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The bacteria Aeromonas hydrophila, which causes motile Aeromonas septicemia (MAS), is dangerous to aquaculture because it affects the fish's well-being and production. As the aquaculture industry seeks sustainable and effective methods to enhance fish immunity and growth, natural supplements such as marine algae have gained attention. This study explored the potential benefits of incorporating the green marine algae Chaetomorpha aerea into the fish diet, focusing on disease resistance, growth, feed utilization, and hematological and immunological responses. Five diets were prepared, varying concentrations of C. aerea (0 control, T1: 1 g/kg; T2: 2 g/kg: T3: 5 g/kg: and T4: 10 g/kg) and administered to fish over 30 days. Following the feeding trial, the fish were exposed to A. hydrophila, and their survival rates were observed for the next 14 days. The findings demonstrated that the final weight, weight gain, relative growth rate, specific growth rate, and daily growth rate were all positively impacted by a diet containing 5 g/kg of C. aerea. Additionally, fish in the 5 g/kg C. aerea group demonstrated improved feed conversion efficiency compared to the control group. While there were no significant changes in red and white blood cell counts on the initial day, serum lysozyme activity and overall resistance to infection were enhanced in fish receiving C. aerea at 2 and 5 g/kg. These results imply that C. aerea supplementation with fish supplements may be a useful immunostimulant, boosting improved health and growth in sustainable aquaculture practices., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Dairy cows experiencing heat stress (HS) during the precalving portion of the transition period give birth to smaller calves and produce less milk and milk protein. Supplementation of rumen-protected methionine (RPM) has been shown to modulate protein, energy, and placenta metabolism, making it a potential candidate to ameliorate HS effects. We investigated the effects of supplementing RPM to transition cows under HS induced by electric heat blanket (EHB) on cow-calf performance. Six weeks before expected calving, 53 Holstein cows were housed in a tiestall barn and fed a control diet (CON, 2.2% Met of MP) or a CON diet supplemented with SmartamineM (MET, 2.6% Met of MP, Adisseo Inc., France). Four weeks precalving, all MET and half CON cows were fitted with an EHB. The other half of the CON cows were considered thermoneutral (TN), resulting in 3 treatments: CONTN (n = 19), CONHS (n = 17), and METHS (n = 17). Respiratory rate (RR), skin temperature (ST), and rectal temperature (RT) were measured thrice weekly and core body temperatures recorded biweekly. Postcalving BW and BCS were recorded weekly, and DMI was calculated and averaged weekly. Milk yield was recorded daily and milk components were analyzed every third DIM. Biweekly AA and weekly nonesterified fatty acids (NEFA), BHB, insulin, and glucose were measured from plasma. Calf birth weight and 24 h growth, thermoregulation, and hematology profile were measured and apparent efficiency of absorption (AEA) of immunoglobulins was calculated. Data were analyzed using the MIXED procedure of SAS with 2 preplanned orthogonal contrasts: CONTN versus the average of CONHS and METHS (C1) and CONHS versus METHS (C2). Relative to TN, EHB cows had increased RT during the postcalving weeks and increased RR and ST during the entire transition period. Body weight, BCS, DMI, and milk yield were not affected by the EHB or RPM. However, protein percentage and SNF were lower in CONHS, relative to METHS cows. At calving, METHS dams had higher glucose concentrations, relative to CONHS, and during the postcalving weeks, the EHB cows had lower NEFA concentrations than TN cows. Calf birthweight and AEA were reduced by HS, whereas RR was increased by HS. Calf withers height tended to be shorter and RT were lower in CONHS, compared with METHS heifers. Overall, RPM supplementation to transition cows reverts the negative effect of HS on blood glucose concentration at calving and milk protein percentage in the dams and increases wither height while decreasing RT in the calf., (The Authors. Published by Elsevier Inc. on behalf of the American Dairy Science Association®. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).)

Early lactation is a critical period for dairy cows, as energy requirements rapidly increase with the onset of lactation; however, early-lactation DMI in pasture-based systems are under measured. The objectives of this study were (1) to measure and profile total DMI (TDMI) and animal performance of dairy cows during early lactation in a pasture-based system, (2) to investigate early-lactation energy balance in pasture-based systems, and (3) to examine production efficiencies, including TDMI and milk solids production per 100 kg of BW. Eighty spring-calving dairy cows were allocated to a grazing group as they calved over a 2-yr period (2021 and 2022). Cows were offered a daily herbage allowance to achieve a postgrazing sward height of 4 cm, with silage supplementation when necessary due to inclement weather. Total DMI was measured using the n-alkane technique over a 12-wk period from February 1, 2021, to April 23, 2022. Total DMI and daily milk yield were significantly affected by parity with both variables being greatest for third-parity animals (17.7 kg of DM and 26.3 kg/cow per day, respectively), lowest for first parity (13.2 kg of DM and 19.6 kg/cow per day, respectively) and intermediate for second-parity animals (16.8 kg of DM and 24.1 kg/cow per day, respectively). Peak TDMI was reached on wk 10 for first-parity animals (14.6 kg of DM), wk 11 for second parity animals (19.3 kg of DM) and wk 12 for third-parity animals (19.9 kg of DM). Parity also had a significant effect on unité fouragère lait (UFL; feed units for milk) feed balance as first-parity animals experienced a greater degree of negative energy balance (-3.2 UFL) compared with second- and third-parity animals (-2.3 UFL). Breed and parity had an effect on production efficiencies during the first 12 wk of lactation as Jersey × Holstein Friesian cows had greater TDMI/100 kg of BW and milk solids/100 kg of BW compared with Holstein Friesian cows., (The Authors. Published by Elsevier Inc. on behalf of the American Dairy Science Association®. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).)

Botanical extracts (BE; Apex, Adisseo) have demonstrated enhanced DMI and improved gut health, whereas direct-fed microbials (DFM), such as Lactobacillus acidophilus fermentation product (Excell [EX], Pacer Technology Inc.), have demonstrated improved gut health and growth performance of growing Holstein calves. The hypothesis was this combination may be synergistic to neonatal calf growth performance and intestinal health. Eighty 2- to 5-d-old Holstein bull calves were blocked by BW and randomly assigned to 1 of 8 treatments arranged in a 2 × 4 factorial using a randomized complete block design. The main factors were milk replacer (MR) without (control) and with EX added at 5 g/d fed and calf starter (CS). The CS containing no additives (control); CS containing BE at 496 mg/kg; CS containing EX at 2.50 g/kg; and CS containing BE and EX at the same inclusion rates. The MR were fed 2×/d at 0630 and 1800 h along with free choice CS (amounts and orts weighed daily) and water. Weaning occurred after d 42 for the 56-d experiment. No MR by CS main effects interactions were detected for BW, ADG, CS intake, total DMI, feed efficiency, or body frame gain parameters. The BW gain (38.0 and 39.3 kg for control and EX, respectively) for MR main effect was similar for calves fed both MR treatments, whereas CS main effects (38.7, 39.7, 39.2, and 37.2 kg for control, BE, EX, and BE+EX, respectively) was similar among all CS. Gains in body length (10.6 and 10.8 cm), hip width (4.5 and 4.5 cm), withers height, (10.5 and 10.6 cm), heart girth (18.6 and 19.9 cm), and body length (9.1 and 7.9 cm) were similar for calves fed both MR, while CS main effects for hip height (10.5, 10.2, 10.3, and 10.9 cm), hip width (4.7, 4.6, 4.4, and 4.3 cm), withers height (10.7, 10.9, 10.3 and 10.6 cm), heart girth (19.9, 18.9, 18.9, and 19.4 cm), and body length (11.7, 9.1, 8.3, and 8.4 cm) were similar. Total days of a fecal score = 0 was greater for calves fed control MR and BE CS compared with calves fed control MR and the combination of BE+EX, with calves fed the remaining treatments being intermediated and similar. This study demonstrated little calf growth performance and health benefits when feeding a BE or EX alone or in combination compared with calves fed control treatments., (The Authors. Published by Elsevier Inc. on behalf of the American Dairy Science Association®. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).)

The study investigated the effects of dietary protein level and the inclusion of hydroponic barley sprouts (HB) on lactation performance, blood biochemistry and N use efficiency in mid-lactation dairy cows. Treatments were arranged in a 2 × 2 factorial design with 2 CP levels (16.8% and 15.5% of DM), with HB (4.8% of DM, replacing 4.3% of alfalfa hay and 0.5% of distillers dried grains with solubles [DDGS]) or without HB. Forty-eight multiparous Holstein dairy cows (146 ± 15 DIM, 40 ± 5 kg/d of milk) were randomly allocated to 1 of 4 diets: high-protein diet (16.8% CP, HP), HP diet with HB (HP+HB), low-protein diet (15.5% CP, LP), or LP diet with HB (LP+HB). An interaction between CP × HB on DMI was detected, with DMI being unaffected by HB inclusion in cows fed the high-protein diets, but was lower in cows fed HB when the low-protein diet was fed. A CP × HB interaction was also observed on milk and milk protein yield, which was higher in cows fed HB with HP, but not LP. Inclusion of HB also tended to reduce milk fat content, and feeding HP resulted in a higher milk protein and MUN content, but lower milk lactose content. Feed efficiency was increased by feeding HP or HB diets, whereas N use efficiency was higher for cows fed LP or HB diets. There was an interaction on the apparent total-tract digestibility of DM and CP, which was higher when HB was fed along with HP, but reduced when fed with LP, whereas the digestibility of ADF was increased by feeding low-protein diets. In conclusion, feeding a low-protein diet had no adverse effect on cow performance, while feeding HB improved milk and milk component yield, and N efficiency when fed with a high-CP diet, but compromised cow performance with a low-CP diet., (The Authors. Published by Elsevier Inc. on behalf of the American Dairy Science Association®. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).)

The aim of this study was to evaluate the effects of different anti-mycotoxin feed additives on the concentration of mycotoxins in milk, urine, and blood plasma of dairy cows fed diets artificially contaminated with mycotoxins. Secondarily, performance, total-tract apparent digestibility of nutrients, and blood parameters were evaluated. Twelve multiparous cows (165 ± 45 DIM, 557 ± 49 kg BW, and 32.1 ± 4.57 kg/d milk yield at the start of the experiment) were blocked according to parity, milk yield, and DIM and used in a 4 × 4 Latin square design experiment with 21-d periods, where the last 7 d were used for sampling and data analysis. Treatments were (1) mycotoxin group (MTX), basal diet (BD) without anti-mycotoxin feed additives; (2) hydrated sodium calcium aluminosilicate (HSCA), HSCA added to the BD at 25 g/cow per day; (3) mycotoxin deactivator (MD; Mycofix Plus, dsm-firmenich) added to the BD at 15 g/cow per day (MD15); and (4) MD added to the BD at 30 g/cow per day (MD30). Cows from all treatments were challenged with a blend of mycotoxins containing 404 μg of aflatoxin B 1 , 5,025 μg of deoxynivalenol (DON), 8,046 μg of fumonisins (FUM), 195 μg of T2 toxin (T2), and 2,034 μg of zearalenone (ZEN) added daily to the BD during the last 7 d of each period. Neither performance (milk yield and composition) nor nutrient digestibility was affected by treatments. All additives reduced aflatoxin M 1 (AFM1) concentration in milk, whereas MD15 and MD30 group had lower excretion of AFM1 in milk than HSCA. Deoxynivalenol, FUM, T2, or ZEN were not detected in milk of MD15 and MD30. Concentrations in milk of DON, FUM, T2, and ZEN were similar between MTX and HSCA. Except for AFM1, none of the analyzed mycotoxins were detected in urine of MD30 group. Comparing HSCA to MD treatments, the concentration of AFM1 was greater for HSCA, whereas MD30 was more efficient at reducing AFM1 in urine than MD15. Aflatoxin M1, DON, FUM, and ZEN were not detected in the plasma of cows fed MD30, and DON was also not detected in MD15 group. Plasma concentration of FUM was lower for MD15, similar plasma FUM concentration was reported for HSCA and MTX. Plasma concentration of ZEN was lower for MD15 than MTX and HSCA. Serum concentrations of haptoglobin and hepatic enzymes were not affected by treatments. Blood concentration of sodium was lower in HSCA compared with MD15 and MD30 groups. In conclusion, the mycotoxin deactivator proved to be effective in reducing the secretion of mycotoxins in milk, urine, and blood plasma, regardless of the dosage. This reduction was achieved without adverse effects on milk production or total-tract digestibility in cows fed multi-mycotoxin-contaminated diets over a short-term period. Greater reductions in mycotoxin secretion were observed with full dose of MD., (The Authors. Published by Elsevier Inc. on behalf of the American Dairy Science Association®. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).)

