Mid-lactation Holstein Cows Research Articles

Associations of serum fatty acids, serum urea nitrogen, and ruminal ammonia nitrogen with residual feed intake in lactating dairy cows.

Feed efficiency is critical in dairy farming, impacting production costs and environmental sustainability. The development of the trait residual feed intake (RFI) has provided an opportunity to select dairy cows that are more efficient in converting nutrients into milk. Note that RFI requires individual daily intake records, which are typically collected on a limited number of research farms. In this context, the identification of biomarkers that can be used to identify and select more feed-efficient cows is of great interest. As such, this study aimed to identify ruminal and serum biomarkers associated with RFI in mid-lactation Holstein cows. A selected subset of 24 out of 454 Holstein cows was used in this study. This subset was strategically selected to represent extremes of least feed-efficient (LFE, n = 12, RFI = 2.44) and most feed-efficient (MFE, n = 12, RFI = -2.69) cows with no difference in the 3 energy sinks, namely body weight (BW) change, metabolic BW, and energy secreted in milk. Rumen fluid and serum samples were collected between 60 and 90 d in milk (DIM). Rumen fluid samples were collected using an oresophageal tubing procedure. Serum samples were used to measure fatty acids using a 2-step assay. The fatty acid methyl ester was assessed using solid phase extraction and quantified using chromatographic peak area and internal standard-based calculations. Ruminal ammonia nitrogen was measured using a phenol-hypochlorite assay, while serum urea was measured using a commercial ELISA assay. Cows in the MFE group had higher ruminal ammonia nitrogen concentrations than cows in the LFE group. There were no differences in serum urea concentration between MFE and LFE cows. Serum fatty acids concentration differed between groups, with myristic acid (C14:0), palmitic acid (C16:0), cis-heptadecenoic acid (cis9-17:1), stearic acid (C18:0), and total saturated fatty acids (SFA) having greater concentrations in the MFE group than in the LFE group. Total polyunsaturated fatty acids (PUFA) concentration was lower in the MFE group than in the LFE group. A model incorporating C14:0, C16:0, palmitoleic acid (trans9-C16:1), ante iso heptadecanoic acid plus palmitoleic acid (C17:0+trans13-C16:1), oleic acid (cis9-C18:1), cis-vaccenic acid (cis11-C18:1), petroselinic acid (cis12-C18:1), C18:0, linoleic acid (C18:2n-6), dihomo-γ-linolenic acid (C20:3n-6), cis-monounsaturated fatty acids, omega 6 PUFA, total PUFA, total SFA, other and unknown fatty acids was used to assess goodness-of-fit for RFI and showed an adjusted R2 of 0.74. When ruminal ammonia nitrogen was added to the previous model, adjusted R2 improved to 0.84. Our findings provide evidence that ruminal ammonia nitrogen and serum fatty acids are associated with RFI, thus suggesting that these metabolites might be helpful in identifying more feed-efficient dairy cows.

Effects of calcium and buffer sources on lactational performance, ruminal fermentation, nutrient digestibility, and metabolism of dairy cows

The objective was to investigate the effects of calcium and rumen buffer sources on lactational performance, ruminal fermentation, enteric gas emissions, apparent total-tract digestibility of nutrients, and blood variables of lactating dairy cows. A replicated 3 × 3 Latin square design experiment was conducted with 9 primi- and 9 multiparous mid-lactation Holstein cows. Cows were fed the same basal diet, except for the inclusion (as % DMI) of the following minerals: (1) CON, 0.80% limestone and 0.55% NaCl; (2) BICARB, 0.80% limestone and 0.80% NaHCO3; and (3) ARAG, 0.80% aragonite and 0.55% NaCl. Dry matter intake was decreased by ARAG compared with CON but not with BICARB. Treatments did not affect milk yield, ECM, or yields of milk components. Feed efficiency was increased by BICARB compared with CON but not with ARAG (i.e., 1.72, 1.64, and 1.70, respectively). Compared with CON, milk fat was increased by both BICARB and ARAG (3.32% vs. 3.58%, respectively). Milk protein concentration tended to be slightly decreased (i.e., 2%) by ARAG compared with CON but not with BICARB. Milk total solids and MUN concentrations were increased by BICARB and tended to be increased by ARAG, compared with CON. Treatments did not affect ruminal fermentation variables, except that ARAG increased butyrate molar proportion, compared with CON but not with BICARB. Additionally, ruminal NH3 concentration was greater for ARAG than for BICARB. Enteric gas emission and apparent total-tract digestibility of nutrients were not affected by treatments in the current study. Blood pH was not affected by treatments, and blood ionized Ca concentration was greater for ARAG than for BICARB, but neither was different from CON. Treatments did not affect plasma haptoglobin, β-hydroxybutyrate, or urea N concentrations. Overall, rumen buffering capacity of ARAG appears to be similar to that of BICARB, which was supported by increased milk fat, compared with CON. Compared with BICARB, ARAG increased ionized Ca concentration because of decreased base excess and HCO3- concentrations in blood.

Genetic Characterisation of Feeding Patterns in Lactating Holstein Cows and Their Association With Feed Efficiency Traits.

