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Abstract Background In the context of the RABOLA project, which aimed to identify operational practices that lead to the reduction of antibiotic use in dairy cattle farming, lyophilised Aloe arborescens was administered orally to cows during the dry-off and peripartum periods. In this specific paper we wanted to examine whether oral administration of Aloe arborescens, in combination with the topical application of a teat sealant could exert an effect on the microbial populations of three cow microbiomes (rumen, milk, rectum), between dry-off and peripartum. Dry-off and peripartum are critical physiological phases of the cow’s life, where both the mammary gland and the gastrointestinal tract undergo dramatic modifications, hence the relevance of evaluating the effects of dietary treatments. Methods Thirty multiparous dairy cows were randomly allocated to three groups: Control (antibiotic treatment and internal teat sealant), Sealant (only internal teat sealant) and Aloe (internal teat sealant and Aloe arborescens homogenate administered orally). For 16S rRNA gene sequencing, rumen, rectum and milk samples were collected, not synchronously, at the most critical timepoints around dry-off and calving, considering the physiological activity of each biological site. Results The rumen microbiome was predominantly characterized by Bacteroidetes and Firmicutes followed by Proteobacteria, while the rectum exhibited a prevalence of Firmicutes and Bacteroidetes. The milk microbiome mainly comprised Firmicutes, Proteobacteria, Actinobacteria and Bacteroidetes. Alistipes spp., Ruminococcaceae UCG-10 group, Prevotellaceae UCG-001 group, and Bacteroides spp., involved in cellulose and hemicellulose degradation, enhancement of energy metabolism, and peptide breakdown, showed increment in the rectum microbiome with Aloe supplementation. The rectum microbiome in the Aloe group exhibited a significant increase in the Firmicutes to Bacteroidetes ratio and alpha-diversity at seven days after dry-off period. Beta-diversity showed a significant separation between treatments for the rectum and milk microbiomes. Aloe supplementation seemed to enrich milk microbial composition, whereas the Sealant group showed greater diversity compared to the Control group, albeit this included an increase in microorganisms frequently associated with mastitis. Conclusions Aloe arborescens administration during the dry-off period did not demonstrate any observable impact on the microbial composition of the rumen, a finding further supported by volatilome analysis. Instead, the oral Aloe supplementation at dry-off appears to significantly influence the composition of the dairy cow rectum and milk microbiomes in the following lactation.

Dairy goat’s lactation persistence forces farmers at limiting nutrient supply to reduce yield at dry-off. Omitting dry period could be a solution, but metabolic effects of this practice have never been tested. Eight Alpine (AL) and 12 Saanen (SA) goats approaching their second kidding blocked by breed and number of kids carried (single - SIN - or double - DOU) were allocated to one out of two groups (4 AL and 6 SA; 5 SIN and 5 DOU in each group). At −42 ± 7 days from kidding (DFK), they were either dried off (DR) or milked continuously until kidding (CL). Body condition score (BCS) was assessed, and blood samples were collected at −10, −3, 5, 12, and 29 DFK to determine metabolic profile. Milk yield and composition were assessed at −56, 7, 31, 62, and 97 DFK. Compared with DR, CL had higher plasma reactive oxygen metabolites and liver enzymes. Compared with DR counterparts, AL-CL had higher nonesterified fatty acids (NEFA) at −10, whereas SA-CL had lower NEFA at −3 DFK. CL goats had lower BCS, higher plasma beta-hydroxybutyrate (BHB) and urea before kidding, but higher glucose at −3 and 5, lower NEFA at 5 and 12, and higher BCS at 29 DFK. CL goats had lower haptoglobin and myeloperoxidase at −3 and 5 DFK, paired with higher albumin, cholesterol, and paraoxonase at 12 DFK. Omitting dry period mitigated the inflammatory condition around kidding in dairy goats, possibly accounting for an improved energy balance in early lactation, despite body reserve mobilisation prepartum was greater under continuous lactation.

Abstract Background Various blood metabolites are known to be useful indicators of health status in dairy cattle, but their routine assessment is time-consuming, expensive, and stressful for the cows at the herd level. Thus, we evaluated the effectiveness of combining in-line near infrared (NIR) milk spectra with on-farm (days in milk [DIM] and parity) and genetic markers for predicting blood metabolites in Holstein cattle. Data were obtained from 388 Holstein cows from a farm with an AfiLab system. NIR spectra, on-farm information, and single nucleotide polymorphisms (SNP) markers were blended to develop calibration equations for blood metabolites using the elastic net (ENet) approach, considering 3 models: (1) Model 1 (M1) including only NIR information, (2) Model 2 (M2) with both NIR and on-farm information, and (3) Model 3 (M3) combining NIR, on-farm and genomic information. Dimension reduction was considered for M3 by preselecting SNP markers from genome-wide association study (GWAS) results. Results Results indicate that M2 improved the predictive ability by an average of 19% for energy-related metabolites (glucose, cholesterol, NEFA, BHB, urea, and creatinine), 20% for liver function/hepatic damage, 7% for inflammation/innate immunity, 24% for oxidative stress metabolites, and 23% for minerals compared to M1. Meanwhile, M3 further enhanced the predictive ability by 34% for energy-related metabolites, 32% for liver function/hepatic damage, 22% for inflammation/innate immunity, 42.1% for oxidative stress metabolites, and 41% for minerals, compared to M1. We found improved predictive ability of M3 using selected SNP markers from GWAS results using a threshold of > 2.0 by 5% for energy-related metabolites, 9% for liver function/hepatic damage, 8% for inflammation/innate immunity, 22% for oxidative stress metabolites, and 9% for minerals. Slight reductions were observed for phosphorus (2%), ferric-reducing antioxidant power (1%), and glucose (3%). Furthermore, it was found that prediction accuracies are influenced by using more restrictive thresholds (−log10(P-value) > 2.5 and 3.0), with a lower increase in the predictive ability. Conclusion Our results highlighted the potential of combining several sources of information, such as genetic markers, on-farm information, and in-line NIR infrared data improves the predictive ability of blood metabolites in dairy cattle, representing an effective strategy for large-scale in-line health monitoring in commercial herds.

Abstract Background Metabolic disturbances adversely impact productive and reproductive performance of dairy cattle due to changes in endocrine status and immune function, which increase the risk of disease. This may occur in the post-partum phase, but also throughout lactation, with sub-clinical symptoms. Recently, increased attention has been directed towards improved health and resilience in dairy cattle, and genomic selection (GS) could be a helpful tool for selecting animals that are more resilient to metabolic disturbances throughout lactation. Hence, we evaluated the genomic prediction of serum biomarkers levels for metabolic distress in 1353 Holsteins genotyped with the 100K single nucleotide polymorphism (SNP) chip assay. The GS was evaluated using parametric models best linear unbiased prediction (GBLUP), Bayesian B (BayesB), elastic net (ENET), and nonparametric models, gradient boosting machine (GBM) and stacking ensemble (Stack), which combines ENET and GBM approaches. Results The results show that the Stack approach outperformed other methods with a relative difference (RD), calculated as an increment in prediction accuracy, of approximately 18.0% compared to GBLUP, 12.6% compared to BayesB, 8.7% compared to ENET, and 4.4% compared to GBM. The highest RD in prediction accuracy between other models with respect to GBLUP was observed for haptoglobin (hapto) from 17.7% for BayesB to 41.2% for Stack; for Zn from 9.8% (BayesB) to 29.3% (Stack); for ceruloplasmin (CuCp) from 9.3% (BayesB) to 27.9% (Stack); for ferric reducing antioxidant power (FRAP) from 8.0% (BayesB) to 40.0% (Stack); and for total protein (PROTt) from 5.7% (BayesB) to 22.9% (Stack). Using a subset of top SNPs (1.5k) selected from the GBM approach improved the accuracy for GBLUP from 1.8 to 76.5%. However, for the other models reductions in prediction accuracy of 4.8% for ENET (average of 10 traits), 5.9% for GBM (average of 21 traits), and 6.6% for Stack (average of 16 traits) were observed. Conclusions Our results indicate that the Stack approach was more accurate in predicting metabolic disturbances than GBLUP, BayesB, ENET, and GBM and seemed to be competitive for predicting complex phenotypes with various degrees of mode of inheritance, i.e. additive and non-additive effects. Selecting markers based on GBM improved accuracy of GBLUP.

ABSTRACT: Residual Feed Intake (RFI) is defined as the difference between measured and predicted intake. Understanding its biological regulators could benefit farm profit margins. The most-efficient animals (M-Eff) have observed intake smaller than predicted resulting in negative RFI, whereas the least-efficient (L-Eff) animals have positive RFI. Hence, this observational study aimed at retrospectively comparing the blood immunometabolic profile in calves with divergent RFI during the preweaning period. Twenty-two Italian Simmental calves were monitored from birth through 60 d of age. Calves received 3 L of colostrum from their respective dams. From 2 to 53 d of age, calves were fed a milk replacer twice daily, whereas from 54 to 60 d (i.e., weaning) calves were stepped down to only one meal in the morning. Calves had ad libitum access to concentrate and intakes were recorded daily. The measurement of BW and blood samples were performed at 0, 1, 7, 14, 21, 28, 35, 45, 54, and 60 d of age. Calves were ranked and categorized as M-Eff or L-Eff according to the median RFI value. Median RFI was −0.06 and 0.04 kg of DMI/d for M-Eff and L-Eff, respectively. No evidence for group differences was noted for colostrum and plasma IgG concentrations. Although growth rate was not different, as expected, (0.67 kg/d [95% CI = 0.57–0.76] for both L-Eff and M-Eff) throughout the entire preweaning period (0–60 d), starter intake was greater in L-Eff compared with M-Eff calves (+36%). Overall, M-Eff calves had a greater gain-to-feed ratio compared with L-Eff calves (+16%). Plasma ceruloplasmin, myeloperoxidase, and reactive oxygen metabolites concentrations were greater in L-Eff compared with M-Eff calves. Compared with L-Eff, M-Eff calves had an overall greater plasma concentration of globulin, and γ-glutamyl transferase (indicating a better colostrum uptake) and Zn at 1 d. Retinol and urea were overall greater in L-Eff. The improved efficiency in nutrient utilization observed in M-Eff was paired with a lower grade of oxidative stress and systemic inflammation. L-Eff may have had greater energy expenditure to support the activation of the immune system.

