restricted for 16 h to induce hyperlipidemia in an attempt to model the periparturient period. Blood was sampled at routine intervals. Changes in circulating total fatty acid concentrations and inflammatory parameters were measured. Plasma samples were analyzed using untargeted lipidomics and metabolomics. Endotoxin increased circulating serum amyloid A, LPS-binding protein, and cortisol concentrations. Endotoxin administration decreased plasma lysophosphatidylcholine (LPC) concentrations and increased select plasma ceramide concentrations. These outcomes suggest modulation of the immune response and insulin action. Lipopolysaccharide decreased the ratio of phosphatidylcholine to phosphatidylethanomanine, which potentially indicate a decrease in the hepatic activation of phosphatidylethanolamine N-methyltransferase and triglyceride export. Endotoxin administration also increased plasma concentrations of pyruvic and lactic acids, and decreased plasma citric acid concentrations, which implicate the upregulation of glycolysis and downregulation of the citric acid cycle (i.e., the Warburg effect), potentially in leukocytes. Acute intravenous LPS administration decreased circulating LPC concentrations, modified ceramide and glycerophospholipid concentrations, and influenced intermediary metabolism in dairy cows experiencing hyperlipidemia.

Fermented Chinese herbal preparation: Impacts on milk production, nutrient digestibility, blood biochemistry, and antioxidant capacity of late-lactation cows under heat stress

The objective of this study was to investigate the effects of dietary supplementation of fermented Chinese herbal preparation (CHP) on the milking performance, nutrient digestibility, blood biochemistry, and antioxidant capacity in cows under heat stress (HS). A total of 40 late-lactation Holstein cows (Average body weight: 560 ± 51.0 kg, milk per day: 16 ± 3.0 kg) were randomly assigned to 4 groups (10 cows per group). The cows in treatment groups were supplied with 25, 50, or 100 g CHP in diet per cow per day, and the control group was no supplied with CHP. All cows were raised at a natural HS environment (Average THI: 74.5) during 42-d experimental period. Milk, fecal samples, and blood samples were collected on 14, 28, and 42 d for measuring milk production, nutrient apparent digestibility, and serum biochemical parameters, heat shock protein involving HSP70, HSP90, and HSF. The antioxidant enzymes (T-AOC, SOD, CAT, GPx, and PRDX2) activities in blood and its genes expression level were also measured on 14, 28, and 42 d of experiment. The results showed that the dietary supplementation of CHP demonstrated an increased tendency in DMI (P = 0.08), and 50 or 100 g/d CHP increased milk yield, milk composition (fat and protein) content (all P < 0.05) for the entire 42-d period, compared with the control group. However, there was no difference (P > 0.05) on nutrient digestibility among all groups. From blood biochemistry, the HSP90 or HSP70 level in serum exhibited a decrease (P < 0.05) with CHP supplements. Also, the serum triiodothyronine and glucose content were increased (P ≤ 0.02) by CHP, while the prolactin content demonstrated an increasing trend (P = 0.08). For antioxidant capacity, the doses of CHP increased serum T-AOC (P < 0.05), SOD (P = 0.02), and CAT (P < 0.01) concentration, and the mRNA levels of both SOD and CAT genes were also up-regulated (P ≤ 0.05) for groups with CHP. Besides, there was a positively correlated between milking yield or milk quality (milk protein and milk fat) with DMI (P < 0.01), various of antioxidant enzymes concentrations in serum, or mRNA levels of SOD and CAT (P < 0.05). However, the negative correlation between milking performance and HSP70 or HSP90 was also exhibited (P < 0.05). Therefore, the CHP supplements may improve blood biochemistry and antioxidant capacity under HS to increase milking performance in dairy cows.

