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Dairy cows have to face several nutritional challenges during the transition period, and live yeast supplementation appears to be beneficial in modulating rumen activity. In this study, we evaluated the effects of live yeast supplementation on rumen function, milk production, and metabolic and inflammatory conditions. Ten Holstein multiparous cows received either live Saccharomyces cerevisiae (strain Sc47; SCY) supplementation from −21 to 21 d from calving (DFC) or a control diet without yeast supplementation. Feed intake, milk yield, and rumination time were monitored until 35 DFC, and rumen fluid, feces, milk, and blood samples were collected at different time points. Compared with the control diet, SCY had increased dry matter intake (16.7 vs. 19.1 ± 0.8 kg/d in wk 2 and 3) and rumination time postpartum (449 vs. 504 ± 19.9 min/d in wk 5). Milk yield tended to be greater in SCY (40.1 vs. 45.2 ± 1.7 kg/d in wk 5), protein content tended to be higher, and somatic cell count was lower. In rumen fluid, acetate molar proportion was higher and that of propionate lower at 21 DFC, resulting in increased acetate:propionate and (acetate + butyrate):propionate ratios. Cows in the SCY group had lower fecal dry matter but higher acetate and lower propionate proportions on total volatile fatty acids at 3 DFC. Plasma analysis revealed a lower degree of inflammation after calving in SCY (i.e., lower haptoglobin concentration at 1 and 3 DFC) and a likely better liver function, as suggested by the lower γ-glutamyl transferase, even though paraoxonase was lower at 28 DFC. Plasma IL-1β concentration tended to be higher in SCY, as well as Mg and P. Overall, SCY supplementation improved rumen and hindgut fermentation profiles, also resulting in higher dry matter intake and rumination time postpartum. Moreover, the postcalving inflammatory response was milder and liver function appeared to be better. Altogether, these effects also led to greater milk yield and reduced the risk of metabolic diseases. [ABSTRACT FROM AUTHOR]

The dry-off of dairy cows represents an important phase of the lactation cycle, influencing the outcome of the next lactation. Among the physiological changes, the severity of the inflammatory response can vary after the dry-off, and this response might have consequences on cow adaptation in the transition period. The plasma protein profile is a diagnostic tool widely used in humans and animals to assess the inflammatory status and predict the outcome of severe diseases. The albumin-to-globulin ratio (AG) can represent a simple and useful proxy for the inflammatory condition. In this study, we investigated the relationship between AG before dry-off and inflammation, metabolic profile, and performance of 75 Holstein dairy cows. Blood samples were collected from −62 (7 d before dry-off) to 28 d relative to calving (DFC) to measure metabolic profile biomarkers, inflammatory variables, and liver function. Daily milk yield in the first month of lactation was recorded. Milk composition, body condition score, fertility, and health status were also assessed. The AG calculated 1 wk before dry-off (−62 DFC) was used to retrospectively group cows into tertiles (1.06 ± 0.09 for HI, 0.88 ± 0.04 for IN, and 0.72 ± 0.08 for LO). Data were subjected to ANOVA using the PROC MIXED program in SAS software. Differences among groups observed at −62 DFC were almost maintained throughout the period of interest, but AG peaked before calving. According to the level of acute-phase proteins (haptoglobin, ceruloplasmin, albumin, cholesterol, retinol-binding protein), bilirubin, and paraoxonase, a generally overall lower inflammatory condition was found in HI and IN than in the LO group immediately after the dry-off but also after calving. The HI cows had greater milk yield than LO cows, but no differences were observed in milk composition. The somatic cell count reflected the AG ratio trend, with higher values in LO than IN and HI either before dry-off or after calving. Fertility was better in HI cows, with fewer days open and services per pregnancy than IN and LO cows. Overall, cows with high AG before dry-off showed an improved adaptation to the new lactation, as demonstrated by a reduced systemic inflammatory response and increased milk yield than cows with low AG. In conclusion, the AG ratio before dry-off might represent a rapid and useful proxy to evaluate the innate immune status and likely the ability to adapt while switching from the late lactation to the nonlactating phase and during the transition period with emphasis on early lactation. [ABSTRACT FROM AUTHOR]

Mechanisms controlling immune function of dairy cows are dysregulated during heat stress (HS). Methyl donor supply—methionine (Met) and choline (Chol)—positively modulates innate immune function, particularly antioxidant systems of polymorphonuclear leukocytes (PMN). The objective of this study was to investigate the effect of Met and Chol supply in vitro on mRNA abundance of genes related to 1-carbon metabolism, inflammation, and immune function in short-term cultures of PMN isolated from mid-lactating Holstein cows in response to heat challenge. Blood PMN were isolated from 5 Holstein cows (153 ± 5 d postpartum, 34.63 ± 2.73 kg/d of milk production; mean ± SD). The PMN were incubated for 2 h at thermal-neutral (37°C; TN) or heat stress (42°C; HS) temperatures with 3 levels of Chol (0, 400, or 800 μg/mL) or 3 ratios of Lys:Met (Met; 3.6:1, 2.9:1, or 2.4:1). Supernatant concentrations of IL-1β, IL-6, and tumor necrosis factor-α were measured via bovine-specific ELISA. Fold-changes in mRNA abundance were calculated separately for Chol and Met treatments to obtain the fold-change response at 42°C (HS) relative to 37°C (TN). Data were subjected to ANOVA using PROC MIXED in SAS (SAS Institute Inc., Cary, NC). Orthogonal contrasts were used to determine the linear or quadratic effect of Met and Chol for mRNA fold-change and supernatant cytokine concentrations. Compared with PMN receiving 0 μg of Chol/mL, heat-stressed PMN supplemented with Chol at 400 or 800 μg/mL had greater fold-change in abundance of CBS , CSAD , GSS , GSR , and GPX1. Among genes associated with inflammation and immune function, fold-change in abundance of TLR2 , TLR4 , IRAK1 , IL1B , and IL10 increased with 400 and 800 μg of Chol/mL compared with PMN receiving 0 μg of Chol/mL. Fold-change in abundance of SAHH decreased linearly at increasing levels of Met supply. A linear effect was detected for MPO , NFKB1 , and SOD1 due to greater fold-change in abundance when Met was increased to reach Lys:Met ratios of 2.9:1 and 2.4:1. Although increasing Chol supply upregulated BAX , BCL2 , and HSP70 , increased Met supply only upregulated BAX. Under HS conditions, enhancing PMN supply of Chol to 400 μg/mL effectively increased fold-change in abundance of genes involved in antioxidant production (conferring cellular processes protection from free radicals and reactive oxygen species), inflammatory signaling, and innate immunity. Although similar outcomes were obtained with Met supply at Lys:Met ratios of 2.9:1 and 2.4:1, the response was less pronounced. Both Chol and Met supply enhanced the cytoprotective characteristics of PMN through upregulation of heat shock proteins. Overall, the modulatory effects detected in the present experiment highlight an opportunity to use Met and particularly Chol supplementation during thermal stress. [ABSTRACT FROM AUTHOR]

The use of modern prolific lines of rabbit does in intensive production systems leads to an increase in productivity but also causes a rise in several problems related to the does' health status. Hence, the aim of this study was to investigate the effect of the litter size on the metabolic, inflammatory and plasma amino acid profile in rabbit does. The blood of 30 pregnant does was sampled on the 27th day of pregnancy. The does were retrospectively grouped according to the number of offspring into a high litter size group (HI, does with ≥ 12 kits; n = 16) and a low litter size group (LO, does with ≤ 11 kits; n = 14). Data were subjected to Pearson's correlation analysis. Further, data were analysed in agreement to a completely randomized design in which the main tested effect was litter size. The linear or quadratic trends of litter size on parameters of interests were post hoc compared by using orthogonal contrasts. In addition, compared with the LO group, the HI group had lower levels of glucose (−5%; P < 0.01), zinc (−19%; P < 0.05), albumin (−6%; P < 0.05) and total cholesterol (−13%; P < 0.07), but the total bilirubin level was higher in the HI group (+14%; P < 0.05). Regarding the plasma amino acids, the HI group had lower concentrations of threonine (−15%), glycine (−16%), lysine (−16%) and tryptophan (−26%) and a higher level of glutamic acid (+43%; P < 0.05) compared with the LO group. The exclusively ketogenic amount of amino acids was lower (P < 0.06) in the HI (55.8 mg/100 ml) does compared with the LO does (56.8 mg/100 ml). These results show that a few days before delivery, rabbit does that gave birth to a higher number of offspring had a metabolic profile and an inflammatory status that was less favourable with respect to does who gave birth to a lower number of offspring. Moreover, the plasma amino acid profile points out that there was an enhanced catabolic condition in the rabbit does with a high number of gestated foetuses; it was likely related to the greater energy demand needed to support the pregnancy and an early inflammatory response. [ABSTRACT FROM AUTHOR]