ABSTRACT: The legalization of industrial hemp by the 2018 Farm Bill in the United States has driven a sharp increase in its cultivation, including for cannabinoid extraction. Spent hemp biomass (SHB), produced from the extraction of cannabinoids, can potentially be used as feed for dairy cows; however, it is still illegal to do so in the United States, according to the US Food and Drug Administration Center for Veterinary Medicine, due to the presence of cannabinoids and the lack of data on the effect on animals. To assess the safety of this byproduct as feed for dairy cows, late-lactation Jersey cows (245 ± 37 d in milk; 483 ± 38 kg body weight; 10 multiparous and 8 primiparous) received a basal total mixed ration (TMR) diet plus 13% alfalfa pellet (CON) or 13% pelleted SHB for 4 wk (intervention period [IP]) followed by 4 wk of withdrawal period (WP), where all cows received only the basal TMR during WP. The dry matter intake (DMI), body weight, body condition score, milk yield, milk components, and fatty acid profile, blood parameters, N metabolism, methane emission, and activity were measured. Results indicated that feeding SHB decreased DMI mainly due to the low palatability of the SHB pellet, as the cows consumed only 7.4% of the total TMR with 13.0% SHB pellet offered in the ration. However, milk yield was not affected during the IP and was higher than CON during the WP, leading to higher milk yield/DMI. Milk components were not affected, except for a tendency in decreased fat percentage. Milk fat produced by cows fed SHB had a higher proportion of oleate and bacteria-derived fatty acids than CON. The activity of the cows was not affected, except for a shorter overall lying time in SHB versus CON cows during the IP. Blood parameters related to immune function were not affected. Compared with CON, cows fed SHB had a lower cholesterol concentration during the whole experiment and higher β-hydroxybutyric acid during the WP, while a likely low-grade inflammation during the IP was indicated by higher ceruloplasmin and reactive oxidative metabolites. Other parameters related to liver health and inflammatory response were unaffected, except for a tendency for higher activity of alkaline phosphatase during IP and a lower activity of gamma-glutamyl transferase during WP in the SHB group versus CON. The bilirubin concentration was increased in cows fed SHB, suggesting a possible decrease in the clearance ability of the liver. Digestibility of the dry matter and protein and methane emission were not affected by feeding SHB. The urea, purine derivatives, and creatinine concentration in urine was unaffected, but cows fed SHB had higher N use efficiency and lower urine volume. Altogether, our data revealed a relatively low palatability of SHB affecting DMI with minimal biological effects, except for a likely low-grade inflammation, a higher N use efficiency, and a possible decrease in liver clearance. Overall, the data support the use of SHB as a safe feed ingredient for lactating dairy cows.

The high energy, protein, and fiber contents of whole cottonseed make it a potential candidate for the inclusion in calf starters to promote the rumen development. This study aimed at assessing whether the inclusion of whole cottonseed in the starter would affect performance, metabolic profile, and rumination time in Holstein dairy calves. From 2 to 55 d of age, 12 heifer calves were fed a constant amount of milk replacer twice daily (8 L/d), whereas from 56 to 65 d (weaning) milk replacer was gradually reduced (from 4 to 1 L/d) and fed in a single meal. Calves were blocked by birth body weight and % Brix of colostrum received and randomly assigned to 1 out of 2 dietary treatments: (1) control starter (CTR); (2) starter with 8% inclusion of whole cottonseed (WCS). Treatments were fed for ad libitum intake. From d 56, TMR and hay were offered ad libitum. At 0, 2, 7, 21, 65, and 80 d, BW was measured and blood samples were collected. Feed intake and rumination time were automatically recorded. Compared with CTR, WCS calves were heavier at weaning and after weaning, and consumed more starter from 59 to 72 d. In the immediate postweaning (from 66 to 72 d of age), rumination time increased more in WCS calves. Overall, WCS calves had greater plasma glucose, β-carotene, and retinol concentrations, whereas ceruloplasmin and myeloperoxidase were lower. Calves in the WCS group had greater glucose concentration at 21 and 65 d and lower urea at 65 d (weaning). Plasma β-carotene concentration was greater at 65 and 80 d in calves of the WCS group compared with CTR calves. At 80 d, WCS calves had lower plasma alkaline phosphatase and greater β-hydroxybutyrate, paraoxonase, and tocopherol. These results suggest that inclusion of WCS in the calf starter might be beneficial for rumen development, leading to greater feed intake and BW. Moreover, WCS inclusion was associated with lower oxidative stress and inflammation, improved energy metabolism and liver functionality, and likely quicker rumen development, as might be indicated by the higher plasma β-hydroxybutyrate and rumination time. These changes occurred mainly after weaning, when no differences in starter intake and average daily gain were detected, suggesting a better efficiency of nutrient utilization at this age.

ABSTRACT: The causes of variation in the milk mineral profile of dairy cattle during the first phase of lactation were studied under the hypothesis that the milk mineral profile partially reflects the animals' metabolic status. Correlations between the minerals and the main milk constituents (i.e., protein, fat, and lactose percentages), and their associations with the cows' metabolic status indicators were explored. The metabolic status indicators (MET) that we used were blood energy-protein metabolites [nonesterified fatty acids, β-hydroxybutyrate (BHB), glucose, cholesterol, creatinine, and urea], and liver ultrasound measurements (predicted triacylglycerol liver content, portal vein area, portal vein diameter and liver depth). Milk and blood samples, and ultrasound measurements were taken from 295 Holstein cows belonging to 2 herds and in the first 120 d in milk (DIM). Milk mineral contents were determined by ICP-OES; these were considered the response variable and analyzed through a mixed model which included DIM, parity, milk yield, and MET as fixed effects, and the herd/date as a random effect. The MET traits were divided in tertiles. The results showed that milk protein was positively associated with body condition score (BCS) and glucose, and negatively associated with BHB blood content; milk fat was positively associated with BHB content; milk lactose was positively associated with BCS; and Ca, P, K and S were the minerals with the greatest number of associations with the cows' energy indicators, particularly BCS, predicted triacylglycerol liver content, glucose, BHB and urea. We conclude that the protein, fat, lactose, and mineral contents of milk partially reflect the metabolic adaptation of cows during lactation and within 120 DIM. Variations in the milk mineral profile were consistent with changes in the major milk constituents and the metabolic status of cows.

Milk has become a staple food product globally. Traditionally, milk quality assessment has been primarily focused on hygiene and composition to ensure its safety for consumption and processing. However, in recent years, the concept of milk quality has expanded to encompass a broader range of factors. Consumers now also consider animal welfare, environmental impact, and the presence of additional beneficial components in milk when assessing its quality. This shifting consumer demand has led to increased attention on the overall production and sourcing practices of milk. Reflecting on this trend, this review critically explores such novel quality parameters, offering insights into how such practices meet the modern consumer’s holistic expectations. The multifaceted aspects of milk quality are examined, revealing the intertwined relationship between milk safety, compositional integrity, and the additional health benefits provided by milk’s bioactive properties. By embracing sustainable farming practices, dairy farmers and processors are encouraged not only to fulfill but to anticipate consumer standards for premium milk quality. This comprehensive approach to milk quality underscores the necessity of adapting dairy production to address the evolving nutritional landscape and consumption patterns.

Abstract Background Blood metabolic profiles can be used to assess metabolic disorders and to evaluate the health status of dairy cows. Given that these analyses are time-consuming, expensive, and stressful for the cows, there has been increased interest in Fourier transform infrared (FTIR) spectroscopy of milk samples as a rapid, cost-effective alternative for predicting metabolic disturbances. The integration of FTIR data with other layers of information such as genomic and on-farm data (days in milk (DIM) and parity) has been proposed to further enhance the predictive ability of statistical methods. Here, we developed a phenotype prediction approach for a panel of blood metabolites based on a combination of milk FTIR data, on-farm data, and genomic information recorded on 1150 Holstein cows, using BayesB and gradient boosting machine (GBM) models, with tenfold, batch-out and herd-out cross-validation (CV) scenarios. Results The predictive ability of these approaches was measured by the coefficient of determination (R2). The results show that, compared to the model that includes only FTIR data, integration of both on-farm (DIM and parity) and genomic information with FTIR data improves the R2 for blood metabolites across the three CV scenarios, especially with the herd-out CV: R2 values ranged from 5.9 to 17.8% for BayesB, from 8.2 to 16.9% for GBM with the tenfold random CV, from 3.8 to 13.5% for BayesB and from 8.6 to 17.5% for GBM with the batch-out CV, and from 8.4 to 23.0% for BayesB and from 8.1 to 23.8% for GBM with the herd-out CV. Overall, with the model that includes the three sources of data, GBM was more accurate than BayesB with accuracies across the CV scenarios increasing by 7.1% for energy-related metabolites, 10.7% for liver function/hepatic damage, 9.6% for oxidative stress, 6.1% for inflammation/innate immunity, and 11.4% for mineral indicators. Conclusions Our results show that, compared to using only milk FTIR data, a model integrating milk FTIR spectra with on-farm and genomic information improves the prediction of blood metabolic traits in Holstein cattle and that GBM is more accurate in predicting blood metabolites than BayesB, especially for the batch-out CV and herd-out CV scenarios.

Dairy cows face several challenges during the transition period, and the administration of live yeast might be useful to mitigate this stressful condition. In the current study, the effects of live yeast administration on milk production, feed intake, and metabolic and inflammatory conditions were evaluated. Multiparous Holstein cows were enrolled in this randomized controlled trial and received either a control diet (CTR, n = 14) or the control diet plus 4 g/d of live Saccharomyces cerevisiae yeast (LSC, n = 14) from −21 to 56 days relative to calving. Dry matter intake, milk yield and composition, and rumination time were monitored daily. Blood samples were collected at −21, −7, 3, 14, 28, 42, and 56 days relative to calving to evaluate the metabolic profile. Fecal samples were collected at 56 days relative to calving to measure volatile fatty acids and feed digestibility. No differences between groups were observed in dry matter intake. Compared with CTR, rumination time was lower in LSC in after calving. Although there were no differences in milk components between groups, LSC had greater milk yield in the last three weeks of the study than CTR. No differences were observed in inflammatory markers or other plasma metabolites, except for β-hydroxybutyrate, which was higher in LSC, and reactive oxygen metabolites (ROMs), which were lower in LSC. Overall, these outcomes suggest that live yeast supplementation had some positive effects on milk yield and oxidative status.