Feeding behaviour traits, such as number, duration or intake per feeder visit, have been associated with feed efficiency in dairy cattle. Those traits, however, do not fully capture cows' feeding patterns throughout the day. The goal of this study was to propose a new phenotype for characterising within-day feeding patterns and estimate its heritability and genetic correlations with dry matter intake (DMI), secreted milk energy, metabolic body weight and residual feed intake. Feeding patterns were evaluated using 4.8 million bunk visits from 1684 midlactation Holstein cows collected from 2009 to 2023 with an Insentec system. Feed efficiency traits were available from 6099 lactating Holstein cows at six research stations across the United States. Daily bunk visits were ordered, with Time 0 designated as the time of first feed delivery. Intake proportions were calculated by visit for each cow by dividing feed intake per visit by the total intake of the cow for that day. Feeding patterns were characterised by the area under the curve of cumulative feed intake proportions for each cow throughout the day. The feeding pattern phenotype per cow was defined as the average of areas under the curve across days, whereas consistency of feeding pattern was calculated as the natural logarithm of variance of daily area under the curve values. Estimates of heritability and genetic correlations were performed using Bayesian inference with an animal model, considering lactation, days in milk and cohort (trial-treatment) as fixed effects and animal as a random effect. Heritability estimates for average area under the curve and variance of daily area under the curve were 0.35 ± 0.05 and 0.16 ± 0.05, respectively. The genetic correlation between average area under the curve and secreted milk energy was -0.30 ± 0.14. Genetic correlations between average area under the curve and DMI, metabolic body weight and residual feed intake were not statistically significant. Variance of daily area under the curve was genetically correlated with DMI (0.47 ± 0.15), secreted milk energy (0.40 ± 0.17) and metabolic body weight (0.28 ± 0.13). The genetic correlation between variance of daily area under the curve and residual feed intake was not significant. Overall, we provided a reliable method to truly characterise feeding patterns in midlactation dairy cows. Feeding pattern and its consistency were heritable, indicating that a significant proportion of phenotypic variation is explained by additive genetic effects. Genetic correlation estimates indicate that cows with more consistent daily feeding patterns have lower DMI, lower secreted milk energy and lower metabolic body weight.

Revealing host genome–microbiome networks underlying feed efficiency in dairy cows

Ruminants have the ability to digest human-inedible plant materials, due to the symbiotic relationship with the rumen microbiota. Rumen microbes supply short chain fatty acids, amino acids, and vitamins to dairy cows that are used for maintenance, growth, and lactation functions. The main goal of this study was to investigate gene-microbiome networks underlying feed efficiency traits by integrating genotypic, microbial, and phenotypic data from lactating dairy cows. Data consisted of dry matter intake (DMI), net energy secreted in milk, and residual feed intake (RFI) records, SNP genotype, and 16S rRNA rumen microbial abundances from 448 mid-lactation Holstein cows. We first assessed marginal associations between genotypes and phenotypic and microbial traits through genomic scans, and then, in regions with multiple significant hits, we assessed gene-microbiome-phenotype networks using causal structural learning algorithms. We found significant regions co-localizing the rumen microbiome and feed efficiency traits. Interestingly, we found three types of network relationships: (1) the cow genome directly affects both rumen microbial abundances and feed efficiency traits; (2) the cow genome (Chr3: 116.5 Mb) indirectly affects RFI, mediated by the abundance of Syntrophococcus, Prevotella, and an unknown genus of Class Bacilli; and (3) the cow genome (Chr7: 52.8 Mb and Chr11: 6.1–6.2 Mb) affects the abundance of Rikenellaceae RC9 gut group mediated by DMI. Our findings shed light on how the host genome acts directly and indirectly on the rumen microbiome and feed efficiency traits and the potential benefits of the inclusion of specific microbes in selection indexes or as correlated traits in breeding programs. Overall, the multistep approach described here, combining whole-genome scans and causal network reconstruction, allows us to reveal the relationship between genome and microbiome underlying dairy cow feed efficiency.

Different sources of alfalfa hay alter the composition of rumen microbiota in mid-lactation Holstein cows without affecting production performance.

Alfalfa hay is a commonly used and important feed ingredient in dairy production. To better expand the alfalfa supply market, it is of great significance to explore the impact of alfalfa hay from different sources on dairy cow production performance. This study compared the effects of imported alfalfa hay from America (AAH) and Spain (SAH) on lactation performance and rumen microbiota of cows. Three hundred and sixty healthy mid-lactation Holstein cows with similar body weight, milk yield, and parity were randomly divided into two groups fed diets based on AAH or SAH for a 70-day experimental period. Each group was composed of four pens, with 45 cows in each pen. Daily records were kept for MY per cow and dry matter intake per pen. Twelve randomly selected cows per group were sampled to collect milk, feces, rumen fluid, and blood. The findings revealed no significant differences between the two groups in terms of production performance, nutrient apparent digestibility, serum biochemical indices, or rumen fermentation parameters. However, rumen microbial composition differed significantly between the two groups of cows based on β-diversity. On the genus level, the relative abundance of Prevotella, Succinivibrionaceae_UCG-002 increased while that of NK4A214_group, Ruminococcus, norank_f_F082 and Lachnospiraceae_NK3A20_group decreased in the SAH group compared with AAH group. There was no significant correlation between these core differential bacteria and the molar proportions of acetate and propionate, the concentration of total volatile fatty acids, and milk yield. In conclusion, the feeding effects of SAH were similar to those of AAH. These findings provided a reference for the application of alfalfa hay from different sources and for the improvement of the economic benefit of dairy farms.