Interventions targeting the gut microbiota, such as fecal microbiota transplantation, prove effective in repairing the intestinal barrier and facilitating the recovery of its function and metabolism. However, the regulatory mechanisms governing the remodeling of rumen epithelial morphology and function, rumen metabolism, and host metabolism in cows of SARA remain poorly understood. Here, we explored the changes in rumen epithelial morphology and transcriptome, rumen metabolome, and blood biochemical parameters in SARA cows following rumen content transplantation (RCT). The entire experiment consisted of 2 periods: the SARA induction period and the RCT period. During the SARA induction period, 12 ruminally cannulated lactating Holstein cows were randomly allocated into 2 groups, fed either a conventional diet (CON; n = 4; 40% concentrate, DM basis) or a high-grain diet (HG; n = 8; 60% concentrate, DM basis). Following the SARA induction period, the RCT period started. The HG cows were randomly assigned to 2 groups: the donor-recipient (DR) group and the self-recipient (SR) group. Rumen contents were entirely removed from both groups before RCT. For the DR group, cows were administered 70% rumen content from the CON cows, paired based on comparable BW; for the SR group, each cow received 70% self-derived rumen content. The results revealed no significant differences in the thicknesses of the stratum corneum, granulosum, and spinosum or basale layers, as well as the total depth of the epithelium between the SR and DR groups. All these measurements exhibited a decreasing trend and fluctuations over time after the transfer. Notably, these fluctuations tended to stabilize at 13 or 16 d after RCT in the SR group, whereas they tended to stabilize after 8 or 13 d of transfer for the DR group. Transcriptome sequencing revealed that a total of 277 differentially expressed genes (DEG) were identified between the 2 groups. Enrichment analysis showed that the DEG were significantly enriched in 11 Gene Ontology biological processes and 14 Kyoto Encyclopedia of Genes and Genomes pathways. The DEG corresponding to almost any of these 11 biological process terms and 14 pathways showed mixed up- or downregulation following RCT. Metabolomics analysis indicated that a total of 33 differential metabolites were detected between the SR and DR groups, mainly enriched in 5 key metabolic pathways, including plant polysaccharides and starch degradation, lipid metabolism, amino-sugar and nucleotide-sugar metabolism, purine metabolism, and Krebs cycle. Among them, the levels of differential metabolites associated with the degradation of plant polysaccharides and starches, metabolism of amino sugars and nucleotides, and purine metabolism pathways were significantly elevated in the DR cows. The results of blood biochemical parameters showed that the triglyceride concentration of the DR cows was increased than that of the SR cows, comparable to the level observed in the CON cows during the SARA induction period. Generally, our findings indicated that RCT facilitated the recovery of rumen epithelial morphological structure but did not promote its function recovery. Moreover, RCT enhanced rumen plant polysaccharide and starch degradation, amino-sugar and nucleotide-sugar metabolism, as well as purine metabolism. Additionally, it further promoted the recovery of plasma metabolites related to lipid metabolism., (The Authors. Published by Elsevier Inc. on behalf of the American Dairy Science Association®. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).)

In modern breeding systems, cows are subjected to many stress factors. Animals fed a high-grain diet may have a decreased rumen pH, which would lead to subacute ruminal acidosis syndrome. The aim of this study was to investigate the evolution of microbial community composition in cows undergoing a dietary stress challenge. Twelve cows were subjected to a challenge period that consisted of a rapid change of ration, from a normal (45.4:54.6 forage:concentrate) to a high-grain content diet (24.8:75.2 forage:concentrate) to induce subacute ruminal acidosis. Individual rumen fluid content samples were collected before (T0) and during the challenge (T3, T14, T28). The DNA from rumen contents was extracted, purified, and sequenced to evaluate bacterial populations, and sequencing was performed on Illumina MiSeq. The effect of animal conditions on rumen microbial community was quantified through a linear mixed model. The acidogenic diet created 2 main clusters: ruminal hypomotility (RH) and milk fat depression (MFD). The microbial composition did not differ in T0 between the 2 groups, whereas during the challenge Ruminococcus spp., Treponema spp., Methanobrevibacter spp., and Methanosphaera spp. concentrations increased in RH cows; and Succinivibrio spp. and Butyrivibrio spp. concentrations increased in MFD cows. Prevotella spp. and Ruminococcus spp. were negatively correlated, whereas the Christenellaceae family was positively correlated with both Methanobrevibacter spp. and Methanosphaera spp. Moreover, the same diet affected cows' microbiota composition differently, underlying the impact of the host effect. Other studies are necessary to deepen the relationship between microbiota composition and host., (The Authors. Published by Elsevier Inc. on behalf of the American Dairy Science Association®. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).)

Cow milk microbiota has received increased attention in recent years, not only because of its importance for human health but also because of its effect on the quality and technological properties of milk. Several studies, therefore, have investigated the effect of various production factors on the microbial composition of milk. However, most of the previous studies considered a limited number of animals from experimental or single farm, which could have biased the results. Therefore, this study aimed to understand the effect of different alpine production systems in Italy on the compositional and microbiological quality of milk, considering commercial herds with different feeding intensities and cattle breeds. The results obtained in this work indicated that the month and season of sampling (July for summer or February for winter) more than farm, breed, and cow diet exerted significant effects on cow milk parameters and microbiota. In particular, significant differences were observed for urea content in milk between sampling seasons. Differences in milk fat were mainly related to breed specific effects. From a microbiological point of view, statistically significant differences were found in presumptive lactic acid bacteria counts. Based on a culture-independent method, milk obtained in February harbored the highest number of Firmicutes (e.g., Lactobacillus) and the lowest number of Actinobacteria (e.g., Corynebacterium). Moreover, bacterial richness and diversity were higher in July during alpine pasture season indicating a significant effect of pasture feed on the growth of bacterial communities. The results of this study highlighted the effect of month or season mainly related to differences in feeding management (e.g., access to pasture during vegetation period, concentrates supplementation) on composition and microbiota in milk., (The Authors. Published by Elsevier Inc. on behalf of the American Dairy Science Association®. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).)

Dietary carbohydrate manipulation can be used to reduce enteric CH 4 emission, but few studies have focused on the interaction of the different types of carbohydrates that can affect feed intake and ruminal fermentation. Understanding this interaction is necessary to make the most out of CH 4 mitigation feeding strategies using different dietary carbohydrates. The aim of this study was to test the effect on enteric CH 4 emission, feed intake, and milk production response when cows were fed either grass-clover silage (GCS) or corn silage (CS) as the sole forage source (55% of dry matter, DM), in combination with either barley (BAR) or dried beet pulp (DBP) as a concentrate (21.5% of DM). A total of 24 (half first-parity and half second-parity) cows were used in a crossover design with 2 periods of 21 d each, receiving 2 of 4 diets obtained from a 2 × 2 factorial arrangement of the experimental diet. Feed intake, CH 4 emission metrics, and milk production were recorded at the end of the experimental periods. The diets had NDF concentrations between 258 and 340 g/kg of DM and starch concentrations between 340 and 7.45 g/kg of DM (CS-BAR and GCS-DBP, respectively). The effects of silage and concentrate on dry matter intake (DMI) were additive, with the highest feed intake in cows fed CS-BAR, followed by cows fed CS-DBP, GCS-BAR, and GCS-DBP (21.2, 19.9, 19.1, and 18.3 kg/d, respectively). Energy corrected milk (ECM) yield was not affected by silage source in first parity cows, but it was higher for cows fed CS than cows fed GCS in second parity. The effects of silage and concentrate on CH 4 production (g/d), yield (g/kg of DMI), and intensity (g/kg of ECM) were not additive, as cows fed GCS had similar responses regardless of the concentrate used, but cows fed CS had lower CH 4 production, yield, and intensity when fed BAR instead of DBP. The lower CH 4 production, yield, and intensity in cows fed CS-BAR compared with other diets could be partially explained by the nonlinear relationship between ruminal VFA and carbohydrates (NDF and starch) concentration reported in the literature; however, we observed a linear relationship between the acetate/propionate ratio and CH 4 yield, suggesting possible other effects. The effects of silage and concentrate on the ruminal VFA were additive in first parity cows, but not in second parity cows. The interaction between dietary carbohydrate type and parity might indicate an effect of feed intake or the energy balance of the cow. Feeding cows silage and concentrate both rich in starch can result in the lowest enteric CH 4 emission., (The Authors. Published by Elsevier Inc. on behalf of the American Dairy Science Association®. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).)

We determined the effects of increasing the dietary inclusion of whole cottonseed (WCS) on nutrient digestibility and the milk production responses of high-producing dairy cows. Twenty-four multiparous Holstein cows (mean ± SD; 52.7 ± 2.63 kg/d milk; 104 ± 23 DIM) were randomly assigned to treatment sequences in a replicated 4 × 4 Latin square design with 21-d periods. Treatments were increasing doses of WCS at 0%, 8%, 16%, and 24% DM, with WCS replacing soybean meal and hulls to maintain similar diet nutrient composition (% DM) of NDF (32%), forage NDF (21%), starch (27%), and CP (17%). Total fatty acid (FA) content of each treatment was 1.70%, 2.96%, 4.20%, and 5.40% DM, respectively. Three preplanned contrasts were used to test the linear, quadratic, and cubic effects of increasing dietary WCS. Increasing dietary WCS from 0% to 24% DM quadratically influenced the intake of DM and NDF, with the highest value being the inclusion of 8% WCS, and intakes of 16- and 18-carbon, and total FA, with maximum values obtained up to 24% WCS. Increasing dietary WCS affected digestibility of DM (cubic) and NDF (quadratic), with the lowest values being the inclusion of 8% WCS. Increasing WCS increased 16-carbon digestibility (quadratic) but decreased digestibility of 18-carbon and total FA (both quadratic), with highest and lowest values for the inclusion of 24% WCS, respectively. Increasing dietary WCS quadratically increased absorbed 16- and 18-carbon, and total FA, with maximum values obtained for 24% WCS. Increasing dietary WCS quadratically increased yields of milk, milk fat, milk protein, milk lactose, 3.5% FCM, and ECM, and linearly increased BW gain. The source of milk FA was affected by dietary WCS, with a quadratic decrease in the yield of de novo and mixed milk FA and a quadratic increase in preformed milk FA. Increasing dietary WCS linearly increased trans-10 C18:1 milk FA content. As dietary WCS increased, plasma insulin linearly decreased, and plasma gossypol levels linearly increased. Despite the decrease in total FA digestibility, increasing dietary WCS from 0% to 24% DM increased FA absorption. Increasing the dietary inclusion of WCS up to 16% DM increased milk production responses and DM intake. Under the current dietary conditions, high-producing dairy cows benefited best from a diet containing an inclusion of 8% to 16% WCS in DM., (The Authors. Published by Elsevier Inc. on behalf of the American Dairy Science Association®. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).)

Understanding the extent to which genetics × environment plays a role in shaping individual strategies to environmental challenges is of considerable interest for future selection of more resilient animals. The objective of this study was to evaluate the metabolic responses of goats, selected for functional longevity, to a nutritional challenge. This evaluation was based on plasma metabolites and was repeated across 2 experimental farms over 2 years. We carried out 6 different experimental trials from 2018 to 2022 (4 trials on site Bourges, 2018-2021 and 2 trials on site Grignon, 2021-2022) in which 267 first kidding goats, daughters of Alpine bucks divergently selected for functional longevity, longevity plus (n = 137), and longevity minus (n = 130), were exposed to a 2-d nutritional challenge in early lactation. The experiments consisted of a 5 or 7-d control period (prechallenge) on a standard lactation diet followed by a 2-d nutritional challenge with straw-only feeding, and then a 7- or 10-d recovery period on a standard lactation diet, for site Bourges and Grignon, respectively. During the challenge plasma metabolite composition was measured. Linear mixed models were used to analyze all traits, considering the individual as a random effect and the 2 × 2 treatments (i.e., genetic line and year nested in site) and litter size as fixed effects. The linear mixed-effects model using a piecewise arrangement was used to analyze the response/recovery profiles to the nutritional challenge. The estimates of the random effect for each individual, using the mixed models without the fixed effects of genetic line, were used in a sparse partial least square discriminant analysis (sPLS-DA) to compare the goat metabolism response to the challenge on a multivariate scale. The plasma glucose, BHB, nonesterified fatty acids (NEFA), and urea concentrations significantly responded to the 2-d nutritional challenge. Selection for functional longevity did not affect plasma glucose, NEFA, BHB, and urea response/recoveries to a 2-d nutritional challenge. However, site, trial, and litter size significantly affected these responses. Moreover, the plasma metabolites seem not to fully recover to prechallenge levels after the recovery phase. The sPLS-DA analysis did not discriminate between the 2 longevity lines. We observed meaningful between-individuals variability in plasma BHB, especially on the prechallenge and rate of response and rate of recovery from the 2-d nutritional challenge (CV = 26.2%, 36.1%, and 41.2%, repeatability = 0.749, 0.322, and 0.741, respectively). Plasma NEFA recovery from challenge also demonstrated high between-individuals variability (CV = 16.4%, repeatability = 0.323). Selection for functional longevity did not affect plasma metabolites responses to a 2-d nutritional challenge in dairy goats. Plasma NEFA and BHB response and recovery presented high between-individuals variability, indicating that individual adaptative characteristics to nutritional challenges are not related to the environmental conditions but to inherent individual characteristics., (The Authors. Published by Elsevier Inc. on behalf of the American Dairy Science Association®. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).)

The objective of this study was to evaluate the effect of energy and protein in the diet on the recovery of milk ethanol stability (MES) induced by feed restriction. Twelve Holstein and Holstein × Jersey crossbred cows with an average of 146 ± 50 DIM, 575.4 ± 70 kg of BW, and 18.93 ± 5.46 kg/d of milk yield were distributed in a 3 × 3 Latin square design with 3 treatments and 3 experimental periods. Each experimental period lasted 24 d, comprising 3 phases: a 13-d adaptation phase (100E+100P), a 4-d induction phase for milk ethanol instability (50E+50P), and a 7-d recovery phase for MES (3 treatments). The 3 treatments during the recovery phase consisted of 3 diets aiming to meet the requirements of energy and protein (100E+100P), only energy (100E+50P), or only protein (50E+100P). The diet during the adaptation and induction phases was common for all cows. The energy and protein levels to meet each cow's requirements were based on the group average. Restriction of energy and protein reduced DM, CP, and total digestive nutrient intake for cows fed 100E+50P and 50E+100P. The lowest BW was observed for cows fed 50E+100P, with no difference for BCS. During the induction phase, MES was reduced by 9 percentage units. Cows fed 100E+100P recovered MES in the first days of the recovery phase, whereas 100E+50P slightly improved MES, and 50E+100P had a constant decrease in MES. Cows fed 100E+50P and 50E+100P produced, respectively, 3.6 and 5.9 kg less milk than those fed 100E+100P. The 50E+100P treatment exhibited the highest milk fat content and SCS, along with the lowest milk lactose content. Protein content was higher in the 100E+100P treatment. Cows fed 50E+100P showed higher serum albumin levels compared with those on the 100E+100P treatment, not differing from the 100E+50P treatment. We concluded that the complete recovery of MES in cows with feed restrictions is possible only by supplying both the energy and protein requirements in the cows' diet. However, restricting energy intake poses a greater limitation on MES recovery compared with restricting protein., (The Authors. Published by Elsevier Inc. on behalf of the American Dairy Science Association®. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).)