Thirty-six multiparous Holstein mid-lactating cows were housed in 6 pens (6 cows/pen) and allocated in two groups (3 pens/group), receiving a control total mixed ration (CTR), or the same diet supplemented with a feed additive (TRT). Between 15 and 35 days from enrolment (DFE) the dry matter intake (DMI) of each pen, and the individual milk yield (MY), rumination time (RT), the compositions of milk and rumen fluid and the metabolic profile of plasma were monitored regularly. At 35 DFE, cows started a 1-week wash-up period prior to the changeover. Data were analysed with a mixed model using repeated measures. Cows receiving the additive had higher DMI and MY (p

With an increased knowledge of the mechanism of action of Fusarium mycotoxins, the concept that these substances are deleterious only for monogastric species is obsolete. Indeed, most mycotoxins can be converted into less toxic compounds by the rumen microflora from healthy animals. However, mycotoxin absorption and its conversion to more toxic metabolites, as well as their impact on the immune response and subsequently animal welfare, reproductive function, and milk quality during chronic exposure should not be neglected. Among the Fusarium mycotoxins, the most studied are deoxynivalenol (DON), zearalenone (ZEN), and fumonisins from the B class (FBs). It is remarkable that there is a paucity of in vivo research, with a low number of studies on nutrient digestibility and rumen function. Most of the in vitro studies are related to the reproductive function or are restricted to rumen incubation. When evaluating the production performance, milk yield is used as an evaluated parameter, but its quality for cheese production is often overlooked. In the present review, we summarize the most recent findings regarding the adverse effects of these mycotoxins with special attention to dairy cattle.

The aim of the current study was to evaluate the effect of the starter restriction and of the ad libitum TMR (total mixed ration) inclusion on intake, growth performance, rumination time (RT), and health condition of Holstein dairy calves during weaning. We randomly assigned thirty female Holstein calves (with an average weight of 38.5 ± 1.96 kg at birth) to one of three treatments. From 21 days of age, the calves were fed one of three treatments as follows: a control diet (CTR) with an ad libitum calf starter but without TMR; Treatment 1 diet (TRT1) with both an ad libitum calf starter and ad libitum TMR; Treatment 2 diet (TRT2) with ad libitum TMR and a restricted amount of a calf starter (50% of the intake recorder in the control group day by day). Calves in the TRT2 group, between 56 and 63 days of age, had a lower body weight (80.1; 79.5; 75.6 kg for the CTR, TRT1, and TRT2 groups, respectively) compared with CTR and TRT1 calves. This outcome is ascribed to the average daily gain (0.759; 0.913; 0.508 kg/day for the CTR, TRT1, and TRT2 groups, respectively), resulting also in TRT2 being lower than CTR or TRT1 calves. The inclusion of ad libitum TMR increased the rumination time, especially after weaning (15.28 min/h, 18.38 min/h, and 18.95 min/h for the CTR, TRT1, and TRT2 groups, respectively). Concerning the rumen metabolism and inflammometabolic response, overall, no differences were observed between the three dietary treatments. In conclusion, the results indicated that a TMR could partially replace a calf starter in weaning dairy calves, since neither growth performance nor health status were impaired. In addition, providing TMR (with or without concentrate restriction) led to a better rumen development and likely a better rumen fermentation efficiency in post-weaning.

ABSTRACT: Cheese-making traits in dairy cattle are important to the dairy industry but are difficult to measure at the individual level because there are limitations on collecting phenotypic information. Mid-infrared spectroscopy has its advantages, but it can only be used during monthly milk recordings. Recently, in-line devices for real-time analysis of milk quality have been developed. The AfiLab recording system (Afimilk) offers significant benefits as phenotypes can be collected from each cow at each milking session. The objective of this study was to assess the potential of integrating AfiLab real-time milk analyzer measures with the stacking ensemble learning technique using heterogeneous base learners for the in-line daily monitoring of cheese-making traits in Holstein cattle with a view to developing a precision livestock farming system for monitoring the technological quality of milk. Data and samples for wet-laboratory analyses were collected from 499 Holstein cows belonging to 2 farms where the AfiLab system was installed. The traits of concern were 9 milk coagulation traits [3 milk coagulation properties (MCP), and 6 curd firming traits (CFt)], and 7 cheese-making traits [3 cheese yield (CY) traits, and 4 milk nutrient recovery in the curd (REC) traits]. The near-infrared AfiLab spectral data and on-farm information (days in milk and parity) were used to assess the predictive ability of different statistical methods [elastic net (EN), gradient boosting machine (GBM), extreme gradient boosting (XGBoost), and artificial neural network (ANN)] across different cross-validation scenarios. These statistical methods were considered the base learners, which were then combined in a stacking ensemble learning. Results indicate that including information on the cows (days in milk and parity) in the AfiLab infrared prediction increased its accuracy by 10.3% for traditional MCP, 13.8% for curd firming, 9.8% for CY, and 11.2% for REC traits compared with those obtained from near-infrared AfiLab alone. The statistical approaches exhibited high prediction accuracies (R2) averaged across the cross-validation scenarios for traditional MCP (0.58 for ANN, 0.55 for EN and GBM, 0.52 for XGBoost, and 0.62 for stacking ensemble), CFt (0.55 for ANN, 0.54 for EN and GBM, 0.53 for XGBoost, and 0.61 for stacking ensemble), and similar R2 averages for CY and REC (0.55 for ANN, 0.54 for EN and GBM, 0.53 for XGBoost, and 0.61 for stacking ensemble). The ANN approach was more accurate than the other base learners (EN, GBM, and XGBoost) and improved accuracy across cross-validation scenarios on average by 7% for traditional MCP, 5% for CFt, 8% for CY, and 7% for REC. The stacking ensemble method improved prediction accuracy by 3% to 31% for traditional MCP, 2% to 26% for CFt, 1% to 38% for CY traits, and 2% to 27% for REC traits compared with the base learners. The prediction accuracies of the different approaches evaluated tended to decrease from the 10-fold cross-validation to the independent validation scenario, although there was a smaller reduction in prediction accuracy with the stacking ensemble learning technique across all the cross-validation scenarios. Our results show that combining in-line on-farm information with stacking ensemble machine learning represents an effective alternative for obtaining robust daily predictions of milk cheese-making traits.

Abstract Precision livestock farming technologies are used to monitor animal health and welfare parameters continuously and in real time in order to optimize nutrition and productivity and to detect health issues at an early stage. The possibility of predicting blood metabolites from milk samples obtained during routine milking by means of infrared spectroscopy has become increasingly attractive. We developed, for the first time, prediction equations for a set of blood metabolites using diverse machine learning methods and milk near-infrared spectra collected by the AfiLab instrument. Our dataset was obtained from 385 Holstein Friesian dairy cows. Stacking ensemble and multi-layer feedforward artificial neural network outperformed the other machine learning methods tested, with a reduction in the root mean square error of between 3 and 6% in most blood parameters. We obtained moderate correlations (r) between the observed and predicted phenotypes for γ-glutamyl transferase (r = 0.58), alkaline phosphatase (0.54), haptoglobin (0.66), globulins (0.61), total reactive oxygen metabolites (0.60) and thiol groups (0.57). The AfiLab instrument has strong potential but may not yet be ready to predict the metabolic stress of dairy cows in practice. Further research is needed to find out methods that allow an improvement in accuracy of prediction equations.

Fusarium mycotoxins are inactivated by rumen flora; however, a certain amount can pass the rumen and reticulum or be converted into biological active metabolites. Limited scientific evidence is available on the impact and mitigation of Fusarium mycotoxins on dairy cows’ performance and health, particularly when cows are exposed for an extended period (more than 2 months). The available information related to these mycotoxin effects on milk cheese-making parameters is also very poor. The objective of this study was to evaluate a commercially available mycotoxin mitigation product (MMP, i.e., TOXO® HP-R, Selko, Tilburg, The Netherlands) in lactating dairy cows fed a Fusarium mycotoxin-contaminated diet, and the repercussions on the dry matter intake, milk yield, milk quality, cheese-making traits and health status of cows. The MMP contains smectite clays, yeast cell walls and antioxidants. In the study, 36 lactating Holstein cows were grouped based on the number of days of producing milk, milk yield, body condition score and those randomly assigned to specific treatments. The study ran over 2 periods (March/May–May/July 2022). In each period, six animals/treatment were considered. The experimental periods consisted of 9 days of adaptation and 54 days of exposure. The physical activity, rumination time, daily milk production and milk quality were measured. The cows were fed once daily with the same total mixed ration (TMR) composition. The experimental groups consisted of a control (CTR) diet, with a TMR with low contamination, high moisture corn (HMC), and beet pulp; a mycotoxins (MTX) diet, with a TMR with highly contaminated HMC, and beet pulp; and an MTX diet supplemented with 100 g/cow/day of the mycotoxin mitigation product (MMP). The trial has shown that the use of MMP reduced the mycotoxin’s negative effects on the milk yield and quality (protein, casein and lactose). The MTX diet had a lower milk yield and feed efficiency than the CTR and MMP HP-R diets. The MMP limited the negative effect of mycotoxin contamination on clotting parameters, mitigating the variations on some coagulation properties; however, the MMP inclusion tended to decrease the protein and apparent starch digestibility of the diet. These results provide a better understanding of mycotoxin risk on dairy cows’ performances and milk quality. The inclusion of an MMP product mitigated some negative effects of the Fusarium mycotoxin contamination during this trial. The major effects were on the milk yield and quality in both the experimental periods. These results provide better insight on the effects of mycotoxins on the performance and quality of milk, as well as the cheese-making traits. Further analyses should be carried out to evaluate MMP’s outcome on immune–metabolic responses and diet digestibility.

Maternal metabolic disruptions, such as ketosis, can have adverse effects on fetal development and influence postnatal factors. Twelve Holstein calves were randomly enrolled in this study at birth and monitored until 8 weeks of age. The study was conducted from fall 2018 until spring 2019. After completing the data collection period, calves were classified according to their respective dams ketotic condition after parturition. This classification was based on dam blood β-hydroxybutyrate < 1.4 mmol/L nonketotic (NONKET; n = 6 calves) or ≥1.4 mmol/L subclinical-ketotic (SK; n = 6 calves). SK calves had greater birth body weight (p = 0.05) but exhibited a slower growth rate compared to NONKET calves from 1 to 8 weeks (p = 0.02). At birth, SK calves had lower (p < 0.01) levels of non-esterified fatty acids and bilirubin compared to NONKET calves. Analysis of feces alpha diversity indicates that by 3 weeks, NONKET calves had greater diversity, richness, and evenness. Butyricicoccus pullicaecorum and Gallibacterium anatis were more abundant in SK calves (p < 0.05) at 3 weeks. In contrast, NONKET calves had a greater (p < 0.05) abundance of Sharpae azabuensis at 3 weeks. These findings suggest that subclinical ketosis in cows can impact the in-utero development, postnatal growth, and maturing gut microbiome of their offspring.

