Weaning represents a critical transition in dairy calves, involving simultaneous dietary, social, and environmental changes. This study investigated how the weaning affects immune-related gene expression in circulating leukocytes of Holstein calves and whether post-weaning growth performance is associated with distinct immunometabolic responses. Forty female calves were retrospectively classified into two groups based on average daily gain (ADG) during the 10 days following complete weaning (from 60 to 70 days of age). Calves were fully weaned at 60 days of age using a step-down approach. Animals were categorized as high ADG (HIGH, n = 20) or low ADG (LOW, n = 20) Blood samples collected at 60 and 70 days were analyzed for plasma metabolites and leukocyte transcriptomic profiles targeting 37 genes involved in innate immunity, inflammation, oxidative stress, and metabolic regulation. Weaning induced a marked activation of innate immune pathways in all calves, as evidenced by increased expression of CD14, NFKB1, IL1β, TNFα, and genes related to cell adhesion and trafficking. However, LOW calves exhibited greater transcriptional responses, with higher expression of TLR2, CASP1, IL18, CD16, and markers of oxidative (SOD2, HSPA5) at 70 days, indicating a metabolic and transcriptional profile consistent with a greater physiological burden. Additionally, at 70 days, LOW calves had lower plasma glucose, insulin, triglycerides, and cholesterol, along with higher urea and creatinine, while β-hydroxybutyrate did not differ between groups, suggesting reduced energy availability and increased reliance on amino acid catabolism. These findings suggest that calves with lower post-weaning growth exhibit greater immune activation and altered metabolic status, highlighting variability in resilience to weaning stress. Understanding such differences may support the development of targeted nutritional or management strategies to improve early-life adaptation and long-term productivity in dairy systems.

Heat stress (HS) disrupts oxidative homeostasis and impairs immune function in dairy cows. In this context, plant polyphenols, when included in the diet, may modulate antioxidant defenses and immune response, including the antioxidant ability of leukocytes. This study evaluated whether supplementation with rumen-protected standardized dry grape extract (Nor-Grape BP-O; Nor-Feed) modulates antioxidant and immune-related gene expression in circulating leukocytes of mid-lactating Fleckvieh cows under natural HS. Thirty cows were randomly assigned to a control diet (CTR; n = 15) or to the same diet supplemented with 470 mg/cow per day of Nor-Grape BP-O (NG-BPO; n = 15) and monitored for 35 d during summer. Blood samples were collected at 0, 17, and 35 d for leukocyte isolation and quantitative PCR analysis. Relative gene expression was calculated using LinReg software and then transformed into ln fold change (lnFC) relative to 0 d. Data were subjected to PROC GLIMMIX procedure of SAS version 9.4 (SAS Institute). Overall, NG-BPO cows showed higher lnFC expression of glutathione peroxidase 1 (GPX1) and glutathione-disulfide reductase (GSR) than CTR cows. The lnFC superoxide dismutase 1 (SOD1) expression tended to be greater in NG-BPO cows compared with CTR cows, whereas superoxide dismutase 2 (SOD2) expression was unaffected by the supplementation. Among immune-related genes, interleukin-1β (IL1B) had an overall greater lnFC expression in NG-BPO cows, particularly at d 35, when a heat wave occurred, compared with CTR cows. A similar pattern was observed for lnFC interleukin-10 (IL10) expression, which was greater at 35 d in NG-BPO cows compared with CTR cows. In contrast, toll-like receptor 2 (TLR2), myeloid differentiation primary response 88 (MYD88), interleukin-1 receptor (IL1R), and glucose transporter 3 (GLUT3) were not affected by supplementation. The early upregulation of GPX1 and GSR at 17 d suggests a transitory activation of oxidative detoxification pathway driven by polyphenols, followed by downregulation at 35 d. Concomitantly, the increased IL10 and IL1B expression at 35 d in NG-BPO cows may suggest an enhanced capacity to regulate inflammatory signaling and maintain immune homeostasis, by preventing the negative effect of excessive immune activation. Overall, rumen-protected standardized dry grape extract supplementation was associated with a positive modulation of leukocyte genes involved in antioxidant defense system and regulation of inflammation, providing new molecular insight supporting previously reported improvements in physiological and immunometabolic responses during HS.

