Emerging Parameters Justifying a Revised Quality Concept for Cow Milk.

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.

Methods of Evaluating the Potential Success or Failure of Transition Dairy Cows.

Early monitoring of the failure of metabolic adaptation to calving, represents the most effective measure for allowing a prompt intervention on transition dairy cows. This prevents deleterious effects on animal performance, health, and welfare, which are driven by multiple disorders during the following lactation. Applying metabolic profiling could (1) provide a deeper view on the cause of any pathologic condition affecting transition cows, aimed at increasing the effectiveness and timely application of any treatment and (2) provide detailed feedback on the management practices adopted in a farm during this challenging phase based on animal responses.

Effect of Different Anticoagulant Agents on Immune-Related Genes in Leukocytes Isolated from the Whole-Blood of Holstein 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.

Effects of an Intravenous Infusion of Emulsified Fish Oil Rich in Long-Chained Omega-3 Fatty Acids on Plasma Total Fatty Acids Profile, Metabolic Conditions, and Performances of Postpartum Dairy Cows During the Early Lactation.

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.

Gene network expression of whole blood leukocytes in dairy cows with different milk yield at dry-off.

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.

Changes of Plasma Analytes Reflecting Metabolic Adaptation to the Different Stages of the Lactation Cycle in Healthy Multiparous Holstein Dairy Cows Raised in High-Welfare Conditions.

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 ß-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.

Role of nutraceuticals during the transition period of dairy cows: a review.

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.

Effects of Aloe arborescens Whole Plant Homogenate on Lipid Metabolism, Inflammatory Conditions and Liver Function of Dairy Cows during the Transition Period.

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.

Administration of an Immune Stimulant during the Transition Period Improved Lipid Metabolism and Rumination without Affecting Inflammatory Status.

Omnigen-AF (OAF) increases leukocyte functions in immunosuppressed animal models and reduces incidence of infectious diseases in early lactating dairy cows, although its mode of action is still unclear. This study aims to provide a wider perspective of the metabolic effect of OAF to test its potential as a strategy to address metabolic disorders of the transition period. A group of 10 Holstein dairy cows were divided into 2 groups: The treated group (IMS; 5 cows) received 32.5 g of OAF twice a day (65 g d-1) as top-dress in the morning and afternoon feeds from -55 to 42 days from calving (DFC), whereas the control group (CTR; 5 cows) received no supplementation. From -62 to 42 DFC, body condition score, body weight, dry matter intake, rumination time and milk yield were measured; blood samples were collected weekly to assess a wide hematochemical profile and to test white blood cell functions by ex-vivo challenge assays. At 30 DFC, rumen fluid was collected and analyzed for pH, volatile fatty acids composition, urea nitrogen, and lactate contents. Data were submitted to ANOVA using a mixed model for repeated measures, including treatment, time, and their interaction as fixed effects. OAF decreased blood nonesterified fatty acids and beta hydroxybutyrate concentrations and increased rumination time in early lactation. Leukocytes from IMS cows had lower lactate production and lower glucose consumption after ex-vivo stimulation. OAF did not reduce the acute phase response indicators and reduced the blood concentrations of albumin and antioxidants after calving, suggesting impairment of hepatic functions related to protein synthesis and antioxidant management. Nevertheless, the lack of effect on bilirubin and liver enzymes refutes the possibility of severe liver damage occurring with OAF supplementation. Positive effects in reducing mobilization of body fats and ketogenesis and in increasing rumination time after calving suggest OAF effectiveness in preventing metabolic disorders of the transition period.

What's the norm in normalization? A frightening note on the use of RT-qPCR in the livestock science.

Reverse-Transcription quantitative PCR (RT-qPCR) provides a valuable tool to study gene expression with exquisite sensitivity. To retain its inferential power, user-introduced technical variability must be reduced and accounted for. Selecting a set of stably expressed internal control genes (ICG), validated for each experimental condition/sample set, is widely accepted as a reliable way to normalize RT-qPCR data and account for said variability. Despite significant efforts in establishing standardized and resource-efficient normalization approaches, numerous recent reports have underlined deficiencies in the state of RT-qPCR normalization. Livestock science has benefitted tremendously from the use of RT-qPCR; however, the issue of lack of proper normalization likely affects this discipline as well. We thus decided to determine whether this is true, and to which extent. We conducted an in-depth analysis of all (225) RT-qPCR articles published in the six most prominent livestock journals in the field from 2013 to 2017. A quantitative scale was constructed, and values were assigned to each article based on the number of ICG used, the use of a publicly available algorithm to assess the reliability of ICG, and the reporting of pertinent information related to ICG (ranges from 0 = total noncompliance - to 100 = total compliance). Out of the surveyed group, only 10.7% of the publications obtained a score of 100, while the largest group (n = 158) was represented by articles that scored 0. Subdividing articles based on whether an algorithm to validate ICG was used (YAL) or not (NAL) revealed the use of a larger number of ICG to normalize RT-qPCR in the YAL group compared to NAL (1.4-fold more, 95% C.I.: 1.11-1.84) and was closer to the "gold standard" of three ICG. Using an algorithm also increased the diversity of ICG and significantly reduced the use of RNA18S, whose suitability as ICG has been thoroughly debated. These remarkably low normalization standards are likely to generate questionable results that can severely hinder the advance of transcriptomic studies in livestock science and related fields.