Recent research on the transition period (TP) of dairy cows has highlighted the pivotal role of immune function in affecting the severity of metabolic challenges the animals face when approaching calving. This suggests that the immune system may play a role in the etiology of metabolic diseases occurring in early lactation. Several studies have indicated that the roots of immune dysfunctions could sink way before the “classical” TP (e.g., 3 weeks before and 3 weeks after calving), extending the time frame deemed as “risky” for the development of early lactation disorders at the period around the dry-off. Several distressing events occurring during the TP (i.e., dietary changes, heat stress) can boost the severity of pre-existing immune dysfunctions and metabolic changes that physiologically affect this phase of the lactation cycle, further increasing the likelihood of developing diseases. Based on this background, several operational and nutritional strategies could be adopted to minimize the detrimental effects of immune dysfunctions on the adaptation of dairy cows to the new lactation. A suitable environment (i.e., optimal welfare) and a balanced diet (which guarantees optimal nutrient partitioning to improve immune functions in cow and calf) are key aspects to consider when aiming to minimize TP challenges at the herd level. Furthermore, several prognostic behavioral and physiological indicators could help in identifying subjects that are more likely to undergo a “bad transition”, allowing prompt intervention through specific modulatory treatments. Recent genomic advances in understanding the linkage between metabolic disorders and the genotype of dairy cows suggest that genetic breeding programs aimed at improving dairy cows’ adaptation to the new lactation challenges (i.e., through increasing immune system efficiency or resilience against metabolic disorders) could be expected in the future. Despite these encouraging steps forward in understanding the physiological mechanisms driving metabolic responses of dairy cows during their transition to calving, it is evident that these processes still require further investigation, and that the TP—likely extended from dry-off—continues to be “the final frontier” for research in dairy sciences.

Abstract Metabolic disorders, including hepatic lipidosis and ketosis, severely affect animal health status and welfare with a large economic burden in dairy herds. The gold standard for diagnosing hepatic lipidosis is the liver biopsy, which is impractical and invasive for the screening at farm level. Ultrasound (US) imaging is a promising technique for identifying liver dysfunction, but standardized specifications in physiological conditions are needed. Herein, we described the features of four US measurements, namely the liver predicted triacylglycerol (pTAG) content, liver depth (LD), and portal vein area (PVA) and depth (PVD) and we investigated their associations with a set of hematochemical (HC) indicators in 342 clinically healthy Holstein Friesian dairy cows. Liver pTAG content was negatively associated with hematocrit and positively with globulin, whereas PVA was negatively associated with thiol group levels, and LD positively with ceruloplasmin. We found significant interactions between some HC parameters and parity: in particular, creatinine, thiol groups and globulin for PVA, and aspartate aminotransferase, paraoxonase and ceruloplasmin for PVD. This study offers new insights on variations in liver function occurring after calving and pave the way for the potential use of minimally invasive techniques for prompt detection of metabolic disorders in dairy herds.

Vitamin B12 plays a role in the remethylation of homocysteine to Met, which then serves as a substrate for Met adenosyltransferase (MAT) to synthesize S-adenosylmethionine (SAM). We investigated effects of feeding two cobalt sources [Co-glucoheptonate (CoPro) or CoPectin, Zinpro Corp.], an experimental ruminally-available source of folic acid (FOA), and rumen-protected Met (RPM) on performance and hepatic one-carbon metabolism in peripartal Holstein cows. From −30 to 30 d around calving, 72 multiparous cows were randomly allocated to: CoPro, CoPro + FOA, CoPectin + FOA, or CoPectin + FOA + RPM. The Co treatments delivered 1 mg Co/kg of DM (CoPro or CoPectin), each FOA group received 50 mg/d FOA, and RPM was fed at 0.09% of DM intake (DMI). Milk yield and DMI were not affected. Compared with other groups, the percentage of milk protein was greater after the second week of lactation in CoPectin + FOA + RPM. Compared with CoPro or CoPro + FOA, feeding CoPectin + FOA or CoPectin + FOA + RPM led to a greater activity of MAT at 7 to 15 d postcalving. For betaine–homocysteine S-methyltransferase, CoPro together with CoPectin + FOA + RPM cows had greater activity at 7 and 15 d than CoPro + FOA. Overall, supplying FOA with CoPectin or CoPectin plus RPM may enhance S-adenosylmethionine synthesis via MAT in the liver after parturition. As such, these nutrients may impact methylation reactions and liver function.

Abstract Background Methionine (Met) supply during late-pregnancy enhances fetal development in utero and leads to greater rates of growth during the neonatal period. Due to its central role in coordinating nutrient and one-carbon metabolism along with immune responses of the newborn, the liver could be a key target of the programming effects induced by dietary methyl donors such as Met. To address this hypothesis, liver biopsies from 4-day old calves (n = 6/group) born to Holstein cows fed a control or the control plus ethyl-cellulose rumen-protected Met for the last 28 days prepartum were used for DNA methylation, transcriptome, metabolome, proteome, and one-carbon metabolism enzyme activities. Results Although greater withers and hip height at birth in Met calves indicated better development in utero, there were no differences in plasma systemic physiological indicators. RNA-seq along with bioinformatics and transcription factor regulator analyses revealed broad alterations in ‘Glucose metabolism’, ‘Lipid metabolism, ‘Glutathione’, and ‘Immune System’ metabolism due to enhanced maternal Met supply. Greater insulin sensitivity assessed via proteomics, and efficiency of transsulfuration pathway activity suggested beneficial effects on nutrient metabolism and metabolic-related stress. Maternal Met supply contributed to greater phosphatidylcholine synthesis in calf liver, with a role in very low density lipoprotein secretion as a mechanism to balance metabolic fates of fatty acids arising from the diet or adipose-depot lipolysis. Despite a lack of effect on hepatic amino acid (AA) transport, a reduction in metabolism of essential AA within the liver indicated an AA ‘sparing effect’ induced by maternal Met. Conclusions Despite greater global DNA methylation, maternal Met supply resulted in distinct alterations of hepatic transcriptome, proteome, and metabolome profiles after birth. Data underscored an effect on maintenance of calf hepatic Met homeostasis, glutathione, phosphatidylcholine and taurine synthesis along with greater efficiency of nutrient metabolism and immune responses. Transcription regulators such as FOXO1, PPARG, E2F1, and CREB1 appeared central in the coordination of effects induced by maternal Met. Overall, maternal Met supply induced better immunometabolic status of the newborn liver, conferring the calf a physiologic advantage during a period of metabolic stress and suboptimal immunocompetence.

A group of 10 multiparous Italian Holstein cows were housed in individual tied stalls and infused with 150 ml of saline (CTR; 5 cows), or of 10% solution rich in long-chained omega-3 fatty acids (n3FA; 5 cows) at 12, 24, and 48 h after calving. From −7 to 21 days from calving (DFC), the body condition score, body weight, dry matter intake (DMI), and milk yield were measured, blood samples were collected to assess the plasma fatty acids (FA) and metabolic profiles, and milk samples were collected to assess the milk composition. Data underwent a mixed model for repeated measurements, including the treatment and time and their interactions as fixed effects. Plasma FA profile from n3FA cows had lower myristic and higher myristoleic proportions, higher cis-11,14-eicosadienoic acid and monounsaturated FA proportions at 3 DFC, and lower cis-10-pentadecanoic proportion at 10 DFC. Besides these, n3FA cows had higher eicosapentaenoic (EPA) and docosahexaenoic (DHA) proportions (1.09 vs. 0.71 and 0.33 vs. 0.08 g/100 g), confirming the effectiveness of the infusion in elevating plasma availability of these FA. The plasma metabolic profile from n3FA cows revealed a tendency toward a lower concentration of reactive oxygen metabolites at 1 DFC and lower haptoglobin at 2 and 3 DFC, reflecting a mitigated inflammatory state. Furthermore, n3FA cows had a higher DMI during the first week of lactation. Higher DMI of n3FA could account for the changes detected on their plasma FAs, the higher milk yield they had at 1 and 2 DFC, the reduced lactose and urea nitrogen content in their milk. Higher DMI could also account for the lower plasma urea that n3FA cows had at 1 and 2 DFC, suggesting a lower amount of endogenous amino acids deserved to gluconeogenic fate. Milk from n3FA cows had lower rennet clotting time and higher curd firmness, which is probably driven by a higher EPA and DHA inclusion in the milk fat. Together, these outcomes suggest that the infusion exerts a short-term anti-inflammatory action on dairy cows at the onset of lactation.

An unsupervised machine learning approach (ML) of rumination time (RT) data was used to evaluate metabolic and lactation performance in a cohort of Simmental dairy cows (29) around parturition. The k-means clustering (unsupervised ML) was used to generate clusters based on RT (Hr-Tags; SCR by Allflex) over 24 h from 1 to 7-d post-calving. Two large clusters were obtained: high (High-RT, n = 13) and low (Low-RT, n = 12). Milk production was recorded until 42 d in lactation and milk samples collected weekly. Blood samples were collected from −21 to +28 d relative to calving. Data were subjected to PROC MIXED of SAS. Compared with Low-RT, High-RT cows had lower plasma BHB (3, 7, and 14 d), haptoglobin (3 and 7 d), and non-esterified fatty acids (1 and 3 d). High-RT cows had greater tocopherol from 3 to 28 d, fructosamine and albumin at 7 d, and retinol at 3 and 7 d after calving than Low-RT cows. Compared with multiparous Low-RT, milk production was greater in multiparous High-RT cows, but no differences were detected for primiparous. Only for multiparous, High-RT cows displayed a greater LFI than Low-Rt cows. The close relationship between RT and the physiological state at the onset of lactation supports the use of RT as an indicator of metabolic and inflammatory adaptations to the negative energy balance of cows after parturition. At the farm level, these outcomes provide information to farmers that can be helpful in management decisions for cow health, complementing traditional methods.HIGHLIGHTS The unsupervised ML applied was able to group cows with different RT increase rates after calving based on differences in plasma biomarkers of energy metabolism, inflammatory response, and liver functionality, particularly in multiparous cows. A quicker increase in RT after calving was associated with a lower inflammatory response, lower lipid mobilisation, and greater milk production. At the farm level, the fine-tuning of specific algorithm in the actual sensors considering the rate of increase of RT after calving can be helpful in management decisions for cow health, complementing traditional methods to better monitor early lactation dairy cows.