We evaluated the milk biomarkers cathelicidin (Cath), haptoglobin (Hp), milk serum amyloid A (M-SAA), N-acetyl-β-D-glucosaminidase (NAGase), lysozyme (LZ), and the half-udder milk somatic cell count (H-SCC), for their potential association with intramammary infections (IMI) in dairy goats across lactation stages. Half-udder milk from 105 goats was collected during early (39 ± 13 days in milk, DIM), mid (136 ± 12 DIM), and late (269 ± 12 DIM) lactation stages and analyzed for bacteriological culture (BC), H-SCC, and potential biomarker reactivity. Predictive performance was evaluated with BC as the outcome, and Dairy Herd Improvement records provided contextual production data. Cath positivity, assessed by western blot, showed the strongest association with positive BC in early lactation and performed well also in mid-lactation. A dynamic H-SCC threshold was reliably correlated with positive BC in early and mid-lactation. Hp positivity, assessed by Western blot, was associated with BC in mid-lactation, while NAGase, assessed by enzymatic assay, showed no consistent association. M-SAA, assessed by ELISA, suffered technical limitations due to a lack of dilution linearity, and LZ, assessed by enzymatic assay, showed no association with BC. Our data suggest stage-dependent associations and support further validation of Cath, Hp, and SCC as potential components of combined diagnostic panels, particularly in early and mid-lactation. However, a semi-quantitative Western blot assessment for Cath and Hp, as well as technical limitations for M-SAA, warrant cautious interpretation and further analytical validation.

Drying-off high-producing dairy cows can induce inflammatory responses associated with mammary involution, particularly when milk yield remains high at milking cessation. Nutrient restriction is often used to reduce milk yield to approximately 15 kg/d before dry-off, a level considered favorable for mammary involution. However, restriction can also alter metabolic status during late lactation and the effects of this strategy on systemic inflammatory status and liver function during the subsequent transition period remain unclear. The objective of this study was to determine whether a severe reduction in dietary nutrient supply before dry-off affects systemic inflammatory status, liver function, and selected immune-related indicators during dry-off and the subsequent transition period. Thirty-two Holstein cows arranged in 16 pairs were initially enrolled and assigned at the pair level to either receive ryegrass hay ad libitum as the sole feed for 7 d before dry-off (NR) or to continue receiving the lactation diet (CTR). After exclusion of cows that did not meet the inclusion criteria, 26 cows producing more than 15 kg/d remained in the final data set (NR, n = 13; CTR, n = 13). Treatments were applied at the pair level, whereas biological measurements were collected at the individual-cow level. After dry-off, carried out 55 d before the expected calving, both groups received only grass hay for 7 d, and free access to water was always provided. Blood samples were collected at -7, -3, 0, 1, 4, 7, 14, and 28 d from dry-off (DFD), and -14, -3, 3, 7, 14, and 28 d from calving (DFC). Milk samples were collected at ‒7, ‒3, and 0 DFD and 7, 14, and 28 DFC for SCC determination, bacteriological culture, and flow cytometry analysis. Restriction reduced milk yield by 62% before dry-off. Dry-off significantly altered blood cell count and plasma concentrations of inflammatory biomarkers. Milk SCC increased during restriction but was similar afterward. Overall differences between treatments in blood cell counts were limited. Similarly, no differences in CD3+, CD4+, CD8+, and CD11b+ milk cell populations were detected. Compared with CTR, in NR monocyte count was lower and eosinophils higher at 0 DFD. Lower haptoglobin and ROM concentrations at 4 DFD suggested a different temporal pattern of the systemic response after dry-off, potentially consistent with an earlier onset of involution-related changes. Nevertheless, after calving, NR cows had lower albumin and paraoxonase, and higher globulin, haptoglobin, bilirubin, AST-GOT, and myeloperoxidase at key time points, consistent with a less favorable inflammatory and hepatic biomarker profile. These findings, together with the previously reported reduction in DMI in NR cows, suggest that severe nutrient restriction before dry-off may alter the timing of dry-off-related responses and impair metabolic and inflammatory adaptation during the subsequent lactation. Further research is needed to decipher the mechanism underlying these long-term effects.