The purpose of this experiment was to determine if nicotinic acid (NA) effects on dairy cows and rumen microbial characteristics are forage-type dependent (corn silage, CS; grass silage, GS). Four late-lactation (DIM = 225 ± 12 d) Holstein cows were used in a 4 × 4 Latin square design with a 2 × 2 factorial arrangement of treatments. The main effects were a CS (66.10% CS) based diet or a GS (79.59%) based diet with or without 12 g/d NA. Each experimental period lasted for 28 d. Milk production and milk components, blood metabolites, apparent total-tract nutrient digestibilities, minutes rumen pH were below 5.8 as an indicator of ruminal acidosis, and body temperature changes were analyzed as indicators of heat stress. Nicotinic acid supplementation did not improve apparent total-tract nutrient digestibility. Feeding a GS-based diet improved NDF and hemicellulose digestibility. Feeding a CS-based diet increased the apparent total-tract digestibility of fat, and minutes rumen pH below 5.8 for a greater proportion of the time. The CS-based diet also improved milk yield, milk fat and protein yields, and ECM yield; however, somatic cell count and BHB were also increased. Supplementing NA tended to decrease nonesterified fatty acids, especially when combined with GS where DMI was low. There was a trend for the total protozoa population to increase when GS and NA were fed but decreased when CS and NA were fed. Nicotinic acid tended to decrease rumen protozoal populations of Dasytricha, but increased populations of Ophryoscolex and Diplodiniinae with GS diets and decreased with CS diets. Entodiniinae were increased with CS but NA had no effect. Body temperature was increased when a CS-based diet was fed when compared with a GS-based diet. More research is needed to determine how NA can affect rumen microbial protein synthesis and what kind of diets will provide the optimum effect., (The Authors. Published by Elsevier Inc. on behalf of the American Dairy Science Association®. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).)

Most trace minerals (TM) are fed above dairy cow requirements in commercial herds but their fate and effects on dairy cows have not been well documented. In this study, we evaluated the effects of feeding short-term sulfate TM above recommendations on apparent total-tract digestibility of nutrients, rumen fermentation characteristics, serum concentrations, and milk yield and composition, as well as milk, fecal, and urinary TM excretion in midlactation dairy cows. Eight multiparous Holstein cows with an average body weight (± SD) of 684 ± 29 kg at 82 ± 10 DIM in a quadruple 2 × 2 crossover design were fed a basal diet, differing in sulfate TM supplement concentrations, to provide either 0.11, 17, and 63 (control; CON) or 0.95, 114, and 123 (high trace minerals; HTM) mg of dietary Co, Mn, and Zn per kilogram of DM, respectively. Each experimental period had a 21-d adaptation to the diet, followed by a 10-d sample collection period. Feed ingredients and total feces and urine were collected during 4 consecutive d and rumen fluid was collected 0, 1, 2, 4, and 6 h relative to feeding. Milk yield was recorded daily, and milk samples were collected on 4 consecutive milkings. Ingestion of Co, Mn, and Zn was higher for the HTM group compared with the CON group by 216%, 233%, and 93%, respectively. Dry matter intake averaged 25.0 (SE = 0.6) kg/d, and apparent total-tract digestibility of major nutrients was similar between treatments. High trace minerals had no measurable effect on ruminal pH, major volatile fatty acids, and protozoa counts. Isovalerate molar proportion was 9.4% greater for the HTM group compared with the CON group. Neither milk yield (43.5 kg/d; SE = 0.8) nor milk fat and protein concentrations differed between treatments. Milk urea nitrogen concentration was significantly higher for HTM (11.7 mg/dL) compared with CON (9.7 mg/dL; SE = 0.7). Fecal excretion of Co, Mn, and Zn increased by 223%, 198%, and 75%, respectively, for the HTM group compared with the CON group. Urinary excretions of TM were marginal compared with feces, and only urinary Co and Mn were significantly higher for HTM cows than CON cows, as was similarly obtained for serum Co and Mn concentrations. Milk TM yields were not modified by treatments. In summary, short-term dietary sulfate TM supply over the recommendation did not improve cow performance but significantly increased fecal TM excretion, which could affect TM accumulation in soils where manure is applied and could potentially result in leaching into nearby watersheds. Further studies are needed to evaluate the impact of high fecal TM excretion on the environment using the One Health approach. Moreover, the effects of TM oversupply on milk production and cow health should be evaluated by long-term experiments., (Copyright © 2024 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.)

Our objective was to compare abomasal infusions of linoleic (18:2n-6) and α-linolenic (18:3n-3) acids on the enrichment of n-6 and n-3 fatty acids (FA) into the plasma lipid fractions of lactating dairy cows and evaluate their potential carryover effects in plasma lipid fractions postinfusion. Six rumen-cannulated multiparous Holstein cows (252 ± 33 DIM) were fed the same diet and assigned to 1 of 2 treatments in a completely randomized design with repeated measures. Treatments were abomasal infusions (67 g/d total FA) of (1) n-6 FA blend (N6) to provide approximately 43 g/d 18:2n-6 and 8 g/d of 18:3n-3 or (2) n-3 FA blend (N3) providing 43 g/d 18:3n-3 and 8 g/d 18:2n-6. Treatments were dissolved in ethanol, and the daily dose for each treatment was divided into 4 equal infusions, occurring every 6 h. The treatment period lasted from d 1 to 20, and the carryover period lasted from d 21 to 40. Results are presented as FA contents within each of the 4 main plasma lipid fractions: cholesterol esters (CE), phospholipids (PL); triglycerides, and plasma nonesterified fatty acids. Concentrations of individual lipid fractions in plasma were not quantified. Plasma CE and PL had the highest content of PUFA during both the treatment and carryover periods. In plasma PL, N3 increased the contents of total n-3 FA (134%), 18:3n-3 (267%), and eicosapentaenoic acid (96.3%, 20:5n-3), and decreased total n-6 FA (8.14%) and 18:2n-6 (8.16%) from d 4 to 20 compared with N6. In plasma CE, N3 increased the contents of total n-3 FA (191%) from d 4 to 20, 18:3n-3 from d 2 to 20 (178%), and 20:5n-3 from d 6 to 20 (59.9%), while N3 decreased total n-6 FA from d 4 to 20 (11.2%) and 18:2n-6 from d 2 to 20 (10.5%) compared with N6. In addition, compared with N6, N3 decreased arachidonic acid (20:4n-6) at d 2 (45%) and from d 10 to 20 (14.7%) in PL and tended to decrease 20:4n-6 without interacting with time for CE. Phospholipids were the only lipid fraction with detectable levels of docosahexaenoic acid (22:3n-6) in all samples, but we did not observe differences between treatments. In plasma trigylcerides, N3 increased the contents of total n-3 FA (135%) and 18:3n-3 (146%) from d 4 to 20, increased 20:5n-3 from d 12 to 20 (89%), decreased or tended to decrease total n-6 FA content from d 6 and 8 (26.9%), and tended to decrease 18:2n-6 at d 8 compared with N6. A similar pattern was observed for plasma nonesterified FA. We observed positive carryover effects for both N3 and N6 at different degrees in all lipid fractions, with N3 promoting more consistent outcomes and increasing total n-3 FA throughout the carryover period (from d 22 to 40) in both PL (52.8%) and CE (68.6%) compared with N6. It is important to emphasize that the higher magnitude responses observed for n-3 FA are also influenced by the content of n-3 FA being much lower than those of n-6 FA in all lipid fractions. Although these data provide important and robust information, future research quantifying changes in concentrations of individual lipid fractions in plasma and the entry and exit rates of specific FA will further enhance our understanding. In conclusion, abomasally infusing N3 and N6 increased the contents of n-3 and n-6 FA, respectively, in all plasma lipid fractions. These responses were more evident in PL and CE. We also observed positive carryover effects in all lipid fractions, where N3 had more consistent outcomes than N6. Our results indicate that dairy cows have a robust mechanism to conserve essential FA, with a pronounced preference for n-3 FA., (The Authors. Published by Elsevier Inc. on behalf of the American Dairy Science Association®. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).)

The objectives of this study were to determine the effects of 2 dietary microbial additives on productive performance and feed efficiency when supplemented to diets of Holstein cows. One hundred seventeen Holstein cows were enrolled at 61 d (31-87 d) postpartum in a randomized complete block design experiment. Cows were blocked by parity group, as nulliparous or multiparous cows and, within parity, by pretreatment ECM yield. Within block, cows were randomly assigned to one of 3 treatments administered as top-dress for 140 d. Treatments consisted of either 100 g of corn meal containing no microbial additive (CON; 15 primiparous and 25 multiparous), 100 g of corn meal containing 5 g of a mixture of Clostridium beijerinckii and Pichia kudriavzevii (G1; 4 × 10 7 cfu of C. beijerinckii and 1 × 10 9 cfu of P. kudriavzevii; 14 primiparous and 24 multiparous), or 100 g of corn meal containing 5 g of a mixture of C. beijerinckii, P. kudriavzevii, Butyrivibrio fibrisolvens, and Ruminococcus bovis (G2; 4 × 10 7 cfu of C. beijerinckii, 1 × 10 9 cfu of P. kudriavzevii, 1 × 10 8 cfu of B. fibrisolvens, and 1 × 10 8 cfu of R. bovis; 15 primiparous and 24 multiparous). Intake of DM, milk yield, and BW were measured daily, whereas milk composition was analyzed at each milking 2 d a week, and body condition was scored twice weekly. Milk samples were collected on d 60 and 62 in the experiment and analyzed for individual fatty acids. The data were analyzed with mixed-effects models with orthogonal contrast to determine the effect of microbial additive (MA; CON vs. 1/2 G1 + 1/2 G2) and type of microbial additive (TMA; G1 vs. G2). Results are described in sequence as CON, G1, and G2. Intake of DM (22.2 vs. 22.4 vs. 22.4 kg/d), BW (685 vs. 685 vs. 685 kg) and the daily BW change (0.40 vs. 0.39 vs. 0.39 kg/d) did not differ among treatments; however, feeding MA tended to increase BCS (3.28 vs. 3.33 vs. 3.36). Supplementing MA increased yields of milk (39.9 vs. 41.3 vs. 41.5 kg/d), ECM (37.9 vs. 39.3 vs. 39.9 kg/d), fat (1.31 vs. 1.37 vs. 1.40 kg/d), TS (4.59 vs. 4.75 vs. 4.79 kg/d), and ECM per kg of DMI (1.72 vs. 1.76 vs. 1.80 kg/kg). Furthermore, cows fed MA increased yields of preformed fatty acids in milk fat (>16C; 435 vs. 463 vs. 488 g/d), particularly UFA (367 vs. 387 vs. 410 g/d), such as linoleic (C18:2 cis-9,cis-12; 30.9 vs. 33.5 vs. 35.4 g/d) and α-linolenic acids (C18:3 cis-9,cis-12,cis-15; 2.46 vs. 2.68 vs. 2.82 g/d) on d 60 and 62 in the experiment. Collectively, supplementing G1 and G2 improved productive performance of cows with no differences between the 2 MA., (The Authors. Published by Elsevier Inc. on behalf of the American Dairy Science Association®. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).)

Animals vary in the way in which they use energy due to diet, genetics, and management. Energy consumed by the animal supports milk production, but considerable variation among animals in energy utilization is thought to exist. The study objective was to estimate the among-animal variance in energy utilization in data collected from Jersey cows using indirect calorimetry. Individual animal-period data from 15 studies (n = 560) were used. The dataset included 115 animals from 44 to 410 DIM producing 11.5 to 39.1 kg/d of milk. On average, the 63 treatments in the dataset ranged 14.8% to 19.5% CP, 21.4% to 43.0% NDF, 16.2% to 33.3% starch, and 2.21% to 6.44% crude fat. Data were analyzed with the GLIMMIX procedure of SAS (9.4) with random effects of cow, treatment nested within period, square, and experiment. The percentage of among-animal, dietary treatment, and experimental variance was calculated as the variance associated with each fraction divided by the sum of variance from animal, dietary treatment, experiment, and residual, which was considered the total variance. The percentage of among-animal variance was characterized as high or low when the value was greater than or less than the mean value of 29.2%. Among-animal variance explained approximately 29.3% to 42.5% of the total variance in DMI, gross energy (GE), digestible energy (DE), ME, and NE L in megacalories per day. When energetic components of feces, urine, heat, and lactation in megacalories per day were expressed per unit of DMI, the among-animal variance decreased by 20.4, 4.82, 9.55, and 3.80 percentage units, respectively. However, among-animal variance explained 4.80 and 5.02 percentage units more of the total variation for methane energy and tissue energy in megacalories per day when expressed per unit of DMI. Variance in energetic efficiencies of DE/GE, ME/GE, and ME/DE were explained to a lesser extent by among-animal variance (averaging 17.8 ± 1.95%). The among-animal contribution to total variance in milk energy was 28.8%. Milk energy was a large proportion of the energy efficiency calculation, which included milk energy plus corrected tissue energy over net energy intake, which likely contributed to the 22.2% of total among-animal variance in energy efficiency. Results indicate that among-animal variance explains a large proportion of the total variation in DMI. This contributes to the variance observed for energy fractions as well as energy components when expressed in megacalories per day. Variation in energetic loss associated with methane was primarily explained by differences among animals and was increased when expressed per unit of DMI, highlighting the role of inherent animal differences in these losses., (The Authors. Published by Elsevier Inc. on behalf of the American Dairy Science Association®. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).)

