Multiparous Holstein Cows Research Articles

Replacing soybean meal with high-oil pumpkin seed cake in the diet of lactating Holstein dairy cows modulated rumen bacteria and milk fatty acid profile

This research aimed to investigate the effects of replacing soybean meal with high-oil pumpkin seed cake (HOPSC) on ruminal fermentation, lactation performance, milk fatty acid, and ruminal bacterial community in Chinese dairy cows. Six multiparous Chinese Holstein cows at 105.50 ± 5.24 d in milk (mean ± standard deviation) and 36.63 ± 0.74 kg/d of milk yield were randomly allocated, in a 3 × 3 Latin square design, to 3 dietary treatments in which HOPSC replaced soybean meal. Group 1 was the basal diet with no HOPSC (0HOPSC); group 2 was a 50% replacement of soybean meal with HOPSC and dried distillers grains with solubles (DDGS; 50HOPSC), and group 3 was a 100% replacement of soybean meal with HOPSC and DDGS (100HOPSC). We found no difference in the quantity of milk produced or milk composition among the 3 treatment groups. Feed efficiency tended to increase linearly as more HOPSC was consumed. In addition, rumen fermentation was not influenced when soybean meal was replaced with HOPSC and DDGS; the relative abundance of ruminal bacteria at the phylum and genus levels was altered. We also observed that as the level of HOPSC supplementation increased, the relative abundance of Firmicutes and Tenericutes linearly increased, whereas that of Bacteroidetes decreased. However, with increasing HOPSC supplementation, the relative abundance of Ruminococcus decreased linearly at the genus level in the rumen, and the relative abundance of Prevotella showed a linear downward tendency. Changes in dietary composition and rumen bacteria had no significant effect on the fatty acid composition of milk. In conclusion, our results indicated that replacing soybean meal with a combination of HOPSC and DDGS can meet the nutritional needs of high-yielding dairy cows without adversely affecting milk yield and quality; however, the composition of rumen bacteria could be modified. Further study is required to investigate the effects of long-term feeding of HOPSC on rumen fermentation and performance of dairy cows.

Medium chain fatty acid supplementation improves animal metabolic and immune status during the transition period: A study on dairy cattle.

The transition period is the stage of the high incidence of metabolic and infectious diseases in dairy cows. Improving transition dairy cows' health is crucial for the industry. This study aimed to determine the effects of dietary supplementation medium-chain fatty acids (MCFAs) on immune function, metabolic status, performance of transition dairy cows. Twenty multiparous Holstein cows randomly assigned to two treatments at 35 d before calving. 1) CON (fed the basal 2) MCFA treatment (basal diet was supplemented at an additional 20 g MCFAs mixture every day) until 70 d after calving. The results showed that the serum amyloid A myeloperoxidase concentrations in the blood of cows in MCFA treatment significantly decreased during the early lactation (from 1 d to 28 d after calving) 0.03, 0.04, respectively) compared with the CON, while the tumor necrosis factor concentration was significantly decreased at 56 d after calving (P = 0.02). In addition, the concentration of insulin in the pre-calving (from 21 d before calving to calving) blood of cows in MCFA treatment was significantly decreased (P = 0.04), and concentration of triglyceride also showed a downward trend at 28 d after calving 0.07). Meanwhile, MCFAs supplementation significantly decreased the concentrations of lithocholic acid, hyodeoxycholic acid, and hyocholic acid in the blood at 1 d calving (P = 0.02, < 0.01, < 0.01, respectively), and the level of hyocholic acid taurocholic acid concentrations (P < 0.01, = 0.01, respectively) decreased dramatically at 14 d after calving. However, compared with the CON, the pre-calving dry matter intake and the early lactation milk yield in MCFA treatment were significantly decreased (P = 0.05, 0.02, respectively). In conclusion, MCFAs supplementation transition diet could improve the immune function and metabolic status of dairy cows, and the health of transition cows might be beneficial from the endocrine status.

Replacing alfalfa hay with industrial hemp ethanol extraction byproduct and Chinese wildrye hay: Effects on lactation performance, plasma metabolites, and bacterial communities in Holstein cows.

This trial was designed to investigate the effects of industrial hemp ethanol extraction byproduct (IHEEB) and Chinese wildrye hay (CWH) replacement of alfalfa hay (AH) on digestibility, and lactation performance, plasma metabolites, ruminal fermentation, and bacterial communities in Holstein dairy cows. Nine healthy multiparous Holstein cows (parity = 3) with similar body weights (584 ± 12.3 kg), days in milk (108 ± 11.4), and milk yields (30 ± 1.93 kg; all mean ± standard deviation) were used in a replicated 3 × 3 Latin square design with 3 periods of 21 d. During each period, each group consumed 1 of 3 diets: (1) 0% IHEEB (0IHEEB); (2) 6.0% IHEEB and 1.7% Chinese wildrye hay (6IHEEB); (3) 10.8% IHEEB and 4.3% Chinese wildrye hay (11IHEEB). The diets in each group were isocaloric and isonitrogenous, with similar contents of concentrate and silage but different ratios of IHEEB and CWH to replace AH. The results showed that increasing the substitute did not affect the total-tract apparent nutrient digestibility. There was no difference in lactation performance of dairy cows fed the three diets, except for the cows' somatic cell count (SCC), which decreased with the increase in the amount of the substitute. Cannabidiol and tetrahydrocannabinol were not detected in milk samples of dairy cows in the different treatment groups. 6IHEEB and 11IHEEB-fed cows showed a linear decrease in total volatile fatty acids (VFA) and butyrate compared to the 0IHEEB cows. Plasma IL-1β content quadratically decreased with feeding IHEEB and CWH, and other blood parameters were unaffected. The rumen fluid's relative abundances of Bacteroidota, Fibrobacterota, and Prevotellaceae quadratically increased, while Firmicutes tended to decrease quadratically as the substitution increased. Feeding IHEEB and CWH linearly increased the relative abundances of Firmicutes, Lachnospiraceae, Monoglobaceae, and Butyricicoccaceae in the feces. As the substitution increased, the cost of dairy farming was reduced. In summary, substituting AH with IHEEB and CWH in diets did not affect the total-tract apparent nutrient digestibility, improved milk composition, and plasma immune indices. It changed the bacterial composition in rumen fluid and feces and improved dairy farming benefits.