The transition period in dairy cows is characterized by temporary decline in immune efficiency, increasing susceptibility to metabolic and infectious diseases. Pro-inflammatory cytokines (PIC) play central role in coordinating immune responses and initiating the acute phase reaction (APP). This study aimed to determine whether calving itself influences PIC release and to explore the association between cytokine dynamics, metabolic changes, and development of clinical disorders. Blood samples were collected from 12 Holstein Friesian cows from -7 to 7 days relative to calving and analysed for plasma concentrations of interleukin-1β (IL-1β), interleukin-6 (IL-6), and tumor necrosis factor-α (TNF-α). Dry matter intake and health status were recorded daily, and cows were categorized as healthy (n=7) or diseased (n=5; showing ketosis, milk fever, mastitis, and diarrhoea within the first week postpartum). A comprehensive metabolic profile was also assessed. When considering all cows, calving had no significant impact on PIC release. However, diseased cows displayed elevated TNF-α concentrations (P<0.05) from -7 to 0.5 days prepartum, a trend for higher IL-6 concentrations from -1 day, reaching significance (P<0.05) at +1 day, and increased IL-1β concentrations from -2 to +3 days. Compared with healthy, Diseased cows also showed reduced DMI and a stronger APP, reflected by significantly (P<0.05) increased haptoglobin and ceruloplasmin, reduced albumin, and elevated zinc concentrations. In this exploratory cohort, cows that developed postpartum disorders showed higher prepartum cytokine concentrations. These findings support the hypothesis that increased prepartum inflammatory tone is associated with poorer transition adaptation, but larger prospective studies are required before predictive claims.

Early life is a critical phase for dairy calves, influencing their growth, metabolism, and overall health. This study aimed to evaluate the effects of feeding either whole milk (WM) or milk replacer (MR) on growth performance, metabolic markers, and hematological parameters from birth until weaning at 60 d of age in Fleckvieh dairy. Eighteen heifer calves were assigned to one of 2 dietary treatments: WM (n = 9; DM basis, 29.1% protein, 27.3% fat, and 37.3% lactose; 5.25 Mcal/kg of DM) or MR (n = 9; DM basis, 22.9% protein, 19.1% fat, and 51.73% lactose; 4.76 Mcal/kg DM; Elvor, Maen Roch, France). The MR was prepared at 130 g/L to match solids content in WM. Calves were housed in individual hutches, fed 3 L of milk twice daily until 53 d and then 3 L of milk once daily until weaning (60 d). Calves had ad libitum access to starter and intake was recorded daily. Growth performance was assessed through BW and period-specific ADG (gain per day between successive weighings), whereas metabolic and hematological markers were analyzed through plasma biomarkers and blood cell counts, respectively. Data were subjected to ANOVA using the GLM procedure for single measures and the GLIMMIX procedure for repeated measures. Despite similar total DMI, calves fed WM exhibited an overall greater BW and greater period-specific ADG (0.77 vs. 0.85 kg/d in MR and WM, respectively), and from 21 to 60 d, WM calves were 3.8 kg heavier than MR-fed calves. From the metabolism standpoint, feeding WM resulted in overall greater plasma concentrations of glucose, urea, cholesterol, alkaline phosphatase, and Ca, but lower bilirubin. Specifically, WM calves had greater paraoxonase activity at 35, 45, and 60 d (weaning), greater ceruloplasmin levels at 15, 21, and 28 d, and greater Mg at 60 d of age. Among hematological parameters, WM calves had greater circulating total leukocytes at 21 and 28 d of age due to enhanced neutrophils. Although WM calves consumed milk with greater proportions of fat and protein and lower lactose content, the intake of milk and starter DM and total energy was similar compared with MR calves. However, considering the better growth rate of WM calves (especially in the second week of life), the better balance between fat, protein, and lactose suggests that feeding WM can enhance bioavailability of nutrients. Further research is needed to explore the long-term implications of these feeding strategies on calf health and productivity, particularly in relation to sustainability and economic feasibility.