Isoacid supplementation influences feed sorting, chewing behaviors, and enteric methane emissions differentially in mid-lactation dairy cows depending on dietary forage level

Past studies have shown that isoacids (ISO) improve dairy cow performance, with effects varying based on dietary forage levels, leading us to speculate that ISO supplementation may also differentially affect enteric methane (CH4) emissions depending on dietary forage levels. Therefore, our primary objective was to examine the effects of ISO supplementation on enteric CH4 emissions in lactating dairy cows fed 2 forage NDF levels (FL), along with monitoring feed particle sorting and chewing behaviors to assess any potential interactions. Sixty-four (64) mid-lactation Holstein cows were utilized in a 10-wk long randomized complete block design trial. Parity, DIM, and prior milk yield (MY) for multiparous cows or genetic merit for primiparous cows were used as blocking factors. Cows were randomly assigned to 1 of 4 diets (n = 16 per diet) with a 2 × 2 factorial arrangement of treatment combinations, including 2 FL, 17 (LF) and 21% forage NDF (HF), without or with ISO supplementation (7.85 mmol/kg DM for isobutyrate and 3.44 mmol/kg DM for 2-methylbutyrate, respectively). Enteric CH4 and chewing activity (rumination and eating time) were measured using the GreenFeed system and sensor-based ear tag system, respectively. The particle size of each diet and ort from individual cows was measured using the Penn State Particle Separator, and a sorting index was calculated. A sorting index of 100 indicates no sorting, while values above or below 100 indicate sorting for or against, respectively. Data were analyzed using a mixed model including FL, ISO, and FL × ISO as fixed effects and block as a random effect (lme4 in R). Our result shows that ISO increased sorting index for long particle in LF (96.1 vs. 109; P < 0.01) but decreased it in HF (100.8 vs. 92.5; P = 0.04). In contrast, ISO did not affect the physically effective particle sorting index (P = 0.51) or intake (P = 0.27) regardless of FL. In alignment with the long particle sorting index, ISO decreased eating and chewing time in the HF but increased them in the LF diet (P < 0.01). In contrast, rumination time was comparable between FL (P = 0.70) and ISO levels (P = 0.19). In the LF diet, ISO supplementation reduced daily CH4 production (g/d) by 9% and intensity (g/kg of MY) by 18% (P < 0.01). In the HF diet, ISO supplementation led to a 10% increase in daily CH4 (P < 0.01) but did not change CH4 intensity (P = 0.17; g/kg of MY) due to improved milk production. Overall, ISO altered feed sorting, feeding behaviors and enteric CH4 emissions depending on FL.

Supplementation of isoacids to lactating dairy cows fed low or high forage diets: Effects on performance, digestibility, and milk fatty acid profile

Our objective was to determine the effects of isoacids (ISO) on the lactation performance, digestibility, and milk fatty acid (FA) profile of Holstein cows fed 2 forage NDF levels (FL). The study was 10-wk long (including 2-wk for covariate) utilizing a randomized complete block design. Sixty-four mid-lactating Holstein cows [662 ± 71 kg BW, 119 ± 51 DIM, 2 ± 0.9 parity] were blocked by parity, DIM, and prior milk yield (MY) for multiparous cows or genetic merit for primiparous cows, and randomly assigned to 1 of the 4 diets (n = 16). Diets were arranged as a 2 × 2 factorial, with 2 FL containing 21 (HF) and 17% forage NDF (LF) without (WIA) or with ISO supplementation (IA, 7.85 mmol/kg DM and 3.44 mmol/kg DM for isobutyrate and 2-methylbutyrate, respectively). Diets were balanced for similar NEL (1.58 Mcal/kg DM), CP (16.0%) and total NDF (27.2%). Feed intake and MY were recorded daily. Nutrient digestibility for each cow was determined using indigestible NDF as a marker, and fecal samples were collected at 8-time points (4 h intervals between samples). Individual cow milk samples composited over a 10-wk period were analyzed using gas chromatography for FA profile. The statistical model included FL, ISO, and FL × ISO as fixed effects and block as a random effect (lme4 in R). The ISO did not affect DMI (P = 0.13), while LF had greater DMI than HF diets (27.8 vs. 26.0 kg/d; P < 0.01). However, ISO increased MY (34.7 vs. 37.2 kg/d; P < 0.01) and ECM (41.9 vs. 39.0 kg/d; P < 0.01) by 7% in HF but not in the LF diet, suggesting FL × ISO interaction (P = 0.04). Interestingly, ISO increased ADG (0.4 kg/d) but decreased MUN by 9% only in LF diet as indicated by FL × ISO interaction (P < 0.01). Additionally, ISO increased DM, OM, NDF, and CP digestibility by 10-24% in HF (P < 0.01), but not in LF (FL × ISO; P > 0.05). As expected, ISO increased milk odd chain FA profiles in the IA groups irrespective of FL, e.g., the IA had greater C15:0 (1.87 vs. 1.54 g/100g FA; P = 0.03) and a tendency to be greater C17:0 levels (0.86 vs. 0.76 g/100g FA; P = 0.05) compared with WIA groups. Overall, ISO improved MY and nutrient digestibility in the HF whereas it increased ADG and decreased MUN in LF diet. Additionally, ISO increased milk odd chain FA (C15:0 and C17:0) regardless of FL.