Effects of tanniferous sainfoin and Acacia mearnsii extract on urinary N excretion and ammonia volatilization from the slurry of dairy cows

This study examined whether the effects of the tanniferous extract of Acacia mearnsii on N partitioning in dairy cows and ammonia (NH3) volatilization from their excreta are additive to that of the tanniferous legume sainfoin (Onobrychis viciifolia) and remain consistent when supplemented to different silage types with varying crude protein (CP) content. In a 6 × 6 Latin Square arrangement, six multiparous Holstein cows (milk yield: 36.6 ± 3.9 kg/d; 70 ± 13 d in milk) were assigned randomly to six treatments. The experiment included a 14-d adaptation and a 7-d data collection period, where intake, milk yield, and composition were recorded daily and excreta were collected. Cows had ad libitum access to one of six total mixed rations containing (dry matter [DM] basis) 750 g/kg DM silage. The silages were rich in either sainfoin (180 g/kg CP of DM), ryegrass (118 g/kg CP) or clover (220 g/kg CP). Each silage was supplemented with 20 g/kg DM of Acacia mearnsii extract or straw meal. The effects on N partitioning, ruminal volatile fatty acids (VFA) and microbiota, and NH3 volatilization from slurry reconstituted from fresh urine and feces were determined. The sainfoin-based diet reduced the apparent digestibility of organic matter, acid detergent fiber, and N compared with the ryegrass- and clover-based diets (P < 0.001). Acacia reduced the DM intake (P < 0.05) and apparent digestibility of organic matter and N across all silage types (P < 0.01). The digestibility of neutral detergent fiber was reduced with sainfoin, and ryegrass with Acacia (Silage × Acacia interaction P < 0.05). Acacia reduced the ruminal acetate proportion (P < 0.05) but did not affect ruminal propionate and n-butyrate or the microbiota composition. With ryegrass, the rumen fluid had lower acetate (P < 0.001) and greater n-butyrate proportions (P < 0.01) compared to sainfoin and clover, lower propionate proportions as compared with sainfoin (P < 0.05), and the lowest blood urea N concentration (P < 0.001). Acacia reduced energy-corrected milk yield (P < 0.05). Milk protein content was reduced with the sainfoin diet (P < 0.05). Sainfoin (compared to clover) and Acacia supplementation shifted the N excretion from urine to feces and decreased urinary urea N excretion and NH3 volatilization from the slurry (P < 0.05). The effects of the two tannin sources were widely additive in mitigating urinary urea N and NH3 volatilization. The results illustrate that combinatory N-abatement measures based on different CT sources are efficient, but supplementing Acacia resulted in decreasing dry matter intake and milk production, while sainfoin supplementation resulted in lower organic matter digestibility.

Is adenosine deaminase (ADA) activity in saliva and serum a more accurate disease detection tool than traditional redox balance parameters in early-lactating dairy cows?

Enzyme adenosine deaminase (ADA) is a marker of inflammation in domestic animals, but it is unclear whether it is a reliable marker of oxidative stress, especially in the transition period in dairy cows. This study aims to assess if ADA and redox status measurements in saliva provide the same utility to detect disease condition as that obtained from serum. Sixty-eight multiparous Holstein cows, between 2 and 3 weeks postpartum were selected. Five study groups were established: control (healthy), and cows with ketosis, mastitis, laminitis, and metritis. The parameters measured were ADA activity, total oxidants (TOS), antioxidants (TAC), and OSi ratio.Regarding redox status, no significant differences arise in both saliva and serum being the correlations negative and not significant. In saliva, ADA activity in healthy cows differs from those with pathological processes, having the lowest activities. In serum, ADA activity is similar in the healthy and ketosis cows, showing the lowest activities meanwhile animals with mastitis, laminitis, or metritis have significantly higher activities. In conclusion, the measurement of ADA activities and redox status in saliva does not give consistent results, being preferable to measure them in serum during the transition period.

Evaluation of fecal sampling time points to estimate apparent nutrient digestibility in lactating Holstein dairy cows.