Recent research has highlighted the importance of dietary fatty acid profile of fatty acid supplements on production responses of high-producing dairy cows. Conventional soybeans contain ∼15% oleic acid and ∼50% linoleic acid whereas high oleic acid soybeans (HOSB) contain ∼70% oleic acid and ∼7% linoleic acid. We determined the effect of increasing dietary inclusion of roasted and ground HOSB on production responses of high-producing dairy cows. Twenty-four multiparous Holstein cows (mean ± SD; 50.7 ± 4.45 kg/d of milk; 122 ± 57 DIM) were randomly assigned to treatment sequences in a replicated 4 × 4 Latin square design with 21-d periods. Treatments were increasing doses of HOSB at 0%, 8%, 16%, and 24% of DM. The HOSB replaced conventional soybean meal and hulls to maintain similar diet nutrient composition (% of DM) of 27.4% to 29.4% NDF, 20.6% forage NDF, 27.5% starch, and 15.9 to 16.5% CP. Total fatty acid content of treatments was 1.65%, 3.11%, 4.52%, and 5.97% of DM, respectively. Preplanned polynomial orthogonal contrasts included the linear, quadratic, and cubic effects of increasing HOSB. Increasing dietary inclusion of HOSB linearly decreased DMI and MUN and increased yields of milk, 3.5% FCM, ECM, and milk fat, and quadratically increased milk protein. The increased response to milk fat was due to an increase in preformed milk fatty acids. Due to the increase in milk component yields and decrease in DMI, there was an increase in feed efficiency. Increasing HOSB inclusion linearly decreased plasma BUN concentration and tended to decrease plasma insulin. Increasing HOSB had no effect on BW change or BCS change. In summary, increasing dietary inclusion of HOSB up to 24% of DM increased production responses of high-producing dairy cows and did not affect body reserves., (The Authors. Published by Elsevier Inc. on behalf of the American Dairy Science Association®. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).)

The objective of this study was to evaluate the effects of amino resin-treated soybean meal (SBM) on ruminal fermentation, nutrient digestion, and N partitioning. The treatments were (1) untreated solvent-extracted SBM, (2) amino resin-treated SBM (AR-SBM), and (3) heat-treated SBM (HT-SBM). The experimental design was arranged as a replicated 3 × 3 Latin square with 6 fermentors in a dual-flow continuous culture system. Treatments were randomly assigned to fermentors within a Latin square for each period. Each fermentor was fed 106 g/d of diet DM equally distributed in 2 feeding times daily at 0800 and 1800 h. Diets were formulated to contain 16% CP, 30% NDF, and 30% starch across treatments. The experiment consisted of 3 experimental periods, each lasting for 10 d. The first 7 d of each period were considered adaptation, and the last 3 d were used for sampling and data collection. On d 8 and 9, samples were collected for analysis of diurnal variation in concentrations of NH 3 -N, pH, and VFA during the first 8 h after feeding. On d 8, 9, and 10, samples were collected from the liquid and solid effluents accumulated over 24 h for analysis of daily averages of NH 3 -N and VFA pools, and true ruminal digestibility estimates. Data were analyzed using the MIXED procedure of SAS, and significance was declared when P ≤ 0.05. The model included the fixed effect of treatment and random effects of square, period, and fermentor within square, whereas time and interaction treatment × time were included for analyses of diurnal variation, with time as repeated measures. Compared with SBM, the cultured ruminal contents of AR-SBM and HT-SBM had lower NH 3 -N concentrations, indicating lower microbial fermentation of protein. Molar proportions of isovalerate and isobutyrate were greater in SBM than AR-SBM and HT-SBM, with greater molar proportion of isobutyrate for SBM, particularly during the first 2 h after feeding. The flow of NH 3 -N was greater for SBM compared with AR-SBM and HT-SBM, whereas NAN flow, bacterial N flow, and N efficiency were greater for AR-SBM and HT-SBM compared with SBM. Our results indicate that both the amino resin and heat treatments of SBM allow for similar decreases in microbial degradation of CP without limiting microbial protein synthesis in diets with 16% CP. Amino resin treatment may be effective in reducing microbial fermentation of protein in the rumen without adverse effects on digestibility or fermentation parameters as compared with SBM., (The Authors. Published by Elsevier Inc. on behalf of the American Dairy Science Association®. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).)

Dietary forage levels contribute to the performance of non-nutritive oral behaviors (NNOB) in cattle, yet the impact of varying forage levels on these behaviors is unknown. To evaluate the impact of dietary corn stalk inclusion (CSI) levels on NNOB, rumination time, and activity, pre-dominantly British-continental crossbred drylot-housed steers (n=27) were blocked by weight and randomly assigned to one of three dietary treatments (5%, 10%, or 15%) of CSI on a DM basis. Animals were fitted with a rumination collar upon arrival that measured rumination time and activity and video recorded. Cattle that spent more time bar licking had greater DMI, tended to have greater ADG and be more active. CSI in this study did influence NNOB performance; however, the impacts observed were not as expected. Cattle fed the 10% CSI performed the most bar licking and tongue rolling. This pilot investigation suggest that these CSI were insufficient to have a meaningful impact on NNOBs. Cattle spending more time bar licking and bar licked more frequently may be more orally motivated as reflected in their increased DMI and activity levels.

Nutrition and physiological state affect hepatic metabolism. Our objective was to determine whether feeding flaxseed oil (∼50% C18:3n-3 cis), high oleic soybean oil (∼70% C18:1 cis-9), or milk fat (∼50% C16:0) alters hepatic expression of PC, PCK1, and PCK2 and the flow of carbons from propionate and pyruvate into the tricarboxylic acid (TCA) cycle in preruminating calves. Male Holstein calves (n = 40) were assigned to a diet of skim milk with either: 3% milk fat (MF; n = 8), 3% flaxseed oil (Flax; n = 8), 3% high oleic soybean oil (HOSO; n = 8), 1.5% MF + 1.5% high oleic soybean oil (MF-HOSO; n = 8), or 1.5% MF + 1.5% flaxseed oil (MF-Flax; n = 8) from d 14 to d 21 postnatal. At d 21 postnatal, a liver biopsy was taken for gene expression and metabolic flux analysis. Liver explants were incubated in [U- 13 C] propionate and [U- 13 C] pyruvate to trace carbon flux through TCA cycle intermediates or with [U- 14 C] lactate, [1- 14 C] palmitic acid, or [2- 14 C] propionate to quantify substrate oxidation to CO 2 and acid-soluble products. Compared with other treatments, plasma C18:3n-3 cis was 10 times higher and C18:1 cis-9 was 3 times lower in both Flax (Flax and MF-Flax) treatments. PC, PCK1, and PCK2 expression and flux of [U- 13 C] pyruvate as well as [U- 13 C] propionate were not different among treatments. PC expression was negatively correlated with the enrichment of citrate M+5 and malate M+3, and PCK2 was negatively correlated with citrate M+5, suggesting that when expression of these enzymes is increased, carbon from pyruvate enters the TCA cycle via PC-mediated carboxylation, and then oxaloacetate is converted to phosphoenolpyruvate via PCK2. Acid-soluble product formation and PC expression were reduced in HOSO (MF-HOSO and HOSO) treatments compared with Flax (MF-Flax and Flax), indicating that fatty acids regulate PC expression and carbon flux, but that fatty acid flux control points are not connected to PC, PCK1, or PCK2. In conclusion, fatty acids regulate hepatic expression of PC, PCK1, and PCK2, and carbon flux, but the point of control is distinct., (The Authors. Published by Elsevier Inc. on behalf of the American Dairy Science Association®. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).)

Milk production and overall dairy farm economics depend on rearing dairy heifers. This study investigated the presence of a genotype by environment interaction in Holstein (HOL), Nordic Red dairy cattle (RDC), and their F 1 crossbreeds (HOL × RDC) when provided different feed rations. The aim of our study was to assess how different energy concentrations in feed rations affect growth, BCS, feed intake, and feed efficiency in the 3 groups during the prepubertal period. The 3 breed groups were randomly allocated to receive either a standard or a low-energy feed ration. Holstein heifers exhibited reduced growth and a lower BCS when they were fed the low-energy feed ration. In contrast, the RDC heifers demonstrated similar growth rates with the different feed rations and maintained similar BCS irrespective of feed energy concentration. The HOL × RDC crossbred heifers performed as an intermediate between the HOL and RDC groups. Significant differences were observed in DMI and energy intake in the HOL and HOL × RDC groups depending on feed ration treatment. The RDC heifers had similar feed intake irrespective of treatment. There were no significant differences in the feed conversion ratio among breeds and feed treatments. These results indicate the presence of a genotype by environment interaction in prepubertal HOL and RDC heifers in response to differences in feed ration treatment. Due to the influence of prepubertal growth on future milk production, reproduction, and health status, it is important to be aware of breed-specific requirements during the prepubertal period, particularly in mixed breed and crossbred groups, to optimize growth rates and production potential., (The Authors. Published by Elsevier Inc. on behalf of the American Dairy Science Association®. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).)

Schizochytrium sp., a feed additive, positively affects the quality of animal meat. In this study, the molecular mechanisms through which dietary Schizochytrium sp. affects the meat quality characteristics of Tan lambs were investigated using transcriptomic techniques. The findings demonstrate that the lambs supplemented with Schizochytrium sp. had a larger loin eye area and a higher average daily gain and intramuscular fat content (P < 0.05). They also had lower drip loss (at 24 and 48 h) and shear force (P < 0.05). Further, 745 genes were differentially expressed between lambs supplemented with Schizochytrium and the control group. Moreover, KEGG pathway analysis showed that the ECM-receptor interaction pathway, which is related to muscle generation and intramuscular fat deposition, was significantly enriched in the lambs administered a diet containing Schizochytrium sp. Herein, we identified some pivotal genes linked to muscular system development and lipid metabolism. Thus, using Schizochytrium sp. may boost the meat quality of Tan lambs by modifying the expression of genes related to hub pathways. The results supply a new basis to determine the molecular mechanisms through which Schizochytrium sp. supplementation regulates the meat quality characteristics of sheep., Competing Interests: Declaration of competing interest None., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Supplemental dietary rumen available fats show promise as enteric methane (eCH 4 ) mitigators for lactating dairy cows. However, concerns include variability in eCH 4 response and possible negative effects on dairy cow performance. Successful implementation of this mitigation option requires better prediction of responses specifically to rumen available fatty acids (FA) as well as understanding the modulating effects of other dietary and animal characteristics. Using meta-analytic and meta-regression techniques, 35 published studies with diet definition were used to assess changes in eCH 4 emissions and lactation performance associated with supplemental fat, specific supplemental rumen available FA types, and other dietary characteristics. Enteric CH 4 (g/d) was reduced by 3.77% per percentage unit of supplemental rumen available ether extract (EE). Supplemental rumen available PUFA (C18:2 and C18:3) and UFA (C18:1, C18:2, C18:3) mitigated eCH 4 (g/d) emissions in dairy cows by 6.88 and 4.65% per percentage unit increase, respectively. The anti-methanogenic effects of PUFA, MUFA and MCFA increased with correspondingly greater basal dietary levels of each FA type. Higher rumen-degradable starch (RDS; >18% DM) in the basal diet promoted greater reductions in eCH 4 yield (eCH 4 /DMI, g/kg) with supplemental rumen available PUFA and UFA. Both milk fat percentage and yield (kg/d) were reduced with rumen available fat supplementation with a reduction of 7.8% and 6.0%, respectively, relative to control diets. Our results highlight the importance of determining basal levels of the rumen available FA before providing supplemental rumen available FA as an option for enteric eCH 4 mitigation. Dairy nutritionists can use estimates generated from this analysis to predict changes in eCH 4 emissions and dairy cow performance associated with dietary supplementation of rumen available EE and specific rumen available FA types for the purpose of eCH 4 mitigation., (The Authors. Published by Elsevier Inc. on behalf of the American Dairy Science Association®. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).)

Increasing nutrient supply to dairy calves has well known benefits; however, the effects of milk replacer (MR) composition when supplied in higher amounts are not fully understood, particularly in the first weeks of life. To better understand the metabolism of macronutrient supply in young calves (21 d old), we investigated diurnal patterns of heat production and substrate oxidation in young calves fed MR with an incremental supply of fat, lactose, or protein. Thirty-two groups of 3 mixed-sex Holstein-Friesian newborn calves (3.4 ± 1.6 d of age), were randomly assigned to one of 4 dietary treatments and studied for 21 d. Diets consisted of a basal MR (23.3% CP, 21.2% EE, and 48.8% lactose of DM) fed at 550 kJ/kg BW 0.85 per day (CON; n = 24), or the basal MR incrementally supplied with 126 kJ of digestible energy/BW 0.85 per day as milk fat (+FAT; n = 23), lactose (+LAC; n = 24), or milk protein (+PRO; n = 23). Calves were fed MR in 2 daily meals and had ad libitum access to water, but were not supplied with any calf starter nor forage. After 2 weeks of adaptation to the diets, groups of 3 calves were placed for 1 wk in an open-circuit respiration chamber for nitrogen and energy balance measurements (lasting 7 d). On d 3, glucose oxidation kinetics was estimated by using [U- 13 C]glucose. Measurements included total heat production (total energy [HP], activity [H act ] expenditure, resting metabolic rate [RMR]), respiration quotient (RQ), carbohydrate (COX) and fat oxidation (FOX) in 10 min. intervals and averaging these values per hour over days. Incremental supply of lactose and fat increased body fat deposition, with observed patterns in RMR indicating that this increase occurred primarily after the meals. Specifically, the average daily RMR was highest in the +PRO group and lowest in the CON treatment. The HP was higher in the +PRO group and throughout the day, hourly means of HP were higher in this treatment mainly caused by an increase in H act . The recovery of 13 CO 2 from oral pulse-dosed [U- 13 C]glucose was high (77%), and not significantly different between treatments, indicating that ingested lactose was oxidized to a similar extent across treatments. Increasing lactose supply in young calves increased fat retention by reduction in fatty oxidation. Calves fed a MR with additional protein or fat raised RMR persistently throughout the day, whereas the extra lactose supply only affects RMR after the meal. Dietary glucose was almost completely oxidized (77% based on ( 13 C) glucose measurement) regardless of nutrient supplementation. Extra protein supply increased HP and FOX compared with similar intakes of fat and lactose. Fasting heat production (FHP) of young, group-housed calves is comparable to literature values and unaffected by energy intake. Overall, these findings deepen our understanding of how different nutrients affect metabolic processes, fat retention, and energy expenditure in young dairy calves., (The Authors. Published by Elsevier Inc. on behalf of the American Dairy Science Association®. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).)