Finding a rapid and simple method of serum IgG determination in lambs is essential for monitoring failure of passive transfer of immunity. The aim of this study was to assess the ability of capillary electrophoresis (CE), an instrument mainly used in blood serum protein analysis, to estimate IgG content in serum of newborn lambs through determination of only total Ig percentage by comparing the results with those obtained with radial immunodiffusion (RID), the reference method for serum IgG quantification. Serum samples were collected at 24 h after birth from 40 Sarda lambs. The IgG concentration measured by RID and serum total Ig concentration measured by CE were (mean ± standard deviation) 29.8 ± 16.1 g/L and 37.8 ± 15.0%, respectively. Data provided by RID and CE analysis showed a polynomial trend (RID = 0.02CE2 − 0.04CE + 4.13; coefficient of determination, R2 = 0.96), displaying a strong relationship between these 2 methods. Applying the polynomial equation, the IgG values were predicted. Predicted IgG values were highly correlated (r = 0.98) and related (R2 = 0.96) to IgG values obtained by RID assay. These data were subjected to Bland–Altman analysis, revealing a high level of agreement between CE and RID methods with a bias that was not different from 0 (−0.04 g/L) and agreement limits of −6.38 g/L (low) and +6.30 g/L (high). In addition, the linear regression analysis between differences (dependent variable) and average of IgG concentration by CE and RID (independent variable) did not show proportional bias (R2 = 0.01). In conclusion, CE is a reliable instrument for a lamb health monitoring program, where Bland–Altman analysis also confirmed that the CE method can be a suitable alternative to RID. [ABSTRACT FROM AUTHOR]

The aim of this study was to investigate changes in the abundance of genes involved in leukocyte function between cows highly specialized for milk production (Holstein, n = 12) and cows selected for meat and milk (Simmental, n = 13). Blood was collected on d 3 after calving in PAXgene tubes (Preanalytix, Hombrechtikon, Switzerland) to measure mRNA abundance of 33 genes. Normalized gene abundance data were subjected to MIXED model ANOVA using SAS (SAS Institute Inc. Cary, NC). Simmental cows had greater transcript abundance of proinflammatory cytokines and receptor genes (IL1B , TNF , IL1R , TNFRSF1A), cell migration- and adhesion-related genes (CX3CR1 , ITGB2 , CD44 , LGALS8), and the antimicrobial IDO1 gene. In contrast, compared with Holstein cows, Simmental cows had lower abundance of the toll-like receptor (TLR) recognition-related gene TLR2 , the antimicrobial-related gene LTF , and S100A8 , which is involved in cell maturation, regulation of inflammatory processes, and immune response. These results revealed that breed plays an important role in the modulation of genes involved in immune adaptation and inflammatory response, and the immune system of Simmental cows could potentially have a more acute response in early lactation. In turn, this might be beneficial for mounting a more efficient response after calving and allow for a smoother homeorhetic adaptation to lactation. [ABSTRACT FROM AUTHOR]

Neutrophils are the most important polymorphonuclear leukocytes (PMNL), representing the front-line defense involved in pathogen clearance upon invasion. As such, they play a pivotal role in immune and inflammatory responses. Isolated PMNL from 5 mid-lactating Holstein dairy cows were used to evaluate the in vitro effect of methionine (Met) and choline (Chol) supplementation on mRNA expression of genes related to the Met cycle and innate immunity. The target genes are associated with the Met cycle, cell signaling, inflammation, antimicrobial and killing mechanisms, and pathogen recognition. Treatments were allocated in a 3 × 3 factorial arrangement, including 3 Lys-to-Met ratios (L:M, 3.6:1, 2.9:1, or 2.4:1) and 3 levels of supplemental Chol (0, 400, or 800 μg/mL). Three replicates per treatment group were incubated for 2 h at 37°C and 5% atmospheric CO 2. Both betaine-homocysteine S -methyltransferase and choline dehydrogenase were undetectable, indicating that PMNL (at least in vitro) cannot generate Met from Chol through the betaine pathway. The PMNL incubated without Chol experienced a specific state of inflammatory mediation [greater interleukin-1β (IL1B), myeloperoxidase (MPO), IL10 , and IL6 ] and oxidative stress [greater cysteine sulfinic acid decarboxylase (CSAD), cystathionine gamma-lyase (CTH), glutathione reductase (GSR), and glutathione synthase (GSS)]. However, data from the interaction L:M × Chol indicated that this negative state could be overcome by supplementing additional Met. This was reflected in the upregulation of methionine synthase (MTR) and toll-like receptor 2 (TLR2); that is, pathogen detection ability. At the lowest level of supplemental Chol, Met downregulated GSS , GSR , IL1B , and IL6 , suggesting it could reduce cellular inflammation and enhance antioxidant status. At 400 µg/mL Chol, supplemental Met upregulated PMNL recognition capacity [higher TLR4 and L-selectin (SELL)]. Overall, enhancing the supply of methyl donors to isolated unstimulated PMNL from mid-lactating dairy cows leads to a low level of PMNL activation and upregulates a cytoprotective mechanism against oxidative stress. Enhancing the supply of Met coupled with adequate Chol levels enhances the gene expression of PMNL pathogen-recognition mechanism. These data suggest that Chol supply to PMNL exposed to low levels of Met effectively downregulated the entire repertoire of innate inflammatory-responsive genes. Thus, Met availability in PMNL during an inflammatory challenge may be sufficient for mounting an appropriate biologic response. [ABSTRACT FROM AUTHOR]

The supply of methionine (Met) in late pregnancy can alter mRNA abundance of genes associated with metabolism and immune response in liver and polymorphonuclear leukocytes (PMN) of the neonatal calf. Whether prenatal supply of Met elicits postnatal effects on systemic inflammation and innate immune response of the calf is not well known. We investigated whether enhancing the maternal supply of Met via feeding ethyl-cellulose rumen-protected Met (RPM) was associated with differences in calf innate immune response mRNA abundance in PMN and systemic indicators of inflammation during the first 50 d of life. Calves (n = 14 per maternal diet) born to cows fed RPM at 0.09% of diet dry matter per day (MET) for the last 28 ± 2 d before calving or fed a control diet with no added Met (CON) were used. Blood for biomarker analysis and isolation of PMN for innate immune function assays and mRNA abundance was harvested at birth (before colostrum feeding) and at 7, 21 and 50 d of age. Whole blood was challenged with enteropathogenic bacteria (Escherichia coli 0118:H8) and phagocytosis and oxidative burst of neutrophils and monocytes were quantified via flow cytometry. Although concentration of haptoglobin and activity of myeloperoxidase among calves from both maternal groups increased markedly between 0 and 7 d of age followed by a decrease to baseline at d 21 the responses were lower in MET compared with CON calves. Nitric oxide concentration decreased markedly between 0 and 7 d regardless of maternal group but MET calves tended to have lower overall concentrations during the study. In vitro phagocytosis in stimulated neutrophils increased markedly over time in both CON and MET calves but responses were overall greater in MET calves. Oxidative burst in both neutrophils and monocytes increased over time regardless of maternal treatment. The mRNA abundance of lactate dehydrogenase (LDHA) signal transducer and activator of transcription 3 (STAT3) and S100 calcium binding protein A8 (S100A8) in PMN was overall greater in MET calves. Overall data suggest that increasing the maternal supply of Met during late pregnancy could affect the neonatal calf inflammatory status and innate immune response. Although changes in mRNA abundance could play a role in coordinating the immune response the exact mechanisms merit further study [ABSTRACT FROM AUTHOR]

In the present study, we aimed to investigate the side effects of pegbovigrastim, injected approximately 7 d before parturition and on the day of calving, on a panel of plasma biomarkers to evaluate energy, inflammatory, oxidative, and liver function status. We also addressed treatment responses in different breeds during the transition period. Holstein and Simmental cows were randomly assigned into 2 groups based on expected calving date and according to parity: the treated group (PEG; 14 Holstein and 12 Simmental cows) received pegylated recombinant bovine granulocyte colony stimulating factor (pegbovigrastim, Imrestor; Elanco Animal Health, Greenfield, IN), and the control group (CTR; 14 Holstein and 14 Simmental cows) received saline solution. The PEG or CTR treatments were administered via subcutaneous injection in the scapular region at approximately 7 d (mean 7.80 ± 5.50 d) before expected parturition and within 24 h after calving. Blood samples were collected at −21, −7 (before injection), 1, 3, and 28 d relative to calving. Milk production was recorded at 7, 15, 21, 30, and 42 d. A mixed model with repeated measures was fitted to the normalized data using Proc MIXED of SAS (SAS Institute Inc., Cary, NC). Simmental PEG cows showed higher plasma protein concentrations at 1 and 3 d after calving compared with Simmental CTR and Holstein PEG cows, whereas no differences were detected between Holstein PEG and CTR cows. Albumin was greater at 1 d in Simmental PEG compared with Simmental CTR cows. In contrast, γ-glutamyl transferase was higher overall (across breed) in PEG than in CTR. The PEG group had higher values throughout the postcalving period compared with CTR. Cows treated with pegbovigrastim had also higher alkaline phosphatase (ALP) activity at 1 and 3 d after calving. The Holstein PEG group had higher ALP activity at 3 d compared with the Holstein CTR and Simmental PEG groups, and higher ALP at 1 d compared with the Simmental CTR group. The PEG group had higher levels of IL-6 at 3 and 28 d but higher IL-1β only at 28 d after calving compared with the CTR group. Overall, Holstein cows were characterized by a greater response in the production of inflammation biomarkers (cytokines, haptoglobin, and ceruloplasmin). In addition, PEG cows had higher values of zinc at 1 and 3 d after calving compared with CTR cows. The response observed in plasma biomarkers for energy metabolism and liver functionality after pegbovigrastim treatment in Simmental and Holstein cows was not different from that in control cows. However, our data shed light on the different metabolic adaptations during the transition period between Simmental and Holstein cows, characterized by different energy, inflammatory, and oxidative pattern responses. For the first time, we have highlighted the effect of pegbovigrastim in maintaining stable cytokine levels during the first month after parturition, reflecting greater regulation of neutrophil recruitment, trafficking, and maturation during the inflammatory response. These results provide evidence of the immunomodulatory action of pegbovigrastim around parturition, when dairy cows are highly immunosuppressed. At the same time, these data demonstrate that increasing release of cytokines after parturition is not linked to exacerbation of a systemic inflammation evaluated based on haptoglobin and ceruloplasmin levels. [ABSTRACT FROM AUTHOR]

