Elevated Milk Research Articles

Dietary thiamine supplementation modulates ruminal microbiota and partly restores lactation performance in lactating Hu ewes under heat-stress conditions

This study aimed to investigate the impact of dietary thiamine supplementation on milk production, rumen microbiota profile and inflammatory response of Hu ewes under heat-stress conditions. Twenty-seven multiparous pregnant and lactating Hu ewes of 71.6 ± 1.4 kg body weight, 11 ± 0.5 months gestation age, 31 ± 1 days in milk, 1.68 ± 0.15 L/d milk yield, and 2.14 ± 0.12 kg of daily dry matter intake (DMI), were used in the study. Following an acclimation period of 7 days, ewes were divided into 3 blocks of 9 each (one Hu ewe per pen), in a complete randomized design and assigned to one of three environmental temperature conditions for 15 days: thermoneutrality without supplementation (CON), heat-stressed without supplementation (HS) or heat-stressed with dietary thiamine supplementation (200 mg/kg of DMI; HST). The results indicated that the HS group experienced notable declines in milk protein, fat, and lactose content from day 10–15 (P < 0.05) compared to the CON group. In contrast, thiamine supplementation significantly elevated milk protein content from day 13–15 and raised milk fat and lactose content on days 14 and 15 (P < 0.05) compared to the HS group, thereby enhancing milk quality to a certain extent. Additionally, thiamine supplementation (HST group) significantly increased the circulating thiamine and Immunoglobulin A (IgA) concentrations accompanied by a significant decline in LPS-binding protein, interleukin-1 (IL-1), and serum amyloid A protein (SAA) levels compared to the HS group (P < 0.05). Rumen microbial composition analysis revealed that thiamine supplementation increased the abundance of Pyramidobacter, Butyrivibrio, Bacteroides, and Saccharofermentans. In conclusion, heat exposure during lactation in Hu ewes lowered milk production and quality, affecting rumen microbes. Dietary thiamine supplementation could improve rumen fermentation, mitigate heat stress-induced inflammation, and restore milk production in Hu ewes under heat stress conditions.

Mastitis has a cumulative and lasting effect on milk yield and lactose content in dairy cows

Milk lactose content (LC) physiologically decreases with parity order in dairy cows, but also after udder health inflammation(s) and/or in presence of elevated milk SCC in subclinical cases. Therefore, the progressive decrease in milk LC observed along cows' productive life can be attributed to a combination of factors that altogether impair the epithelial integrity, resulting in weaker tight junctions, e.g., physiological aging of epithelium, mechanical epithelial stress due to milking, and experienced clinical or subclinical mastitis. Mastitis is known to affect the udder synthesis ability too, so our intention through this study was to evaluate if there is a cumulative and lasting effect of mammary gland inflammation(s) on milk yield (MY) and LC. For this purpose, we used diagnoses of clinical mastitis and milk data of Austrian Fleckvieh cows to evaluate the effect of cumulative mastitis events on LC and MY. Only mastitis diagnoses recorded by trained veterinarians were used. Finally, we investigated if cumulative mastitis is a heritable trait and whether it is genetically correlated with either LC or MY. Estimates were obtained using univariate and bivariate linear animal models. A significant reduction in LC and MY was observed in cows that suffered from mastitis compared with those that did not experience udder inflammation. The h2 of cumulative mastitis is promising and much greater (0.09) than the h2 of the binary event itself (≤0.03). The genetic correlations between cumulative mastitis with LC and MY were negative, suggesting that cows with a great genetic merit for MY and LC are expected to be more resistant to repeated inflammations and less recidivist. When we used number of lifetime SCC peaks (≥200,000 or 400,000 cells/mL) to calculate cumulative inflammation events, h2 was even higher (up to 0.38), implying that also subclinical mastitis has a relevant negative impact on both LC and MY. Finally, the present study demonstrated how repeated mastitis events can permanently affect the mammary gland epithelial integrity and synthesis ability, and that the number of cumulative mastitis is a promising phenotype to be used in selection index in combination with other indicator traits toward more resistant and resilient mammary glands.