Methionine supplementation during a hydrogen peroxide challenge alters components of insulin signaling and antioxidant proteins in subcutaneous adipose explants from dairy cows

Enhanced postruminal supply of methionine (Met) during the peripartal period alters protein abundance of insulin, AA, and antioxidant signaling pathways in subcutaneous adipose tissue (SAT). Whether SAT is directly responsive to supply of Met and can induce molecular alterations is unknown. Our objective was to examine whether enhanced Met supply during an oxidative stress challenge in vitro alters insulin, AA, inflammation, and antioxidant signaling-related protein networks. Four late-lactation Holstein cows (average 27.0 kg of milk per day) were used for SAT collection. Tissue was incubated in duplicate for 4 h in a humidified incubator with 5% CO2 at 37°C according to the following experimental design: control medium with an "ideal" profile of essential AA (CTR; Lys:Met 2.9:1), CTR plus 100 μM H2O2 (HP), or CTR with greater Met supply plus 100 μM H2O2 (HPMET; Lys:Met 2.5:1). Molecular targets associated with insulin signaling, lipolysis, antioxidant nuclear factor, erythroid 2 like 2 (NFE2L2), inflammation, and AA metabolism were determined through reverse-transcription quantitative PCR and western blotting. Data were analyzed using the MIXED procedure of SAS 9.4 (SAS Institute Inc.). Among proteins associated with insulin signaling, compared with CTR, HP led to lower abundance of phosphorylated AKT serine/threonine kinase (p-AKT) and solute carrier family 2 member 4 (SLC2A4; insulin-induced glucose transporter). Although incubation with HPMET restored abundance of SLC2A4 to levels in the CTR and upregulated abundance of fatty acid synthase (FASN) and phosphorylated 5'-prime-AMP-activated protein kinase (p-AMPK), it did not alter p-AKT, which remained similar to HP. Among proteins associated with AA signaling, compared with CTR, challenge with HP led to lower abundance of phosphorylated mechanistic target of rapamycin (p-MTOR), and HPMET did not restore abundance to CTR levels. Among inflammation-related targets studied, incubation with HPMET led to greater protein abundance of nuclear factor kappa B subunit p65 (NFKB-RELA). The response in NFKB observed with HPMET was associated with a marked upregulation of the antioxidant transcription regulator NFE2L2 and the antioxidant enzyme glutathione peroxidase 1 (GPX1). No effects of treatment were detected for mRNA abundance of proinflammatory cytokines or antioxidant enzymes, underscoring the importance of post-transcriptional regulation. Overall, data indicated that short-term challenge with H2O2 was particularly effective in reducing insulin and AA signaling. Although a greater supply of Met had little effect on those pathways, it seemed to restore the protein abundance of the insulin-induced glucose transporter. Overall, the concomitant upregulation of key inflammation and antioxidant signaling proteins when a greater level of Met was supplemented to oxidant-challenged SAT highlighted the potential role of this AA in regulating the inflammatory response and oxidant status. Further studies should be conducted to assess the role of postruminal supply of Met and other AA in the regulation of immune, antioxidant, and metabolic systems in peripartal cow adipose tissue.

Providing heat-stress abatement to late-lactation Holstein cows affects hormones, metabolite blood profiles, and hepatic gene expression but not productive responses

Our objective was to evaluate the effects of providing shade and shade combined with evaporative cooling on production, cow activity, metabolism, and hepatic gene expression of late-lactation Holstein dairy cows under moderate heat-stress conditions. Forty-eight multiparous Holstein cows were used in a completely randomized block-design trial and randomly assigned to 1 of 3 treatments: control (CTL), without access to shade; access to artificial shade (SH); and shade combined with evaporative cooling (SHplus). Results were analyzed using a mixed procedure with repeated measures. No differences were observed in DMI. Milk yield was not different among treatments, but lactose concentration was greater in SHplus. Treatments CTL and SH had greater BW losses than SHplus. Control cows spent less time grazing. The CTL and SH had higher p.m. rectal temperatures than SHplus, whereas CTL had the highest p.m. respiration rate. Control cows had greater serum insulin levels. Control and SH had greater BHB and urea concentrations and lower glucose concentration compared with SHplus. The hepatic expression of PCK1 , PDK4 , and HP genes was downregulated in SH and SHplus relative to control. Hepatic expression of NFKB1 was downregulated, whereas SOCS2 was upregulated, for SHplus compared with CTL. Despite the absence of treatment effects on productive variables, changes in blood profiles and hepatic expression of target genes were observed among treatments. These results suggest that the provision of shade combined with evaporative cooling was effective in alleviating the negative effects of heat stress.