We assessed effects of overfeeding energy to nonlactating and nonpregnant Holstein cows during a length of time similar to a typical dry period on body lipid storage and the abundance of genes related to insulin signaling, inflammation, and ubiquitination in subcutaneous adipose tissue (SAT) in vitro challenged with insulin and recombinant bovine tumor necrosis factor-a. Fourteen cows were randomly assigned to either a highenergy (OVE; net energy for lactation = 1.60 Mcal/kg of dry matter; n = 7) or control (CON; net energy for lactation = 1.30 Mcal/kg of dry matter; n = 7) diet for 6 wk. Immediately after slaughter, liver, kidneys, and mammary gland were separated and weighed. The adipose tissue mass in the omental, mesenteric, and perirenal depots was dissected and weighed. Subcutaneous adipose tissue was collected from the tail-head region and was used as follows: control, bovine insulin (INS) at 1 µmol/L, tumor necrosis factor-a at 5 ng/mL (TNF), and their combination. Despite a lack of difference in final body condition score, OVE cows had greater energy intake and were heavier than CON cows. Furthermore, overfeeding led to greater mass of mesenteric and perirenal adipose, liver, and mammary gland. Overall, SAT incubated with INS had an upregulation of insulin receptor (INSR), interleukin 10 (IL10), small ubiquitin-like modifier 3 (SUMO3), and ubiquitin conjugating enzyme E2I (UBC9), whereas TNF upregulated peroxisome proliferator-activated receptor gamma (PPARG), diacylglycerol O-acyltransferase 2 (DGAT2), interleukin 6 (IL6), nuclear factor kappa B subunit 1 (NFKB1), small ubiquitin-like modifier 2 (SUMO2), and UBC9. Regardless of in vitro treatment, feeding OVE upregulated PPARG, fatty acid synthase (FASN), and insulin induced gene 1 (INSIG1). Abundance of PPARG was greater in SAT of OVE cows cultured individually with INS and TNF. The interaction between diet and in vitro treatment revealed that sterol regulatory element binding transcription factor 1 (SREBF1) had greater abundance in SAT from the CON group in response to culture with INS, whereas SAT from OVE cows had greater SREBF1 abundance in response to culture with TNF. The mRNA abundance of IL6 and NFKB1 was greater in response to TNF treatment and overall in CON cows. Furthermore, SAT from these cows had greater IL10 abundance when cultured with INS and TNF. Overall, data highlighted that overfeeding energy increases adipose tissue mass in part by stimulating transcription of key genes associated with insulin signaling, adipogenesis, and lipogenesis. Because SAT thickness or mass was not measured, the lack of effect of overfeeding on body condition score limits its use to predict overall body lipid storage. An overt inflammatory response in SAT after a 6-wk period of over-consumption of energy could not be discerned. [ABSTRACT FROM AUTHOR]

The present experiment aimed to evaluate the potential of the Hr-Tag rumination-monitoring system ( Hr-Tag ; SCR Engineers Ltd., Netanya, Israel) to describe the variation in rumination activity behavior in pre-weaned calves from 2 until 35 days of age. The study was carried out in a commercial farm and involved 9 Italian Simmental calves (3 females and 6 males). All calves were provided ad libitum access to water and starter throughout the study and were offered 6 L of bulk whole milk/d from an open-bucket starting at day 2 from birth and through 5 weeks. The Hr-Tag system continuously recorded the rumination time ( RT ) by Hr-Tag loggers positioned on the left side of the neck, and data were summarized in 2-h intervals. RT recorded by Hr-Tag loggers increased ( P  < 0.0001) through the first 4 weeks, reaching a plateau at wk 5 and the pattern of daily RT was similar along the weeks. Around milk feeding the observed 2-h intervals of RT drastically decreased ( P  < 0.0001) compared to the 2-h intervals before. These data shed light on the daily pattern of rumination activity in pre-weaned calves, showing a distinct daily behavior, as observable in adult cattle. Data of this study support the use of the Hr-Tag system for studying rumination activity behavior in the pre-weaning phase of calves. [ABSTRACT FROM AUTHOR]

Estimation of individual intakes in dairy cows would be beneficial to select animals according to efficiency of milk production. The n-alkanes technique has been used to estimate intake by dairy cows in free stall barns, limiting faecal samples to one or two per day, but diurnal variation in the ratio of dosed to natural alkanes in faeces represents a source of error. We aimed to adapt this technique to estimate feed intake on a large number of lactating dairy cows housed on commercial dairy farms. Using a change-over design, six lactating cows fed a TMR diet were given a dose of 0.5 g/day of C32 n-alkane administered once a day as a drench (DRE) or top fed (TP). Feed offered and refused were weighed daily. After the lead-in phases, faeces were collected at 0700 hours, 1400 hours and 1900 hours on the last 5 days of each dosing period. Dry matter intake (DMI) was estimated from C31, C33 and C32 concentrations in feeds and in faeces for each dosing system and sampling time. DMI estimated from C31 : C32 ratio was better correlated with measured individual DMI than estimates from C33 : C32 ratio. For DRE, the best correlation was for sampling at 1400 hours (R² = 0.96, P < 0.001), but estimated DMI for the 1400 hours sampling was much higher than the actual DMI. For TP, the best correlation was for sampling at 0700 hours (R² = 0.83, P < 0.001). Feeding C[sub 32] n-alkane and collecting faeces once a day at the same time (0700 hours) represents a valuable protocol to measure individual intake on a large number of animals. To select dairy cows according to efficiency of milk production it would be beneficial to estimate individual intakes, which can be predicted using n-alkanes as inert markers. A simplified procedure based on once-daily dosing of C[sub 32] alkane and spot sampling of faeces was tested. Dosing the marker in a top-fed concentrate and collecting faeces at morning feeding time represents a valuable protocol to accurately estimate individual feed intake on a large number of animals. [ABSTRACT FROM AUTHOR]

• Response of weaning at an early age on rumen pH, rumen fermentation, dietary intake, growth rate in the post-weaning phase, and rumination behavior employing the Hr-Tag rumination-monitoring system (Hr-Tag; SCR Engineers Ltd, Netanya, Israel) in Simmental calves. • Weaning Simmental calves at 45 d of age compared with 60 d, fed whole milk in the pre-weaning period, did not impair rumen fermentation after weaning, and overall the rate of VFAs clearance have a major impact on ruminal pH. • Daily rumination time measured by Hr-Tags positively correlated to pH and negatively correlated to concentration of total VFAs, acetate, and propionate. A positive linear relationship was also found between daily rumination time and the molar proportion of acetate:propionate ratio. • The use of Hr-Tags could give quick information on rumen health and thus to identify in a timely fashion those calves at a greater risk (i.e. marked decrease of rumen pH) during this critical phase, from liquid to only solid feeds. The present experiment aimed to investigate (1) the effect of weaning age on rumen fermentation, dietary intake, growth, and rumination behavior at 20 d after weaning in Simmental calves, and (2) the relationship between rumination time measured by the Hr-Tag rumination-monitoring system (Hr-Tag ; SCR Engineers Ltd, Netanya, Israel) and rumen fermentation variables. Ten Simmental calves were randomly assigned at birth to be weaned at 45 (early weaning, EW) or 60 d of age (normal weaning, NW). EW calves were offered 6 L of bulk whole milk/d from d 2 to 38 and 3 L/d from d 39 to 45, while NW were offered 6 L of bulk whole milk/d from d 2 to 53 and 3 L/d from d 54 to 60. Individual calf starter intake was recorded daily and average daily gain (ADG) was calculated taken into account the BW at 6 d after weaning and final BW at 20 d after weaning. On d 20 after weaning, samples of rumen fluid were obtained via an esophageal tube. These samples were used to determine pH and volatile fatty acids (VFAs) concentration. Throughout the entire experiment, the Hr-Tag system continuously recorded the rumination time (RT) by Hr-Tag loggers positioned on the left side of the neck. No treatment differences were detected for total VFAs concentration and molar proportion of acetate, propionate, and butyrate (P > 0.05). Overall in the period -25 to 20 d relative to weaning time, daily RT did not differ between NW and EW calves and it increased around weaning in both groups. Hr-Tags revealed no differences in daily RT pattern (2h-RT) during the 3 d before rumen collection, while during the day (24 h) 2h-RT significantly varied overall in both EW and NW groups. Daily RT measured by Hr-Tags positively correlated to pH (r = 0.98; P < 0.001), and negatively correlated to concentration of total VFAs (r = −0.90; P < 0.001), acetate (r = −0.85; P < 0.001), and propionate (r = −0.90; P < 0.001). On the contrary, a positive linear relationship was found between daily RT and the molar proportion of acetate:propionate ratio (R2 = 0.76; P = 0.002). Results herein provide evidence that early weaning (45 d) in Simmental calves fed whole milk in the pre-weaning period did not impair rumen fermentations 20 d after weaning and BW gain. Surprisingly, the use of Hr-Tags could prove interesting for the prediction of rumen fermentation characteristics and patterns in calves after weaning. The collected data could be used to obtain quick information on rumen functionality allowing the identification in a timely fashion of those calves at a greater risk (i.e. marked decrease of rumen pH) during this critical transition phase, from liquid to only solid feeds, when rumen of calves is not completely developed and efficient. The provision of these results from Hr-Tags in calves may give some input for further research involving a greater number of calves to better establish correlations with phenotypic variables. Moreover, this device could be adopted in studies focused on the effect of different pre- and post-weaning diets or weaning management. [ABSTRACT FROM AUTHOR]