The transition period is characterized by major metabolic and immunological changes that increase disease susceptibility in dairy cows. Genetic background may influence the regulation of immune responses during this phase. The aim of this study was to compare the expression of immune-related genes in circulating leukocytes of Brown Swiss and Holstein cows across the transition period. Twenty-two pregnant heifers (10 Brown Swiss and 12 Holstein) were managed under identical conditions and sampled at -21, +7, and +28 days relative to calving. Whole blood was collected using PAXgene tubes, and gene expression was analyzed by quantitative PCR. Target genes were associated with immune recognition, leukocyte migration and adhesion, antimicrobial activity, oxidative stress, and inflammatory response. Gene expression profiles differed between breeds over time. Holstein cows showed higher expression of genes involved in pathogen recognition before calving, including CD14 and TLR4. In contrast, Brown Swiss cows showed higher expression of genes related to leukocyte migration (ITGAL, CCR2), antimicrobial activity (MMP9, MPO), inflammatory response (TNFA, IL18), and oxidative stress (ALOX5, ALOX15), particularly after calving. These results indicate breed-related differences in leukocyte gene expression and suggest that genetic background influences immune adaptation during the transition period.

This study evaluated the effects of supplementing dairy calves with Saccharomyces cerevisiae fermentation-derived postbiotics (SCFP) on growth, metabolism, immune status, and first lactation performance. Eighteen Holstein heifer calves were blocked by birth body weight and serum total protein and randomly assigned to control (CTR; n = 9; no supplementation) or SCFP (n = 9; 1 g/d SmartCare® in milk replacer until weaning plus 5 g/d NutriTek® until 70 d; Diamond V™, USA). Calves were weaned at 60 d and monitored until 160 d. Feed intake did not differ between groups. SCFP calves had greater post-weaning average daily gain from 71 to 100 d (0.93 vs. 0.60 kg/d, SCFP and CTR, respectively) and body weight from 100 to 160 d. They tended to have greater plasma β-hydroxybutyrate at 60 (0.32 vs. 0.27 mmol/L, SCFP and CTR, respectively) and 70 d (0.46 vs. 0.42, SCFP and CTR, respectively) and urea at 70 d (4.89 vs. 4.33 mmol/L, SCFP and CTR, respectively) and had greater acetate (515 vs. 384 μmol/L, SCFP and CTR, respectively) and propionate (33.13 vs. 22.4 ± 4.86 μmol/L, SCFP and CTR, respectively) at 60 d. SCFP calves also had lower nonesterified fatty acids at 21 d (0.23 vs. 0.38 mmol/L, SCFP and CTR, respectively), suggesting reduced energy mobilization during the most critical pre-weaning stage. Plasma myeloperoxidase was greater at 70 d (340 vs. 262 U/L, SCFP and CTR, respectively), as was phagocytosis by polymorphonuclear neutrophils at 60 (+10.4%) and 70 d (+8.2%). Feeding SCFP increased rumen activity and plasma volatile fatty acid concentrations, likely due to enhanced nutrient absorption and reduced weaning stress. SCFP calves exhibited a better immune response to lipopolysaccharide stimulation, as indicated by leukocyte gene expression, MPO, and PMN phagocytosis. Metagenomic analyses showed minor but significant changes in early-life microbiota composition at 7, 21, and 42 d. During first lactation, SCFP cows produced 2.1 kg/d more milk in the first 100 days in milk compared with CTR. In conclusion, early supplementation with SCFP supported rumen development, improved metabolic and immune function, and may enhance future productivity in dairy cows.