Validation of an in vivo dual permeability marker technique to characterize regional gastrointestinal tract permeability in mid-lactation Holstein cows during short-term feed restriction

This study evaluated the impact of short-term feed restriction in lactating dairy cows on regional permeability of the gastrointestinal tract (GIT), and the recovery of DMI, ruminal pH, and milk yield. In addition, sampling methods for a novel dual marker technique to characterize total GIT and post ruminal permeability were validated. Six ruminally cannulated lactating Holstein cows were blocked by parity (3 primiparous, 3 multiparous; 189 DIM ± 25.2) and enrolled in a crossover design. Experimental periods included a 5-d baseline phase (BASE), 5-d challenge phase (CHAL), and 2 weeks of recovery (REC1 and REC2). During CHAL cows received either 100% ad libitum feed intake (AL) or 40% of ad libitum feed intake (FR). To assess, total-tract and post-ruminal permeability, equimolar doses of Cr-EDTA and Co-EDTA were infused on d 3 of CHAL into the rumen and abomasum (0.369 mmol/kg BW). Following infusions, total urine and feces were collected every 8 h over 96 h, and blood samples were collected at h 0, 1, 2, 3, 4, 6, 8, 12, 16, 20, 24, 32, 40, 48, and 64. The plasma area under the curve (AUC) for Cr and Co were calculated. By design, DMI for FR was reduced by 60% during CHAL and remained 19% lower than AL during REC1 but was not different from AL in REC2. Mean ruminal pH for FR was greatest during CHAL and the least during REC1, with no differences detected between AL and FR in REC2. The duration that pH was < 5.8 was the least for FR during CHAL and greatest during REC1 which were different from AL and were no longer different between treatments in REC2. Milk yield was the least for FR during CHAL and REC1 and no longer different from AL in REC2. Feed restriction reduced milk fat, protein, and lactose yields by 26, 31% and 31%, respectively. Plasma Cr AUC was 34% greater and Co AUC tended to be 35% greater for FR than AL on d 3 of CHAL. Urinary Cr recovery after 48-h was not affected by treatment; however, urinary Co recovery was 36% greater for FR than AL. Positive correlations between plasma AUC and urinary recovery for Cr and Co were detected. It was determined that blood samples collected at h 2, 8, 20, 40, and 48 could predict the total plasma Cr and Co AUC within 1.9% and 6.2%, respectively. In summary, short-term FR in lactating dairy cows increases permeability of the total GIT and may increase permeability of the post-ruminal regions with more than 60% of the permeability occurring post-ruminally. After FR, cows experienced low ruminal pH and a sustained reduction in milk yield. When utilizing Cr- and Co-EDTA to evaluate regional GIT permeability, plasma AUC can be used as an alternate to urinary Cr and Co excretion. In addition, blood samples collected at h 2, 8, 20, 40, and 48 result in adequate prediction accuracy, at least when comparing GIT permeability for lactating dairy cows exposed to AL and FR.

Biofermentation of aquatic plants: Potential novel feed ingredients for dairy cattle production

The study assessed the impacts of aquatic plant silages on feeding efficiency and dairy cattle health as an alternative to conventional corn silage under high altitude conditions. Mid-lactation Holstein cows were assigned to treatment groups according to a randomized complete block design of parity, previous 105-d milk yield, and body weight. Cows (n = 8 per group) were fed with aquatic plant silage inoculated with Bacillus subtilis (BS), Yeast (YS), or conventional corn silage without inoculants (control) in addition to [standard grain feed] for 75 consecutive days. BS and YS had higher protein contents than control silage (111.20 ± 7.68, 112.10 ± 6.83 vs 76.94 ± 3.48 g/kg DM), while feeding efficiency was comparable between treatments (1.07, 0.99, and 0.90, respectively). In addition, the addition of aquatic plant silage in ruminant diets enhanced immunity and antioxidant capacity when compared with control group. Metagenomic analysis showed similar composition in rumen microbiota between YS and control groups, with higher enrichment for energy and nitrogen utilization pathways in YS-treated cows. This study highlights the use of aquatic plant silage as an alternative feed for dairy cattle with higher protein than corn silage. Our results suggest YS or BS could potentially boost immune and antioxidant functions, improving adaptation to high-altitudes and reducing demand for high input corn production on the Qinghai-Tibetan Plateau.

Comparison of lipidome profiles in serum from lactating dairy cows supplemented with Acremonium terrestris culture based on UPLC-QTRAP-MS/MS

This study evaluated the effects of supplementing the diet of lactating cows with Acremonium terrestris culture (ATC) on milk production, serum antioxidant capacity, inflammatory indices, and serum lipid metabolomics. Over 90 days, 24 multiparous Chinese Holstein cows in mid-lactation (108 ± 10.4 days in milk, 637 ± 25 kg body weight, 30.23 ± 3.7 kg/d milk yield) were divided into either a control diet (CON) or a diet supplemented with 30 g of ATC daily. All the data were analyzed using Student’s t test with SPSS 20.0 software. The results showed that compared with CON feeding, ATC feeding significantly increased milk yield, antioxidant capacity, and immune function. Lipidome screening identified 143 lipid metabolites that differed between the two groups. Further analysis using “random forest” machine learning revealed three glycerophospholipid serum metabolites that could serve as lipid markers with a predictive accuracy of 91.67%. This study suggests that ATC can be a useful dietary supplement for improving lactational performance in dairy cows and provides valuable insights into developing nutritional strategies to maintain metabolic homeostasis in ruminants.