The aim of this study was the evaluation of nutrient excretion patterns in samples of feces collected every 2 h to define the best sampling protocol for estimation of apparent digestibility. Four multiparous mid-lactation Holstein cows, housed in a tie stall barn and milked twice a day (0800; 1900 h), were enrolled. Dry total mixed ration (TMR) without silages was fed once (0800 h) per day. Feces were sampled every 2 h for 72 h. Each sample was divided in 3 portions: hourly sample sample (8, 10, 12, 14, 16, 18, 20, 22, 00, 2, 4, 6), 8-h composite sample (00-06, 08-14 and 16-22), and a 24-h composite sample. Complete chemical analyses were performed and total tract nutrient digestibility was calculated using undegraded neutral detergent fiber at 240 h of in vitro fermentation (uNDF240h) as a marker. Feeding and rumination patterns were also recorded during the trial. For some parameters, excretion was not constant throughout the day: neutral detergent fiber (aNDFom), undegraded neutral detergent fiber at 24 h of in vitro fermentation (uNDF24h), uDNF240h, total tract crude protein digestibility (TTCPD), total tract neutral detergent fiber digestibility (TTaNDFomD), total tract potentially degraded neutral detergent fiber at 240 h of in vitro fermentation digestibility (TTpdNDF240hD) with minimal values after new TMR delivery and maximal values 12 h after feed delivery. Feeding and ruminating behavior seemed to have an important role in the excretion pattern, due to the pushing and evacuating effect they have. Considering our results, two fecal samples at 12 and 24 h after the TMR delivery are suggested. For one daily sample, 12 h post time of most stable and constant rumination 0000-0600 h, which is also 8 h post feed delivery is suggested.

Effect of withdrawing an immunomodulatory feed additive from lactating cow diets on peripheral blood mononuclear cell proliferation.

Supplementation of immunomodulatory feed additives, such as OmniGen AF (OG), helps to maintain immune competency; however, it is unknown if immune benefits persist in lactating cows after OG is removed from the diet. The aim of this trial was to evaluate the effect of withdrawing OG from the diet on peripheral blood mononuclear cells (PBMC) proliferation of midlactation dairy cows. Multiparous Holstein cows (N = 32), blocked by parity (2.7 ± 0.8) and days in milk (153 ± 39 d) were randomly assigned to one of the two dietary treatments within each block: diets were top dressed with either OG (56 g/d/cow) or placebo (CTL, 56 g/d/cow). Cows were housed in the same free-stall pen and individually fed once per day through Calan gates. All cows were fed the same diet containing OG for at least 1 yr before the onset of treatments. Cows were milked three time per day and milk yield was recorded at each milking. Milk samples were collected from three consecutive milkings weekly and composition analyzed. Body weight (BW) and condition score were measured weekly. Blood samples were collected at -1, 1, 3, 5, and 7 wk relative to the onset of treatments for the isolation of PBMC. The PBMC were cultured with concanavalin A (ConA) and lipopolysaccharides (LPS) for 72 h in vitro to determine proliferative responses. Prior to the experiment, cows in both treatments had similar disease incidence. During the experiment, cows did not show symptoms of disease. Withdrawing OG from the diet did not affect (P ≥ 0.20) milk yield or composition, intake, or BW. Compared with CTL, feeding OG maintained greater body condition score (2.83 vs. 2.92, P = 0.04). Regardless of time, relative to CTL, PBMC isolated from cows fed with OG had a greater proliferative rate when stimulated with LPS (stimulation index: 1.27 vs. 1.80, P = 0.05) and tended to have greater proliferation when stimulated with ConA (stimulation index: 5.24 vs. 7.80, P = 0.08). In conclusion, withdrawing OG from the diet of midlactation cows reduced proliferative response of PBMC suggesting that the immunomodulatory role of OG is lost as early as 1 wk after its withdrawal from the diet of lactating dairy cows.

One-carbon metabolism and related pathways in ruminal and small intestinal epithelium of lactating dairy cows.

Physiological and environmental stresses such as the transition into lactation and heat load contribute to gastrointestinal tract (GIT) dysfunction. The nonruminant gastrointestinal tract has mechanisms to cope with pro-oxidant and pro-inflammatory stressors arising from the gut lumen or within intestinal cells. One-carbon metabolism (OCM) contributes to anti-oxidant capacity via the production of glutathione (GSH) and taurine, and the synthesis of phospholipid, creatine, and the osmolyte glycinebetaine among others. A multipronged approach was used to assess the biological relevance of OCM and closely-related pathways on GIT function in dairy cows. Ruminal papillae (Rum) and scrapings from duodenum (Duo), jejunum (Jej), and ileum (Ile) were collected at slaughter from eight multiparous Holstein cows averaging 128 ± 12 d in milk and producing 39 ± 5 kg/d. A MIXED model ANOVA with preplanned orthogonal contrasts was used for statistical analysis. Methionine adenosyl transferase 1 activity (MAT) was ~10-fold greater (P < 0.01) and cystathionine β-synthase activity doubled in Rum vs. small intestine. Total glutathione peroxidase (GPX) activity was greatest (P = 0.03) in Ile, but similar to Rum. Activity and mRNA abundance of betaine-homocysteine S-methyltransferase were undetectable. There was a 2.5-fold greater protein abundance of GPX1 (P < 0.01) and a ~2-fold greater abundance of GPX3 (P < 0.01) in Rum vs. small intestine. Among the various amino acids (AA) with roles in OCM or closely-related pathways (e.g. creatine synthesis), concentrations of arginine, aspartate, glutamine, methionine, and serine were lower (P < 0.01) in Rum vs. small intestine. Unlike AA, concentrations of OCM-related intermediates S-5'-adenosyl-homocysteine (SAH), glycinebetaine, carnitine, creatine (CRE), and cysteinesulfinic acid were greater (P < 0.01) while taurine was lower in Rum vs. small intestine. Intermediates of the folate cycle were undetectable. The fact that S-adenosylmethionine (SAM) was undetectable while MAT activity and SAH were greater in Rum suggested that availability of SAM (a methyl donor) is a key determinant of flux through the folate and methionine cycles in the GIT. Except for adenosine, concentrations of glutamate, glycine, α-ketoglutarate, hypotaurine, and GSH were lowest in Ile. Together, the data underscored unique differences in activity of one-carbon metabolism and related pathways across sections of the GIT.