BACKGROUND:Rapeseed (Brassica napus L. var. oleifera) is a globally significant vegetable oilseed crop. Its seeds are sources of oil and protein for industrial applications and food. This study shows how rapeseed cultivar, and environmental and fertilization conditions influence the agronomic parameters, fatty acid composition, and polyphenol content of rapeseed. RESULTS:Seasonality was the factor affecting grain yield the most, with variations attributed to sowing dates, rainfall patterns, and temperature trends. In 2020/21, earlier sowing (6 October) and higher precipitation during flowering (56 mm in April), resulted in a grain yield of 2.16 Mg ha-1, whereas delayed sowing (21 October), reduced rainfall (-52% in April), and higher temperature (+4.2 °C during vegetative growth) in 2022/23 caused a 79% yield decline. Higher nitrogen rates (140 and 190 kg N ha-1) increased the dry biomass at flowering and the grain yield at harvest, with intermediate rates (110 kg N ha-1), achieving comparable results to higher rates by optimizing timing, particularly in pre-sowing and early spring. Nitrogen fertilization also significantly impacted fatty acid composition and accumulation of polyphenols, with higher nitrogen rates related to increased levels of specific fatty acids. Phenolic compounds analysis revealed significant fluctuations depending on the year, with anthocyanins, flavanols, and flavonols being the most susceptible. In 2020/21, anthocyanins were the most abundant polyphenols, whereas in 2022/23, the highest levels were observed for fatty acids, specifically α-linolenic acid (10.9%) and eicosenoic acid (1.17%). CONCLUSION:The results highlight the importance of strategic planning, environmental monitoring, and targeted nitrogen fertilization in optimizing rapeseed yield and quality under a changing climate, providing valuable insights for sustainable rapeseed cultivation. © 2025 Society of Chemical Industry.

Heat stress (HS) in dairy cows disrupts homeostasis and thermoregulation, negatively affecting milk production, health status, and metabolism. Dietary supplementation with phytoextracts such as polyphenols may offer an effective dietary strategy by stimulating the immune system, reducing oxidative stress, and ultimately enhancing welfare, metabolic function, and milk performance. This study aimed to evaluate the effects of supplementing mid-lactating Fleckvieh cows with rumen-protected grape extract (Nor-Grape BP-O, Nor-Feed) on milk performance, rectal temperature (RT), behavior, immunometabolism, rumen fermentation, and innate immune response under naturally occurring HS conditions (temperature-humidity index >72). Thirty cows were balanced by days in milk, milk yield, and parity, and randomly assigned to receive either 470 mg/cow per day of Nor-Grape BP-O supplementation (NG-BPO, n = 15) or a control diet without supplementation (CTR, n = 15) and monitored for 35 d. Both groups were cooled daily using forced ventilation and fans. Temperature-humidity index, milk yield, heavy breathing (HB), rumination, and eating time were recorded daily. Milk, blood, and rumen fluid samples were collected at different time points, and RT was measured. The NG-BPO cows exhibited greater milk production, fat-corrected milk, energy-corrected milk, and yields of fat and protein, alongside lower SCC compared with CTR cows. The NG-BPO cows exhibited improved thermoregulatory responses, reflected by significantly lower RT and reduced HB. Supplementation with Nor-Grape BP-O lowered inflammatory status and enhanced innate immune responses, as indicated by higher plasma zinc and myeloperoxidase levels, lower haptoglobin concentrations, and increased phagocytic activity of neutrophils and monocytes compared with CTR cows. These findings were supported by higher levels of proinflammatory cytokines TNF-α and IL-1 β during peak HS periods, compared with CTR cows. In addition, the NG-BPO group had lower circulating platelet and eosinophil counts compared with the CTR group. In conclusion, supplementation with a low dose of rumen-protected dry grape extract-rich in low-molecular-weight polyphenols (oligomeric and monomeric flavanols, anthocyanins)-effectively supported milk production and quality in heat-stressed dairy cows. Moreover, supplementation improved thermotolerance during heat waves and enhanced innate immune functions while reducing inflammatory responses. These findings suggest Nor-Grape BP-O as an effective dietary strategy to mitigate the negative effects of HS by enhancing cow performance and welfare during periods of elevated environmental temperatures.