Managing body condition in dairy cows during the close-up period could alter the availability of nutrients to the fetus during the final growth stages in utero. We investigated how maternal body condition score (BCS) in late pregnancy affected calf whole-blood mRNA abundance and IL-1β concentrations after ex vivo lipopolysaccharide (LPS) challenge. Thirty-eight multiparous Holstein cows and their calves from a larger cohort were retrospectively grouped by prepartal BCS as normal BCS (≤3.25; n = 22; NormBCS) and high BCS (≥3.75; n = 16; HighBCS). Calf blood samples collected at birth (before receiving colostrum, d 0) and at ages 21 and 42 d (at weaning) were used for ex vivo whole-blood challenge with 3 µg/mL of LPS before mRNA isolation. Target genes evaluated by real-time quantitative PCR were associated with immune response, antioxidant function, and 1-carbon metabolism. Plasma IL-1β concentrations were also measured. Responses in plasma IL-1β and mRNA abundance were compared between LPS-challenged and nonchallenged samples. Statistical analyses were performed at all time points using a MIXED model in SAS 9.4. Neither birth body weight (NormBCS = 43.8 ± 1.01 kg; HighBCS = 43.9 ± 1.2 kg) nor colostrum IgG concentration (NormBCS = 70 ± 5.4 mg/mL; HighBCS = 62 ± 6.5 mg/mL) differed between groups. At birth, whole blood from calves born to HighBCS cows had greater mRNA abundance of IL1B , NFKB1 , and GSR and lower GPX1 and CBS abundance after LPS challenge. The longitudinal analysis of d 0, 21, and 42 data revealed a BCS × age effect for SOD2 and NOS2 due to lower mRNA abundance at 42 d in the HighBCS calves. Regardless of maternal BCS, mRNA abundance decreased over time for genes encoding cytokines (IL1B , IL6 , IL10 , TNF), cytokine receptors (IRAK1 , CXCR1), toll-like receptor pathway (TLR4 , NFKB1), adhesion and migration (CADM1 , ICAM1 , ITGAM), and antimicrobial function (MPO). Concentration of IL-1β after LPS challenge was also markedly lower at 21 d regardless of maternal BCS. Overall, results suggested that maternal BCS in late prepartum influences the calf immune system response to an inflammation challenge after birth. Although few genes among those studied were altered due to maternal BCS, the fact that genes related to oxidative stress and 1-carbon metabolism responded to LPS challenge in HighBCS calves underscores the potential role of methyl donors (e.g., methionine, choline, and folic acid) in the early-life innate immune response. [ABSTRACT FROM AUTHOR]

Although choline requirements for cows are unknown, enhanced postruminal supply may decrease liver triacylglycerol and increase flux through the Met cycle to improve immunometabolic status during a negative nutrient balance (NNB). Our objectives were to investigate the effects of postruminal choline supply during a feed restriction–induced NNB on (1) hepatic activity cystathionine β-synthase and transcription of enzymes in the transsulfuration pathway and Met cycle; (2) hepatic metabolites in the Met cycle and the transsulfuration pathway, bile acids, and energy metabolism; and 3) plasma biomarkers of liver function, inflammation, and oxidative stress. Ten primiparous rumen-cannulated Holstein cows (158 ± 24 d postpartum) were used in a replicated 5 × 5 Latin square design with 4-d treatment periods and 10 d of recovery (14 d/period). Treatments were unrestricted intake with abomasal infusion of water, restricted intake (R; 60% of net energy for lactation requirements) with abomasal infusion of water, or R plus abomasal infusion of 6.25, 12.5, or 25 g/d choline ion. Liver tissue was collected on d 5 after infusions ended, and blood was collected on d 1, 3, and 5. Statistical contrasts were A0 versus R0 (CONT1), R versus the average of choline doses (CONT2), and tests of linear and quadratic effects of choline dose. Activity of cystathionine β-synthase was lower with R (CONT1) and decreased linearly with choline. Hepatic glutathione was not different with R or choline, but taurine tended to be greater with choline (CONT2). Betaine and carnitine were greater with R (CONT1) and further increased with choline (CONT2). Concentrations of NAD+ were greater with choline (CONT2). Cholic and glycol-chenodeoxycholic acids were decreased by R and choline, while taurocholic and tauro-chenodeoxycholic acids were not altered. Plasma aspartate aminotransferase and bilirubin were greater with R (CONT1) but decreased with choline (CONT2). Paraoxonase was lower with R and increased with choline (CONT2). Data suggest that enhanced supply of choline during NNB decreases entry of homocysteine to the transsulfuration pathway, potentially favoring remethylation to Met by acquiring a methyl group from betaine. As such, Met may provide methyl groups for synthesis of carnitine. Along with production data indicating that 12.5 g/d choline ion improved milk yield and liver fatty acid metabolism during NNB, the changes in blood biomarkers also suggest a beneficial effect of choline supply on liver function and oxidative stress. [ABSTRACT FROM AUTHOR]

Capillary electrophoresis (CE) is a technique routinely used in clinical laboratories that allows the separation and quantification of blood serum proteins in a rapid, precise, accurate, and inexpensive manner. Recently, CE has been proposed to separate and measure colostral proteins, but an evaluation of the agreement between CE and radial immunodiffusion (RID) method, currently used to quantify IgG in colostrum, is still lacking. The purpose of this study was to test the ability of a CE instrument, normally used in blood serum protein analysis, to realize the correct quantification of total Ig concentration in ewe colostrum, using RID assay as reference. Colostrum samples (n = 68) were collected from 35 multiparous Sarda ewes at first milking (n = 33) and at 24 h postpartum (n = 35). The mean ± standard deviation of IgG concentration measured by RID and whey colostrum total Ig concentration measured by CE were 54.76 ± 41.82 g/L and 54.70 ± 41.43 g/L, respectively. Lin's concordance correlation coefficient (r = 0.993; 95% confidence interval = 0.989 to 0.996) and linear regression analysis results (RID = 1.0022CE - 0.063; R2 = 0.986) showed an excellent agreement between these 2 methods. Bland-Altman analysis confirmed that CE method can be a suitable alternative to RID: the mean of the differences between CE and RID was -0.055 ± 4.95 g/L (95% confidence interval = -1.25 to 1.14 g/L) and the agreement limits were -9.75 to 9.60 g/L (low limit 95% confidence interval = -11.82 to -7.68 g/L; high limit 95% confidence interval = 7.57 to 11.72 g/L). In conclusion, the current study indicates that CE method may be a reliable tool for the quantification of the total Ig concentration in ewe colostrum. [ABSTRACT FROM AUTHOR]

• To reduce antibiotic usage in the dairy sector, selective dry-cow therapy is spreading. • Dry-off and calving altered time patterns of plasma biomarkers. • Cows treated or not with antibiotics in addition to the teat sealant at dry-off had similar performances. • No significant effects of different dry-cow therapies on udder health and milk composition were detected. • Immunometabolic profile was not affected by different dry-cow therapies. Pressure aimed at reducing the use of antibiotics in livestock is steadily increasing. In 2022, the prophylactic use of antibiotics for preventive purposes will be banned in the European Union (EU), including blanket therapy at dry-off. The objective of this study was to evaluate the short- and long-term effects of selectively treating cows with low somatic cell count (SCC) at dry-off using internal teat sealant with or without antibiotic therapy on udder health, milk production, metabolic, and inflammatory conditions through the next lactation. Fifteen Holstein dairy cows without intramammary infections and with SCC less than 200×103 cells/mL at dry-off were enrolled in the study. Cows were abruptly dried off and treated either with antibiotic plus teat sealant (AB) or with teat sealant only (TS). Milk and blood samples were collected on scheduled days from 10 days before dry-off to 28 days after calving. Milk composition and both inflammatory and metabolic profiles were assessed. Fertility and milk production were recorded during the previous and subsequent lactation. Rumination time was monitored from three weeks before dry-off to four weeks after calving. Data were analyzed with PROC MIXED and PROC GLM of SAS software. Total milk production, reproductive performance, rumination time, and mastitis incidence did not differ between groups. Milk yield during the first 120 days after calving was not affected by treatment. Milk composition and SCC were not affected during the first month. Compared with AB, TS had lower plasma calcium at -47 days from calving, gamma-glutamyl transferase at –50, –47, and –42, tocopherol at –65, –50, and –47, and alkaline phosphatase at 3 days from calving. Overall, both metabolic and inflammatory statuses were similar between AB and TS cows with only small transient differences. With the perspective of reducing antibiotic usage in dairy farms, internal teat sealant could be used alone in healthy cows with low SCC with no relevant effects on udder health and immunometabolic profile in the subsequent lactation. [ABSTRACT FROM AUTHOR]