Asparagopsis taxiformis inclusion in grass silage-based diets fed to Norwegian red dairy cows: Effects on ruminal fermentation, milk yield, and enteric methane emission

Efforts to mitigate enteric methane (CH4) emission from ruminants by using the red macroalgae Asparagopsis taxiformis (AT) have demonstrated promising results both in vitro and in vivo. We assessed the effects of wild harvested and freeze-dried AT inclusion in the grass silage-based diets of Norwegian Red dairy cows on feed intake, milk yield and composition, rumen fermentation and CH4 emission. Fifteen (9 rumen cannulated and 6 intact) multiparous cows with an average (±SD) covariate period milk yield (MY) of 33.0 (3.8) kg/day were used in the experiment. The cows were divided into three groups (n = 5) after blocking for cannulation, and initial MY. After feeding on a common diet for 21 days (i.e., Covariate period), the groups were randomly allocated to three treatments: Control (no seaweed), 0.125%AT (Control + 0.125%AT, on organic matter (OM) basis) and 0.25%AT (Control + 0.25%AT, on OM basis). The cows were adapted to their diets for 13 days, followed by 39 experimental days for data collection. Inclusion of AT at 0.125% level did not affect dry matter intake (DMI, kg/d) but signicantly reduced both MY and energy corrected milk yield (ECM, kg/d) (P < 0.05) with a tendency for lower CH4 production (g/d) relative to the control group. Dry matter intake, ECM and CH4 production were reduced (P < 0.05) by 0.25%AT inclusion relative to the Control group. Apparent total tract nutrient digestibility (%), ECM per kg DMI, CH4 yield (g CH4/kg DMI) and CH4 intensity (g CH4/kg ECM) were not affected by AT inclusion. Inclusion of AT, at both levels, significantly reduced (P < 0.05) ruminal fluid total short chain fatty acids and molar proportions of actate, while increasing (P < 0.05) the molar proportion of propionate and isovalerate. Inclusion of AT decreased contents (%) of milk fat (P = 0.018) and protein (P = 0.010) resulting in significanly lower fat and protein yields. Lactose content (%) was higher for the AT groups (P = 0.021) but this did not compensate for lower MY resulting in lower milk lactose yield. Milk iodine content increased in a dose dependent manner with AT inclusion. In conclusion, AT inclusion at 0.25% level reduced CH4 emission from dairy cows with a grass silage-based diets. However, the reduced DMI for the 0.25%AT group along with reduced MY and elevated milk iodine content for both AT groups, for this particular harvest of AT, would suggest possible constraints for large scale inclusion of AT in dairy cow production.

In-Line Registered Milk Fat-to-Protein Ratio for the Assessment of Metabolic Status in Dairy Cows.

This study endeavors to ascertain alterations in the in-line registered milk fat-to-protein ratio as a potential indicator for evaluating the metabolic status of dairy cows. Over the study period, farm visits occurred biweekly on consistent days, during which milk composition (specifically fat and protein) was measured using a BROLIS HerdLine in-line milk analyzer (Brolis Sensor Technology, Vilnius, Lithuania). Clinical examinations were performed at the same time as the farm visits. Blood was drawn into anticoagulant-free evacuated tubes to measure the activities of GGT and AST and albumin concentrations. NEFA levels were assessed using a wet chemistry analyzer. Using the MediSense and FreeStyle Optium H systems, blood samples from the ear were used to measure the levels of BHBA and glucose in plasma. Daily blood samples were collected for BHBA concentration assessment. All samples were procured during the clinical evaluations. The cows were categorized into distinct groups: subclinical ketosis (SCK; n = 62), exhibiting elevated milk F/P ratios without concurrent clinical signs of other post-calving diseases; subclinical acidosis (SCA; n = 14), characterized by low F/P ratios (<1.2), severe diarrhea, and nondigestive food remnants in feces, while being free of other post-calving ailments; and a healthy group (H; n = 20), comprising cows with no clinical indications of illness and an average milk F/P ratio of 1.2. The milk fat-to-protein ratios were notably higher in SCK cows, averaging 1.66 (±0.29; p < 0.01), compared to SCA cows (0.93 ± 0.1; p < 0.01) and healthy cows (1.22). A 36% increase in milk fat-to-protein ratio was observed in SCK cows, while SCA cows displayed a 23.77% decrease. Significant differences emerged in AST activity, with SCA cows presenting a 26.66% elevation (p < 0.05) compared to healthy cows. Moreover, SCK cows exhibited a 40.38% higher NEFA concentration (p < 0.001). A positive correlation was identified between blood BHBA and NEFA levels (r = 0.321, p < 0.01), as well as a negative association between BHBA and glucose concentrations (r = -0.330, p < 0.01). Notably, AST displayed a robust positive correlation with GGT (r = 0.623, p < 0.01). In light of these findings, this study posits that milk fat-to-protein ratio comparisons could serve as a non-invasive indicator of metabolic health in cows. The connections between milk characteristics and blood biochemical markers of lipolysis and ketogenesis suggest that these markers can be used to check the metabolic status of dairy cows on a regular basis.