The mismatch between the nutrient intake from the diet and the output by the mammary gland causes a negative energy balance in transition dairy cows that, if excessive, can promote several metabolic disorders. Other relevant phenomena occur during transition, such as inflammation at calving and changes in immunocompetence, redox balance, and mineral metabolism. Despite previous efforts, some aspects of the adaptive mechanisms observed in the transition period still need to be clarified. For instance, alterations of physiological responses even before the dry-off or during the dry period can affect the success of the whole transition period in certain cows. In this context, the mechanism regulating the inflammatory response around calving may play a pivotal role, as suggested by the variety of factors influencing it and its consequences, particularly feed intake depression, that can amplify and anticipate the negative energy balance. When this mechanism derails is still unclear, but detecting the triggers of diverted or abnormal physiological responses and their origins (e.g., liver, rumen and gut epithelia, uterus, or mammary gland) will help to discover the weak points in the immune system and the possible ways of restoring it. Furthermore, the postpartum healthy cow appears to have an acute phase response at the liver level, despite a decrease in circulating proinflammatory cytokines. What is physiological and what is pathological in this context? To understand the latter, finding markers of an unsuccessful transition period that go beyond the energy deficit would be advisable. Future efforts should be dedicated to clarifying the causes of the acute phase response at calving, exploiting the potential of the system biology. Moreover, it would be helpful, for both basic and applied research, to define biomarkers associated with pathological responses (i.e., cytokines and acute phase proteins) and to introduce in the genetic selection phenotypes related to the ability of cows to adapt to the immunometabolic stress typical of the transition period.

The objective of this study was to evaluate the adaptation of mid-lactating Holstein cows regarding their introduction at the automatic milking system (AMS) and its effect on behaviour and milk performance. From a herd of 400 milking cows, a group of twenty-eight mid-lactating Holstein cows were monitored in the transition from the conventional milking system (twice daily) to the free-traffic automatic milking system. After 37 days from the full transition to the AMS, cows were retrospectively divided into two groups according to the average number of milking visits during the first week of AMS: 1) milking visits < 2 (MV-LOW) and 2) milking visits < 2 (MV-HIGH). Number of milking visits, milk yield, milk quality and rumination and eating time were recorded. In the first week, MV-HIGH cows had an average of milking visits higher compared to the MV-LOW cows (2.44 vs. 1.67 ± 0.06 n°/d, respectively). Starting from the similar milk production in the pre-AMS period, milk yield after the introduction of AMS (from 0 to 37 days) was higher in the MV-HIGH than in the MV-LOW cows (36.91 vs. 30.94 ± 2.11 kg/d, respectively), but with lower fat, protein and lactose percentage. The first week of AMS was characterized by a marked increase in milk yield in the MV-HIGH cows (+30.98 %) and a decrease in the MV-LOW cows (-11.60 %) compared to the pre-AMS. The time spent for eating was also greater in the MV-HIGH compared to the MV-LOW cows. Even though MV-LOW cows increased the milk production over the first week of AMS, these cows had always a lower increase in milk yield compared with the MV-HIGH cows until 37 days of AMS. These results suggest that the adaptability of the cows to the automatic milking system may also depend on the individual behaviour of cows and not only on the environment around them. Therefore, it is necessary to apply new livestock management strategies, when AMS (as a free-traffic system) is going to displace the milking parlour in order to better fine-tune the adaptation of cows (especially for heifers) and also to select cows that respond best to the robotic milking system.