Milk yield and composition are modified by level and chemical characteristics of dietary energy and protein. Those factors determine nutrient availability from a given diet, and once absorbed, they interact with the endocrine system and together determine availability of metabolites to the mammary gland. Four multiparous dairy cows in early lactation and subsequently in late lactation were fed 2 diets for 28 d in a changeover design that provided, within the same stage of lactation, similar amounts of rumen fermentable feed with either high (HS) or low starch (LS). All diets had similar dietary crude protein (15.5% dry matter) and rumen-undegradable protein (∼40% of crude protein) content. Profiles of AA were calculated to be similar to that of casein. On d 28, [1-13C] Leu was infused into one jugular vein with blood samples taken at 0, 2, 4, 6, and 8 h, and cows milked at 0, 2, 4, 5, 6, 7, and 8 h from start of infusion. Isotopic enrichments of plasma Leu, keto-isocaproic acid, and milk casein were determined for calculation of Leu kinetics. Data were subjected to ANOVA using the MIXED procedure of SAS (SAS Institute Inc.), with time as repeated factor and cow as the random effect. Dry matter intake within each stage of lactation was similar between groups. Feeding LS resulted in lower blood glucose and greater ratio of bovine somatotropin to insulin. This response was associated with greater blood concentrations of nonesterified fatty acids and β-hydroxybutyrate, which might have contributed to greater milk fat content in LS-fed cows. Except for His, average concentrations of all AA in blood were higher in late than early lactation. Diet did not alter average plasma concentrations of AA. However, for most of the essential AA (particularly branched-chain), the HS diet led to a marked decrease in concentrations after the forage meal, resulting in significant differences between dietary groups in early lactation. In early-lactating cows fed HS, a greater reduction in plasma concentrations at 8 h relative to pre-feeding values (time zero) was observed for Met, Lys, and His, resulting in decreases of 27.9%, 33.6%, and 38.5%, respectively. A higher bovine somatotropin/insulin ratio in early lactation and in cows fed LS could possibly have led to greater breakdown and, consequently, higher AA flux from peripheral tissues. In LS-fed cows, higher mobilization of body fat and protein was confirmed by the greater body weight loss in both stages of lactation. Higher irreversible loss of [1-13C] Leu in early lactation suggested lower protein retention in peripheral tissues during early compared with late lactation. Milk yield, protein output, and composition were similar between groups at both stages of lactation, whereas milk coagulation was faster (lower curd firming rate) and with higher curd firmness in response to feeding HS in late lactation. Overall, data indicated that rate of carbohydrate fermentability in the rumen can modify the availability of metabolites to the mammary gland and consequently modify milk protein coagulation. [ABSTRACT FROM AUTHOR]

We investigated how prepartal body condition score (BCS) alters key hepatic enzymes associated with 1-carbon, carnitine, and glutathione metabolism and the related biomarkers in liver tissue and plasma of periparturient dairy cows. Twenty-six multiparous Holstein dairy cows were retrospectively selected according to BCS at 4 wk prepartum and divided into high (HighBCS, BCS ≥ 3.50) and normal (NormBCS, BCS ≤ 3.25) BCS groups (n = 13 each). Blood plasma samples were obtained at −30, −10, 7, 15, and 30 d relative to calving. Liver tissue biopsies were performed at −15, 7, and 30 d relative to calving, and samples were used to assess protein abundance via Western blot assay. Cows in the HighBCS group lost ∼1 unit of BCS between −4 and 4 wk around calving, while NormBCS cows lost ∼0.5 unit in the same period. Prepartal dry matter intake (DMI, kg/d) did not differ between groups. Compared with NormBCS cows, HighBCS cows had higher postpartal DMI and milk yield (+5.34 kg/d). In addition, greater overall plasma concentrations of fatty acids and activity of the neutrophil-enriched enzyme myeloperoxidase were observed in HighBCS compared with NormBCS cows. Despite similar reactive oxygen metabolite concentrations in both groups at 30 d, HighBCS cows had lower overall concentrations of β-carotene and tocopherol, explaining the lower (BCS × Time) antioxidant capacity (ferric reducing ability of plasma). The HighBCS cows also had greater liver malondialdehyde concentrations and superoxide dismutase activity at 30 d. Overall, compared with NormBCS cows, HighBCS cows had lower hepatic protein abundance of the 1-carbon metabolism enzymes cystathionine-β-synthase, betaine-homocysteine methyltransferase, and methionine adenosyltransferase 1 A (MAT1A), as well as the glutathione metabolism-related enzymes glutathione S-transferase α 4 and glutathione peroxidase 3 (GPX3). A lower protein abundance of glutathione S-transferase mu 1 (GSTM1) at −15 and 7 d was also observed. Regardless of BCS, cows had increased abundance of GSTM1 and GPX3 between −15 and 7 d around calving. A marked decrease of gamma-butyrobetaine dioxygenase 1 from −10 to 7 d in HighBCS compared with NormBCS cows suggested a decrease in de novo carnitine synthesis that was partly explained by the lower abundance of MAT1A. Overall, data suggest biologic links between BCS before calving, milk yield, immune response, and hepatic reactions encompassing 1-carbon metabolism, carnitine, and antioxidant synthesis. [ABSTRACT FROM AUTHOR]

Peripartal cows mobilize not only body fat but also body protein to satisfy their energy requirements. The objective of this study was to determine the effect of prepartum BCS on blood biomarkers related to energy and nitrogen metabolism, and mRNA and protein abundance associated with AA metabolism and insulin signaling in subcutaneous adipose tissue (SAT) in peripartal cows. Twenty-two multiparous Holstein cows were retrospectively classified into a high BCS (HBCS; n = 11, BCS ≥ 3.5) or normal BCS (NBCS; n = 11, BCS ≤ 3.17) group at d 28 before expected parturition. Cows were fed the same diet as a total mixed ration before parturition and were fed the same lactation diet postpartum. Blood samples collected at −10, 7, 15, and 30 d relative to parturition were used for analyses of biomarkers associated with energy and nitrogen metabolism. Biopsies of SAT harvested at −15, 7, and 30 d relative to parturition were used for mRNA (real time-PCR) and protein abundance (Western blotting) assays. Data were subjected to ANOVA using the MIXED procedure of SAS (v. 9.4; SAS Institute Inc., Cary, NC), with P ≤ 0.05 being the threshold for significance. Cows in HBCS had greater overall plasma nonesterified fatty acid concentrations, due to marked increases at 7 and 15 d postpartum. This response was similar (BCS × Day effect) to protein abundance of phosphorylated (p) protein kinase B (p-AKT), the insulin-induced glucose transporter (SLC2A4), and the sodium-coupled neutral AA transporter (SLC38A1). Abundance of these proteins was lower at −15 d compared with NBCS cows, and either increased (SLC2A4, SLC38A1) or did not change (p-AKT) at 7 d postpartum in HBCS. Unlike protein abundance, however, overall mRNA abundances of the high-affinity cationic (SLC7A1), proton-coupled (SLC36A1), and sodium-coupled amino acid transporters (SLC38A2) were greater in HBCS than NBCS cows, due to upregulation in the postpartum phase. Those responses were similar to protein abundance of p-mTOR, which increased (BCS × Day effect) at 7 d in HBCS compared with NBCS cows. mRNA abundance of argininosuccinate lyase (ASL) and arginase 1 (ARG1) also was greater overall in HBCS cows. Together, these responses suggested impaired insulin signaling, coupled with greater postpartum AA transport rate and urea cycle activity in SAT of HBCS cows. An in vitro study using adipocyte and macrophage cocultures stimulated with various concentrations of fatty acids could provide some insights into the role of immune cells in modulating adipose tissue immunometabolic status, including insulin resistance and AA metabolism. [ABSTRACT FROM AUTHOR]