Effects of feeding frequency and oil supplementation on feeding behavior, ruminal fermentation, digestibility, blood metabolites, and milk performance in late-lactation cows fed a high-forage diet

Many dairy producers are keen to feed low-producing late-lactation cows only once per day (1×) to reduce production costs. This study examined effects of feeding frequency (FF: thrice versus once daily) on behavioral patterns, ruminal fermentation, and milk production performance of cows and supplementation of yellow grease oil (YO) rich in 18:2n-6 as a potential strategy to alleviate the possible negative effects of 1× daily feeding. Twenty-four late-lactation Holstein cows (215 ± 53.8 DIM) housed in tiestalls were assigned to 4 treatments according to a 2 × 2 factorial arrangement with 2 FF [3 times daily (3×) at 0800, 1400, and 2000 h; or 1× at 0800 h] and 2 high-forage total mixed rations (TMR), without (CON) or with 25 g/kg of dry matter of YO (YGO), in a randomized complete block design. Treatments were applied for 21 d. Feeding behavior was recorded every 5 min over a 24-h period on d 19. Fresh TMR and orts were sampled (d 15 to 21) and separated using a 3-screen (19, 8, and 1.18 mm) Penn State Particle Separator for sorting activity. Ruminal fluid samples were collected using oral stomach tubing on d 21. Cows on 1×-CON spent more time eating during the first 6 h after feeding at 0800 h than did cows on 3×-CON or 1×-YGO. Decreasing FF increased meal length and tended to increase meal size for CON cows, but supplementing YO increased meal bouts and reduced meal length and size for cows fed 1×. Cows on 1×-CON had the greatest ruminating time between 2000 and 0800 h compared with other treatments. Total and daytime distribution of lying time did not vary by treatments. Sorting activity was higher for cows fed frequently, and the extent of sorting was increased by oil supplementation. In the morning ruminal fluid samples, pH was not different among treatments, but in the evening samples 1× daily feeding reduced ruminal pH compared with 3×. In the morning and evening samples, ratios of acetate to propionate were the lowest for 1×-CON cows compared with other treatments. Dry matter intake and milk yield were similar among the groups. Milk fat content and yield decreased with 1×-CON treatment, but supplementing YO numerically increased milk fat for cows fed 1×. These results suggest that decreasing FF from 3× to 1× daily increases meal length, particularly after feed delivery, in cows fed high-forage diets; but supplementation of plant oil changes feeding patterns and may improve ruminal pH and milk fat in cows fed once a day.

Technical note: A novel approach to estimate dry matter intake of lactating dairy cows through multiple on-cow accelerometers

The objective of this study was to evaluate the feasibility of using multiple 3-dimensional accelerometers to estimated individual dry matter intake (DMI) of lactating dairy cows. Twenty-four Holstein cows in late lactation were assigned into 2 groups, a calibration group (n = 12) and a validation group (n = 12). All cows were fitted with 3 sensors that recorded 3-dimensional acceleration (i.e., x, y, and z) at 10-s intervals, 1 on the lateral side of the left hind leg and 2 attached directly to a halter over the nose and jaw area on the left side. Then, 3 accelerations were generated from each accelerometer (e.g., Leg-X, Leg-Y, and Leg-Z). Six new variables were created based on the change in acceleration in the nose and jaw accelerometers between 2 consecutive time points (e.g., LagJaw-X). For both groups (i.e., calibration and validation), cows were continuously video recorded while data on acceleration and intake of total mixed ration were collected for 10 consecutive days. Cows were fed once daily using an individual gate system, and individual refusals were recorded next day before morning feeding. Cows were fed a common lactating cow diet (17.9% crude protein; 1.70 Mcal/kg of dry matter). In the calibration group, individual eating bouts were obtained based on video recordings and merged with the corresponding accelerometer data. Then, a stepwise regression analysis was conducted using the REG procedure of SAS (SAS Institute, Cary, NC) to determine the ranges in acceleration that accounted for the highest variation in DMI (highest R2) in each acceleration variable. All 32,767 potential acceleration combinations were tested in the validation group using the acceleration ranges predetermined in the calibration group. The CORR procedure of SAS was used to test the Pearson correlation coefficient (r) between the type of DMI [i.e., based on accelerations (DMIaccel) or actual DMI (DMIactual)]. The MIXED procedure of SAS was used to perform a repeated-measures analysis with type (DMIaccel vs. DMIactual), day, and their interaction (T × D) in the model. From this analysis, 8 candidate acceleration models were selected based on high r and similarity (P > 0.15) in terms of T and T × D between DMIaccel and DMIactual. A simulated effect on DMIactual was artificially created in the validation group by dividing this group (n = 12) into high and low intake cows (n = 6/group; DMI of 24.1 vs. 18.7 kg/d), and the candidate models were tested to determine whether they were sensitive enough to detect this effect. From these candidate models, AEN (Leg-X + Jaw-Z + LagJaw-Z) showed a weak correlation (r = 0.36) between DMIaccel and DMIactual, but DMIaccel and DMIactual were highly similar (21.2 vs. 21.4 kg/d of DMI). In addition, this was the only model that could detect the simulated effect on DMIactual (22.1 vs. 20.3 kg/d of DMI) in the validation group. The fact that the simulated effect on DMIactual was detected based only on accelerations is highly significant, and models such as AEN could be substantially improved if they were derived from a greater sample size and included different physiological stages in dairy cows.