Anticoagulants, such as ethylenediaminetetraacetic acid (EDTA), sodium citrate (Na-citrate), or heparin are normally used in hematological clinical tests to prevent coagulation. Although anticoagulants are fundamental for the correct application of clinical tests, they produce adverse effects in different fields, such as those involving specific molecular techniques; for instance, quantitative real time polymerase chain reactions (qPCR) and gene expression evaluation. For this reason, the aim of this study was to evaluate the expression of 14 genes in leukocytes that were isolated from the blood of Holstein cows, and which were collected in Li-heparin, K-EDTA, or Na-citrate tubes; then, they were analyzed using qPCR. Only the SDHA gene showed a significant dependence (p ≤ 0.05) on the anticoagulant that was used with the lowest expression; this was observed in Na-Citrate after being compared with Li-heparin and K-EDTA (p < 0.05). Although a variation in transcript abundance with the three anticoagulants was observed in almost all the investigated genes, the relative abundance levels were not statistically significant. In conclusion, the qPCR results were not influenced by the presence of the anticoagulant; thus, we had the opportunity to choose the test tube that was used in the experiment without interfering effects impacting the gene expression levels caused by the anticoagulant.

This study evaluated the effects of feeding a commercial yeast culture on blood biomarkers and polymorphonuclear leukocyte (PMNL) gene expression in dairy cows during the transition period until 50 d postpartum. Forty Holstein dairy cows were used in a randomized complete block design from −30 to 50 d. At −30 d, cows were assigned to a basal diet plus 114 g/d of top-dressed ground corn (control; n = 20) or 100 g/d of ground corn and 14 g/d of a yeast culture product (YC; n = 20). Blood samples were collected at various time points from −30 to 30 DIM to evaluate blood biomarkers and PMNL gene expression related to inflammation, liver function, and immune response. Liver function biomarkers, gamma-glutamyl transferase (GGT) and albumin were greater and lower, respectively, in YC cows in comparison to control. However, these biomarkers remained within physiological levels, indicating an active inflammatory process. Genes in PMNL expression related to inflammation (NFKB1, TNFA, TRAF6), anti-inflammation (IL10), and cell membrane receptors (SELL) were upregulated in the YC group in comparison to control. These results suggest that YC could stimulate a more active inflammatory response with signs of a resolution of inflammation in transition cows.

Our study investigates metabolic changes occurring at dry-off and the contribution of milk yield (MY) in such alterations. Thirteen Holsteins were dried off at 55 days from expected calving day (assumed as 0 days from dry-off, DFD) and divided in two groups according to their average daily MY in the last week of lactation, assuming a cut-off of 15 kg·d−1: low MY (7 cows) and high MY (6 cows). From −7 to 34 DFD dry matter intake (DMI) and rumination time were measured. Blood samples were collected at −7, 2, 7, 27 and 34 DFD to assess an haematological and metabolic profile and at −7, 7 and 34 DFD to test functions of circulating white blood cell (WBC) through ex vivo challenges. Data were included in a mixed model for repeated measures assuming MY at dry-off, time and their interaction as fixed effects. After dry-off, DMI was reduced and rumination time was increased in all the animals. High MY cows had greater DMI and rumination time than low MY cows. In blood, WBC counts decreased at 7 DFD and increased the production of pro-inflammatory cytokines at 7 and 34 DFD. Plasmatic concentrations of liver enzymes indicators, positive acute phase proteins (APPs); and nitrogen species increased after dry-off. Conversely, negative APPs and antioxidant species decreased. Those alterations were more marked in high MY animals. This study suggests that dry-off decreased liver function, triggered a systemic inflammation and depleted antioxidant systems, especially in the group of cows with high MY at dry-off.Highlights Inflammation, liver dysfunctions and altered redox balance has been detected after dry-off in all the animals. Cows with highest milk yield before halting of milk removal faced the most severe metabolic challenges. Such founding indicates the management of dry-off as a key point for dairy cows health.

Abstract The transition period of dairy cattle is characterized by a number of metabolic, endocrine, physiologic, and immune adaptations, including the occurrence of negative energy balance, hypocalcemia, liver dysfunction, overt systemic inflammatory response, and oxidative stress status. The degree and length of time during which these systems remain out of balance could render cows more susceptible to disease, poor reproductive outcomes, and less efficient for milk production and quality. Studies on both monogastrics and ruminants have reported the health benefits of nutraceuticals (e.g. probiotics, prebiotics, dietary lipids, functional peptides, phytoextracts) beyond nutritional value, interacting at different levels of the animal’s physiology. From a physiological standpoint, it seems unrealistic to disregard any systemic inflammatory processes. However, an alternate approach is to modulate the inflammatory process per se and to resolve the systemic response as quickly as possible. To this aim, a growing body of literature underscores the efficacy of nutraceuticals (active compounds) during the critical phase of the transition period. Supplementation of essential fatty acids throughout a 2-month period (i.e. a month before and a month after calving) successfully attenuates the inflammatory status with a quicker resolution of phenomenon. In this context, the inflammatory and immune response scenario has been recognized to be targeted by the beneficial effect of methyl donors, such as methionine and choline, directly and indirectly modulating such response with the increase of antioxidants GSH and taurine. Indirectly by the establishment of a healthy gastrointestinal tract, yeast and yeast-based products showed to modulate the immune response, mitigating negative effects associated with parturition stress and consequent disorders. The use of phytoproducts has garnered high interest because of their wide range of actions on multiple tissue targets encompassing a series of antimicrobial, antiviral, antioxidant, immune-stimulating, rumen fermentation, and microbial modulation effects. In this review, we provide perspectives on investigations of regulating the immune responses and metabolism using several nutraceuticals in the periparturient cow.

Abstract Background Ketosis in dairy cows is associated with body fat mobilization during the peripartal period. Sub-clinical and clinical ketosis arise more frequently in cows that are overfed energy during the entire dry (last 50 to 45 days prior to parturition) or close-up period (last ~ 28 days prepartum). Methods A retrospective analysis was performed on 12 cows from a larger cohort that were fed a higher-energy diet [1.54 Mcal/kg of dry matter (DM); 35.9% of DM corn silage and 13% of DM ground corn] during the close-up dry period, of which 6 did not develop clinical ketosis (OVE, 0.83 mmol/L plasma hydroxybutyrate; BHB) and 6 were diagnosed with clinical ketosis (KET, 1.4 mmol/L BHB) during the first week postpartum. A whole-transcriptome bovine microarray (Agilent Technologies) and metabolomics (GC-MS, LC-MS; Metabolon® Inc.) were used to perform transcript and metabolite profiling of liver tissue harvested at − 10 days relative to parturition which allowed to establish potential associations between prepartal transcriptome/metabolome profiles and susceptibility to clinical ketosis postpartum. Results Cows in KET had greater (P = 0.01) overall body weight between − 2 and 1 week around parturition, but similar body condition score than OVE. Although dry matter intake (DMI) did not differ prepartum, KET cows had lower (P

Some pasture species are rich in phytochemicals, able to improve milk yield and quality and to reduce the environmental impacts of livestock farming. The phytochemicals interact with the different gene networks within the animal, such as nuclear factor erythroid 2-related factor 2 (NRF2), but their overall impact on animal health remains to be fully understood. The objective of this study was to identify the effects of pasture Legumes and non-leguminous Forbs containing high bioactive compounds on metabolism and activity of the liver, antioxidant response, kidney function, and inflammation of dairy cows using a large array of blood parameters associated with metabolism and the innate immune system. For this purpose, 26 parameters and the concentration of certain bioactive compounds were assessed in blood plasma, collected from the Jersey cows grazing either Grass, Legume, or Forb-based pastures. In addition, serum collected from all the cows was utilized to detect the changes in NRF2 activation in bovine mammary alveolar cells (MACT) and hepatocytes. Compared with Grass, the cows that grazed both Forb and Legume pastures had lower β-hydroxybutyric acid (BHB) and creatinine and larger vitamin E and the ferric reducing ability of the plasma, supporting an improved antioxidative status for these animals. Compared with both Grass and Legume, the cows that graze Forb pasture had lower urea and urea to creatinine ratio, and lower creatinine, indicating a better kidney function. The cows grazing Legume pasture had greater hematocrit, bilirubin, cholesterol, albumin, β-carotene, retinol, and thiol groups but lower ceruloplasmin, paraoxonase, and myeloperoxidase (MPO) than those grazed Grass and Forb pastures, indicating a positive effect of Legume pasture on the liver, oxidative stress, and red blood cells. The plasma of cows in the various pastures was enriched with various isoflavonoids, especially the cows grazed on Forb and Legume pastures, which likely contributed to improving the antioxidative status of those cows. However, this effect was likely not due to the higher activation of NRF2. Overall, these results indicate that Forb and Legume pastures rich in secondary metabolites do not strongly affect the metabolism but can improve the status of the liver and the kidney and improve the efficiency of N utilization and antioxidant response, compared with the Grass pasture.

The aim of this study was to evaluate the use of Fourier transform mid-infra-red (FT-MIR) spectrometry to analyse blood biochemical parameters of the horse. For this purpose, mid infrared transmission spectra were acquired from plasma samples from 72 healthy horses. Each sample was also analysed using reference clinical chemical methods, and these results were used as calibrating values to develop predictive models by partial least squares method. The validation was carried out using external validation method. The coefficient of determination (R2) and the ratio of prediction to deviation (RPD) showed high values for parameters regarding energy and protein metabolism. Among energy parameters, an excellent prediction model was developed for total cholesterol (R2 = 0.94; RPD = 4.40) and triglycerides (R2 = 0.96; RPD = 5.0) while fair results were obtained for cholesterol fractions (R2 range: 0.75–0.80; RPD range: 2.0–2.3). Among protein metabolism parameters, excellent prediction models were developed for total protein, albumin, globulin (R2 range: 0.96–0.99; RPD range: 5.40–9.30) and good prediction model for urea (R2 = 0.90; RPD = 3.2), confirming previous results with the plasma of dairy cows. Our results highlight that FT-MIR spectrometry offers an accurate measurement of important plasma biomarkers for the evaluation of energy (cholesterol and triglycerides) and protein metabolism (urea), as well as for health status (albumin/globulin ratio). Our results may open an interesting perspective of a more cost-effective approach to monitoring the metabolic status and health conditions of the horse, with the future possibility to predict some blood biomarkers by the practitioner in field.Highlights FT-MIR potential to measure blood parameters in horses was explored; Infra-red spectrometry can be used in horse’s clinical chemistry; Fast and cost-effective metabolic status evaluation in horses; Accurate FT-MIR predictions for plasma protein and lipid fractions in horses.