Sixty Holstein cows were enrolled at −76 days from calving (DFC) and classified based on the daily SCC during the previous week from an automated milking system. The separation thresholds for low (L, n = 46) and high (H, n = 14) classifications were 100 K/mL for primiparous and 200 K/mL for multiparous cows. Cows were then assigned to two homogeneous groups to receive diets supplemented with 19 g/d of a Saccharomyces cerevisiae fermentation product (TRT; NutriTek, Diamond V, Cedar Rapids, IA, USA) or without supplementation (CTR) until 60 DFC. Cows were dried off at −56 DFC and monitored for disease incidence, milk yield and composition, plasma metabolic profile, and whole blood count from −76 to 60 DFC. Data were analyzed utilizing ANOVA and mixed models for repeated measures. During the dry period, TRT cows had greater plasma thiol and albumin compared to CTR. TRT-L cows had greater plasma protein and globulin than CTR-L. TRT-H cows had heightened hematocrit; reduced plasma globulin and haptoglobin; and higher albumin, albumin to globulin ratio, and thiol than CTR-H. TRT-H cows had greater concentrations of leukocytes and lymphocytes and lower plasma protein and ceruloplasmin at −54 DFC; lower reactive oxygen species to ferric ion-reducing antioxidant power ratios at −44 DFC; and greater concentrations of lymphocytes and plasma gamma glutamyl transferase at −7 DFC than CTR-H. After calving, TRT cows had a lower incidence of mastitis and higher butterfat, as well as greater plasma haptoglobin and aspartate amino transferase (AST) and reduced Mg compared to CTR. TRT cows had lower SCC between 1 and 7 DFC and a greater ECM between 41 and 60 DFC compared to CTR. TRT-H cows had lower SCC between 1 and 7 DFC and greater hemoglobin and plasma AST than CTR-H. Ameliorated immune system functions due to Saccharomyces cerevisiae fermentation product administration lowered the SCC in TRT-H cows and prevented the onset of new intramammary infections across both L and H SCC groups, supporting the improved productive performance of dairy cows.

The early lactation period in dairy cows is characterized by complex interactions among energy balance (EB), disease, and alterations in metabolic and inflammatory status. The objective of this study was to cluster cows based on EB time profiles in early lactation and investigate the association between EB clusters and inflammatory status, metabolic status, oxidative stress, and disease. Holstein-Friesian dairy cows (n = 153) were selected and monitored for disease treatments during wk 1 to 6 in lactation. Weekly EB was calculated based on energy intake and energy requirements for maintenance and milk yield in wk 1 to 6 in lactation. Weekly plasma samples were analyzed for metabolic variables in wk 1 to 6, and inflammatory and oxidative stress variables in wk 1, 2, and 4 in lactation. Liver activity index (LAI) was computed from plasma albumin, cholesterol, and retinobinding protein concentration. First, cows were clustered based on time profiles of EB, resulting in 4 clusters (SP: stable positive; MN: mild negative; IN: intermediate negative; SN: severe negative). Cows in the SN cluster had higher plasma nonesterified fatty acids and BHB concentrations, compared with cows in the SP cluster, with the MN and IN clusters being intermediate. Cows in the SN cluster had a higher milk yield, lower DMI in wk 1, lower insulin concentration compared with cows in the SP cluster, and lower glucose and IGF-1 concentration compared with cows in the SP and MN clusters. Energy balance clusters were not related to plasma haptoglobin, cholesterol, albumin, paraoxonase, and LAI. Second, cows were grouped based on health status: IHP, cows with treatment for inflammatory health problem (endometritis, fever, clinical mastitis, vaginal discharge or retained placenta); OHP, cows with no IHP but treatment for other health problem (milk fever, cystic ovaries, claw and leg problems, rumen and intestine problems, or other diseases); and NHP, cows with no treatments, in the first 6 wk after calving. Energy balance was not different among health status groups. The IHP cows had lower nonesterified fatty acids and greater insulin concentration in plasma compared with OHP cows. The IHP cows had lower plasma albumin concentration, lower LAI, and higher haptoglobin concentration compared with OHP and NHP. Overall, EB time profiles were associated with the metabolic status of dairy cows in early lactation, but were only limitedly related to markers of inflammation and oxidative stress status. Inflammatory and metabolic status were related to disease events in early lactation and caused prolonged effects on liver metabolism.