Effects of starch sources varying in particle sizes on ruminal fermentation, nutrient flow, starch digestibility, and lactation performance of dairy cows.

Cereal grains are the predominant starch source (SS) for dairy cows; however, starch digestibility varies greatly depending on source, grain processing, and potentially interactions between these factors. The objective was to study the effects of the interactions between SS, and particle sizes (PS) on ruminal fermentation, nutrient flow, starch digestibility, and lactation performance of dairy cows. Four ruminally cannulated multiparous Holstein cows were used in a 4 × 4 Latin square design with a 2 × 2 factorial arrangement of treatments. Two SS (corn or sorghum) used in this study were either finely or coarsely ground (using a 1- or 4-mm screen sieve). Digesta flow was quantified using the reticular sampling technique, applying the triple-marker method. Data were analyzed using the GLIMMIX procedure of SAS version 9.3 (SAS Institute Inc., Cary, NC, USA). For ruminal pH, data were analysed with time as repeated measure. There were no interactions between SS and PS on production or intake, flow, and digestibility of nutrients. Dry matter intake was greater for the corn diet compared to the sorghum diet (25.15 vs. 21.98 kg/d), which consequently affected nutrient intake, however, PS did not affect intake. Milk yield was not affected by SS; however, it was greater for cows fed fine grains than cows fed coarser grains (25.32 vs. 23.16 kg/d). Milk fat and milk protein were not affected by SS or PS. Interactions (SS × PS) were observed for ruminal pH, reticular pH, and volatile fatty acids (VFA) concentrations but not for ruminal NH3-N concentration. Ruminal and reticular pH were greater for sorghum when coarsely ground and the total VFA concentration was decreased, compared to coarse corn and fine sorghum; however, coarsely grinding corn did not affect ruminal or reticular pH nor VFA concentration. Acetate concentration was lower for corn when finely ground; however, finely grinding sorghum did not affect acetate. Decreasing PS increased ruminal digestibility of starch (87.18% vs. 83.43%), reduced the flow of starch to the reticulum (0.79 vs. 0.96 kg/d) but decreased neutral detergent fiber digestibility in the rumen (30.23% vs. 34.88%). Although SS were differently affected by processing, the effects of PS on production, intake, flow, and digestibility of nutrients were observed regardless of the SS. Furthermore, the effects of decreasing PS on pH and VFA concentrations were more pronounced in sorghum compared to corn.

In utero choline exposure alters growth, metabolism, feed efficiency, and carcass characteristics of Holstein x Angus cattle from weaning to slaughter.

Feeding rumen protected choline (RPC) to late gestation dairy cows has potential to affect growth in offspring. The objective of this study was to evaluate the effects of in utero choline exposure on the growth, feed efficiency (FE), metabolism, and carcass quality of Angus x Holstein cattle. Multiparous Holstein cows pregnant with male (n=17) or female (n=30) Angus-sired calves were enrolled 21 d prepartum and randomly assigned to one of four dietary treatments varying in quantity and formulation of rumen-protected choline (RPC). The treatments included a control with 0g/d supplemental RPC (CTL), supplemental RPC fed at the recommended dose (RD) of 15g/d from either an established RPC product (RPC1RD; ReaShure; Balchem Corp) or choline ion from a concentrated RPC prototype (RPC2RD; Balchem Corp), or a high dose (HD) of RPC2 fed at 22g/d (RPC2HD). From 2 to 6 mo of age, calves were group housed and offered 2.3kg grain/hd/d (42% CP) with ad libitum grass hay, and stepped up to a complete finishing diet by 7 mo (12.0% CP; 1.34 Mcal/kg NEg). Weight and height were measured monthly. Animal FE was measured in individual pens for 35 d at 8 mo. Feed intake was measured daily, and blood was obtained on d 18 during the FE period. Afterwards, cattle were group housed and offered a free-choice finishing diet until slaughter, where carcass yield and quality characteristics were measured. Mixed models were used in PROC MIXED (SAS, 9.4) with the fixed effects of treatment, sex, time, their interactions, and the random effect of calf. Month was the repeated measure, and preplanned contrasts were used. Blood and FE data were analyzed with the fixed effect of dam choline treatment, calf sex, and the interaction. Increasing dose of RPC tended to increase weight over the entire study period. Feeding any RPC increased hip and wither height compared with CTL, and increasing RPC dose linearly increased hip and wither height. Treatment and sex interacted on DMI whereby increasing RPC intake linearly increased DMI for males but not females. Compared with control, feeding any RPC decreased plasma insulin, glucose, and an insulin sensitivity index (RQUICKI). In utero choline exposure increased kidney-pelvic-heart fat and marbling score. Mechanisms of action for intrauterine choline exposure on offspring growth, metabolism, and carcass characteristics should be explored as they have direct implications for profitability for cattle growers and feeders.

Intrauterine infusion of a pathogenic bacterial cocktail is associated with the development of clinical metritis in postpartum multiparous Holstein cows.