Epigallocatechin-3-gallate protects bovine ruminal epithelial cells against lipopolysaccharide-induced inflammatory damage by activating autophagy

BackgroundSubacute ruminal acidosis (SARA) causes an increase in endotoxin, which can induce immune and inflammatory responses in the ruminal epithelium of dairy cows. In non-ruminants, epigallocatechin-3-gallate (EGCG), a major bioactive ingredient of green tea, is well-known to alleviate inflammation. Whether EGCG confers protection against SARA-induced inflammation and the underlying mechanisms are unknown.ResultsIn vivo, eight ruminally cannulated Holstein cows in mid-lactation were randomly assigned to either a low-concentrate (40%) diet (CON) or a high-concentrate (60%) diet (HC) for 3 weeks to induce SARA (n = 4). Cows with SARA had greater serum concentrations of tumor necrosis factor (TNF)-α and interleukin-6, and epithelium had histological signs of damage. In vitro, immortalized bovine ruminal epithelial cells (BREC) were treated with lipopolysaccharide (LPS) to imitate the inflammatory damage caused by SARA. Our data revealed that BREC treated with 10 µg/mL LPS for 6 h successfully induce a robust inflammatory response as indicated by increased phosphorylation of IκBα and nuclear factor kappa-B (NF-κB) p65. Pre-treatment of BREC with 50 µmol/L EGCG for 6 h before LPS challenge promoted the degradation of NLR family pyrin domain containing 3 (NLRP3) inflammasome through activation of autophagy, which further repressed activation of NF-κB pathway targeting Toll-like receptor 4 (TLR4). Analyses also revealed that the ECGG upregulated tight junction (TJ) protein expression upon incubation with LPS.ConclusionsSubacute ruminal acidosis causes ruminal epithelium injury and systemic inflammation in dairy cows. However, the anti-inflammatory effects of EGCG help preserve the integrity of the epithelial barrier through activating autophagy when BREC are exposed to LPS. Thus, EGCG could potentially serve as an effective therapeutic agent for SARA-associated inflammation.Graphical

Effects of feeding enriched-olive cake on milk quality, metabolic response, and rumen fermentation and microbial composition in mid-lactating Holstein cows

The aim of this study was to evaluate the effects of supplementing OC enriched in polyphenols (EOC) to mid-lactating Holstein cows on milk quality, metabolic response, and rumen fermentation and microbiota. Twenty multiparous Holstein dairy cows (10 cows/group) were enrolled for 30 d in two homogeneous groups according to milk yield, days in milking, parity, and body condition score. The control group (CTR) received no supplemented concentrate, whereas the EOC group received concentrate with a 7% inclusion of EOC. Milk and blood samples were collected at 0 d and 30 d of treatment, while rumen fluid samples were collected after 30 d of treatment. Overall, the dietary supplementation of EOC did not affect milk yield and its quality, rumen pH, and rumen VFA. However, milk of EOC cows had lower C10:0, C12:0, C14:0, and C16:0 content, and greater C18:0, C18:1 cis-9, and C18:2 n6 content, compared with CTR cows. Regarding the rumen microbiota composition, EOC diet increased the relative abundance of Bacteroidales_RF16_group, Absconditabacteriales_(SR1), and Fibrobacteracea in the rumen. On cow’s metabolism, EOC cows showed lower blood haptoglobin and greater ferric reducing antioxidant power (FRAP), cholesterol, and Zn levels compared with CTR cows. The supplementation of EOC led to a milder low-grade inflammatory response of cows and a positive modulation of cholesterol synthesis (probably at both intestinal and hepatic level). Supplementing EOC modulated rumen microbiota and improved the lipid profile of milk. Thereby, OC may be considered a valid feed to be included in the dairy cow’s diet leading to a positive effects on dairy cattle metabolism (less low-grade inflammatory response and oxidative stress) and at the same time giving an added value to the milk with a better profile of the fatty acids.

Lactational performance, ruminal fermentation, and enteric gas emission of dairy cows fed an amylase-enabled corn silage in diets with different starch concentrations

This study investigated the effects of feeding an amylase-enabled corn silage (ACS) on the performance and enteric gas emissions in lactating dairy cows. Following a 2-wk covariate period, 48 mid-lactation Holstein cows were assigned to 1 of 3 treatments in a 10-wk randomized complete block design experiment. Treatments were diets containing the same proportion of corn silage (40% of dietary DM) as follows: (1) a conventional hybrid corn silage control (CON), (2) ACS replacing the control silage (ADR), and (3) the ADR diet replacing soybean hulls with ground corn grain to achieve the same dietary starch concentration as CON (ASR). Control corn silage and ACS were harvested on the same day and contained 40.3% and 37.1% DM and (% of DM): 37.2% and 41.0% NDF and 37.1% and 30.0% starch, respectively. Enteric gas emissions were measured using the GreenFeed system. Two cows were culled due to health-related issues during the covariate period. Ruminal fluid was collected from 24 cows (8 per treatment) using the orogastric ruminal sampling technique. When compared with CON, cows fed ADR had increased DMI during experimental wk 3, 4, and 9, but treatment did not affect milk or ECM milk yields (39.0 kg/d on average; SEM = 0.89). Compared with CON, feed efficiency (per unit of milk, but not ECM) tended to be lower for ADR, whereas milk true protein concentration (a tendency) and yield were lower for ASR. Milk urea N was decreased by both ADR and ASR diets relative to CON. Compared with CON, daily CH4 emission and emission intensity were increased by ADR but not ASR. Total protozoal count tended to be increased by both diets formulated with ACS when compared with control corn silage. Total-tract digestibility of dietary NDF was greater for ASR, and that of ADF was greater for both ADR and ASR versus CON. The molar proportion of acetate (a tendency) and acetate-to-propionate ratio were increased by ADR, but not ASR, when compared with CON. Replacement of CON with ACS (having lower starch concentration) in the diet of dairy cows increased DMI during the initial weeks of the experiment, maintained ECM, tended to decrease feed efficiency, and increased enteric CH4 emissions, likely due to increased intake of digestible fiber, compared with CON.