The objective of this study was to evaluate whether dairy sheep during the transition period are affected by their parity numbers with regard to (1) body weight (BW), body condition score (BCS), and production performance (milk yield and composition) and (2) metabolic, inflammation, and stress biomarkers. For this purpose, 30 Sarda dairy ewes [15 primiparous (PRP) and 15 multiparous (MUP) ewes] were recruited on d 90 of gestation. Each group was homogeneous according to age, BW, and BCS. Sampling was carried out at −60, −30, −7, 0, +30, and +60 d from lambing. The MUP ewes showed a higher BW (46.32 vs. 38.71 kg) and larger litter size (1.45 vs. 1.06 kg) but a lower BCS (2.47 vs. 2.70) than the PRP ewes. Furthermore, the MUP ewes had lower concentrations of glucose (3.49 vs. 4.27 mol/L), cholesterol (1.63 vs. 1.81 mmol/L), free fatty acids (0.47 vs. 0.62 mmol/L), and triglycerides (0.22 vs. 0.25 mmol/L) compared with PRP ewes. With regard to inflammation and oxidative stress parameters, the PRP group had higher haptoglobin (0.48 vs. 0.18 g/L) and paraoxonase (187.90 vs. 152.11 U/L) activity than the MUP group. Overall, the MUP ewes were characterized by greater milk production performance and greater feed intake, resulting in a better energy balance, than the PRP ewes. Interestingly, these findings highlighted a different metabolic and inflammatory response over the transition period between PRP and MUP ewes, with the latter displaying lower concentrations of inflammatory-related biomarkers. [ABSTRACT FROM AUTHOR]

Dairy cows with high body condition score (BCS) in late prepartum are more susceptible to oxidative stress (OS). Nuclear factor erythroid 2-like 2 (NFE2L2) is a major antioxidant transcription factor. We investigated the effect of precalving BCS on blood biomarkers associated with OS, inflammation, and liver function, along with mRNA and protein abundance of targets related to NFE2L2 and glutathione (GSH) metabolism in s.c. adipose tissue (SAT) of periparturient dairy cows. Twenty-two multiparous Holstein cows were retrospectively classified into a high BCS (HBCS; n = 11, BCS ≥3.5) or normal BCS (NBCS; n = 11, BCS ≤3.17) on d 28 before parturition. Cows were fed a corn silage- and wheat straw-based total mixed ration during late prepartum, and a corn silage- and alfalfa hay-based total mixed ration postpartum. Blood samples obtained at −10, 7, 15, and 30 d relative to parturition were used for analyses of biomarkers associated with inflammation, including albumin, ceruloplasmin, haptoglobin, and myeloperoxidase, as well as OS, including ferric reducing ability of plasma (FRAP), reactive oxygen species (ROS), and β-carotene. Adipose biopsies harvested at −15, 7, and 30 d relative to parturition were analyzed for mRNA (real-time quantitative PCR) and protein abundance (Western blotting) of targets associated with the antioxidant transcription regulator nuclear factor, NFE2L2, and GSH metabolism pathway. In addition, concentrations of GSH, ROS and malondialdehyde were measured. High BCS cows had lower prepartum dry matter intake expressed as a percentage of body weight along with greater BCS loss between −4 and 4 wk relative to parturition. Plasma concentrations of ROS and FRAP increased after parturition regardless of treatment. Compared with NBCS, HBCS cows had greater concentrations of FRAP at d 7 postpartum, which coincided with peak values in those cows. In addition, NBCS cows experienced a marked decrease in plasma ROS after d 7 postpartum, while HBCS cows maintained a constant concentration by d 30 postpartum. Overall, ROS concentrations in SAT were greater in HBCS cows. However, overall mRNA abundance of NFE2L2 was lower and cullin 3 (CUL3), a negative regulator of NFE2L2, was greater in HBCS cows. Although HBCS cows had greater overall total protein abundance of NFE2L2 in SAT, ratio of phosphorylated NFE2L2 to total NFE2L2 was lower, suggesting a decrease in the activity of this antioxidant system. Overall, mRNA abundance of the GSH metabolism-related genes glutathione reductase (GSR), glutathione peroxidase 1 (GPX1), and transaldolase 1 (TALDO1), along with protein abundance of glutathione S-transferase mu 1 (GSTM1), were greater in HBCS cows. Data suggest that HBCS cows might experience greater systemic OS after parturition, while increased abundance of mRNA and protein components of the GSH metabolism pathway in SAT might help alleviate tissue oxidant status. Data underscored the importance of antioxidant mechanisms at the tissue level. Thus, targeting these pathways in SAT during the periparturient period via nutrition might help control tissue remodeling while allowing optimal performance. [ABSTRACT FROM AUTHOR]

Synthetic zeolites are used to control the availability of dietary minerals (e.g., Ca, Mg, and P) in dairy cows. Due to calcium demand increasing with lactation onset, most cows become hypocalcemic immediately postpartum, which likely contributes to poorer immune function because calcium is important for immune cell signaling. To overcome postpartum hypocalcemia, we fed transition cows synthetic zeolite A (sodium aluminosilicate) precalving and hypothesized that it would alter calcium and thus neutrophil function during the transition period. Multiparous Holstein-Friesian cows in late gestation were randomly allocated to an untreated control group (n = 10) or a treatment group in which each cow received 500 g of zeolite A daily (n = 10) for 14 d prior to the expected calving date (actual duration = 17 ± 3 d prepartum). The cows grazed pasture, and each was supplemented with 2 kg/d of maize silage (dry matter basis), with or without zeolite, until calving. Blood samples for neutrophil isolation and analysis of plasma indicators of mineral status, energy status, liver function, and inflammation were collected pretreatment (covariate; d −19); on d −14 and −7 precalving; on the day of calving (d 0); and on d 1, 4, 7, and 28 postcalving. Neutrophils were isolated and gene expression was analyzed using microfluidic gene expression arrays. Neutrophil respiratory burst was assessed using stimulation with phorbol 12-myristate 13-acetate and flow cytometry. Plasma calcium and phosphorus revealed a treatment by time interaction; cows offered zeolite had greater plasma calcium concentrations at d 0, 1, and 4 postcalving and plasma phosphorus concentrations were lower in zeolite-treated cows during the precalving period until d 1 postcalving compared with control animals. Zeolite treatment downregulated neutrophil gene expression of CXCR4 and S100A8 and tended to lower gene expression for other immune mediators (CXCR1 , IFNG , S100A12 , and S100A9) compared with the control. Zeolite treatment did not affect neutrophil respiratory burst or expression of the other genes investigated. Plasma concentrations of cytokine IL-6 were reduced with zeolite treatment, which was most evident immediately postcalving (d 0, 1, and 7). Overall, feeding zeolite precalving had few effects on neutrophil gene expression and function; however, the lower gene expression of neutrophil inflammatory mediators may be due to altered availability of dietary minerals prepartum and indicates that zeolite A may control inflammation during the transition period. [ABSTRACT FROM AUTHOR]

In this study, Caciocavallo, a typical cheese produced in Sicily Island (Italy), was obtained from the milk of dairy cows fed with and without enriched olive cake (ECO and CTR, respectively) in order to evaluate nutritional, microbiological, volatile, and sensory differences in cheeses. ECO cheese showed greater ( p < 0.05) MUFA and PUFA and polyphenols content and lower SFA content than CTR cheese. Microbiological analyses revealed the absence of Salmonella, Listeria monocytogenes , Escherichia coli , and E. coli O157, and no significant differences in the viable counts of the remaining microbial groups analyzed, between samples. Thermophilic lactococci were more prevalent in ECO cheese. The implementation of a culture-independent method, such as PCR-DGGE analyses, revealed the presence of a more diverse microbial population in both cheeses. Regarding the volatile compounds profile, long-chain free fatty acids were more abundant in the ECO cheese, resulting in a healthier free fatty acid profile. This study also showed that, especially for their appearance and taste, consumers mostly appreciated the ECO cheese. The results show that using enriched olive cake could enhance the sustainability and the quality of Ragusano cheese, improving not only the health of its consumers but also positively influencing tastes and acceptability.

Animal health is affected during heat stress as a result of impaired immune responses, increased production of reactive oxygen species, and/or a deficiency of antioxidants. This leads to an imbalance between oxidants and antioxidants and results in oxidative stress. Heat stress is usually measured in dairy cattle via the temperature-humidity index (THI). In the present study, we aimed at assessing the influence of incremental THI on the balance between oxidative markers and the antioxidant defence system in the plasma of Modicana cows. Twenty-four multiparous, mid-lactating dairy cows were divided into two groups on the basis of different levels of mean THI reached in the period of the previous week up until the day of blood and milk sampling (April THI 1 :55, May THI 2 :68, June THI 3 :71, July THI 4 :80). The blood samples were collected to measure reactive oxygen metabolites (ROM) and antioxidant defense markers (ferric reducing ability of plasma (FRAP), paraoxonase (PON), advanced oxidation protein products (AOPP), plasma thiol groups (SHp), as well as lipid-soluble antioxidant pro-vitamin (β-carotene) and vitamins (tocopherol and retinol). Milk characteristics, haematological values, and plasma biochemical metabolites were also evaluated. Results showed a significant increase in ROM ( p < 0.05) and a significant decrease in PON ( p < 0.05), AOPP ( p < 0.05), and β-carotene ( p < 0.001). Incremental THI significantly decreased levels of milk fat content, red and white blood cells, plasma glucose, and non-esterified fatty acids, while significantly increasing monocytes and the concentrations of β-hydroxybutyrate and creatinine, but not fructosamine. The results of the study show that heat stress significantly affects reactive oxygen species production and antioxidant parameters. Carotenoid supplementation should be considered to alleviate the impact of these effects.