Two independent trials were carried out to test the effectiveness of different environmental enrichments in improving Italian heavy pigs’ welfare. Eighty undocked Landrace × Large White barrows (body weight range: 27.1–158.4 kg) were used. In Trial 1, group C1 received a metal chain and group WL wooden logs (placed inside a specifically designed metal rack). In trial two, the enrichments proposed were a hanging chain (C2), and a vegetal edible block (EB) inside the metal rack. General health, animal behaviour, hair cortisol, blood parameters and growth performance were recorded. In Trial 1, WL pigs interacted with the enrichment at a lower extent than C1 (p

The increased resistance to disease observed after monensin treatment could reflect a reduction of inflammation and oxidative stress. We hypothesised that if monensin supplementation was given before calving, it would help in controlling inflammation, reduce the susceptibility to disease and increase the performance. Fourteen pregnant heifers (PR) and 24 multiparous cows (ML) were randomly assigned to a treated group (Mon) or a control group (Ctr). The Mon group received 32.4 g of monensin in a controlled-release capsule 21 days before calving (expected release rate, 335 mg/d for 95 days). Their health status, rumination activity, and plasma parameters were monitored from –28 to 56 days from calving. The milk yield (MY), milk composition, and cheese-making properties were also monitored. Rumen samples were collected at 30 days from calving to assess the volatile fatty acids composition and investigate immunological parameters. After calving, the Mon group had fewer clinical diseases, an increased rumination time, and a higher MY. Monensin reduced the infiltration of both T and B cells in rumen fluid. In ML, the Mon group had lower levels of β-hydroxybutyrate in the early postpartum period and a lower level of total reactive oxygen species. Of PR, the Mon group had a tendency for lower levels of nonesterified fatty acids, higher levels of ceruloplasmin after the first month of lactation, a tendency for lower levels of paraoxonase, higher levels of γ-glutamyl transferase and higher levels of total reactive oxygen species. Monensin treatment decreased the cheese-making properties in the milk of PR.Highlights Monensin improved the performance of all the animals and decreased the disease incidence in all of them. Monensin heightened the inflammatory and oxidative stress status and reduced the cheese-making properties in pregnant heifers. Although different effects were seen in cows with different parity, dairy cows generally beneficed of monensin administration.

Weaning plays a key role in health status and future performance of calves. The aim of this study was to investigate the effects of weaning age (Wa), early (45 d, EW) or conventional (60 d, CW), on growth performance and metabolic profile of ten Simmental calves (5 EW and 5 CW calves). Daily intake of milk and calf starter was recorded. Blood samples and measurements of body weight (BW), heart girth (HG), and wither height (WH) were collected at −25, −15, 0, 6, and 20 days relative to weaning. Growth performances (BW, HG, WH) were affected by Wa, resulting lower in EW calves compared with CW calves (p < 0.05). Average daily gain was affected by overall Wa and Time but also by the interaction Wa × Time (p < 0.05). EW calves had lower paraoxonase and higher oxidation protein products levels, lower glucose levels in the post-weaning period, lower Ca and cholesterol levels at 20 d after weaning, and higher GGT activity at −25 d from weaning (p < 0.05). A significant interaction effect between Wa and Time was reached for glucose, Ca, cholesterol. In conclusion, weaning Simmental calves at approximately six weeks of age might not affect inflammatory status and liver functionality after weaning. As secondary outcome, even though the low number of animals could represent a limitation, the average daily gain obtained by Simmental calves weaned at 45 d supported this strategy (despite the lower body weight at weaning and after was due only to the age difference of 15 days). Hence, in order to reduce rearing costs, early weaning for Simmental calves (dual-purpose breed, milk and beef) might not jeopardize calf development, as long as calves can reach body gains as reported in the present study.

Dairy cows have high incidences of metabolic disturbances, which often lead to disease, having a subsequent significant impact on productivity and reproductive performance. As the milk fatty acid (FA) profile represents a fingerprint of the cow’s nutritional and metabolic status, it could be a suitable indicator of metabolic status at the cow level. In this study, we obtained milk FA profile and a set of metabolic indicators (body condition score, ultrasound liver measurements, and 29 hematochemical parameters) from 297 Holstein–Friesian cows. First, we applied a multivariate factor analysis to detect latent structure among the milk FAs. We then explored the associations between these new synthetic variables and the morphometric, ultrasonographic and hematic indicators of immune and metabolic status. Significant associations were exhibited by the odd-chain FAs, which were inversely associated with β-hydroxybutyrate and ceruloplasmin, and positively associated with glucose, albumin, and γ-glutamyl transferase. Short-chain FAs were inversely related to predicted triacylglycerol liver content. Rumen biohydrogenation intermediates were associated with glucose, cholesterol, and albumin. These results offer new insights into the potential use of milk FAs as indicators of variations in energy and nutritional metabolism in early lactating dairy cows.

Abstract Background Pregnancy and early life are critical periods of plasticity during which the fetus and neonate may be influenced by environmental factors such as nutrition. Maternal methionine (Met) supply in non-ruminants during pregnancy can affect offspring development and growth. Thus, the objective of this study was to investigate if increasing Met supply during late-pregnancy affects developmental parameters of the calf at birth and if either maternal Met or colostrum from Met-fed cows alters calf growth. Calves born to Holstein cows individually-fed a basal control [CON; 1.47 Mcal/kg dry matter (DM) and 15.3% crude protein] diet with no added Met or CON plus ethylcellulose rumen-protected Met (MET; Mepron® at 0.09% of diet DM; Evonik Nutrition & Care GmbH, Germany) during the last 28 ± 2 d of pregnancy were used. A total of 39 calves were in CON (n = 22 bulls, 17 heifers) and 42 in MET (n = 20 bulls, 22 heifers). At birth, calves were randomly allocated considering dam treatment and colostrum as follows: 1) calves from CON cows and colostrum from CON cows (n = 21); 2) calves from CON cows and colostrum from MET cows (n = 18); 3) calves from MET cows and colostrum from MET cows (n = 22); and 4) calves from MET cows and colostrum from CON cows (n = 20). All calves were housed, managed, and fed individually during the first 9 wk of life. Results Despite greater daily DM intake pre-partum in cows fed MET (15.7 vs. 14.4 ± 0.12 kg/d, P 0.05) due to maternal Met supply or colostrum source. However, fecal scores tended to be lower (P ≤ 0.10) in MET calves regardless of colostrum source. Conclusions Increasing the maternal supply of MET during late-pregnancy enhanced growth in utero as well as during the pre-weaning and early post-weaning periods. Although the ~ 1 kg/d greater DM intake during the last 2–3 wk prior to parturition could explain a portion of the 2 kg extra body mass of MET calves at birth, other mechanisms potentially encompassing nutrient assimilation efficiency likely played a role. Assessing the exact mechanisms sensitive to supply of Met or total amino acid supply during the latter stages of growth in utero merit further research.

Abstract Background Animal studies show that high fat (HF) diet-induced gut microbiota contributes to the development of obesity. Oil composition of high-fat diet affects metabolic inflammation differently with deleterious effects by saturated fat. The aim of the present study was to examine the diversity and metabolic capacity of the cecal bacterial community in C57BL/6 N mice administered two different diets, enriched respectively with coconut oil (HFC, high in saturated fat) or soy oil (HFS, high in polyunsaturated fat). The relative impact of each hypercaloric diet was evaluated after 2 and 8 weeks of feeding, and compared with that of a low-fat, control diet (LF). Results The HFC diet induced the same body weight gain and fat storage as the HFS diet, but produced higher plasma cholesterol levels after 8 weeks of treatment. At the same time point, the cecal microbiota of HFC diet-fed mice was characterized by an increased relative abundance of Allobaculum, Anaerofustis, F16, Lactobacillus reuteri and Deltaproteobacteria, and a decreased relative abundance of Akkermansia muciniphila compared to HFS mice. Comparison of cecal microbiota of high-fat fed mice versus control mice indicated major changes that were shared between the HFC and the HFS diet, including the increase in Lactobacillus plantarum, Lutispora, and Syntrophomonas, while some other shifts were specifically associated to either coconut or soy oil. Prediction of bacterial gene functions showed that the cecal microbiota of HFC mice was depleted of pathways involved in fatty acid metabolism, amino acid metabolism, xenobiotic degradation and metabolism of terpenoids and polyketides compared to mice on HFS diet. Correlation analysis revealed remarkable relationships between compositional changes in the cecal microbiota and alterations in the metabolic and transcriptomic phenotypes of high-fat fed mice. Conclusions The study highlights significant differences in cecal microbiota composition and predictive functions of mice consuming a diet enriched in coconut vs soy oil. The correlations established between specific bacterial taxa and various traits linked to host lipid metabolism and energy storage give insights into the role and functioning of the gut microbiota that may contribute to diet-induced metabolic disorders.

Dairy cows at dry-off undergo several management and physiological changes, resulting in alterations in plasma biomarkers of inflammation, oxidative stress, and immune system. High milk yield at the end of lactation exacerbates these responses. The underlying mechanism of these changes has yet to be elucidated. We hypothesized altered leukocyte gene expression after dry-off and different responses in cows with different milk yield. Thirteen Holstein dairy cows were sampled at the turn of dry-off to investigated whole blood leukocyte gene expression and were grouped according to the average milk yield during the last week of lactation: low (< 15 kg/d) and high milk yield (> 15 kg/d). Blood samples were collected in PAXgene tubes (Preanalytix, Hombrechtikon, Switzerland) at -7, 7, and 34 days from dry-off (DFD) to measure mRNA abundance of 37 genes. Normalized gene abundance data were subjected to MIXED model ANOVA (SAS Institute Inc., Cary, NC). Compared with -7 DFD, at 7 DFD RNA abundance of lipoxygenase genes (ALOX5, ALOX15) and myeloperoxidase (MPO) increased, and that of the antioxidant gene (SOD2) decreased. Meanwhile, genes related to recognition and immune mediation (CD16, MYD88, TLR2), migration and cell adhesion (CX3CR1, ITGAL, ITGB2, TLN1), and the antimicrobial gene MMP9 were downregulated at 7 or 34 DFD, whereas the antimicrobial IDO1 gene was upregulated. Compared with low-producing cows, cows with high milk yield at dry-off cows had upregulated expression of the pro-inflammatory cytokines IL8 and IL18 and a greater reduction in transcript abundance of the toll-like receptor (TLR) recognition-related gene TLR2. Overall, the dry-off confirmed to be a phase of intense changes, triggering an inflammatory response and somewhat suppressing leukocyte immune function. In cows with high milk yield during the week before dry-off, the inflammatory response was exacerbated.