We investigated the short- and long-term effects of different forage types supplemented in preweaning dairy calves on growth performance, blood metabolites, rumen fermentation, bacterial community, and milk production during first lactation. A total of 60 healthy 30-d-old female Holstein calves were blocked by birth date and body weight and randomly assigned to 1 of 3 groups (n = 20): normal milk and pelleted starter feeding (CON), supplemented with chopped oat hay (OAH; 75.0 g/d per calf [DM basis]), or alfalfa hay (ALF; 75.0 g/d per calf [DM basis]). The forage supplementation started when calves were 30 d old (d 1 of the experimental period) and ended when they were 73 d old (d 44 of the experimental period, when calves were weaned). Milk and feed intakes and fecal consistency scores were recorded daily. Growth performance, rumen fluid, and blood samples were collected biweekly. After weaning, all the calves were integrated with the same barn and diets. After calving, the milk production was recorded daily. During the experimental period, the OAH group had greater solid feed and total DM intakes and greater rumen pH than the CON group (P ≤ 0.04), but had lower forage intake and CP digestibility than the ALF group (P ≤ 0.04). The ALF group had higher rumen pH and blood BHB concentration (P ≤ 0.04), lower fecal score (P = 0.02), and greater ether extract digestibility (P = 0.02) than the CON group. The ALF and OAH groups had lower concentrations of ruminal total VFA (P = 0.01). Still, the ALF group had a greater proportion of acetate and a relative abundance of cellulose degradation-related bacteria (Lachnoclostridium&#95;1 and Oribacterium) and a lower relative abundance of inflammation-related bacteria (Erysipelotrichaceae&#95;UCG-009) in the rumen compared with CON. Interestingly, the average milk production from 6 to 200 DIM was greater in the ALF group (P < 0.01), even though no significant effects were found on the rumen fermentation parameters and blood metabolites at 200 DIM. Generally, alfalfa hay supplementation in preweaning dairy calves had positive effects in the short- and long-term for rumen development, health status, and future milk production., (The Authors. Published by Elsevier Inc. on behalf of the American Dairy Science Association®. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).)

The form of fat supplements, degree of saturation, and the fatty acid (FA) profile influence cows' production response. The objective was to determine the effects of supplemental fats in the form of calcium salts (CS) of FA (CSFA) with different ratios between palmitic (PA) and oleic (OA) acids on nutrient digestibility and cow performance. Forty-two dairy cows were assigned to 3 groups and fed (for 13 wk) rations that contained 2.2% CSFA (on a DM basis) with increasing PA-to-OA ratios as follows: (1) CS45:35, 45% PA and 35% OA; (2) CS60:30, 60% PA and 30% OA; and (3) CS70:20, 70% PA and 20% OA. Rumen and fecal samples were taken for VFA and digestibility measurements, respectively. Increasing the PA-to-OA ratio linearly decreased the milk and ECM yields, whereas a quadratic effect was observed for milk fat concentration (3.55%, 3.94%, and 3.87% in the CS45:35, CS60:30, and CS70:20 groups, respectively) and fat yield. Dry matter intake was highest in the CS60:30 group (33.7 kg/d) and lowest in the CS70:20 group (31.6 kg/d), and a tendency of quadratic effect was observed for calculated energy balance with no difference in BW gain among the groups. The milk-to-DMI ratio was decreased, and the ratio of ECM-to-DMI tended to decrease when the PA-to-OA ratio increased. The highest apparent total-tract digestibilities of DM, OM, and protein were observed in the CS60:30 cows, and NDF tended to decrease with increasing PA-to-OA ratio; however, digestibility of the total FA and FA subgroups (16- and 18-carbon FA) were not different among groups. Across treatments, the 18-carbon FA digestibility was higher than the 16-carbon FA digestibility. Under the current study conditions, the CS60:30 cows had the highest fat concentration and fat yield; energy output in milk; DMI; and digestibility of DM, OM, and protein. However, further research is required to fine tune the optimal FA ratio in supplemental fat sources to maximize production and efficiency under various conditions, such as production level, stage of lactation, and diet composition., (The Authors. Published by Elsevier Inc. on behalf of the American Dairy Science Association®. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).)

Adding milk replacer powder (MRP) to whole milk during the entire preweaning period can increase growth but raises concern because of low starter feed intake and slumps in ADG at weaning and postweaning. In the current study, effects of adding MRP to pasteurized whole milk (PWM) during d 10 to 41 or d 10 to 59 of age were investigated in comparison with PWM. Calves (24 females and 21 males; 39.8 ± 1.85 kg BW) were randomly allocated to 1 of 3 treatments: (1) conventional protocol, 5 L/d PWM from d 3 to 56, and 2.5 L/d from d 57 to 59 of age (CONV; TS intake = 31.9 kg), (2) short duration of adding MRP to PWM protocol, 5 L/d PWM from d 3 to 9, 5 L/d PWM + MRP (18% TS) from d 10 to 41, 5 L/d PWM from d 42 to 56, and 2.5 L/d PWM from d 57 to 59 (SHD; TS intake = 42.3 kg), (3) long duration of adding MRP to PWM protocol, 5 L/d PWM from d 3-9, 5 L/d PWM + MRP from d 10-56, 2.5 L/d PWM + MRP from d 57-59 (LD; TS intake = 47.7 kg). The osmolality of PWM and PWM + MRP was 278 and 519 mOsm/L, respectively. Calves were weaned on d 60, and the study terminated on d 75. There was a treatment × time interaction for starter intake, where intake was greater for CONV than other treatments from d 14-41 and was greater in CONV than LD during d 42 to 48 and d 56 to 62 of age. Final BW was lower in CONV calves than LD calves. Weaning BW and overall hip height were lower in CONV calves than other treatments. The CONV calves had lower ADG at d 14 to 27 and d 35 to 41, and SHD calves had lower ADG at d 42 to 48 than other treatments. Calves fed CONV treatment had lower ruminal acetate and greater propionate than SHD calves during preweaning. Calves fed LD treatment had lower total VFA and tended to have greater ruminal pH than other treatments. Calves fed CONV had greater neutrophils and neutrophils/lymphocytes ratio and lower lymphocytes than other treatments. Glucose concentration was greater for LD versus other treatments at d 56, and lower for SHD versus other treatments at d 70 of study. Insulin concentration and homeostatic model assessment of insulin resistance index were greater in LD compared with other treatments during preweaning but were not different postweaning. Serum BHB was greater in CONV than other treatments. Albumin was greater for CONV versus other treatments at d 56, however, it was greater in LD-fed calves at d 70 of study. Results indicate that feeding a PWM + MRP to the calves during the entire preweaning period resulted in lower starter feed intake around weaning, but overall starter intake was similar with a greater final BW and fewer health-related issues throughout the study. Shifting a PWM + MRP to the conventional whole milk at d 40 of age decreased the ADG of calves., (The Authors. Published by Elsevier Inc. on behalf of the American Dairy Science Association®. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).)

Automated measurements of the ratio of concentrations of methane and carbon dioxide, [CH 4 ]:[CO 2 ], in breath from individual animals (the so-called "sniffer technique") and estimated CO 2 production can be used to estimate CH 4 production, provided that CO 2 production can be reliably calculated. This would allow CH 4 production from individual cows to be estimated in large cohorts of cows, whereby ranking of cows according to their CH 4 production might become possible and their values could be used for breeding of low CH 4 -emitting animals. Estimates of CO 2 production are typically based on predictions of heat production, which can be calculated from body weight (BW), energy-corrected milk yield, and days of pregnancy. The objectives of the present study were to develop predictions of CO 2 production directly from milk production, dietary, and animal variables, and furthermore to develop different models to be used for different scenarios, depending on available data. An international dataset with 2,244 records from individual lactating cows including CO 2 production and associated traits, as dry matter intake (DMI), diet composition, BW, milk production and composition, days in milk, and days pregnant, was compiled to constitute the training dataset. Research location and experiment nested within research location were included as random intercepts. The method of CO 2 production measurement (respiration chamber [RC] or GreenFeed [GF]) was confounded with research location, and therefore excluded from the model. In total, 3 models were developed based on the current training dataset: model 1 ("best model"), where all significant traits were included; model 2 ("on-farm model"), where DMI was excluded; and model 3 ("reduced on-farm model"), where both DMI and BW were excluded. Evaluation on test dat sets with either RC data (n = 103), GF data without additives (n = 478), or GF data only including observations where nitrate, 3-nitrooxypropanol (3-NOP), or a combination of nitrate and 3-NOP were fed to the cows (GF+: n = 295), showed good precision of the 3 models, illustrated by low slope bias both in absolute values (-0.22 to 0.097) and in percentage (0.049 to 4.89) of mean square error (MSE). However, the mean bias (MB) indicated systematic overprediction and underprediction of CO 2 production when the models were evaluated on the GF and the RC test datasets, respectively. To address this bias, the 3 models were evaluated on a modified test dataset, where the CO 2 production (g/d) was adjusted by subtracting (where measurements were obtained by RC) or adding absolute MB (where measurements were obtained by GF) from evaluation of the specific model on RC, GF, and GF+ test datasets. With this modification, the absolute values of MB and MB as percentage of MSE became negligible. In conclusion, the 3 models were precise in predicting CO 2 production from lactating dairy cows., (The Authors. Published by Elsevier Inc. on behalf of the American Dairy Science Association®. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).)

The AA profile of MP affects mammary gland metabolism and milk N efficiency of dairy cattle. Further, the frequency of dietary protein supplementation may influence N partitioning leading to reduced N excretion. This study investigated the effect of source and frequency of rumen-protected (RP) protein supplementation on apparent total-tract digestibility, milk production, mammary gland AA metabolism, and N balance of dairy cattle. Twenty-eight Holstein-Friesian cows (2.3 ± 0.9 lactations; 93 ± 27 DIM; mean ± SD) were used in a randomized complete block design and fed a basal TMR consisting of 41% corn silage, 32% grass silage, and 27% concentrate (DM basis) and formulated to meet 100% and 95% of net energy and MP requirements, respectively. Cows were adapted to the basal TMR in a freestall barn for 7 d, moved to individual tiestalls for 13 d of adaptation to dietary treatments, and then moved into climate respiration chambers for a 4-d measurement period. Treatments consisted of the basal TMR (CON; 159 g CP/kg DM) or the basal TMR including 1 of 3 iso-MP supplements: (1) 315-g mixture of RP soybean meal and RP rapeseed meal fed daily (ST-RPSR), (2) 384-g mixture of RP His, RP Lys, and RP Met fed daily (ST-RPAA), and (3) 768-g mixture of RP His, RP Lys, and RP Met fed every other day (OS-RPAA). The basal TMR with the addition of treatment supplements was designed to deliver 100% of required MP over a 48-h period. The mixture of His, Lys, and Met was formulated to deliver digestible AA in amounts relative to their concentration in casein. Compared with ST-RPSR, ST-RPAA increased milk protein and fat concentration, increased the arterial concentration of total His, Lys, and Met (HLM), decreased mammary clearance of HLM, and increased clearance of Phe, Leu, and Tyr (tendency for Leu and Tyr). Rumen-protected protein source did not affect N balance, but the marginal use efficiency (efficiency of transfer of RP protein supplement into milk protein) of ST-RPAA (67%) was higher than that of ST-RPSR (17%). Milk protein concentration decreased with OS-RPAA compared with ST-RPAA. Arterial concentration of HLM increased on the nonsupplemented day compared with the supplemented day with OS-RPAA, and there was no difference in arterial HLM concentration across days with ST-RPAA. Mammary uptake of HLM tended to increase on the nonsupplemented day compared with the supplemented day with OS-RPAA. Supplementation frequency of RP AA did not affect N balance or overall milk N efficiency, but the marginal use efficiency of OS-RPAA (49%) was lower compared with ST-RPAA. Overall, mammary glands responded to an increased supply of His, Lys, and Met by reducing efflux of other EAA when RP His, RP Lys, and RP Met were supplemented compared with RP plant proteins. Mammary glands increased sequestration of EAA (primarily HLM) on the nonsupplemented day with OS-RPAA, but supplementing RP AA according to a 24-h oscillating pattern did not increase N efficiency over static supplementation., (The Authors. Published by Elsevier Inc. on behalf of the American Dairy Science Association®. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).)