Investigating relationships between the host genome, rumen microbiome, and dairy cow feed efficiency using mediation analysis with structural equation modeling

The rumen microbiome is crucial for converting feed into absorbable nutrients used for milk synthesis, and the efficiency of this process directly impacts the profitability and sustainability of the dairy industry. Recent studies have found that the rumen microbial composition explains part of the variation in feed efficiency traits, including dry matter intake, milk energy, and residual feed intake. The main goal of this study was to reveal relationships between the host genome, rumen microbiome, and dairy cow feed efficiency using structural equation models. Our specific objectives were to (i) infer the mediation effects of the rumen microbiome on feed efficiency traits, (ii) estimate the direct and total heritability of feed efficiency traits, and (iii) calculate the direct and total breeding values of feed efficiency traits. Data consisted of dry matter intake, milk energy, and residual feed intake records, SNP genotype data, and 16S rRNA rumen microbial abundances from 448 mid-lactation Holstein cows from 2 research farms. We implemented structural equation models such that the host genome directly affects the phenotype (GP → P) and the rumen microbiome (GM → P), while the microbiome affects the phenotype (M → P), partially mediating the effect of the host genome on the phenotype (G → M → P). We found that 7 to 30% of microbes within the rumen microbial community had structural coefficients different from zero. We classified these microbes into 3 groups that could have different uses in dairy farming. Microbes with heritability <0.10 but significant causal effects on feed efficiency are attractive for external interventions. On the other hand, 2 groups of microbes with heritability ≥0.10, significant causal effects, and genetic covariances and causal effects with the same or opposite sign to feed efficiency are attractive for selective breeding, improving or decreasing the trait heritability and response to selection, respectively. In general, the inclusion of the different microbes in genomic models tends to decrease the trait heritability rather than increase it, ranging from -15% to +5%, depending on the microbial group and phenotypic trait. Our findings provide more understanding to target rumen microbes that can be manipulated, either through selection or management interventions, to improve feed efficiency traits.

Post-milking application of a Lacticaseibacillus paracasei strain impacts bovine teat microbiota while preserving the mammary gland physiology and immunity.

Bovine mastitis (BM) is a major disease in dairy industry. The current approaches- mainly antibiotic treatments- are not entirely effective and may contribute to antimicrobial resistance dissemination, rising the need for alternative treatment. The present study aims to evaluate the impact of post-milking application of Lacticaseibacillus paracasei CIRM BIA 1542 (Lp1542) on the teat skin (TS) of 20 Holstein cows in mid lactation, in order to reinforce the barrier effect of the microbiota naturally present on the teat. Treatment (Lp1542, iodine or no treatment) was applied post-milking twice a day on the 4 teats of healthy animals for 15 days. Blood and milk samples, and TS swabs were collected at day (D)1, D8, D15 and D26 before morning milking and at D15 before evening milking (D15E) to evaluate Lp1542 impact at the microbial, immune and physiological levels. Lp1542 treatment resulted in a higher lactic acid bacteria and total microbial populations on TS and in foremilk (FM) at D15(E) compared with iodine treatment. Metabarcoding analysis revealed changes in the composition of TS and FM microbiota, beyond a higher Lacticaseibacillus abundance. This included a higher abundance of Actinobacteriota, including Bifidobacterium, and a lower abundance of Pseudomonadota on TS of Lp1542 compared with iodine-treated quarters. In addition, Lp1542 treatment did not trigger any major inflammatory response in the mammary gland, except interleukin 8 production and expression which tended to be slightly higher in Lp1542-treated cows compared with the others. Finally, Lp1542 treatment had no impact on the mammary epithelium functionality (milk yield and composition) and integrity (epithelial cell exfoliation into milk and milk Na+/K+ ratio). Altogether, these results indicate that a topical treatment with Lp1542 is safe with regard to mammary gland physiology and immune system, while impacting its microbiota, inviting us to further explore its effectiveness for mastitis prevention.

Optimizing dietary rumen-degradable starch to rumen-degradable protein ratio improves lactation performance and nitrogen utilization efficiency in mid-lactating Holstein dairy cows.