Abstract Background Previous research has revealed a strong inflammatory response within adipose (AT) tissue during the transition into lactation. Whether this effect is a result of oxidative stress induced by lipolysis and fatty acid oxidation associated with differences in prepartum body condition score remains to be determined. The objectives of this study were to investigate systemic biomarkers of energy balance and inflammation and the expression of lipid metabolism- and inflammation-related genes in AT during the transition period in dairy cows. Results Twenty multiparous Holstein cows were retrospectively divided by body condition score (BCS) prior to parturition into two groups (10 cows/group): BCS ≤ 3.25 (LoBCS) and BCS ≥ 3.75 (HiBCS). Subcutaneous adipose tissue was biopsied from the tail-head region at d − 10, 7 and 20 relative to parturition. Plasma was used to evaluate biomarkers of energy balance (EBAL) [free fatty acids (NEFA), glycerol, insulin] and inflammation [IL-1β, haptoglobin, myeloperoxidase, and reactive oxygen metabolites (ROM)]. Although insulin concentration was not affected by BCS, NEFA was overall greater and glycerol lower in HiBCS cows. Greater activity of myeloperoxidase in plasma coincided with increased haptoglobin and IL-1β postpartum in LoBCS cows. Among genes related with oxidative stress, the expression of the cytosolic antioxidant enzyme SOD1 was greater in LoBCS compared to HiBCS. Cows in LoBCS compared with HiBCS had greater overall expression of ABDH5 and ATGL along with ADIPOQ, indicating enhanced basal lipolysis and secretion of adiponectin. Expression of CPT1A, ACADVL, and ACOX1 was greater overall in HiBCS than LoBCS indicating enhanced NEFA oxidation. Although the temporal increase in plasma NEFA regardless of BCS coincided with the profile of CPT1A, the gradual decrease in genes related with re-esterification of NEFA (PCK1) and glycerol efflux (AQP7) coupled with an increase in glycerol kinase (GK) suggested some stimulation of NEFA utilization within adipose tissue. This idea is supported in part by the gradual decrease in insulin regardless of BCS. Although expression of the inflammation-related gene toll-like receptor 4 (TLR4) was greater in HiBCS versus LoBCS cows at −10 d, expression of TLR9 was greater in HiBCS versus LoBCS at 20 d. These profiles did not seem to be associated with concentrations of pro-inflammatory biomarkers or ROM. Conclusions Overall, data indicated that cows with BCS 3.25 or lower before calving experienced greater alterations in systemic inflammation and basal lipolysis without excessive increases in NEFA plasma concentrations. Despite the greater plasma NEFA around parturition, cows with BCS 3.75 or higher seemed to have a more active system for catabolism of NEFA and utilization of glycerol within adipose tissue. A linkage between those pathways and risk of disorders postpartum remains to be determined.

This study aimed to evaluate the effects of early life fecal microbiota transplantation (FMT) on the health and performance of neonatal dairy calves. The donor was selected based on health and production records and fecal material testing negative for infectious pathogens. Sixteen healthy newborn Holstein calves were randomized to either a baseline nutritional program (CON) or 1×/d inoculations with 25 g of fecal donor material (FMT) mixed in the milk replacer (n = 8/TRT) from 8 to 12 days of age. Blood and fecal samples were collected weekly, and calves were weaned at 7 weeks of age. A TRT × Week interaction was observed in haptoglobin, which was reflected in a positive quadratic effect in FMT calves but not in CON. A trend for a TRT × Week interaction was observed in the liver function biomarker paraoxonase, which resulted in greater paraoxonase in FMT calves than CON at three weeks of age. Fecal microbial community analysis revealed a significant increase in the alpha-diversity between week 1 and week 5 for the FMT calves. These results suggest that early life FMT in neonatal calves has positive effects in mediating the inflammatory response and gut microbial maturation.

The objective of the present study was to evaluate the effects of an antioxidant and anti-inflammatory compound found in rosemary plants (Salvia rosmarinus) named carnosic acid during the transition period of dairy cows. From day 1 to 3 after calving, 16 multiparous Holstein cows received a daily intravenous infusion of either 500 mL of saline (NaCl 0.9%; Saline; n = 8) or carnosic acid at a rate of 0.3 mg/kg of BW supplied in 500 mL of saline (CA; n = 8). Blood samples were taken at –7, 2, 5, 7, 14, and 21 d relative to parturition, then analyzed for metabolites related to energy metabolism, muscle mass catabolism, liver function, inflammation, and oxidative stress. CA infusion tended to improve milk performance; however, DMI was unaffected by treatment. At 2 d relative to parturition, CA cows had lower blood concentrations of haptoglobin, paraoxonase, FRAP, and NO2– than saline cows. After treatment infusions, haptoglobin remained lower in CA cows than saline at 5 d relative to parturition. Our results demonstrate that carnosic acid promoted positive responses on inflammation and oxidative stress biomarkers and may promote beneficial effects on lactation performance in peripartal dairy cows.

Breast cancer (BC) is a major public health concern and substantial research has shown that adhering to a healthy dietary pattern, such as the Mediterranean Diet (MD), may prevent the onset of cancer and BC relapses. This study aims at specifically investigating the association of MD with circulating dietary-related biomarkers in a cohort of BC survivors. Eighty patients (mean age of 54.9 ± 10.6) with a histologically confirmed diagnosis of BC who had not received any pharmacological or radiotherapy treatment for at least two months were enrolled. Fasting serum lipid-soluble vitamins (retinol, tocopherol), plant pigments (β-carotene, lutein + zeaxanthin, cryptoxanthin, lycopene), inflammatory and oxidative stress markers (ceruloplasmin; haptoglobin; paraoxonases; reactive oxygen molecule; thiol groups, Ferric reducing antioxidant power), and cardiometabolic parameters (body mass index (BMI); glucose; insulin; HOMA-IR; total cholesterol; LDL-cholesterol; HDL-cholesterol; triglycerides) were analyzed. Adherence to the MD was assessed through the Mediterranean Diet Score (MDS) questionnaire. Fasting blood samples were collected for the evaluation of selected biomarkers. MDS resulted positively correlated with β-carotene (r 0.331; p < 0.01) and lycopene (r 0.274; p < 0.05) and negatively with retinol (r −0.346; p < 0.05). Among the investigated inflammatory biomarkers, MDS was only correlated with antioxidant capacity (r 0.256; p < 0.05), while none of the investigated cardiometabolic parameters were significantly correlated with this index. The strong significant correlation between β-carotene and MDS encourages us to consider this pro-vitamin as a putative biomarker to take into account for evaluating the adherence to the MD.

Fatty acids are known for their regulatory role in inflammation and oxidative stress. The present study investigated 38 calves born from dams, abomasally supplemented with coconut oil, essential fatty acids (EFA), conjugated linoleic acid (CLA) or EFA + CLA, according to immunological traits and the oxidative and anti-oxidative status for the first 5 days of life. On day 2 of life, plasma total bilirubin, cholesterol, interleukin 1-β and ferric ion reducing anti-oxygen power (FRAP) were lower in calves with than without maternal EFA supplementation, and FRAP additionally on day 4. On day 3, the concentrations of reactive oxygen metabolites were higher in calves with than without maternal EFA supplementation and additionally on day 5 together of retinol. Total leucocyte counts were decreased in the EFA group compared to the CLA group on day 5. Lymphocyte proportions decreased from day 1 to 5 only in the EFA + CLA group. On day 2, plasma total protein was higher in CLA and EFA + CLA than in EFA calves. Similarly, CLA calves had higher interleukin 1-β concentrations compared to EFA + CLA calves. FRAP was decreased by CLA on day 4. Overall, the maternal fatty acid supply affected the inflammatory response and the oxidative and anti-oxidative status of the neonatal offspring.

Here, we tested the changes occurring in several plasma analytes during different stages of the lactation cycle of high welfare raised multiparous Holstein cows, and provided reference intervals (RI) for plasma analytes concentrations. Eleven high-welfare farms (HWF) located in Northern Italy were selected and their herds used to recruit 361 clinically healthy cows undergoing the dry (from −30 to −10 days from real calving; DFC), the postpartum (from 3 to 7 DFC), the early lactation (from 28 to 45 DFC) and the late lactation phases (from 160 to 305 DFC). Cows affected by subclinical diseases (SCD) were retrospectively excluded, and a subset of 285 cows was selected. Data of plasma analytes underwent ANOVA testing using physiological phases as predictors. The individual effect of each phase was assessed using a pairwise t-test assuming p ≤ 0.05 as a significance limit. A bootstrap approach was used to define the reference interval (RI) for each blood analyte within physiological phases having a pairwise t-test p ≤ 0.05. The concentration of nonesterified fatty acids, albumin, cholesterol, retinol, paraoxonase and tocopherol changed throughout all the physiological phases, whereas the concentration of K, alkaline phosphatase and thiol groups remained stable. Triglycerides, Zn, and ferric ion reducing antioxidant power in the dry phase and BHB, Ca, myeloperoxidase, haptoglobin, reactive oxygen metabolites and advanced oxidation of protein product in postpartum differed compared with other physiological phases. During the dry phase, Packed cell volume, Cl, and urea concentrations were similar to during the postpartum phase. Similarly, Na, γ-glutamyl transferase and β-carotene concentrations were similar to during the early lactation phase; fructosamine and bilirubin concentrations were similar to during the late lactation phase. During the postpartum phase, fructosamine and P concentrations were similar to during the early lactation phase, and the aspartate transaminase concentration was similar to during the late lactation phase. During the early lactation phase, Mg, creatinine, total protein, globulin and ceruloplasmin concentrations were similar to during the postpartum phase, while the urea concentration was similar to during the late lactation phase. All these plasma analytes differed among the other phases. This study identifies physiological trends affecting plasma analytes concentrations during the different stages of the lactation cycle and provides a guideline for the duration and magnitude of their changes when animals are healthy and raised in optimal welfare conditions.