Enterococci are widely distributed in dairy sector. They are commensals of the gastrointestinal tract of animals, thus, via fecal contamination, could reach raw milk and dairy products. The aims of this study were: 1) to investigate the enterococcal diversity in cow feces and milk samples and 2) to evaluate the antibiotic resistance (AR) of dairy-related enterococci and their ability to transfer resistance genes. E. faecalis (59.9%), E. faecium (18.6%) and E. lactis (12.4%) were prevalent in milk, while E. faecium (84.2%) and E. hirae (15.0%) were dominant in bovine feces. RAPD-PCR highlighted a high number of Enterococcus biotypes (45 from milk and 37 from feces) and none of the milk strains exhibited genetic profiles similar to those of feces biotypes. A high percentage of enterococci isolated from milk (71%) were identified as multidrug resistant and resistance against streptomycin and tetracycline were widespread among milk strains while enterococci from feces were commonly resistant to linezolid and quinupristin/dalfopristin. Only E. faecalis strains were able to transfer horizontally the tetM gene to Lb. delbrueckii subsp. lactis. Our results indicated that Enterococcus biotypes from milk and bovine feces belong to different community and the ability of these microorganisms to transfer AR genes is strain-dependent., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Authors. Published by Elsevier Ltd.. All rights reserved.)

Mineral elements (Ca, Na, K, Mg, Zn, Ti, Sr, Fe, Ni, Ba, Cr, Mn, Cu, Se, Cd, Mo, B, V, As, Pb and Hg) in Provola cheeses obtained from dairy cows fed with two different integrated diets (Biotrak) and without olive cake (Control) were determined to discriminate between the two different cheeses. The results showed that cheeses from the Biotrak group presented higher values of essential elements. Selenium (Se) was found to be the most interesting: in Biotrak cheeses the content of Se was in the range of 0.112 to 0.281 mg/kg, about twice the content of Se in cheeses from the Control group. Among the toxic elements, only Cd was found in the samples, but at low levels (in average lower than 0.11 mg/kg). Therefore, the use of olive cake in animal feed is a good strategy to improve the mineral profile of the product obtained., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier Ltd. All rights reserved.)

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., (The Authors. Published by Elsevier Inc. and Fass Inc. on behalf of the American Dairy Science Association®. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).)

The aim of 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 present study aimed to investigate the effect of olive cake supplementation on faecal microbiota of Holstein (n = 16) and Modicana (n = 16) dairy cows. Although no difference in richness was detected, within breeds and between the two dietary treatment, the PERMANOVA analysis applied to the beta diversity allowed to discriminate samples according to breeds (p < 0.001) and treatment (p < 0.001). In Holstein cows, the olive cake supplementation led to the increase of Pseudobutyrivibrio and Christensenellaceae&#95;R7-group genera (p < 0.05) recognized as health-promoting or associated with feed efficiency. Differently, no difference was detected between control and treated groups for Modicana suggesting a high adaptive capacity to diet changes. In addition, the higher prevalence of Firmicutes phyla in the Modicana microbiota reflected its better capacity to digest the fibrous sources. Our study supports the suitability of olive cake as a feed supplement for cows and could help validating a sustainable livestock system in the Mediterranean area, characterized by a relevant oil production and by a native breeds reared with extensive systems., Competing Interests: Declaration of Competing Interest Nunziatina Russo, Alessandra Pino, Cinzia Caggia and Cinzia Lucia Randazzo declare that they are members of ProBioEtna, a spinoff of the University of Catania, Italy. In addition, the authors declare that they do not have any personal, financial, professional, political, or legal interests with a significant chance of interfering with the perfor-mance of their ethical or legal duties., (Copyright © 2023 The Authors. Published by Elsevier GmbH.. All rights reserved.)

Competing Interests: Authors declare no conflicts of interest exist.

The Comune di Sicilia, a local goat breed from Sicily (Italy), is currently undergoing recognition as a distinct breed. This study aims to characterize the population both phenotypically and genomically to advance its recognition process. A total of 78 subjects from two locations were enrolled, and their phenotypic data, including qualitative traits and morphometric measurements of adult animals, were recorded and statistically analyzed. The goats were genotyped using the Illumina 50 k Goat SNPchip, comparing them with 473 goats from 15 Italian breeds. Population structure, phylogenetic relationships, admixture, and genomic inbreeding were analyzed. Additionally, subjects with different morphological traits were compared using F ST and runs of homozygosity, leading to the identification of potential candidate genes associated with anotia and wattle presence in goats. The Comune di Sicilia breed exhibited distinctive genomic and phenotypic features, setting it apart from other breeds in the same region. However, moderate variability, possibly influenced by selection practices, was also observed. To ensure the breed's preservation and prevent excessive inbreeding, a comprehensive approach considering both morphology and genomic background is recommended. This study contributes valuable insights into the genetic peculiarities of the Comune di Sicilia goat, supporting its recognition as a unique and valuable breed.

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., (© 2023.)

Cold-pressed pomegranate seed oil (PSO) is a product of the extraction of non-edible pomegranate seeds. Its unique chemical composition in terms of both polyunsaturated fatty acids, especially punicic acid (PA), and secondary metabolites, such as phytosterols, tocopherols and phenols, make it an interesting functional ingredient for food enrichment. It is not clear if the biomarkers profile of PSO depends to factors connected to the geographical origin of seeds. This work presents a statistical comparative analysis, concerning biomolecules composition and geographical origin of 32 commercial cold-pressed PSOs, performed by principal component analysis. The study discriminates between Turkish and Italian PSOs, on the base of the fatty acid profile and phytosterols, and not on the tocopherols and phenols. These results confirmed PA as the main characteristic biomarker of oil genuineness and, for the first time, disclosed a statistically relevant variability of phytosterols, which can be proposed as quality biomarkers for discrimination of geographical origins.

Agro-industrial by-products incorporated into livestock feed formulations can positively impact feed costs and promote a circular bio-economy. Italy produces significant amounts of olive cake (OC), a by-product of olive oil extraction, with the potential for incorporation into bovine diets. However, information on its effects on endocrine responses in growing beef calves is lacking. Forty-eight Limousines randomly allocated to dietary treatment (control or 10%-OC or 15%-OC inclusion) were segregated according to sex and body weight. Serum concentrations of TSH, thyroid hormones, and cortisol were measured on day 0, day 56, and at the end of the trial on day 147. Circulating TSH, total (T 3 , T 4 ) and free (fT 3 , fT 4 ) iodothyronines, and cortisol concentrations were all within the normal physiological ranges, with no significant effect imparted by diet. However, the diet × time interaction was significant for T 3 . The cortisol, T 3 , T 4 , and fT 4 registered on day 147 were higher than those of day 56, and cortisol was higher in heifers than bulls. Final body weight was positively correlated with TSH and T 3 and negatively with cortisol concentration. These findings suggest that the inclusion of OC at levels up to 15% in growing/finishing beef diets had no adverse effects on the calves' thyroid and cortical status.

Vitamin B 12 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.

Olive by-products represent a valuable low-price feed supplement for animal nutrition. In the present study, the effect of the dietary destoned olive cake supplementation, on both composition and dynamics of the fecal bacterial biota of cow, was assessed by Illumina MiSeq analysis of the 16S rRNA gene. In addition, metabolic pathways were predicted by using the PICRUSt2 bioinformatic tool. Eighteen lactating cows, according to the body condition score, the days from calving, and the daily milk production were homogeneously allocated into two groups, control or experimental, and subjected to different dietary treatments. In detail, the experimental diet contained, along with the components of the control one, 8% of destoned olive cake. Metagenomics data revealed significant differences in abundance rather than in richness between the two groups. Results showed that Bacteroidota and Firmicutes were identified as the dominant phyla, accounting for over 90% of the total bacterial population. The Desulfobacterota phylum, able to reduce sulfur compounds, was detected only in fecal samples of cows allocated to the experimental diet whereas the Elusimicrobia phylum, a common endosymbiont or ectosymbiont of various flagellated protists, was detected only in cows subjected to the control diet. In addition, both Oscillospiraceae and Ruminococcaceae families were mainly found in the experimental group whereas fecal samples of control cows showed the presence of Rikenellaceae and Bacteroidaceae families, usually associated with the high roughage or low concentrate diet. Based on the PICRUSt2 bioinformatic tool, pathways related to carbohydrate, fatty acid, lipid, and amino acids biosynthesis were mainly up regulated in the experimental group. On the contrary, in the control group, the metabolic pathways detected with the highest occurrence were associated with amino acids biosynthesis and degradation, aromatic compounds degradation, nucleosides and nucleotides biosynthesis. Hence, the present study confirms that the destoned olive cake is a valuable feed supplement able to modulate the fecal microbiota of cows. Further studies will be conducted in order to deepen the inter-relationships between the GIT microbiota and the host., Competing Interests: NR, AP, CC, and CR declare that they are members of ProBioEtna, a spinoff of the University of Catania, Italy. The remaining authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2023 Russo, Floridia, D’Alessandro, Lopreiato, Pino, Chiofalo, Caggia, Liotta and Randazzo.)