The dietary rumen-degradable starch (RDS) to rumen-degradable protein (RDP) ratio, denoted as the RDS-to-RDP ratio (SPR), has been proven to enhance in vitro rumen fermentation. However, the effects of dietary SPR in vivo remain largely unexplored. This study was conducted to investigate the effect of dietary SPR on lactation performance, nutrient digestibility, rumen fermentation patterns, blood indicators, and nitrogen (N) partitioning in mid-lactating Holstein cows. Seventy-two Holstein dairy cows were randomly assigned to three groups (24 head/group), balanced for (mean ± standard deviation) days in milk (116 ± 21.5), parity (2.1 ± 0.8), milk production (42 ± 2.1 kg/d), and body weight (705 ± 52.5 kg). The cows were fed diets with low (2.1, control), medium (2.3), or high (2.5) SPR, formulated to be isoenergetic, isonitrogenous, and iso-starch. The study consisted of a one-week adaptation phase followed by an eight-week experimental period. The results indicated that the high SPR group had a lower dry matter intake compared to the other groups (p < 0.05). A quadratic increase in milk yield and feed efficiency was observed with increasing dietary SPR (p < 0.05), peaking in the medium SPR group. The medium SPR group exhibited a lower milk somatic cell count and a higher blood total antioxidant capacity compared to other groups (p < 0.05). With increasing dietary SPR, there was a quadratic improvement (p < 0.05) in the total tract apparent digestibility of crude protein, ether extract, starch, neutral detergent fiber, and acid detergent fiber. Although no treatment effect was observed in rumen pH, the rumen total volatile fatty acids concentration and microbial crude protein synthesis increased quadratically (p < 0.05) as dietary SPR increased. The molar proportion of propionate linearly increased (p = 0.01), while branched-chain volatile fatty acids linearly decreased (p = 0.01) with increasing dietary SPR. The low SPR group (control) exhibited higher concentration of milk urea N, rumen ammonia N, and blood urea N than other groups (p < 0.05). Despite a linear decrease (p < 0.05) in the proportion of urinary N to N intake, increasing dietary SPR led to a quadratic increase (p = 0.01) in N utilization efficiency and a quadratic decrease (p < 0.05) in the proportion of fecal N to N intake. In conclusion, optimizing dietary SPR has the potential to enhance lactation performance and N utilization efficiency. Based on our findings, a medium dietary SPR (with SPR = 2.3) is recommended for mid-lactating Holstein dairy cows. Nevertheless, further research on rumen microbial composition and metabolites is warranted to elucidate the underlying mechanisms of the observed effects.

Effects of a multistrain Bacillus-based direct-fed microbial on gastrointestinal permeability and biomarkers of inflammation during and following feed restriction in mid-lactation Holstein cows

Objectives were to evaluate the effects of a multistrain Bacillus-based (Bacillus subtilis and Bacillus pumilus blend) direct-fed microbial (DFM) on production, metabolism, inflammation biomarkers and gastrointestinal tract (GIT) permeability during and following feed restriction (FR) in mid-lactation Holstein cows. Multiparous cows (n = 36; 138 ± 53 DIM) were randomly assigned to 1 of 3 dietary treatments: (1) control (CON; 7.5 g/d rice hulls; n = 12), (2) DFM10 (10 g/d Bacillus DFM, 4.9 × 109 cfu/d; n = 12) or 3) DFM15 (15 g/d Bacillus DFM, 7.4 × 109 cfu/d; n = 12). Before study initiation, cows were fed their respective treatments for 32 d. Cows continued to receive treatments during the trial, which consisted of 3 experimental periods (P): P1 (5 d) served as baseline for P2 (5 d), during which all cows were restricted to 40% of P1 DMI, and P3 (5 d), a "recovery" where cows were fed ad libitum. On d 4 of P1 and on d 2 and 5 of P2, GIT permeability was evaluated in vivo using the oral paracellular marker Cr-EDTA. As anticipated, FR decreased milk production, insulin, glucagon, and BUN but increased nonesterified fatty acids. During recovery, DMI rapidly increased on d 1 then subsequently decreased (4.9 kg) on d 2 before returning to baseline, whereas milk yield slowly increased but remained decreased (13%) relative to P1. The DFM10 cows had increased DMI and milk yield relative to DFM15 during P3 (10%). Overall, milk lactose content was increased in DFM cows relative to CON (0.10 percentage units), and DFM10 cows tended to have increased lactose yield relative to CON and DFM15 during P3 (8% and 10%, respectively). No overall treatment differences were observed for other milk composition variables. Circulating glucose was quadratically increased in DFM10 cows compared with CON and DFM15 during FR and recovery. Plasma Cr area under the curve was increased in all cows on d 2 (9%) and 5 (6%) relative to P1. Circulating LPS binding protein (LBP), serum amyloid A (SAA), and haptoglobin (Hp) increased in all cows during P2 compared with baseline (31%, 100%, and 9.0-fold, respectively). Circulating Hp concentrations continued to increase during P3 (274%). Overall, circulating LBP and Hp tended to be increased in DFM15 cows relative to DFM10 (29% and 81%, respectively), but no treatment differences were observed for SAA. Following feed reintroduction during P3, fecal pH initially decreased (0.62 units), but returned to baseline levels whereas fecal starch markedly increased (2.5-fold) and remained increased (82%). Absolute quantities of a fecal Butyryl-CoA CoA transferase (but) gene associated with butyrate synthesis, collected by fecal swab were increased in DFM10 cows compared with CON and DFM15 cows. In summary, FR increased GIT permeability, caused inflammation, and decreased production. Feeding DFM10 increased some key production and metabolism variables and upregulated a molecular biomarker of microbial hindgut butyrate synthesis, while DFM15 appeared to augment immune activation.

Effects of feeding different dietary rates of mixed fodder beet tops-wheat straw silage on the performance of Holstein lactating cows.