The possibility of inclusion of agro-industrial byproducts in the diet of small ruminants represents both an economic and an environmental strategy for reducing waste management by industries and costs of feeding as well as the impact of livestock farming. Large amounts of wastes from the cocoa industry are produced annually, with a considerable part represented by cocoa bean shells, considered a suitable ingredient to be included in the diet of ruminants within the limits established by European legislation. The aim of this study was to assess the effect of including cocoa bean shells in the diet of dairy sheep on the sensory, volatile, and antioxidant properties of cheese. To this purpose, 20 lactating Comisana ewes were randomly assigned to 2 experimental groups: control (CTRL) and cocoa bean shells (CBS), and received alfalfa hay ad libitum and 800 g of conventional (CTRL) or experimental (CBS) concentrate containing 11.7% CBS to partially replace corn and barley of the CTRL concentrate. Bulk milk collected from each group was used to produce a total of 15 cheeses per group, obtained in 5 different days of cheesemaking (3 cheeses a day per group). After 60 d of aging, each cheese of each experimental group was sampled for the analyses. The results on chemical composition revealed a greater content of monounsaturated fatty acids and an increase in the nutritional indices, suggesting a favorable role of cocoa bean shell dietary inclusion on the nutritive value of cheese. The cheese sensory profile was affected by the cocoa bean shell inclusion, with more pronounced appearance, odor, aroma, and taste attributes in the product. The volatile profile showed only a few significant differences, mainly related to the cheese ripening process, and no differences were found in α-tocopherol contents in cheese fat between the 2 groups. Therefore, the inclusion of coca bean shells in the diet of dairy sheep allowed us to obtain a good-quality cheese, without altering the characteristics associated with the typical profiles of sheep cheese. Furthermore, the use of this byproduct could contribute to decreasing feed costs and waste management, representing a good practice for increasing the sustainability of dairy products., (The Authors. Published by Elsevier Inc. on behalf of the American Dairy Science Association®. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).)

The objective of this study was to determine the potential effect and interaction of 3-nitrooxypropanol (3-NOP; Bovaer, DSM-Firmenich Nutrition Products Ltd.) and whole cottonseed (WCS) on lactational performance and enteric methane (CH 4 ) emission of dairy cows. A total of 16 multiparous cows, including 8 Holstein Friesian (HF) and 8 Brown Swiss (BS; 224 ± 36 DIM, 26 ± 3.7 kg milk yield, mean ± SD), were used in a split-plot design, where the main plot was the breed of cows. Within each subplot, cows were randomly assigned to a treatment sequence in a replicated 4 × 4 Latin square design with 2 × 2 factorial arrangements of treatments with four 24-d periods. The experimental treatments were as follows: (1) control (basal TMR), (2) 3-NOP (60 mg/kg TMR DM), (3) WCS (5% TMR DM), and (4) 3-NOP + WCS. The treatment diets were balanced for ether extract, crude protein, and NDF contents (4%, 16%, and 43% of TMR DM, respectively). The basal diets were fed twice daily at 0800 and 1800 h. Dry matter intake and milk yield were measured daily, and enteric gas emissions were measured (using the GreenFeed System, C-Lock Inc.) during the last 3 d of each 24-d experimental period when animals were housed in tiestalls. There was no difference in DMI on treatment level, whereas the WCS treatment increased ECM yield and milk fat yield. No interaction of 3-NOP and WCS occurred for any of the enteric gas emission parameters, but 3-NOP decreased CH 4 production (g/d), CH 4 yield (g/kg DMI), and CH 4 intensity (g/kg ECM) by 13%, 14%, and 13%, respectively. Further, an unexpected interaction of breed by 3-NOP was observed for different enteric CH 4 emission metrics: HF cows had a greater CH 4 mitigation effect compared with BS cows for CH 4 production (g/d; 18% vs. 8%), CH 4 intensity (g/kg milk yield; 19% vs. 3%), and CH 4 intensity (g/kg ECM; 19% vs. 4%). Hydrogen production was increased by 2.85-fold in HF and 1.53-fold in BS cows receiving 3-NOP. Further, a 3-NOP × time interaction occurred for both breeds. In BS cows, 3-NOP tended to reduce CH 4 production by 18% at approximately 4 h after morning feeding, but no effect was observed at other time points. In HF cows, the greatest mitigation effect of 3-NOP (29.6%) was observed immediately after morning feeding, and it persisted at around 23% to 26% for 10 h until the second feed provision, and 3 h thereafter, in the evening. In conclusion, supplementing 3-NOP at 60 mg/kg DM to a high-fiber diet resulted in 18% to 19% reduction in enteric CH 4 emission in Swiss HF cows. The lower response to 3-NOP by BS cows was unexpected and has not been observed in other studies. These results should be interpreted with caution due to the low number of cows per breed. Finally, supplementing WCS at 5% of DM improved ECM and milk fat yield but did not enhance the CH 4 inhibition effect of 3-NOP of dairy cows., (The Authors. Published by Elsevier Inc. on behalf of the American Dairy Science Association®. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).)

This study investigates Cystobasidium benthicum (Cb) probiotic yeast and Cyrtocarpa edulis (Ce) fruit dietary effects, single (0.5 %) or combined (Cb:Ce, 0.25:0.25 %), on growth performance, humoral immunity in serum and skin mucus, and intestinal morphology of Nile tilapia (Oreochromis niloticus) after 14 and 28 days. The Cb group presented the highest (P < 0.05) specific growth rate, weight gain, and absolute growth rate with respect to the control group. Immunological assays indicated that Cb, Ce and Cb:Ce groups increased serum nitric oxide concentration compared to the control group (P < 0.05). Cb and Cb:Ce groups showed the highest serum myeloperoxidase enzyme activity at day 14 and 28, respectively (P < 0.05); whereas, Cb:Ce group had the highest (P < 0.05) myeloperoxidase activity in skin mucus. The superoxide dismutase enzyme activity was unaffected. On day 28, Cb, Ce, and Cb:Ce groups showed higher and lower (P < 0.05) catalase enzyme activity in serum and skin mucus, respectively, compared with the control group. Only the Cb group had higher (P < 0.05) total protein concentration in serum (day 14) and skin mucus (day 14 and 28) with respect to the control group. The lysozyme activity in serum (day 28) and skin mucus (day 14) was higher (P < 0.05) in the Cb group compared to the control group. Only the skin mucus of Ce group showed bactericidal activity against Aeromonas dhakensis (P < 0.05). Histological studies indicated that Cb and Cb:Ce groups increased microvilli height, and Cb, Ce and Cb:Ce augmented goblet cell area at day 14 compared to the control group (P < 0.05). At day 28, microvilli height was higher in all groups and the number of intraepithelial leukocytes increased in Cb and Ce groups with respect to the control group (P < 0.05). The ex vivo assay revealed that A. dhakensis in leukocytes decreased cell viability similar to the control group (P < 0.05). A principal component analysis (PCA) confirmed the results. In conclusion, C. benthicum in the diet was the best supplement to improve the growth and immunity of Nile tilapia., Competing Interests: Declaration of competing interest The authors declare no conflict of interest., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

An experiment was conducted to identify the factors that cause reduced production of cows fed a diet with high content of corn distillers grains with solubles (DDGS). We hypothesized that the factors could be high sulfur (S) content in DDGS, which may directly (S toxicity) or indirectly (DCAD) cause reduced production. We also hypothesized that high PUFA in DDGS could be another major factor. In a randomized complete block design, 60 lactating cows (15 primiparous and 45 multiparous; average ± SD at the beginning of the trial: milk yield, 44.0 ± 6.9 kg/d; DIM, 123 ± 50; BW, 672 ± 82 kg) were blocked, and cows in each block were randomly assigned to 1 of the following treatments: SBM (4.7% fatty acids [FA], 0.22% S, and 178 mEq/kg DM of DCAD), a diet containing soybean meal as the main protein source; DG, with SBM replacing mainly soybean byproducts and supplemental fat with distillers grains at 30% dietary DM (4.7% FA, 0.44% S, and 42 mEq/kg DM of DCAD); SBM+S, SBM with sodium bisulfate for additional dietary S (4.8% FA, 0.37% S, and 198 mEq/kg DM of DCAD); SBM+CO, SBM with corn oil (4.7% FA, 0.23%, and 165 mEq/kg DM of DCAD); and DG+DCAD, DG with increased DCAD (4.7% FA, 0.40% S, and 330 mEq/kg DM of DCAD). Due to the limited number of tiestalls, blocks 1 to 6 started the experiment first as phase 1, and the rest of the blocks, as phase 2, started the experiment after phase 1. All cows were fed the SBM diet for 10 d as a covariate period followed by the experimental period for 35 d. Data were analyzed using PROC MIXED of SAS (Version 9.4, SAS Institute Inc.); block and phase were random effects; and treatments, repeated week, and interaction were fixed effects. We found an interaction of week by treatment for DMI. Although milk yield did not change, milk fat concentration tended to decrease (2.78% vs. 3.34%) for DG compared with SBM. Dry matter, OM, NDF, and CP digestibilities were lower when cows were fed the DG diet compared with SBM. Additionally, cows fed DG had lower blood concentrations of HCO 3 - compared with SBM. The SBM+S diet did not affect production, nutrient digestibility, or blood parameters compared with SBM. The SBM+CO diet decreased milk fat concentration and yield compared with SBM. The DG+DCAD diet tended to increase milk fat yield and concentration (1.24 vs. 1.47 kg/d; 2.78% vs. 3.37%) and increased ECM (40.9 vs. 45.1 kg/d) compared with DG but did not improve nutrient digestibility. However, blood HCO 2 compared with SBM. The SBM+S diet did not affect production, nutrient digestibility, or blood parameters compared with SBM. The SBM+CO diet decreased milk fat concentration and yield compared with SBM. The DG+DCAD diet tended to increase milk fat yield and concentration (1.24 vs. 1.47 kg/d; 2.78% vs. 3.37%) and increased ECM (40.9 vs. 45.1 kg/d) compared with DG but did not improve nutrient digestibility. However, blood HCO 3 - , altering DCAD, along with the high PUFA content in DDGS, appear to be the factors causing reduced production responses to a high DDGS diet. Increasing DCAD to 300 mEq/kg DM in a high DDGS diet can be a feeding strategy to alleviate reduced production responses.2 were greater for DG+DCAD compared with DG. In conclusion, the indirect role of S - , altering DCAD, along with the high PUFA content in DDGS, appear to be the factors causing reduced production responses to a high DDGS diet. Increasing DCAD to 300 mEq/kg DM in a high DDGS diet can be a feeding strategy to alleviate reduced production responses., (The Authors. Published by Elsevier Inc. on behalf of the American Dairy Science Association®. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).)

This study aimed to investigate the impact of feeding patterns on the production performance, lipo-nutritional quality, and gut microbiota of Sunit sheep. A total of 24 sheep were assigned to two groups: confinement feeding (CF) and pasture feeding (PF) groups. After 90 days, the CF group exhibited significantly increased average daily gain, carcass weight, backfat thickness, and intramuscular fat content of the sheep, whereas the PF group showed significantly increased pH 24h and decreased L ∗ value and cooking loss of the longissimus lumborum (LL) muscle (P < 0.05). In the PF group, the contents of linoleic, α-linolenic, and docosahexaenoic acids were considerably higher and the n-6/n-3 polyunsaturated fatty acid ratio was significantly lower (P < 0.05). Furthermore, the triglyceride, cholesterol, and nonesterified fatty acid levels in the serum of the CF group significantly increased, whereas the enzyme contents of fatty acid synthase (FASN) and hormone-sensitive lipase (HSL) in the LL muscle of the PF group were markedly elevated (P < 0.05). The PF group also showed altered expression of lipid metabolism-related genes, including upregulated FASN, HSL, fatty acid binding protein 4 (FABP4), and peroxisome proliferator-activated receptor γ coactivator 1α (PGC-1α) (P < 0.05). Meanwhile, differences were observed in the abundance of key bacteria and microbiota functions between the groups. Correlation analysis revealed that production performance and lipid metabolism may be related to the differential effects of bacteria. In conclusion, the transition in the feeding patterns of Sunit sheep caused changes in the gut microbial community and lipid metabolism level in the muscle as well as differences in fat deposition and meat quality., Competing Interests: Declaration of competing interest The authors declare that there is no conflict of interest in the study., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

This study aimed to evaluate the effects of direct-fed microbials (DFM) on health and growth responses of preweaning Bos indicus × Bos taurus (Gyr × Holstein) crossbred calves. Ninety newborn heifer calves (initial BW of 35 ± 4.0 kg) were used. At birth, calves were ranked by initial BW and parity of the dam and assigned to: (1) whole milk without DFM supplementation (CON; n = 30), (2) whole milk with the addition of 1.0 g/calf per day of a Bacillus-based DFM (BAC; n = 30), or (3) whole milk with the addition of 1.0 g/calf per day of BAC and 1.2 g/calf per day of Enterococcus faecium 669 (MIX; n = 30). Milk was fed individually during the study (77 d), and the BAC and MIX treatments were offered daily throughout the 77-d preweaning period. All calves were offered a starter supplement and corn silage starting on d 1 and 60 of age, respectively. Milk and starter supplement intake were evaluated daily, and BW was recorded on d 0 and at weaning (d 77). Diarrhea and pneumonia were assessed daily, and fecal samples were collected on d 0, 7, 14, 21, and at weaning (d 77) for assessment of the presence of bacterial and protozoal pathogens via qPCR. All data were analyzed using SAS (v. 9.4) with calf as the experimental unit and using single-df orthogonal contrasts (BAC + MIX vs. CON; BAC vs. MIX). Daily feeding of DFM, regardless of type, improved weaning BW. Odds ratio for occurrence of pneumonia was lower for DFM-supplemented calves, but its occurrence did not differ between BAC and MIX calves. No Salmonella spp. or Escherichia coli F41 were detected in any of the calves. The proportion of calves positive for E. coli F17 was greater for DFM calves on d 7 (92% and 96% vs. 81% for BAC, MIX, and CON, respectively), on d 21 (13% and 26% vs. 7% for BAC, MIX, and CON, respectively), and at weaning (48% and 35% vs. 22% for BAC, MIX, and CON, respectively). For Clostridium difficile, more DFM calves were positive on d 7 (65% and 30% vs. 35% for BAC, MIX, and CON, respectively) and 14 (20% and 28% vs. 7% for BAC, MIX, and CON, respectively), but proportion of positive calves was also greater for BAC versus MIX on d 7. More CON calves were positive for Clostridium perfringens on d 14 (14% vs. 3% and 8% for CON, BAC, and MIX, respectively) compared with DFM-fed calves. Incidence of calves positive for C. perfringens was greater in BAC than MIX on d 7 (50% vs. 18%), and greater for MIX than BAC at weaning (9% vs. 0%). For protozoa occurrence, a lower proportion of DFM calves were positive for Cryptosporidium spp. on d 7 (58% and 48% vs. 76% for BAC, MIX, and CON, respectively), but opposite results were observed on d 21 for Cryptosporidium spp. (3% and 11% vs. 0% for BAC, MIX, and CON, respectively) and Eimeria spp. on d 14 (7% and 8% vs. 0% for BAC, MIX, and CON, respectively) and 21 (50% and 59% vs. 38% for BAC, MIX, and CON, respectively). In summary, DFM feeding alleviated the occurrence of pneumonia and improved growth rates, while also modulating the prevalence of bacteria and protozoa in preweaning Gyr × Holstein calves., (© 2024, The Authors. Published by Elsevier Inc. on behalf of the American Dairy Science Association®. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).)