This study aimed to investigate the immune-metabolic status and growth performance of Simmental calves born from cows subjected to pegbovigrastim administration 7 days before calving. Eight calves born from cows subjected to pegbovigrastim administration (PEG group) and 9 calves born from untreated cows (CTR group) were used. Growth measurements and blood samples were collected from birth to 60 d of age. The PEG group had lower body weight from 28 up to 60 d of age (P < 0.01), lower heart girth (P < 0.05), lower weekly and total average daily gain values (P < 0.05) than the CTR group throughout the monitoring period. A decrease in milk replacer (MR) intake was observed in the PEG group compared with the CTR group around 20-28 d of age (P < 0.01). The PEG group had lower values of γ-glutamyl transferase (GGT) at d 1 of age (P < 0.05), Zn at 21 and 28 d of age (P < 0.05), hemoglobin, MCH and MCHC at 54 and 60 d of age (P < 0.01), and higher urea concentration at 21 and 28 d of age (P < 0.05) compared with the CTR group. Lower values of retinol (P < 0.05), tocopherol (P < 0.01), mean myeloperoxidase index (P < 0.05) and higher total reactive oxygen metabolites (P < 0.05) and myeloperoxidase (P < 0.05) were also detected in the PEG group. In light of the results gathered in the current study, it can be speculated that activation of the cow's immune system by pegbovigrastim could have influenced the immune competence, growth performance as well as the balance between oxidant and antioxidant indices of the newborn calf., Competing Interests: Declaration of Competing Interest Authors declare no conflict of interest., (Copyright © 2023 Elsevier Ltd. All rights reserved.)

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.

Introduction: With the purpose to evaluate the effects of dietary olive cake, a source of bioactive phenolic compounds, as feed supplementation of lactating dairy cows on fatty acid composition, volatile organic compounds, and microbiological profiles of Provola cheese, we performed a two-arm study where control and experimental administered cows derived dairy have been compared., Methods: Our panel of analyses include metabolomics, physicochemical detected variables, culture dependent and independent analyses, and a stringent statistical approach aimful at disclosing only statistically significant results., Results and Discussion: Looking at the physicochemical variable's profiles, a higher content of unsaturated fatty acids, polyunsaturated fatty acid, and conjugated linoleic acids as well of proteins were observed in experimental cheese samples, indicating the beneficial effect of dietary supplementation. Furthermore, based on volatilome composition, a clear cluster separation between control and experimental cheeses was obtained, mainly related to terpenes degradation, able of influencing their aroma and taste. Microbiological results showed a decrease of some spoilage related microbial groups in experimental cheeses, probably due to the inhibitory effect exerted by polyphenols compounds, that contrarily did not affect the core taxa of all cheese samples. This paper confirmed the promising utilization of olive by-product in farming practices to obtain more sustainable and safe dairy food products with lower environmental impact, mainly in Sicily and Mediterranean area, where waste disposal poses serious environmental and economic problems., Competing Interests: NR, AP, CC, and CR declare that they are members of ProBioEtna, a spinoff of the University of Catania, Italy. In addition, the authors declare that they do not have any personal, financial, professional, political, or legal interests with a significant chance of interfering with the performance of their ethical or legal duties. The remaining authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2023 Calabrese, Russo, Celano, Pino, Lopreiato, Litrenta, Di Bella, Liotta, De Angelis, Caggia and Randazzo.)

Stress, inflammatory response, and their relationship were investigated in Simmental cows during the transition period (N = 8; 5 multiparous and 3 primiparous) and in their calves (N = 8; 5 heifers and 3 bulls). From cows, blood was collected at days -21 (±4), 0, +1, +7, and +21 days relative to calving. From calves, blood was collected after birth before colostrum intake (0) and then at 1, 7, and 15 days of age. Cortisol, Interleukin 6 (IL-6) and haptoglobin concentration was assessed by ELISA technique; white blood cells (WBC) were assessed using an ADVIA 2120 Hematology System machine. One-way ANOVA showed an effect of time for all the investigated parameters (P < 0.001) except for lymphocytes in peripartal cows. At calving and 1 d after, cortisol concentration was negatively correlated with levels of IL-6, WBC, and monocytes, whereas levels of IL-6 were positively correlated with WBC, neutrophils, and monocytes count. Cortisol, IL-6, haptoglobin, WBC and all leukocyte populations were affected by the age of neonatal calves (P < 0.001) except for neutrophils. A negative correlation between cortisol and IL-6, neutrophils, monocytes and haptoglobin was found at 15 days of age. A positive correlation between IL-6 and haptoglobin at day 15 of age, and with neutrophils and monocytes at days 7 and 15 of age was found. A positive correlation was obtained between cortisol levels measured in cows around calving and those obtained in calves after birth before colostrum intake (r = 0.83), and between IL-6 concentrations obtained from cows at calving and 1 d after and those obtained in calves at day 1 of age, after the colostrum intake (r = 0.93 and 0.79, respectively). The study suggests that immune function of peripartal cows is in an active state and that, in addition to other well-known factors driving the changes of parameters herein investigated, cortisol could have a role in the immune-modulatory adjustment during peripartum in cows. Furthermore, it can be hypothesized that cortisol is transferred from the cow to newborn calf through the placenta only and not through colostrum, whereas IL-6 levels in calves during the 24 h after birth seem to be influenced by IL-6 values measured in cows around calving due to its transfer through colostrum., Competing Interests: Declaration of competing interest The authors declare no competing interests. None of the authors has any financial or personal relationships that could inappropriately influence or bias the content of the paper., (Copyright © 2022 Elsevier Inc. All rights reserved.)

Organic contaminants belonging to various classes (plasticizers, bisphenols, pesticides, PCBs, and PAHs,) were analyzed in samples of provola cheese produced from Friesian dairy cows fed with a conventional diet (group CTR), and an unconventional diet (group BIO) enriched with olive cake (OC). The results show that for most determined contaminants, the differences between the two diets were very slight, indicating that the contamination does not depend on the olive cake integrated in the unconventional diet. The results also indicate that the minimal contamination could result from environmental contamination or the production process. It can be concluded that unconventional provola is as safe for the consumer as conventional provola.

The study aimed to evaluate the effects of different stages of lactation (0 to >  300 d) and pregnancy (0 to >  180 d) on serum amyloid A (SAA), C-reactive protein (CRP), glucose, total cholesterol (TCho), and triglyceride (TG) concentrations in dairy cows of different breeds. Thus, 40 healthy multiparous cows (10 Holstein, 10 Simmental, 10 Brown, and 10 Modicana) were randomly selected, and blood samples were collected once every 60 d for 1 year. Overall, SAA and CRP serum concentrations progressively increased and became more variable along the lactation, peaking at >  240-300 d, and then decreased in the last period ( >  300 d). Along pregnancy, SAA and CRP initially increased, with the highest concentrations at >  60-120 d, and then decreased until the last phase of pregnancy ( >  180 d). However, lactation and gestation phases did not significantly affect SAA and CRP when all the cows were analyzed together. A significant and positive correlation was observed between SAA and CRP both along lactation ( r = 0.89 ; p < 0.0001 ) and pregnancy ( r = 0.91 ; p < 0.0001 ). Breeds only showed differences in CRP levels along gestation ( p = 0.0102 ), due to a peak registered at 0-60 d in Holstein cows. In pregnant cows, glucose was positively correlated with SAA ( r = 0.43 ; p = 0.0017 ) and CRP ( r = 0.42 ; p = 0.0019 ). Hence, these significant and positive relationships reflect the physiological adaptations of the dairy cows along both gestational and lactational dynamics, suggesting that these proteins may also be involved in non-pathological processes. In this perspective, this study established that the obtained response markedly varies among healthy individuals along lactation and gestation and thus that the physiological range of acute-phase proteins (APPs) is wide; this makes it difficult to use these proteins as a marker of different physiological reproductive and productive periods., Competing Interests: The contact author has declared that none of the authors has any competing interests., (Copyright: © 2022 Esterina Fazio et al.)

The study examined the effects of different stages of pregnancy and lactation on thyroid, insulin, and glucose responses in dairy cows. In the present study, 30 dairy cows (10 Holstein, 10 Simmental, and 10 Brown) at 30 ± 20 d of lactation were randomly selected and blood samples were collected once every 60 d for one year to measure circulating thyroid stimulating hormone (TSH), total and free triiodothyronines (T3, fT3) and thyroxines (T4, fT4), insulin, and glucose. Pregnant cows showed higher T4 (p = 0.010) and insulin (p = 0.046) concentrations at >180 d than at >60−120 d of pregnancy and in nonpregnant cows. Along the lactation phase, circulating T4 concentrations showed a biphasic trend, decreasing from 0−60 d to >60−120 d phase, which showed the lowest values, and then increasing until the end of lactation (>300 d) (p = 0.016). Glucose showed the highest concentrations at the start of lactation (0−120 days) and the lowest values at the end (p = 0.008). The monitoring of thyroid hormones, insulin, and glucose changes represents an important tool to evaluate the anabolic and/or catabolic adaptation in response to functional periods in dairy cows, which can potentially predispose the cows to an excessive negative energy balance and related metabolic issues.