Yeast culture increased plasma niacin concentration, evaporative heat loss, and feed efficiency of dairy cows in a hot environment

The supplementation of dairy cows with yeast culture may increase diet digestibility, plasma niacin concentration, heat dissipation, and lactation performance. Our objective was to evaluate the response of Holstein cows in late lactation (234 ± 131 d in milk) to dead yeast culture (YC, 15 g/d, Factor SC, GRASP, Saccharomyces cerevisiae) during Brazilian summer (temperature-humidity index >68 for 92.2% of the time). Thirty-two cows were individually fed a standard total mixed ration for 14 d and control (CTL) or YC treatments for 35 d, in a covariate adjusted complete randomized block design. Response was evaluated in wk 5 or as repeated measures over time. Cows were milked 3 times per day and treatments (YC or placebo) were orally dosed to each cow before each milking. Plasma niacin was 1.50 for CTL and 1.66 µg/mL for YC. The YC reduced rectal temperature, respiration rate, and skin temperature, whereas it tended to increase sweating rate. The proportion of cows with rectal temperature ≥39.2°C on CTL and YC was, respectively, 8 and 0% at 0730 h, 52 and 25% at 1500 h, and 35 and 26% at 2200 h. Plasma glucose was increased by YC. The total-tract apparent digestibility of nutrients, plasma urea N concentration, molar proportion of ruminal VFA, and urinary allantoin excretion were not affected by YC. Cows fed YC were less selective against feed particles >19 mm in the morning, in the afternoon were more selective against long feed particles and in favor of particles <8 mm, and refused short particles at night. Milk yield was not different (30.5 kg/d for CTL and 30.2 kg/d for YC). Feeding YC reduced dry matter intake (20.3 vs. 19.4 kg/d) and the digestible organic matter intake (15.6 vs. 13.9 kg/d). The inclusion of YC increased the ratios of milk to dry matter intake (1.50 vs. 1.64) and energy-corrected milk to dry matter intake (1.81 vs. 1.98). The covariate adjusted body weight (648 kg) and body condition score (3.0) did not differ. Milk solids yields and concentrations, linear somatic cell count, and milk urea N were also similar. The supplementation of YC increased plasma niacin concentration, body heat loss, and feed efficiency of late lactation dairy cows by reducing intake at similar milk yield.

Intravenous lipid infusion affects dry matter intake, methane yield, and rumen bacteria structure in late-lactating Holstein cows

Increasing the dietary fat content of ruminant diets decreases methane (CH4) production. This effect is caused by the toxic properties of fatty acids on rumen microbial populations, coating of feed particles diminishing the accessibility for microbes, and a reduction in dry matter intake (DMI). The latter effect is caused by postabsorptive long-chain fatty acids eliciting anorexic signaling; however, whether circulating long-chain fatty acids affect rumen CH4 production alike is unknown. To approach this question, 5 rumen-cannulated Holstein cows in late lactation received 2 jugular catheters and were kept in respiration chambers to measure CH4 production and DMI for 48 h. In a crossover design, cows were intravenously infused with a 20% lipid emulsion (LIPO) or 0.9% NaCl (CON). The LIPO cows received 2.1 kg of triglycerides/d [0.152 ± 0.007 g of triglycerides/(kg of BW × h)-1] consisting of 12.1% palmitic acid, 4.2% stearic acid, 31.1% oleic acid, and 52.7% linoleic acid. Blood and rumen fluid samples were taken hourly during the day. Results showed that LIPO compared with CON infusion increased plasma triglyceride as well as free fatty acid and serotonin concentrations but reduced the proportion of de novo synthesized milk fatty acids (sum of C6 to C16). Daily CH4 production and DMI were lower, whereas daily CH4 yield (CH4/DMI) was greater in LIPO than CON cows, although CH4 yield decreased from d 1 to d 2 by 2 to 14% in LIPO-infused cows only. This effect was associated with a higher (acetate + butyrate)/propionate ratio, tending lower propionate concentrations between 24 and 34 h of infusion, reduced relative abundances of genera belonging to Succinivibrio, Ruminococcaceae, and Ruminiclostridium, and greater relative Bacteroidetes genus abundances in the rumen.