Rations containing different rates of the mixed fodder beet tops-wheat straw silage (BS), instead of corn silage (CS), were given to 30 mid-lactation Holstein cows (all in parity 2) to measure the effects on feed consumption, milk production efficiency, milk chemistry, urinary purine derivatives (PD), blood chemistry, antioxidant levels, and in vitro methane (CH4) emission. The BS was prepared by mixing the fodder beet tops with wheat straw at a ratio of 9:1 based on fresh weight. The experimental design was completely randomized (one 28-d period with 21-d adaptation) using 30 cows (10 animals/treatment) and 3 treatments. The treatments were 1) a diet containing CS only (25g/100g DM) (CSD), 2) a diet containing 50% CS (12.5g/100g DM) and 50% BS (12.5g/100g DM) (CBSD), and 3) diet containing BS only (25g/100g DM) (BSD). Each animal (as an experimental unit) was housed individually in the tie stall and had ad libitum access to its diet. Dietary replacing 50% of CS with BS showed no significant differences in milk production, fat-corrected milk, fat and protein yields, feed efficiency, and apparent digestibility, however, these variables were less (P < 0.05) in the cows fed with BSD. Cows fed on BSD had less intakes of DM, organic matter, crude protein, and neutral detergent fiber but greater oxalic acid intake and blood urea-N, as compared to the other cows. Milk percentages of fat, protein, lactose, urea N, blood serum glucose, triglyceride, cholesterol, total protein, albumin, globulin, Ca, and P, as well as in vitro ruminal pH, were not affected by the diets. Saturated fatty acids concentration was less and monounsaturated FA and polyunsaturated FA (PUFA) was greater in the milk of cows receiving CBSD, compared to the other groups. The inclusion of both BS rates in the diet decreased the in vitro gas production, protozoa number, and CH4 emission in comparison to the control. Cows fed BSD had decreased levels of urinary allantoin, PD excreted or absorbed, and estimated microbial-N synthesis than the control and CBSD-fed groups. The milk and blood total antioxidant capacity (TAC) of the animals fed CBSD was the maximum among the cows. Overall, under the current experimental conditions, replacing 50% of dietary CS with BS did not affect milk production, but increased milk PUFA, as well as blood and milk TAC, and decreased in vitro CH4 emission, so it's feeding to lactating Holstein cows is recommended.

Host and rumen microbiome contributions to feed efficiency traits in Holstein cows

It is now widely accepted that dairy cow performance is influenced by both the host genome and rumen microbiome composition. The contributions of the genome and the microbiome to the phenotypes of interest are quantified by heritability (h2) and microbiability (m2), respectively. However, if the genome and microbiome are included in the model, then the h2 reflects only the contribution of the direct genetic effects quantified as direct heritability (hd2), and the holobiont effect reflects the joint action of the genome and the microbiome, quantified as the holobiability (ho2). The objectives of this study were to estimate h2, hd2,m2, and ho2 for dry matter intake, milk energy, and residual feed intake; and to evaluate the predictive ability of different models, including genome, microbiome, and their interaction. Data consisted of feed efficiency records, SNP genotype data, and 16S rRNA rumen microbial abundances from 448 mid-lactation Holstein cows from 2 research farms. Three kernel models were fit to each trait: one with only the genomic effect (model G), one with the genomic and microbiome effects (model GM), and one with the genomic, microbiome, and interaction effects (model GMO). The model GMO, or holobiont model, showed the best goodness-of-fit. The hd2 estimates were always 10% to 15% lower than h2 estimates for all traits, suggesting a mediated genetic effect through the rumen microbiome, and m2 estimates were moderate for all traits, and up to 26% for milk energy. The ho2 was greater than the sum of hd2 and m2, suggesting that the genome-by-microbiome interaction had a sizable effect on feed efficiency. Kernel models fitting the rumen microbiome (i.e., models GM and GMO) showed larger predictive correlations and smaller prediction bias than the model G. These findings reveal a moderate contribution of the rumen microbiome to feed efficiency traits in lactating Holstein cows and strongly suggest that the rumen microbiome mediates part of the host genetic effect.

Intravenous infusion of 5-hydroxytryptophan to mid-lactation Holstein cows transiently affects milk production and circulating amino acid concentrations

In dairy cows, the lactating mammary glands synthesize serotonin, which acts in an autocrine-paracrine manner in the glands and is secreted into the periphery. Serotonin signaling during lactation modulates nutrient metabolism in peripheral tissues such as adipose and liver. We hypothesized that the elevation of circulating serotonin during lactation would increase nutrient partitioning to the mammary glands, thereby promoting milk production. Our objective was to elevate circulating serotonin via intravenous infusion of the serotonin precursor 5-hydroxytryptophan (5-HTP) to determine its effects on mammary supply and extraction efficiency of AA, and milk components production. Twenty-two multiparous mid-lactation Holstein cows were intravenously infused with 5-HTP (1 mg/kg body weight) or saline, in a crossover design with two 21-d periods. Treatments were infused via jugular catheters for 1 h/d, on d 1 to 3, 8 to 10, and 15 to 17 of each period, to maintain consistent elevation of peripheral serotonin throughout the period. Milk and blood samples were collected in the last 96 h of each period. Whole-blood serotonin concentration was elevated above saline control for 96 h after the last 5-HTP infusion. Dry matter intake was decreased for cows receiving 5-HTP, and on average they lost body weight over the 21-d period, in contrast to saline cows who gained body weight. Milk production and milk protein yield were lower in cows receiving 5-HTP during the 3 infusion days, but both recovered to saline cow yields in the days after. Although milk fat yield exhibited a day-by-treatment interaction, no significant difference occurred on any given day. Milk urea nitrogen concentration was lower in 5-HTP cows on the days following the end of infusions, but not different from saline cows on infusion days. Meanwhile, plasma urea nitrogen was not affected by 5-HTP infusion. Circulating concentrations of AA were overall transiently decreased by 5-HTP, with concentrations mostly returning to baseline within 7 h after the end of 5-HTP infusion. Mammary extraction efficiency of AA was unaffected by 5-HTP infusion. Overall, both lactation performance and circulating AA were transiently reduced in cows infused with 5-HTP, despite sustained elevation of circulating serotonin concentration.