Natural history and risk factors of IgE-mediated cow's milk protein allergy in infants

Objective: To investigate the natural history and risk factors for continued allergy in infants with IgE-mediated cow's milk protein allergy (CMPA). Methods: This was a prospective cohort study that included 72 infants under 24 months of age diagnosed with IgE-mediated CMPA in the allergy clinic of the Children's Hospital, Capital Institute of Pediatrics from October 2019 to November 2020. General information, clinical manifestations, serum total IgE, cow's milk specific IgE, and cow's milk protein component specific IgE were collected. Follow-ups were conducted at 24 and 36 months of age, and the patients were divided into the persistent allergy group and the tolerance group based on whether they developed cow's milk tolerance at 36 months of age. Mann-Whitney U test, chi-square test, and binary Logistic regression were used for intergroup comparison and multivariate analysis. Results: Among the 72 CMPA children, there were 42 boys and 30 girls, with an age of 10 (7, 15) months at enrollment. Cow's milk protein tolerance was observed in 32 cases (44%) and 46 cases (64%) at 24 and 36 months of age, respectively. There were 26 cases in the persistent allergy group and 46 cases in the tolerance group. The proportion of respiratory symptoms, history of wheezing, positive specific IgE for α-lactalbumin and the total IgE level in the persistent allergy group were higher than that in the tolerance group (7 cases (27%) vs. 0, 6 cases (23%) vs. 2 cases (4%), 67% (14/21) vs. 26% (10/39), 225 (151, 616) vs. 48 (21, 185) kU/L, χ2=10.82, 4.16, 9.57, Z=4.07, all P<0.05). Multivariate Logistic regression analysis showed that anaphylaxis (OR=21.14, 95%CI 2.55-175.14, P=0.005), a history of allergic rhinitis (OR=5.94, 95%CI 1.54-22.86, P=0.005), elevated milk specific IgE (OR=1.04, 95%CI 1.01-1.08, P=0.024), and positive casein specific IgE (OR=6.64, 95%CI 1.39-31.69, P=0.018) were risk factors for continuous CMPA. Conclusions: Most infants with IgE-mediated CMPA can achieve tolerance within 3 years. Anaphylaxis, a history of allergic rhinitis, elevated milk specific IgE levels, and casein sensitization are risk factors for continuous allergy.

Reproductive performance of lactating dairy cows with elevated milk β-hydroxybutyrate levels during first 6 weeks of lactation

Although there is evidence that ketosis negatively affects fertility, the effect of late and early ketosis on the reproductive performance of lactating cows has not been systematically investigated. The aim of this study was to evaluate the association between time and amplitude of elevated milk BHB (EMB) occurring within 42 d in milk (DIM) and subsequent reproductive performance of lactating Holstein cows. The dairy herd information data of 30,413 cows with 2 test-day milk BHB recordings during early lactation periods 1 and 2 (5-14 and 15-42 DIM, respectively) assessed as negative (<0.15 mmol/L), suspect (0.15-0.19 mmol/L), or positive (≥0.2 mmol/L) for EMB were used in this study. Based on the time and amplitude of milk BHB, cows were grouped into 7 groups: (1) healthy cows negative in both periods 1 and 2 were classified as NEG; (2) suspect in period 1 and negative in period 2: EARLY_SUSP; (3) suspect in period 1 and suspect/positive in period 2: EARLY_SUSP_Pro; (4) positive in period 1 and negative in period 2: EARLY_POS; (5) positive in period 1 and suspect/positive in period 2: EARLY_POS_Pro; (6) negative in period 1 and suspect in period 2: LATE_SUSP; and (7) negative in period 1 and positive in period 2: LATE_POS. The overall prevalence of EMB within 42 DIM was 27.4%, with the highest prevalence being EARLY_SUSP (10.49%). Cows in EARLY_POS and EARLY_POS_Pro, but not other EMB categories, had a longer interval from calving to first service compared with NEG cows. For the reproductive parameters, first service to conception interval, days open and calving interval, cows in all EMB groups except EARLY_SUSP had longer intervals compared with NEG cows. These data indicate that there is a negative association between EMB within 42 d and reproductive performance after the voluntary waiting period. The intriguing findings of this study are the unaltered reproductive performance of EARLY_SUSP cows, and the negative association between late EMB and reproductive performance. Hence, monitoring and prevention of ketosis during the first 6 wk of lactation is necessary to optimize reproductive performance of lactating dairy cows.