In Africa, the number of children under 5 years old who suffer from stunting and wasting are, respectively, 61.4 and 12.1 million, and to manage situations like these, emergency food products like RUTF and RUSF (ready-to-use therapeutic/supplementary food) are very useful. The aim of this study was to develop an RUSF biscuit using the low-cost food resources usually present in Sub-Saharan Africa (Burundi and the DRCongo in our case study); we conducted chemical characterization, nutritional evaluation, and a stability trial simulating the usual storage conditions in a rural context to demonstrate that RUSF can be functional also using low-cost ingredients and a simple method of production. The obtained recipes showed good potential in supplying protein integration—17.81% (BUR) and 16.77% (CON) (% as food) were the protein contents—and the protein digestibility values were very high (BUR: 91.72%; CON: 92.01%). Moreover, 30% of the daily requirement was achieved with less than 50 g of both recipes in all the considered ages. Finally, a good shelf-life was demonstrated during the 35-day testing period at 30 °C, considering moisture, texture, and lipid oxidation evolution. Recipes like these, with appropriate changes, could be very useful in all contexts where child malnutrition is a serious problem.

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

BACKGROUND:Ruminal microbial communities enriched on lignocellulosic biomass have shown considerable promise for the discovery of microorganisms and enzymes involved in digesting cell wall compounds, a key bottleneck in the development of second-generation biofuels and bioproducts, enabling a circular bioeconomy. Cardoon (Cynara cardunculus) is a promising inedible energy crop for current and future cellulosic biorefineries and the emerging bioenergy and bioproducts industries. The rumen microbiome can be considered an anaerobic "bioreactor", where the resident microbiota carry out the depolymerization and hydrolysis of plant cell wall polysaccharides (PCWPs) through the catalytic action of fibrolytic enzymes. In this context, the rumen microbiota represents a potential source of microbes and fibrolytic enzymes suitable for biofuel production from feedstocks. In this study, metatranscriptomic and 16S rRNA sequencing were used to profile the microbiome and to investigate the genetic features within the microbial community adherent to the fiber fractions of the rumen content and to the residue of cardoon biomass incubated in the rumen of cannulated cows.RESULTS:The metatranscriptome of the cardoon and rumen fibre-adherent microbial communities were dissected in their functional and taxonomic components. From a functional point of view, transcripts involved in the methanogenesis from CO2 and H2, and from methanol were over-represented in the cardoon-adherent microbial community and were affiliated with the Methanobrevibacter and Methanosphaera of the Euryarchaeota phylum. Transcripts encoding glycoside hydrolases (GHs), carbohydrate-binding modules (CBMs), carbohydrate esterases (CEs), polysaccharide lyases (PLs), and glycoside transferases (GTs) accounted for 1.5% (6,957) of the total RNA coding transcripts and were taxonomically affiliated to major rumen fibrolytic microbes, such as Oscillospiraceae, Fibrobacteraceae, Neocallimastigaceae, Prevotellaceae, Lachnospiraceae, and Treponemataceae. The comparison of the expression profile between cardoon and rumen fiber-adherent microbial communities highlighted that specific fibrolytic enzymes were potentially responsible for the breakdown of cardoon PCWPs, which was driven by specific taxa, mainly Ruminococcus, Treponema, and Neocallimastigaceae.CONCLUSIONS:Analysis of 16S rRNA and metatranscriptomic sequencing data revealed that the cow rumen microbiome harbors a repertoire of new enzymes capable of degrading PCWPs. Our results demonstrate the feasibility of using metatranscriptomics of enriched microbial RNA as a potential approach for accelerating the discovery of novel cellulolytic enzymes that could be harnessed for biotechnology. This research contributes a relevant perspective towards degrading cellulosic biomass and providing an economical route to the production of advanced biofuels and high-value bioproducts.

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.

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.

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.