Recently reviewed development objectives and feeding practices in young dairy calves require an adaptation of nutrient recommendations set for milk replacer (MR) composition. Nutrient requirements of calves younger than 21 d of age, and those of calves fed with high levels of MR are insufficiently quantified. The efficiency at which macronutrients are utilized, particularly protein, substantially diminishes with age, and little data exists for the first weeks of life. In addition, in older preruminant and ruminant calves, protein and energy can be simultaneously limiting for protein gain. Whether this also applies to calves in the first weeks of life is unknown. Therefore, this study aimed to quantify the responses in protein and fat gain to incremental supply of protein, fat, or lactose to MR in very young calves. Thirty-two groups of 3 mixed-sex Holstein-Friesian newborn calves (3.4 ± 1.6 d of age), were randomly assigned to 1 of 4 dietary treatments applied for 19 d: a basal MR (23.3% crude protein, 21.2% crude fat, and 48.8% lactose, percentages of dry matter), provided at 550 kJ/kg of metabolic body weight (BW 0.85 ) per day (CON; n = 24), or the basal MR incrementally supplied with 126 kJ of digestible energy per BW 0.85 per day as milk fat (+FAT; n = 23), lactose (+LAC; n = 24), or milk protein (+PRO; n = 23). Calves were fed MR in 2 daily meals and had ad libitum access to water, but did not have access to calf starter nor any other solid feed. After 2 wk of adaptation to their respective diets, groups of calves were placed for 1 wk in an open-circuit respiration chamber for nitrogen and energy balance measurements (5 d). The incremental nutrient efficiencies indicate what percentage of extra intake of nutrients is retained. In this study, we observed that with every 100-g increase in protein intake, 52% was converted into protein deposition, and 44% contributed to heat production. Similarly, a 100-g increase in fat intake resulted in 67% being stored as fat, 22% being released as heat, and only 5% being retained as protein. Likewise, a 100-g increment in lactose intake led to 49% being stored as fat, with 38% being released as heat. Additional protein intake was not deposited as fat; extra energy intake (fat and additional lactose) increased postabsorptive N efficiency in young calves., (© 2024, The Authors. Published by Elsevier Inc. on behalf of the American Dairy Science Association®. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).)

There is an increasing consumer desire for pasture-derived dairy products, as outdoor pasture-based feeding systems are perceived as a natural environment for animals. Despite this, the number of grazing animals globally has declined as a result of the higher milk yields achieved by indoor TMR feeding systems, in addition to the changing climatic conditions and lower grazing knowledge and infrastructure. This has led to the development of pasture-fed standards, stipulating the necessity of pasture and its minimum requirements as the primary feed source for products advertising such claims, with various requirements depending on the region for which it was produced. This work investigates the differences in the composition and techno-functional properties of butters produced from high, medium and no pasture allowance diets during early, mid, and late lactation. Butters were produced using milks collected from 3 feeding systems: outdoor pasture grazing (high pasture allowance); indoor TMR (no pasture allowance); and a partial mixed ration (medium pasture allowance) system, which involved outdoor pasture grazing during the day and indoor TMR feeding at night. Butters were manufactured during early, mid, and late lactation. Creams derived from TMR feeding systems exhibited the highest milk fat globule size. The fatty acid profiles of butters also differed significantly as a function of diet and could be readily discriminated by partial least squares analysis. The most important fatty acids in such an analysis, as indicated by their highest variable importance projection scores, were CLA C18:2 cis-9,trans-11 (rumenic acid), C16:1n-7 trans (trans-palmitoleic acid), C18:1 trans (elaidic acid), C18:3n-3 (α-linolenic acid), and C18:2n-6 (linoleic acid). Increasing pasture allowances resulted in reduced crystallization temperatures and hardness of butters and concurrently increasing the "yellow" color. Yellow color was strongly correlated with Raman peaks commonly associated with carotenoids. The milk fat globule size of cream decreased with advancing stage of lactation and churning time of cream was lowest in early lactation. Differences in the fatty acid and triglyceride contents of butter as a result of lactation and dietary effects demonstrated significant correlations with the hardness, rheological, melting, and crystallization profiles of the butters. This work highlighted the improved nutritional profile and functional properties of butter with increasing dietary pasture allowance, primarily as a result of increasing proportions of unsaturated fatty acids. Biomarkers of pasture feeding (response in milk proportionate to the pasture allowance) associated with the pasture-fed status of butters were also identified as a result of the significant changes in the fatty acid profile with increasing pasture allowance. This was achieved through the use of 3 authentic feeding systems with varying pasture allowances, commonly operated by farmers around the world and conducted across 3 stages of lactation., (© 2024, The Authors. Published by Elsevier Inc. on behalf of the American Dairy Science Association®. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).)

Lignin is a polyphenolic polymer that is an important factor in limiting fiber digestibility by ruminants. The objective of the current study was to evaluate lignin's effects on whole animal energy utilization in diets similar in NDF content. A low-lignin (LoLig) diet was formulated to contain 32.5% NDF (DM basis) and 9.59% lignin (NDF basis) and the high-lignin (HiLig) diet was formulated to contain 31.0% NDF (DM basis) and 13.3% lignin (NDF basis). These diets were randomly assigned and fed to 12 late-lactation (mean ± SD; 214 ± 14.9 DIM) multiparous Jersey cows (mean ± SD; 435 ± 13.9 kg) in a 2-period crossover design. Cows fed the LoLig treatment consumed more DM than cows on the HiLig diet (mean ± SD; 19.9 vs. 18.7 ± 0.645 kg/d) and the LoLig diet was concurrently of a greater gross energy concentration (mean ± SEM; 4.27 vs. 4.23 ± 0.03 Mcal/kg). As expected, increasing the concentration of lignin resulted in a reduction in total-tract NDF digestibility (45.5% vs. 40.4% ± 0.742%). Increasing lignin also resulted in a reduction in the digestibility of starch (97.7 vs. 96.3 ± 0.420) and CP (65.0 vs. 60.0 ± 0.829). Lignin also decreased the concentration of digestible energy (2.83 vs. 2.63 ± 0.04 Mcal/kg) and ME (2.52 vs. 2.36 ± 0.05 Mcal/kg), but the concentration of NE L was similar (1.81 vs. 1.75 ± 0.06 Mcal/kg). Increasing the concentration of lignin also reduced yields of ECM (33.7 vs. 30.0 ± 0.838 kg/d), milk protein (1.00 vs. 0.843 ± 0.027 kg/d), and milk fat (1.30 vs. 1.19 ± 0.058 kg/d). Decreasing the dietary lignin concentration did not affect daily methane emissions, averaging 391 ± 29.6 L/d. Results of this study indicate that feeding a diet greater in lignin decreases the digestibility of nutrients and provides less energy for production responses and that energy supplied from digestible NDF may be less than predicted by some nutrition models., (© 2024, The Authors. Published by Elsevier Inc. on behalf of the American Dairy Science Association®. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).)

The aim of this study was to determine the individual and combined effects of supplementing fat with whole-cracked rapeseed (FAT), nitrate (NITRATE), and 3-nitrooxypropanol (3-NOP) on compositional and functional properties of milk fat. An 8 × 8 incomplete Latin square design was conducted with 48 lactating Danish Holstein cows over 6 periods of 21 d each. Eight diets were 2 × 2 × 2 factorially arranged: FAT (30 or 63 g crude fat/kg DM), NITRATE (0 or 10 g nitrate/kg DM), and 3-NOP (0 or 80 mg 3-NOP/kg DM), and cows were fed ad libitum. Milk samples were analyzed for general composition, fatty acids (FA) and thermal properties of milk fat. Milk fat content was decreased by supplementing fat but increased by 3-NOP. The changes in FA composition were mainly driven by the FAT × 3-NOP interaction. Fat supplementation shifted milk FA composition toward a lower content of SFA and greater contents of MUFA and PUFA, whereas these effects became smaller in combination with 3-NOP. However, 3-NOP had no effects on SFA, MUFA, or PUFA in low-fat diets. Fat supplementation lowered solid-fat content in milk fat because of the decreased SFA content. The onset crystallization temperature of milk fat was decreased by 3-NOP when supplemented in low-fat diets. According to the FAT × 3-NOP interaction, supplementation of fat without 3-NOP shifted the peak temperature of the low-melting fraction of milk fat toward low temperature as a result of a decreased proportion of C16:0 and increased proportions of C18:1 cis-9, C18:1 trans-11, C18:2 cis-9, and CLA cis-9,trans-11. In conclusion, no additive effects were observed among FAT, NITRATE, and 3-NOP on chemical and thermal properties of milk fat, and fat supplementation largely changed milk FA composition, which in turn affected the thermal properties of milk fat., (© 2024, The Authors. Published by Elsevier Inc. on behalf of the American Dairy Science Association®. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).)

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 × 10 9 cfu/d; n = 12) or 3) DFM15 (15 g/d Bacillus DFM, 7.4 × 10 9 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., (© 2024, The Authors. Published by Elsevier Inc. on behalf of the American Dairy Science Association®. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).)

Dietary methane (CH 4 ) mitigation is in some cases associated with an increased hydrogen (H 2 ) emission. The objective of the present study was to investigate the acute and short-term effects of acceptors for H 2 (fumaric acid, acrylic acid, or phloroglucinol) supplemented via pulse-dosing to dairy cows fed CH 4 mitigating diets (using nitrate or 3-nitrooxypropanol), on gas exchange, rumen gas, and VFA composition. For this purpose, 2 individual 4 × 4 Latin square experiments were conducted with 4 periods of 3 d (nitrate supplementation) and 7 d (3-nitrooxypropanol supplementation), respectively. In each study, 4 rumen-cannulated Danish Holstein cows were used. Each additive for CH 4 mitigation was included in the ad libitum-fed diet within the 2 experiments (exp. 1 and exp. 2), to which the cows were adapted for at least 14 d. Acceptors for H 2 were administered twice daily in equal portions through the rumen fistula immediately after feeding of the individual cow. In exp. 1 (nitrate), the treatments were CON-1 (no H 2 -acceptor), FUM-1 (fumaric acid), ACR-1 (acrylic acid), and FUM+ACR-1 (50% FUM-1 + 50% ACR-1). In exp. 2 (3-nitrooxypropanol), the 3 treatments, CON-2, FUM-2, and ACR-2, were similar to CON-1, FUM-1 and ACR-1 treatments, however the fourth treatment was PHL-2 (phloroglucinol). Gas exchanges were measured in respiration chambers, and samples of rumen liquid and headspace gas were taken in time series relative to feeding and dosing on specific days. Headspace gas was analyzed for gas composition, and rumen liquid was analyzed for VFA composition and dissolved gas concentrations. Headspace gas composition and dissolved gas concentration were only measured in exp. 2. Dry matter intake was reduced upon acrylic acid supplementation. There were no significant effects of any treatments in any experiments on H 2 emission, except for a decrease in hourly H 2 emission rate (g/h) at 1 h after feeding in both experiments. In exp. 2, H 2 headspace proportions increased with ACR-2 supplementation, whereas dissolved concentrations were unaffected. In exp. 1, cows on ACR-1 increased propionate proportion at 1 h after feeding. In exp. 2, both FUM-2 and ACR-2 increased rumen propionate proportion in the hours after feeding and dosing. There was no effect on rumen acetate for cows on PHL-2. There was a strong positive correlation between rumen dissolved CH 4 and headspace CH 4 (r = 0.84), whereas the equivalent correlation was weaker for H 2 (r = 0.41). For the relationship between dissolved concentrations and emissions of CH 4 and H 2 , there was a moderate positive correlation for CH 4 (r = 0.54), whereas it was weak for H 2 (r = 0.28) with zero slope. In conclusion, the results suggested that fumaric acid and acrylic acid to some extent was reduced to propionate without associative effects on measures for H 2 redirection. Furthermore, phloroglucinol seemed not to be metabolized in the rumen in the present study, because no effects on rumen acetate or measures of H 2 were observed. Changes in H 2 headspace and emission may be a poor proxy for actual changes in the rumen fluid concentration of H 2 ., (© 2024, The Authors. Published by Elsevier Inc. on behalf of the American Dairy Science Association®. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).)