Metritis is a uterine disorder common in dairy cattle caused by bacterial infection, with greater incidence in the early postpartum period. The disease causes delayed uterine involution, with a fetid, watery, red-brown discharge, with animals presenting different clinical signs including fever, dullness, inappetence, decreased milk yield, and dehydration. We developed an in vivo model of clinical metritis in Holstein multiparous cows using a pathogenic cocktail of Escherichia coli, Fusobacterium necrophorum, and Trueperella pyogenes. A total of 36 multiparous cows were randomly allocated to 1 of 3 treatment groups of 12 animals each. Cows assigned to the control group received an intrauterine administration of sterile saline solution, those in the low-dose group received a bacterial inoculum containing 106 cfu of Escherichia coli, Trueperella pyogenes, and Fusobacterium necrophorum; and those in the high-dose group received 109 cfu of these same organisms, all within 24 h of parturition. Clinical signs; milk yield; dry matter intake; serum concentration of acute phase proteins, metabolites, and cytokines; and bacterial counts in vaginal discharge were measured from parturition until 14 d in milk. Based on clinical signs, cows in the low-dose group had a greater incidence of metritis (83.3%) compared with the control (9%) or high-dose (25%) group. The low-dose group also had lesser dry matter intake compared with controls and an acute phase protein profile that typified metritis. The low-dose group presented greater relative abundance of bacteria from the genus Fusobacterium in the vaginal discharge compared with control cows, and the high-dose group was similar to the control group. Unexpectedly, intrauterine infusion of cows with a lower dose of bacteria was more effective than a higher dose, indicating that the bacterial load may affect metritis incidence in a nonlinear manner. These findings demonstrate that clinical metritis can be experimentally induced in postpartum multiparous Holstein cows, creating a relevant model for the study of uterine diseases.

Comparative evaluation of major milk quality parameters of Holstein and Simmental cows at different lactation stages under similar environmental conditions

e aim of this study was to evaluate the composition and quality parameters of Simmental and Holstein cows’ milk with different lactation stages under the same environmental conditions. Multiparous Holstein and Simmental cows from different lactation stages (n = 210) were included in the present study. MDA, SCC, composition, and fatty acid analyzes were performed from the collected milk samples. To determine the effect of breed, lactation stage and their interactions, linear mixed models were applied to these parameters. Among the breeds, only milk fat and pH were determined statistically significant as composition parameters. While C15:0 and C17:1 n8 were observed to be statistically significant for the breed factor, C18:2 n6 trans were statistically significant only for the lactation stage factor. Moreover, the fatty acids of C4:0, C6:0, C8:0, C10:0, C11:0, C12:0, C13:0, C16:0, C18:1 n9, C20:0 and C22:6 n3 were determined to be statistically significant in terms of interactions. For the fatty acid indices, SCFA, MCFA, n3 and n6/n3 were statistically significant in terms of interactions while SFA, MUFA, UFA and AI were statistically significant in terms of breeds. On the other hand, new studies are needed to investigate the differences between these breeds at the molecular level for milk quality and fatty acid synthesis.

Variations of some adipokines, pro-inflammatory cytokines, oxidative stress biomarkers, and energy characteristics during the transition period in dairy cows.

Limited information exists about the relationship of adipose tissue with inflammation, oxidative stress, and energy metabolism during the transition period in dairy cows. The objective of this study was to assess the changes and relation of some adipokines, cytokines, oxidative biomarkers, and serum biochemical parameters related to energy balance (EB) in cows during the transition period. Thirty multiparous Holstein cows were selected based on estimated parturition date, and blood samples were collected from jugular vein on one-week prepartum and one and three weeks postpartum and used to measure the parameters. The serum levels of beta-hydroxybutyric acid (BHB), non-esterified fatty acid, cholesterol, high-density lipoprotein (HDL), aspartate aminotransferase, and total antioxidant capacity increased significantly, and glucose, urea, triglyceride (TG), and low-density lipoprotein (LDL) decreased significantly after parturition. The serum values of adiponectin, resistin, leptin, and cytokines including interleukin 6 (IL-6) and tumor necrosis factor (TNF)-α were not changed significantly during the experiment. The results of the Pearson correlation revealed a significant negative correlation between BHB with glucose, albumin, cholesterol, HDL, LDL, and a positive correlation with TG and malondialdehyde. Also, there was a significant direct correlation between insulin and leptin, adiponectin, resistin, IL-6 and TNF-α in the whole experiment period. These emphasize the difficulty of dairy cows to manage the energy requirements during the transition period. It can be stated that adipokines and cytokines may have an essential role in the metabolic status in this period, and control of their production and, or secretion could be helpful in EB during the transition period.

Effects of dietary antioxidant supplementation on metabolism and inflammatory biomarkers in heat-stressed dairy cows