Productive Response of Dairy Cows Fed with Different Levels of Totally Mixed Ration and Pasture

Forty multiparous Holstein cows in early lactation (66 ± 19 days) were assigned to one of four treatments that consisted in different ratios of total mixed rations (TMR) and pasture at 100% TMR (T0), 75:25 (T1) 50:50 (T2) and 25:75 (T3) over 9 weeks in autumn-winter. Measures of rumen parameters and digestion were performed on 4 additional Holstein cows in late lactation (287 days postpartum) fitted with permanent ruminal fistulae and producing 22.6 (±5.4) kg milk in a 4 × 4 Latin Square design. In T1 to T3 cows were taken to the grazing plot once they finished the pre-planned TMR intake and pasture was offered at 2 times the expected forage DM intake. Milk was analyzed for chemical composition, milk fatty acid (FA) profile and antioxidant compounds. The cows were individually weighed and body condition score (BCS) was determined. After the morning milking, blood samples were taken every 2 weeks and plasma was analyzed for glucose, urea, non-esterified fatty acids (NEFA), insulin, growth hormone (GH) and insulin-like growth factor (IGF-I). Herbage mass in pre grazing strips of pasture averaged 2540 ± 343 kg DM/ha. As TMR intake increased, production variables increased linearly excepting milk fat (3.88 g/100g) and milk protein (3.43 g/100g) contents that were not affected. Milk yield (kg/cow/d) resulted maximal in T0 (34.2) and linearly decreased (p 3-N concentration resulted higher in treatments with higher proportion of pasture. Kinetic parameters of DM and NDF digestion from pasture or corn silage were not affected. Milk FA profile and milk antioxidant quality showed unfavorably changes as TMR increased by a decrease in rumenic and linolenic acids and content of antioxidant vitamins, without effect on the atherogenicity index. Results suggested a depressing effect of the pasture on total DM and energy intake probably explained by qualitative deficiencies chemical composition of the forage and/or factors that affect animal behavior that may induce losses in body condition of high yielding dairy cows.

Effect of the inoculation of sugarcane silage with Lactobacillus hilgardii and Lactobacillus buchneri on feeding behavior and milk yield of dairy cows.

Despite its low NDF digestibility, sugarcane is an option for feeding dairy cattle in tropical regions. We evaluated the effect of sugarcane silages inoculated with CCMA 0170 (LH; an epiphytic bacteria isolated from sugarcane) or with NCIMB 40788 (LB; a commercial strain isolated from temperate grasses) on dairy cow performance and feeding behavior. The microbial inoculums were previously grown in the laboratory to obtain 5 log cfu/g of fresh forage. Nine tons of each inoculated silage and a noninoculated control silage (CON) were harvested from the same field and stored for at least 35 d in experimental 20 × 2.1 × 0.4 m bunker silos. Fifteen Holstein cows in late lactation (336 ± 175 days in milk at the start of the experiment) received the treatments in five 3 × 3 Latin squares with 21-d periods. The diets contained 20% of DM of sugarcane silage and 41% of DM of corn silage. Milk yield was increased from 18.0 kg/d for CON to18.8 kg/d for LH, but LB did not elicit a detectable increase in milk yield (18.1 kg/d). The daily yields of fat, protein, lactose, and total solids were increased by LH. Daily DMI and total tract apparent digestibility of nutrients did not differ among treatments. Both inoculated silages reduced acetate and increased butyrate proportions in ruminal VFA, but only LH silage reduced the acetate to propionate ratio (3.0 vs 3.3). First meal duration was shorter for CON compared to LH and LB. The proportion of daily intake between 0700 and 1300 h tended to be increased, and the proportion between 1900 and 0700 h was reduced by LH. The inoculation of sugarcane silage with affected rumen fermentation profile and feeding behavior of late lactation dairy cows, increasing the yield of milk solids.