Finite natural resources, rising human population, and climate change pose challenges to traditional crop production. Hydroponically grown fodder (i.e., sprouted grains) can be an alternative feed source for dairy cows; however, only sprouted barley has been investigated in low-producing cows. We aimed to evaluate the effect of replacing conventional concentrates with sprouted barley or wheat, grown using hydroponics, on milk production, nutrient digestibility, and milk fatty acid profile in high-producing cows. Twenty-four multiparous Holstein cows (3.25 ± 1.33 lactations; 102 ± 23 DIM; 49 ± 4 kg/d of milk) were used in a replicated 3 × 3 Latin square design with 21-d experimental periods. Following a 2-wk covariate period, cows were fed 1 of 3 experimental diets: a TMR (1) without sprouted grains (control), or with (2) 10% sprouted barley, or (3) 10% sprouted wheat on a DM basis. Experimental diets were formulated to be isoenergetic and isonitrogenous with sprouted grains that replaced ground corn, soybean meal, canola meal, and dextrose. Sprouted grains were grown using a semi-automatic hydroponic system and harvested after 6 d of growth. Data and sample collection occurred during the last 3 d of the covariate and experimental periods. Wide ranges were observed for the DM percent of sprouted grains (12.1%-22.9% and 13.3%-25.7% for barley and wheat, respectively) and the ratio of sprouted fodder to seed (0.67-1.07 for both barley and wheat). Feeding sprouted grains did not modify the yield of milk or ECM; however, DMI were lower for barley, relative to control. Feed efficiencies were greater for barley than for control (1.49 ± 0.03 vs. 1.43 ± 0.03 for milk yield/DMI; 1.85 ± 0.03 vs. 1.73 ± 0.04 for ECM/DMI). Yields and concentrations of milk components (i.e., fat, true protein, and lactose) were not affected by treatment. Milk urea N concentrations were greater for wheat, relative to control or barley. Body weight (752 ± 3 vs. 742 ± 3 kg) and BW gains (6.53 ± 2.99 vs. -9.33 ± 2.91 kg/21 d) were higher for wheat than for control. Apparent total-tract digestibility of organic matter was greater for wheat relative to barley. Digestibilities of NDF and starch were higher for wheat and control, relative to barley, and CP digestibility was greater for wheat, relative to barley and control. Rumination and physical activity were not affected by treatment. In summary, replacing traditional concentrates with sprouted grains grown using hydroponics improved milk production efficiency (barley sprouts) or enhanced body weight gain (wheat sprouts). A life-cycle assessment needs to be conducted to determine the net effect of this feeding strategy for the dairy industry., (© 2024, The Authors. Published by Elsevier Inc. on behalf of the American Dairy Science Association®. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).)

Milk protein production is the largest draw on AA supplies for lactating dairy cattle. Prior NRC predictions of milk protein production have been absorbed protein (MP)-based and used a first-limiting nutrient concept to integrate the effects of energy and protein, which yielded poor accuracy and precision (root mean squared error [RMSE] >21%). Using a meta-data set gathered, various alternative equation forms considering MP, absorbed total EAA, absorbed individual EAA, and digested energy (DE) supplies as additive drivers of production were evaluated, and all were found to be superior in statistical performance to the first limitation approach (RMSE = 14%-15%). Inclusion of DE intake and a quadratic term for MP or absorbed EAA supplies were found to be necessary to achieve intercept estimates (nonproductive protein use) that were similar to the factorial estimates of the National Academies of Sciences, Engineering, and Medicine (2021). The partial linear slope for MP was found to be 0.409, which is consistent with the observed slope bias of -0.34 g/g when a slope of 0.67 was used for MP efficiency in a first-limiting nutrient system. Replacement of MP with the supplies of individual absorbed EAA expressed in grams per day and a common quadratic across the EAA resulted in unbiased predictions with improved statistical performance as compared with MP-based models. Based on Akaike's information criterion and biological consistency, the best equations included absorbed His, Ile, Lys, Met, Thr, the NEAA, and individual DE intakes from fatty acids, NDF, residual OM, and starch. Several also contained a term for absorbed Leu. These equations generally had RMSE of 14.3% and a concordance correlation of 0.76. Based on the common quadratic and individual linear terms, milk protein response plateaus were predicted at approximately 320 g/d of absorbed His, Ile, and Lys; 395 g/d of absorbed Thr; 550 g/d of absorbed Met; and 70 g/d of absorbed Leu. Therefore, responses to each except Leu are almost linear throughout the normal in vivo range. De-aggregation of the quadratic term and parsing to individual absorbed EAA resulted in nonbiological estimates for several EAA indicating over-parameterization. Expression of the EAA as g/100 g total absorbed EAA or as ratios of DE intake and using linear and quadratic terms for each EAA resulted in similar statistical performance, but the solutions had identifiability problems and several nonbiological parameter estimates. The use of ratios also introduced nonlinearity in the independent variables which violates linear regression assumptions. Further screening of the global model using absorbed EAA expressed as grams per day with a common quadratic using an all-models approach, and exhaustive cross-evaluation indicated the parameter estimates for BW, all 4 DE terms, His, Ile, Lys, Met, and the common quadratic term were stable, whereas estimates for Leu and Thr were known with less certainty. Use of independent and additive terms and a quadratic expression in the equation results in variable efficiencies of conversion. The additivity also provides partial substitution among the nutrients. Both of these prevent establishment of fixed nutrient requirements in support of milk protein production., (© 2024, The Authors. Published by Elsevier Inc. on behalf of the American Dairy Science Association®. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).)

Selenium is an essential trace mineral for dairy cattle and can be provided in the diet in various forms that may differ in bioavailability. The objective of this study was to determine how source of Se affects animal performance, Se status, retention, and apparent and true absorption. Multiparous Holstein cows (n = 24; 597 ± 49 kg of BW) were blocked by DIM (161 ± 18) and randomly assigned to receive 0.3 mg Se/kg of DM (100% of National Academies of Sciences, Engineering, and Medicine requirements) of either organic Se (ORG; selenized yeast) or inorganic Se (INO; sodium selenite). The Se premix was top dressed on a common TMR fed daily and mixed into the top 15 cm directly before feeding. Following an 11-wk adaptation period, cows received simultaneous infusions of an intraruminal isotope dose of 77 Se in the same chemical form as the premix, and an intravenous dose of 82 Se in an inorganic form. Infusions were followed by a 4-d period of blood and rumen fluid sampling, and total collection of feces, urine, and milk. Daily DMI (23 ± 0.6 kg), milk yield (35 ± 1.2 kg), and serum Se (0.11 ± 0.003 µg/g) were not different between treatments during the adaptation period, but milk Se concentrations were greater for ORG compared with INO. Serum 77 Se maximum concentration and area under the curve (AUC) were not different between treatments for 72 h following infusion, but rumen fluid 77 Se AUC was higher for ORG than INO. Apparent absorption (64% ± 1.4%), and retention (44% ± 1.5%) of the 77 Se dose did not differ between treatments. True absorption was calculated using 82 Se enrichment in serum and feces and was determined to be 69% ± 1.3% and did not differ between treatments. Fecal excretion of the 77 Se dose was not different between treatments (36% ± 1.4%), but ORG had lower urinary excretion and higher milk excretion compared with INO. These results indicate that organic Se resulted in greater Se concentration of milk and lower urinary Se excretion into the environment, but absorption, Se status, and performance of the cow were not affected by Se source at this supplementation level., (© 2024, The Authors. Published by Elsevier Inc. on behalf of the American Dairy Science Association®. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).)

The objective of this study was to evaluate the interaction of dietary carbohydrate profile and soybean meal (SBM) replacement with either Chlorella pyrenoidosa (CHL) or Spirulina platensis (SPI) on in vitro fermentation. This experiment was conducted as a randomized complete block design, with fermentation run (3 runs) considered as blocks. The treatments were arranged in a 2 × 5 factorial design, where the first factor was the carbohydrate profile, which was composed of diets containing 42.5% neutral detergent fiber (NDF) and 26.8% starch (HF-LS) or 26.8% NDF and 40.6% starch (LF-HS), and the second factor was the protein source, in which a control diet (100% SBM), partial replacement of SBM with CHL (1/2CHL) or SPI (1/2SPI), or total replacement of SBM with CHL or SPI were used. All experimental diets were formulated to have 17% crude protein. The ruminal fluid was collected from 2 lactating Holstein cows, buffered with Van Soest medium at a ratio of 1:2 and added to serum bottles containing 0.50 g of the experimental diets. Bottles were incubated at 39°C for 24 and 48 h in triplicate; headspace pressure was measured, along with gas collection for methane (CH 4 ) quantification at 0, 2, 4, 8, 16, 24, 36, and 48 h after incubation. The final medium was used to measure pH, ammonia, and VFA. After incubation, feed bags were recovered and used for estimation of dry matter (DM), NDF, and organic matter (OM) degradability. Statistical analysis was carried out using the MIXED procedure of SAS, with carbohydrate profile, protein source, assay, and their interactions as fixed effects, with run and bottle as random effects. Orthogonal contrasts were used to compare carbohydrate profile, algae species, carbohydrate profile × algae interaction, and linear and quadratic effects of SBM replacement with CHL or SPI. There was no interaction effect between carbohydrate profile and algae source. The LF-HS improved gas production, degradability of nutrients, and VFA, mainly increasing the production of butyrate and propionate. When compared with CHL, SPI had a greater degradability of nutrients and branched VFA, along with reduction in total gas production and tended to reduce total CH 4 yield. The replacement of SBM with algae linearly reduced the degradability of nutrients, along with a linear reduction in gas production. When replacement of SBM with only SPI was evaluated, SPI slightly reduced the degradability of nutrients; however, it promoted a linear reduction in CH 4 yield, as well as reduction in CH 4 yield by unit of degraded DM, NDF, and OM. In summary, there was no interaction of carbohydrate profile and protein source, which means that SBM replacement had a similar effect, regardless of dietary carbohydrate profile. Spirulina may be a more suitable algae source than Chlorella due to the potential to reduce CH 4 ., (© 2024, The Authors. Published by Elsevier Inc. on behalf of the American Dairy Science Association®. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).)

The objective of this randomized controlled trial was to evaluate the efficacy of supplementing bovine colostrum replacer during weaning to reduce intestinal permeability and improve gain. For this experiment, 65 calves were enrolled and housed individually until 70 d of age. Calves were fed milk replacer (150 g/L) 3 times daily with 9, 10.5, 11.25, and 12 L/d offered from d 1 to 7, 8 to 14, 15 to 21, 22 to 56, respectively. Calves were weaned over 8 d from d 57 to 64, receiving a total of 7.8 L in 2 meals per day from d 57 to 60 and 3.8 L/d in 1 feeding from d 61 to 64. At d 57, calves were blocked by birth weight and randomly assigned to 1 of 2 treatments, equal in ME, which were fed once daily during weaning from d 57 to 64: control (CON; n = 31 calves): 3.8 L of milk replacer (150 g/L) fed by nipple bottle, or colostrum supplementation (COL; n = 34): a mixture of 1 L of bovine colostrum replacer (125 g/L) and 3 L of milk replacer (150 g/L) with 3.8 L of of the mixture fed by nipple bottle. Serum IgG was measured within 48 h of birth and BW was taken at d 0, 57, 60, 64, 70, 77, and 84. Starter intake and bovine respiratory disease (BRD) score were measured daily from d 50 to 70, and fecal consistency was examined daily from d 56 to 70. Serum BHB and lung consolidation were evaluated at d 57, 64, and 70, and intestinal permeability was assessed by recovery of chromium-EDTA, lactulose, and d-mannitol from plasma after oral administration at d 56 and 65. Body weight had no difference between treatment groups at the start of weaning, but COL were 2.79 kg (95% CI: 0.90-4.68) and 2.76 kg (95% CI: 0.86-4.65) heavier than CON at d 77 and 84, respectively. Additionally, COL tended to gain 100.00 g/d more than CON calves (95% CI: -10.41-207.13) from d 57 to 84. No differences were observed in any of the other variables measured. Supplementation of bovine colostrum replacer during weaning may improve weight gain, but the mechanism of action is not clear., (© 2024, The Authors. Published by Elsevier Inc. on behalf of the American Dairy Science Association®. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).)

Thanks to improvements in genetics, nutrition, and management, modern dairy cows can still produce large amounts of milk at the end of lactation, with possible negative effects on health and welfare, particularly when milking is stopped abruptly. To limit yield at dry-off, strategies involving different types of dietary restriction have been used worldwide. Thus, we aimed to investigate the effects of a reduced nutrient density at dry-off on milk production, metabolism, the pattern of rumen fermentation, and milk fatty acid profile around dry-off and in the ensuing periparturient period. During the last week before dry-off, 26 Holstein cows were enrolled in pairs according to the expected calving date and either fed ad libitum ryegrass hay (nutrient restricted, NR; 13 cows) or continued to receive lactation diet (control group, CTR; 13 cows). After dry-off, both groups received only grass hay for 7 d, and free access to water was always provided. Blood, milk, and rumen fluid samples were collected from 7 d before dry-off to 28 DIM. Milk production, DMI (during the periparturient period), and rumination times were recorded daily. At dry-off, NR cows had decreased milk yield (-62%) and milk lactose compared with CTR but had higher fat and protein contents. In the subsequent lactation, no significant differences were observed in milk yield and composition. The BCS did not differ between groups during the transition period, but it decreased in NR after dry-off. Before dry-off, NR had decreased glucose, urea, and insulin, but higher creatinine, BHB, and nonesterified fatty acids (NEFA). The day after dry-off, NEFA were lower in NR, but they were higher 7 d after calving. At dry-off, NR had higher rumen pH, lower lactate, urea, and total volatile fatty acid concentrations. Considering volatile fatty acid molar proportions, NR had increased acetate but decreased propionate and butyrate at dry-off. Rumination time dropped 6 d before dry-off in NR and after dry-off in CTR, but no differences were observed in the periparturient period. Milk fatty acid profile revealed a remarkably lower proportion of short-chain fatty acids in NR at dry-off and a higher proportion of medium- and long-chain ones. These results confirmed that decreasing nutrient density reduce milk yield before dry-off. However, metabolism around dry-off was significantly affected, as suggested by plasma, rumen fluid, and milk analyses. Further research is required to investigate the impact of the metabolic effects on the inflammatory response, liver function, and immune system, particularly concerning the mammary gland., (The Authors. Published by Elsevier Inc. on behalf of the American Dairy Science Association®. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).)