Heat-stress-induced inflammation may be ameliorated by antioxidant supplementation due to the purported effects of increased production of reactive oxygen species or oxidative stress on the gastrointestinal tract barrier. Thus, study objectives were to evaluate whether antioxidant supplementation [AGRADO Plus 2.0 (AP); EW Nutrition] affects metabolism and inflammatory biomarkers in heat-stressed lactating dairy cows. Thirty-two mid-lactation multiparous Holstein cows were assigned to 1 of 4 dietary-environmental treatments: (1) thermoneutral (TN) conditions and fed a control diet (TN-CON; n = 8), (2) TN and fed a diet with AP (10 g antioxidant; n = 8), (3) heat stress (HS) and fed a control diet (HS-CON; n = 8), or (4) HS and fed a diet with AP (HS-AP; n = 8). The trial consisted of a 23-d prefeeding phase and 2 experimental periods (P). Respective dietary treatments were top-dressed starting on d 1 of the prefeeding period and continued daily throughout the duration of the experiment. During P1 (4 d), baseline data were collected. During P2 (7 d), HS was artificially induced using an electric heat blanket (Thermotex Therapy Systems Ltd.). During P2, the effects of treatment, day, and treatment-by-day interaction were assessed using PROC MIXED of SAS (SAS Institute Inc.). Heat stress (treatments 3 and 4) increased rectal, vaginal, and skin temperatures (1.2°C, 1.1°C, and 2.0°C, respectively) and respiration rate (33 breaths per minute) relative to TN cows. As expected, HS decreased dry matter intake, milk yield, and energy-corrected milk yield (32%, 28%, and 28% from d 4 to 7, respectively) relative to TN. There were no effects of AP on body temperature indices or production. Milk fat, protein, and lactose concentrations remained unaltered by HS or AP; however, milk urea nitrogen was increased during HS regardless of AP supplementation (26% relative to TN). Circulating glucose remained unchanged by HS, AP, or time. Additionally, HS decreased circulating glucagon (29% from d 3 to 7 relative to TN), but there was no additional effect of AP. There was a tendency for nonesterified fatty acid concentrations to be increased in HS-AP cows throughout P2 (60% relative to TN-CON), whereas it remained similar in all other treatments. Blood urea nitrogen increased for both HS treatments from d 1 to 3 before steadily decreasing from d 5 to 7, with the overall increase being most pronounced in HS-CON cows (27% relative to TN-CON). Further, supplementing AP decreased blood urea nitrogen in HS-AP on d 3 relative to HS-CON (15%). Circulating serum amyloid A tended to be and lipopolysaccharide binding protein was increased by HS, but neither acute-phase protein was affected by AP. Overall, AP supplementation appeared to marginally alter metabolism but did not meaningfully alter inflammation during HS.

The association of subclinical hypocalcemia at 4 days in milk with reproductive outcomes in multiparous Holstein cows

Delayed or persistent episodes of subclinical hypocalcemia (SCH) have been associated with reduced milk production and feed intake and they put cows at risk for additional disease development. However, little is known about the effect of delayed or persistent SCH on reproductive outcomes. The objectives of our study were to describe the association between SCH status at 4 d in milk (DIM) with the odds of pregnancy to first service and time to pregnancy through 150 DIM. Data were collected from multiparous Holstein cows (n = 697) from 4 farms in New York State that did not receive supplemental Ca during the first 5 DIM. Cows were classified into 1 of 2 SCH groups based on blood total Ca (tCa) at 4 DIM: normocalcemic (NC; n = 515) if tCa >2.2 mmol/L at 4 DIM or SCH (n = 182) if tCa ≤2.2 mmol/L at 4 DIM. We created a multivariable logistic regression model to assess the association between SCH group and pregnancy to first service and a time-to-event analysis to evaluate the effect of SCH group on the hazard of pregnancy by 150 DIM. Cows that were SCH at 4 DIM had lower odds of pregnancy to first service [odds ratio = 0.75, 95% confidence interval (CI) = 0.61 to 0.93, incidence = 18.1%] compared with their NC counterparts (incidence = 27.4%). Similarly, SCH cows tended to have a lower hazard of pregnancy by 150 DIM (hazard ratio = 0.82, 95% CI = 0.67 to 1.01, incidence = 65.4%) than NC cows (incidence = 70.7%). Our results indicate that delayed or prolonged reductions in blood Ca in the early postpartum period, resulting in SCH at 4 DIM, were associated with reduced reproductive outcomes in multiparous cows. Our findings, in conjunction with previous reports of the negative associations of delayed or persistent SCH on health and production, suggest that reductions in blood Ca beyond the first day of lactation might be indicative of maladaptation to lactation, and the ramifications can persist beyond the immediate postpartum period.

Effect of Moringa leaf flavonoids on the production performance, immune system, and rumen fermentation of dairy cows.

Unreasonable use of antibiotics in animals is a major concern and will remain so, thus affecting people's health. However, the application of plant extracts can better solve this problem. The purpose of this study was to study the effect of Moringa leaf flavonoids on the production performance, immunity, and rumen fermentation of dairy cows. Nine Holstein multiparous cows (average weight: 550 kg; days of lactation: 150 ± 6 days) were used in the experiment, using a 3 × 3 Latin square design. Cows were divided into three groups, each of which was supplemented with 0, 50, or 100 mg/body weight (BW) Moringa oleifera leaf flavonoids. Each experimental period consisted of three periods of 21 days, and the prefeeding period lasted 15 days. Our results indicated that supplementation with Moringa leaf flavonoids increased the protein content and decreased the number of somatic cells in milk; had little effect on the biochemical indices of blood, the rumen fermentation, and serum biochemical indicators; and improved the activity of antioxidant enzymes, the antioxidant capacity, and immunity. Addition of 50 mg/BW Moringa leaf flavonoids to cow enhanced the antioxidant and immunity capacity in dairy cows but did not affect physiological levels of common biochemical parameters in blood or fermentation parameters in rumen.