Stress in livestock reduces productivity and is a welfare concern. At a physiological level, stress is associated with the activation of inflammatory responses and increased levels of harmful reactive oxygen species. Biomarkers that are indicative of stress could facilitate the identification of more stress-resilient animals. We examined twenty-one metabolic, immune response, and liver function biomarkers that have been associated with stress in 416 Italian Simmental and 436 Italian Holstein cows which were genotyped for 150K SNPs. Single-SNP and haplotype-based genome-wide association studies were carried out to assess whether the variation in the levels in these biomarkers is under genetic control and to identify the genomic loci involved. Significant associations were found for the plasma levels of ceruloplasmin (Bos taurus chromosome 1—BTA1), paraoxonase (BTA4) and γ-glutamyl transferase (BTA17) in the individual breed analysis that coincided with the position of the genes coding for these proteins, suggesting that their expression is under cis-regulation. A meta-analysis of both breeds identified additional significant associations with paraoxonase on BTA 16 and 26. Finding genetic associations with variations in the levels of these biomarkers suggests that the selection for high or low levels of expression could be achieved rapidly. Whether the level of expression of the biomarkers correlates with the response to stressful situations has yet to be determined.

Abstract Background The objective of the study was to evaluate the effect of overfeeding a moderate energy diet and a 2,4-thiazolidinedione (TZD) injection on blood and hepatic tissue biomarkers of lipid metabolism, oxidative stress, and inflammation as it relates to insulin sensitivity. Results Fourteen dry non-pregnant cows were fed a control (CON) diet to meet 100% of NRC requirements for 3 wk, after which half of the cows were assigned to a moderate-energy diet (OVE) and half of the cows continued on CON for 6 wk. All cows received an intravenous injection of 4 mg TZD/kg of body weight (BW) daily from 2 wk after initiation of dietary treatments and for 2 additional week. Compared with CON cows and before TZD treatment, the OVE cows had lower concentration of total protein, urea and albumin over time. The concentration of cholesterol and tocopherol was greater after 2 wk of TZD regardless of diet. Before and after TZD, the OVE cows had greater concentrations of AST/GOT, while concentrations of paraoxonase, total protein, globulin, myeloperoxidase, and haptoglobin were lower compared with CON cows. Regardless of diet, TZD administration increased the concentration of ceruloplasmin, ROMt, cholesterol, tocopherol, total protein, globulin, myeloperoxidase and beta-carotene. In contrast, the concentration of haptoglobin decreased at the end of TZD injection regardless of diet. Prior to TZD injection, the mRNA expression of PC, ANGPTL4, FGF21, INSR, ACOX1, and PPARD in liver of OVE cows was lower compared with CON cows. In contrast, the expression of HMGCS2 was greater in OVE compared with CON cows. After 1 wk of TZD administration the expression of IRS1 decreased regardless of diet; whereas, expression of INSR increased after 2 wk of TZD injection. Cows fed OVE had lower overall expression of TNF, INSR, PC, ACOX1, FGF21, and PPARD but greater HMGCS2 expression. These differences were most evident before and after 1 wk of TZD injection, and by 2 wk of TZD differences in expression for most genes disappeared. Conclusions Based on molecular and blood data, administration of TZD enhanced some aspects of insulin sensitivity while causing contradictory results in terms of inflammation and oxidative stress. The bovine liver is TZD-responsive and level of dietary energy can modify the effects of TZD. Because insulin sensitizers have been proposed as useful tools to manage dairy cows during the transition period, further studies are required to investigate the potential hepatotoxicity effect of TZD (or similar compounds) in dairy cattle.

Aloe has long been used as a traditional medicine for its numerous beneficial properties, which are mainly ascribed to β-polysaccharides and phenolic compounds including anthraquinones, anthrones and chromones. However, few studies on large animals are currently available. The effect of whole leaf Aloe arborescens homogenate on the in vitro rumen fermentative processes was tested using alfalfa hay and barley meal as substrates. The Aloe homogeneate was added at three different concentrations (0.4, 2.0 and 10.0 g L−1 of fermentation liquid). The same homogenate was dosed (200 g) orally and through the rumen cannula to three rumen cannulated heifers and orally to six lactating dairy cows to measure the rumen degradation of aloin and the transfer of aloin from the gut into the blood, respectively. The Aloe homogenate did not affect in vitro rumen fermentations and feed digestibility. The administration of Aloe homogenate did not negatively affect animal feed intake and health neither on the cannulated heifers nor on the dairy cows. Aloin underwent a rapid degradation in the rumen milieu, and became undetectable 2 h after oral dosage. However, when Aloe homogenate was administered to dairy cows, aloin appeared in blood as early as 2 h after administration, reached a maximum after 4 h (6.2 ± 5.8 μg L−1) and progressively decreased thereafter. These results suggest that Aloe compounds can be absorbed into the blood and encourage the study of Aloe as a potential nutraceutical in ruminants. Further studies should determine the most effective in vivo dosage.

Animal health is affected by many factors such as metabolic stress, the immune system, and epidemiological features that interconnect. The immune system has evolved along with the phylogenetic evolution as a highly refined sensing and response system, poised to react against diverse infectious and non-infectious stressors for better survival and adaptation. It is now known that high genetic merit for milk yield is correlated with a defective control of the inflammatory response, underlying the occurrence of several production diseases. This is evident in the mastitis model where high-yielding dairy cows show high disease prevalence of the mammary gland with reduced effectiveness of the innate immune system and poor control over the inflammatory response to microbial agents. There is growing evidence of epigenetic effects on innate immunity genes underlying the response to common microbial agents. The aforementioned agents, along with other non-infectious stressors, can give rise to abnormal activation of the innate immune system, underlying serious disease conditions, and affecting milk yield. Furthermore, the microbiome also plays a role in shaping immune functions and disease resistance as a whole. Accordingly, proper modulation of the microbiome can be pivotal to successful disease control strategies. These strategies can benefit from a fundamental re-appraisal of native cattle breeds as models of disease resistance based on successful coping of both infectious and non-infectious stressors.

The anti-hyperlipidemic and anti-inflammatory effects exerted by Aloe on monogastric mammals suggest it as a potential strategy to address the tremendous metabolic alterations that affect dairy cows during their transition to calving. A group of 20 multiparous Italian Holstein dairy cows were housed in freestalls and allocated into two homogeneous groups to receive either 200 g/d of water (CTR) or 200 g/day of Aloe arborescens Mill. whole plant homogenate through a rumen tube (AAM) between −14 and 14 days from calving (DFC). From −14 to 35 DFC, the BCS, and milk yield were measured, and blood samples were collected to assess the hematochemical profile. Data underwent ANOVA testing using a mixed model for repeated measurements, including the treatment and time and their interactions as fixed effects. Compared to CTR cows, AAM cows had a less pronounced BCS loss in early lactation (p < 0.01), indicating less mobilization of body reserves. Compared to CTR cows, AAM cows had a lower plasma concentration of nonesterified fatty acids and beta hydroxybutyrate (p < 0.01 and = 0.01 respectively) that, paired with the lower butterfat content and fat/protein ratio in their milk (p = 0.03 and < 0.01 respectively), indicates that Aloe reduced the mobilization of body fats. AAM cows had a reduced concentration of myeloperoxidase in plasma and a lower SCC in milk compared to CTR cows (p = 0.02 for both), indicating an anti-inflammatory effect of Aloe. Furthermore, AAM cows had a lower plasma concentration of ceruloplasmin (p < 0.05) and higher plasma concentration of cholesterol, retinol, and paraoxonase compared to CTR cows (p < 0.01, < 0.01 and < 0.05 respectively), indicating Aloe was effective in mitigating the acute phase response in early lactation. Finally, AAM cows had lower plasma creatinine concentrations around calving (p < 0.05), a lower concentration of plasma bilirubin, and a higher concentration of plasma tocopherol compared to CTR cows (p = 0.01 for both). These data suggest Aloe has anti-hyperlipidemic and anti-inflammatory effects on transition dairy cows that could have ameliorated liver and kidney function disruption and increased the availability of body antioxidants in early lactation.

Pegbovigrastim is a commercial long-acting analog of bovine granulocyte colony-stimulating factor (rbG-CSF) that promotes the increased count and functionality of polymorphonuclear cells in dairy cows around the time of parturition. We hypothesized that pegbovigrastim administered to periparturient cows at approximately seven days before parturition and within 24 hours after calving could affect the profiles of gene networks involved in leukocyte function. Blood was collected on Day 3 after calving from treated groups (pegbovigrastim (PEG); 13 Simmental (seven multiparous and six primiparous) and 13 Holstein (seven multiparous and six primiparous) cows) that received pegbovigrastim (Imrestor; Elanco Animal Health) and controls (CTR; 13 Simmental (seven multiparous and six primiparous) and 13 Holstein (six multiparous and seven primiparous) cows) that received saline solution. Blood from all cows was sampled from the jugular vein in a PAXgene Blood RNA System tube (Preanalytix, Hombrechtikon, Switzerland) for RNA extraction. The RT-qPCR analysis was performed to investigate a panel of 34 genes of interest, representing recognition, immune mediation, migration, cell adhesion, antimicrobial strategies, inflammatory cascade, oxidative pattern, and leukotrienes in whole blood leukocytes. Normalized data were subjected to the MIXED model of SAS (ver. 9.4) with treatment, breed, parity, and their interaction as fixed effects. Compared with CTR, whole blood leukocytes of PEG cows had higher expression of genes involved in recognition and immune modulation (CD14, CD16, MYD88, TLR2, and TLR4), cell adhesion (ITGB2, ITGAL, TLN1, SELL, SELPLG, and CD44), antimicrobial activity (MMP9, LTF, and LCN2), and inflammatory cascade (CASP1, TNFRSF1A, IL1B, IL1R, IL18, IRAK1, NLRP3, and S100A8). This suggested an improvement of migration, adhesion, and antimicrobial ability and an enhanced inflammatory response, which in turn could trigger immune cell activation and enhance function. Expression of SOD2 and ALOX5 was also greater in the PEG group. In contrast, compared with CTR cows, PEG led to lower expression of RPL13A, ALOX15, IL8, and TNF. Overall, leukocytes from Simmental compared with Holstein cows had greater expression of IDO1, RPL13A, ALOX5, CD44, CX3CR1, ITGB2, and TNFA, whereas expression of CD16 and TLR2 was lower. Overall, compared with multiparous cows, primiparous cows had higher expression of IL1B, IL18, MYD88, SELL, and TLR2 and lower expression of MMP9. Simmental cows seemed more sensitive to induction of the immune system after calving, as revealed by the greater abundance of genes involved in immune system adaptation, regardless of pegbovigrastim treatment. Primiparous cows undergoing a new stress condition with respect to older cows were characterized by leukocytes with a higher inflammatory response. In conclusion, pegbovigrastim led to higher expression levels of most genes involved in the processes investigated, suggesting a thorough activation of the immune machinery during the critical post-partum period.