Effects of photoperiod modulation and melatonin feeding around drying-off on bovine mammary gland involution

The risk for a dairy cow to acquire new intramammary infections is high during the transition from lactation to the dry period, because of udder engorgement and altered immune functions. Once the gland is fully involuted, it becomes much more resistant to intramammary infections. Therefore, strategies to depress milk yield before drying-off and accelerate the involution process after drying-off could be beneficial for udder health. The objective of this study was to assess the effect of photoperiod manipulation and melatonin feeding from 14 d before to 14 d after drying-off on the speed of the involution process. Thirty Holstein cows in late lactation were randomly allocated to one of the following treatments: (1) a long-day photoperiod (16 h of light: 8 h of darkness), (2) a short-day photoperiod (8 h of light: 16 h of darkness), and (3) a long-day photoperiod supplemented by melatonin feeding (4 mg/kg of body weight). Milk and blood samples were collected on d -26, -19, -12, -5, -1, 1, 3, 5, 7, 10, and 14 relative to the last milking to determine concentrations of mammary gland involution markers and serum prolactin. Additional blood samples were taken around milking on d -15, before the start of the treatments, and on d -1, before drying-off, to evaluate the treatment effects on milking-induced prolactin release. The short-day photoperiod slightly decreased milk production and basal prolactin secretion during the dry period. The milking-induced prolactin surge was smaller on d -1 than on d -15 regardless of the treatments. Lactoferrin concentration, somatic cell count, and BSA concentration as well as matrix metalloproteinase-2 and -9 activities increased in mammary secretions during the first 2 wk of the dry period, whereas milk citrate concentration and the citrate:lactoferrin molar ratio decreased. The rates of change of these parameters were not significantly affected by the treatments. The long-day photoperiod supplemented by melatonin feeding did not affect milk production, prolactin secretion, or mammary gland involution. Under the conditions in this study, photoperiod modulation and melatonin feeding did not appear to affect the rate of mammary gland involution.

Effects of bedding with recycled sand on lying behaviors, udder hygiene, and preference of lactating Holstein dairy cows.

Effects of bedding with recycled sand and season on lying behaviors, hygiene, and preferences of late-lactation Holstein cows were studied. It was hypothesized that recycled sand will decrease lying time and increase hygiene scores due to increased moisture content and organic matter, and thus a preference for the control sand will be evident. Cows (n = 64) were divided into 4 groups (n = 8 per group) per season. In summer (August to September), cows were balanced by days in milk (268.1 ± 11.9 d) and parity (2.0 ± 0.2). In winter (January to February), mean DIM was 265.5 ± 34.1 d. Cows were assigned to 1 of 2 treatments using a crossover design with each treatment lasting 7 d (no-choice phase): bedding with recycled sand (RS; n = 32) or control (CO; clean sand; n = 32). Stocking density was maintained at 100%. The choice phase allowed cows to have access to either treatment with stocking density at 50%. Accelerometers recorded daily lying time, number of lying bouts per day, lying bout duration (min/bout), and total steps per day. Teat swabs, milk, sand samples, and udder hygiene scores were collected on d 0, 3, and 7 of each experimental week. Samples were cultured for streptococci, staphylococci, and gram-negative bacteria. Video data were used to assess bedding preferences. All data were analyzed using the MIXED and GLIMMIX procedures of SAS 9.4 (SAS Institute Inc., Cary, NC). Lying time was not affected by treatment, but cows did take more steps during winter. Bacterial counts were elevated for cows on recycled sand. A preference was observed for clean sand during the summer, but no preference was observed for sand during the winter. Regardless of bedding, the most commonly observed behavior was lying in the stalls, which suggested either bedding might be suitable. Caution should be used with this interpretation of preference, as sand was recycled only once. This limited reclamation was still sufficient to potentially alter the composition of sand, driving the observed preference. If these changes in composition continue, then the strength of the preference may also change. However, considering all variables within the current study, recycled sand is a viable bedding source to use for dairy cows.