Effects of corn silage supplementation strategy and grazing intensity on herbage intake, milk production, and behavior of dairy cows

Effects of corn silage supplementation on milk production of grazing dairy cows depend in part on the substitution rate between the 2 forages, which may be influenced by grazing management. The aim of this study was to compare 2 grazing management strategies for measuring substitution rate between herbage and corn silage, in interaction with grazing intensity. Six treatments were compared, with 2 grazing intensities and 3 supplementation strategies investigated at both grazing intensities. The 2 grazing intensities were severe and light grazing, defined by either (1) herbage allowance (HA) of 15 (severe) or 30 (light) kg dry matter (DM)/cow per d at 3 cm above ground level or (2) postgrazing sward height, depending on the supplementation strategy. The 3 supplementation strategies were as follows: (U) an unsupplemented treatment, (A) 5 kg DM/d of corn silage offered at a similar HA as in U, and (H) 5 kg DM/d of corn silage offered at a similar postgrazing sward height as in U. Thirty-six multiparous Holstein cows were used in a randomized complete block design and divided in 2 groups for the entire experiment, one for each grazing intensity. Within each grazing intensity group, the corn silage supplementation strategy was studied using a 3 × 3 Greco-Latin square design, with 3 periods of 14 d. Supplementing cows with corn silage increased total DM intake only for severe grazing by 1.7 kg DM/d. The substitution rate between corn silage and grazed herbage was lower for severe than for light grazing, averaging 0.63 and 1.23, respectively. Herbage dry matter intake was lower by 1.2 kg/d for strategy H than A, leading to lower substitution rates (0.81 vs. 0.99, respectively), irrespective of grazing intensity. Milk production increased with silage supplementation for severe grazing (+1.0 kg/d milk) and was unaffected by silage supplementation for light grazing (-0.4 kg/d milk). The milk production response to corn silage supplementation averaged +0.23 and -0.08 kg of milk per kg DM of silage for severe and light grazing, respectively. Fat-corrected milk production tended to be lower by 0.4 kg/d for strategy H than A, leading to lower milk production response (+0.00 vs. +0.12 kg of milk per kg DM of silage, respectively). Milk protein concentration increased with silage supplementation for severe grazing (+1.0 g/kg) but decreased with silage supplementation for light grazing (-0.6 g/kg). Milk fat concentration did not differ among treatments. On average, daily grazing time (-47 min/d, i.e., -9%) and herbage intake rate (-4.9 g of DM/min, i.e., -14%) decreased when cows were supplemented, with greater grazing time reduction at severe than light grazing, and greater herbage intake rate reduction at light than severe grazing. In conclusion, the greater substitution rate and the lower 4% fat-corrected milk production when corn silage was provided at a similar postgrazing sward height rather than at a similar HA to those of unsupplemented cows explain why supplementing grazing dairy cows with conserved forages has no strong effect in practice from a production point of view.

Production performance of Holstein cows at 4 stages of lactation fed 4 dietary crude protein concentrations.

Dairy cow responses to dietary crude protein (CP) may depend on stage of lactation. The primary objective of this study was to evaluate the effects of 4 concentrations of dietary CP on dry matter intake (DMI), production performance, net energy for lactation (NEL) output in milk, feed efficiency (FE: milk NEL/DMI), and nitrogen use efficiency (100 × milk protein-N/N intake) when fed to cows grouped as early, mid-early, mid-late, and late lactation. Our secondary objective was to determine the range of CP concentration at which production responses were not negatively affected across days in milk (DIM). Multiparous Holstein cows (n = 64) were stratified by DIM [initial average ± standard deviation: 86 ± 14.9 (early), 119 ± 10.0 (mid-early), 167 ± 22.2 (mid-late), and 239 ± 11.1 (late)] and then randomly assigned within DIM group to receive 1 of 4 total mixed rations containing 13.6, 15.2, 16.7, and 18.3% CP (dry matter basis) according to a 4 × 4 factorial arrangement of treatments. Cows were individually fed a covariate diet for 14 d, followed by 56 d of treatment diets. Milk yield and DMI were recorded daily and milk components were analyzed weekly for 2 consecutive days at 3 daily milkings. Data were analyzed using a categorical mixed-effect model to evaluate the effects of CP concentration and DIM using linear, quadratic, and cubic contrasts, and their interactions. Additionally, a mixed-effect cubic regression model was fit with DIM, dietary CP concentration, and their interaction as continuous independent variables. Dietary CP concentration deemed optimal across DIM was determined as the range of CP for which the dependent responses did not differ from the predicted maximum. With advancing stage of lactation, DMI, milk NEL output, and FE decreased linearly (from 30.4 to 28.4 kg/d for DMI, from 33.2 to 23.3 Mcal/d for NEL output, and from 1.09 to 0.82 Mcal milk NEL/kg DMI for FE for early and late lactation cows, respectively). Responses to dietary CP concentration were linear, quadratic, and cubic with the greatest values observed when cows were fed the 16.7% CP diet across DIM (30.8 kg/d, 31.0 Mcal/d, and 1.01 Mcal/kg for DMI, milk NEL output, and FE, respectively). There was an interaction between dietary CP concentration and stage of lactation for DMI, milk NEL output, milk component yield, and FE, which was due to the decline in response to additional CP as lactation progressed. Compared with the 16.7% CP diet, feeding the 18.3% CP diet decreased milk NEL 0.81 and 5.3 Mcal/d for early and late lactation cows, respectively, indicating that feeding a higher CP concentration in late lactation had a negative effect on cow performance. Nitrogen use efficiency declined linearly with increasing CP concentration and DIM. Regression analysis suggested that dietary CP ranging from 16.3 to 17.4% maintained production in early and mid-early lactation. However, dietary CP could be reduced to between 15.7 and 17.1% in late lactation. This research suggested that there are distinct ranges of dietary CP concentrations that maintain cow performance at each stage of lactation.

Effects of maternal choline supplementation on performance and immunity of progeny from birth to weaning.

The objectives were to investigate whether supplementation with rumen-protected choline (RPC) during late pregnancy in Holstein cows affects offspring immunity and growth, and whether effects are utero-placental, colostrum dependent, or both. A total of 105 multiparous Holstein cows were assigned randomly to a prepartum diet (1.54 Mcal of NEL/kg of DM, and 15.8% CP) without (control) or with added RPC (12.9 g/d of choline ion). Calves (n = 111) were blocked by sex and assigned randomly to colostrum from control cows or colostrum from RPC cows, resulting in 4 treatments in a 2 × 2 factorial arrangement: (1) calves born and fed colostrum from non-supplemented dams (NN; n = 33); (2) calves from non-supplemented dams and fed colostrum from RPC-fed cows (NC; n = 25); (3) calves from RPC-supplemented dams and colostrum from non-supplemented cows (CN; n = 28); and (4) calves from RPC-supplemented dams and colostrum from RPC-fed cows (CC; n = 25). Growth, intakes, and immunity of females were evaluated up to 56 d of age. Growth and intake of male calves was evaluated up to 35 d of age, and physiological and immune responses to intravenous LPS challenge were evaluated from 21 to 35 d of age. Effects of prenatal and colostrum treatments and interactions between treatments were analyzed using mixed models. Calves fed colostrum from RPC-supplemented dams had a 17.4% increase in apparent efficiency of absorption of IgG compared with calves fed colostrum from control dams (27.4 vs. 23.3%). Incidence of fever in the first 21 d of age tended to be less in females born from RPC-supplemented dams compared with females born from control dams (31 vs. 58%). Prenatal RPC females had increased hematocrit and concentrations of red blood cells, leukocytes, neutrophils, and lymphocytes in blood compared with prenatal females born from control dams. Compared with prenatal control females, prenatal RPC females had greater intake of milk replacer (704 vs. 748 ± 9.9 g/d) and starter (45.4 vs. 60.2 ± 5.9 g/d) during the first 21 d of age. In male calves, mean intake of DM was greater (1,074 vs. 976 ± 45 g/d) after the LPS challenge (0 to 8 d) by calves born from dams fed RPC compared with males born from control dams. Calves born from RPC-fed dams also had lower mean rectal temperature (39.0 vs. 39.2°C) and mean respiration rate (35.6 vs. 39.3 breaths/min) compared with males born from control dams. Moreover, serum concentrations of metabolites (i.e., β-hydroxybutyric acid, fatty acids, and glucose), cytokines (i.e., tumor necrosis factor-α) and acute phase proteins (i.e., serum amyloid A) were consistent with less-severe inflammatory response to LPS in males born from dams fed RPC compared with control. Source of colostrum and interaction between prenatal and colostrum treatments had minimal effects on calf responses to LPS. Overall, maternal RPC supplementation during late gestation suggests a positive effect on immunity, in that colostrum from RPC-fed dams increased efficiency of IgG absorption and maternal supplementation with RPC during late gestation, regardless of colostrum source, attenuated responses to LPS.

Fiber digestibility and protein value of pulp silage for lactating dairy cows: Effects of wet fractionation by screw pressing of perennial ryegrass

The aim of the study was to investigate the effects of substituting silage of chopped grass with pulp silage of grass fractionated once or twice in a biorefinery using a screw press on fiber kinetics, protein value, and production of CH4 in dairy cows. Six lactating multiparous Holstein cows in mid-lactation (176 ± 93 d in milk), cannulated in the rumen, duodenum, and ileum, were used in an incomplete 6 × 4 Latin square design with a 2 × 3 factorial arrangement of treatments. Perennial ryegrass was harvested in third regrowth from the same field at early and late developmental stage (35 and 44 d of regrowth, respectively) and subjected to 1 of 3 types of processing within each developmental stage. Grass was either harvested for normal silage making (mowed, wilted, chopped, and ensiled), or harvested fresh and fractionated using a screw press. Half of the pulp from the first fractionation was ensiled, whereas the other half of the pulp was rehydrated, fractionated a second time, and pulp hereof was ensiled. The grass and pulp silages were used with concentrates (65:35 forage to concentrate ratio) to make total mixed rations (TMR) based on either silage of chopped grass (GS), pulp silage of grass fractionated once (1×P), or pulp silage of grass fractionated twice (2×P), harvested either at early (E) or late (L) developmental stage resulting in 6 different TMR treatments (EGS, E1×P, E2×P, LGS, L1×P, L2×P). The TMR were fed for ad libitum intake and samples of intestinal digesta and feces were collected for determination of digestibility. The effect of processing on ash-free neutral detergent fiber (aNDFom) concentration in silages depended on developmental stage, but showed that within each developmental stage, pulp silage of grass fractionated twice had higher aNDFom concentration than pulp silage of grass fractionated once and silage of chopped grass. The 2×P resulted in lower (14.9 ± 0.55 vs. 17.5 ± 0.54 kg/d) dry matter intake (DMI) compared with GS. The effects of processing and developmental stage interacted such that apparent total-tract aNDFom digestibility was higher (784 ± 13 vs. 715 ± 13 g/kg) for L2×P compared with LGS, whereas no difference was found between E2×P and EGS. Moreover, the protein value was higher (106 ± 5 vs. 92 ± 5 g AA digested in the small intestine/kg of DMI) for 2×P compared with GS. Unexpectedly, processing had no effect on fractional rate of digestion of digestible aNDFom or CH4 yield (L/kg of DMI), whereas feeding forages harvested at early compared with late developmental stage resulted in lower CH4 yield. Feeding pulp silage of grass fractionated once generally yielded results intermediate to cows fed silage of chopped grass and pulp silage of grass fractionated twice. This study showed that pulp silage of fractionated grass could serve as feed for dairy cows because the fiber digestibility and protein value improved, but further research investigating effects of physical processing of forage on fiber kinetics is required.