A randomized clinical trial evaluating the effects of administration of acidogenic boluses at dry-off on rumination and activity behavior in the 14 subsequent days

Elevated milk production at dry-off can lead to increased udder pressure and, in turn, increased stress due to pain and discomfort, affecting natural behaviors. Administering acidogenic boluses at dry-off acts by inducing temporary and mild decreases in blood pH. This decreases dry matter intake, reduces milk yield, and increases cow comfort by lessening udder pressure. The objective of this study was to assess the effect of oral administration of acidogenic boluses at dry-off on total daily activity (TDA) and total daily rumination (TDR) behaviors in the first 2 wk of the dry period. This randomized clinical trial was conducted on a single farm and cows were randomly assigned to either treatment (TRT; n = 30) or control (CON; n = 34). The TRT group received 2 acidogenic boluses at dry-off and the CON group received no intervention. All cows received dry-cow therapy (intramammary antibiotic and internal teat sealant). The TDA and TDR data from 7 d before to 14 d after dry-off were measured using ear-mounted activity monitors. Analyses were performed using linear mixed-effects models with repeated measures. We observed a similar TDA in both groups throughout the study follow-up period. Overall, cows in the TRT group spent 17 min/d less time active than cows in the CON group in the first 2 wk after dry-off with the greatest difference observed on the second day of the dry period (TRT = 395 min/d; 95% CI: 370 to 420 vs. CON = 428 min/d; 95% CI: 404 to 451). The TRT group had lower TDR in the first 24 h after bolus administration (TRT = 437 min/d; 95% CI: 414 to 461 vs. CON = 488 min/d; 95% CI: 466 to 510) when compared with the CON group, but no differences were observed when comparing both groups in the 13 subsequent days. Our results indicate that administering acidogenic boluses at dry-off slightly decreased TDA during the first 2 wk of the dry period and decreased TDR on the first day after administration.

Genetic polymorphisms in immune- and inflammation-associated genes and their association with bovine mastitis resistance/susceptibility.

Bovine mastitis, the inflammation of the mammary gland, is a contagious disease characterized by chemical and physical changes in milk and pathological changes in udder tissues. Depressed immunity and higher expression of inflammatory cytokines with an elevated milk somatic cell count can be observed during mastitis in dairy cattle. The use of somatic cell count (SCC) and somatic cell score (SCS) as correlated traits in the indirect selection of animals against mastitis resistance is in progress globally. Traditional breeding for mastitis resistance seems difficult because of the low heritability (0.10-0.16) of SCC/SCS and clinical mastitis. Thus, genetic-marker-selective breeding to improve host genetics has attracted considerable attention worldwide. Moreover, genomic selection has been found to be an effective and fast method of screening for dairy cattle that are genetically resistant and susceptible to mastitis at a very early age. The current review discusses and summarizes the candidate gene approach using polymorphisms in immune- and inflammation-linked genes (CD4, CD14, CD46, TRAPPC9, JAK2, Tf, Lf, TLRs, CXCL8, CXCR1, CXCR2, C4A, C5, MASP2, MBL1, MBL2, LBP, NCF1, NCF4, MASP2, A2M, and CLU, etc.) and their related signaling pathways (Staphylococcus aureus infection signaling, Toll-like receptor signaling, NF-kappa B signaling pathway, Cytokine-cytokine receptor, and Complement and coagulation cascades, etc.) associated with mastitis resistance and susceptibility phenotypic traits (IL-6, interferon-gamma (IFN-γ), IL17, IL8, SCS, and SCC) in dairy cattle.

Associations of secretory activation breast milk biomarkers with breastfeeding outcome measures.

The objective of this study was to explore associations between day 10 postpartum (D10) secretory activation biomarkers and the breastfeeding outcome measures. This prospective longitudinal descriptive study collected antepartum, D10, and day 60 postpartum (D60) questionnaire data and D10 milk samples. Protein, lactose, and citrate were analyzed with enzymatic spectrophotometric assays. Sodium and potassium were analyzed with inductively coupled plasma optical emission spectrophotometry. Group comparison data were analyzed using χ2, Fisher exact, and independent sample t tests, as appropriate, using SPSS for Mac (version 28). Participants (n=92) provided a D10 breastmilk sample and completed D10 questionnaires, and 83 completed D60 questionnaires. Participants with D10 impaired secretory activation sodium (>23.0mM) were more likely to report D10 perceived insufficient milk supply, χ2=7.002, P<.05; and less D10 feeding/pumping frequency a day, P<.05; and partial breastfeeding at D60, P<.05. Additionally, participants with D10 impaired secretory activation sodium-to-potassium ratio (sodium: potassium)>0.8 were more likely to partially breastfeed at D60, P<.05. Elevated milk sodium and sodium: potassium are biomarkers related to variables indicative of low milk supply. Therefore, immediate milk testing can be useful in identifying lactation compromise and improving breastfeeding duration. Because breastfeeding affords maternal and infant health benefits, clinicians should identify ways to measure lactation compromise in conjunction with an examination and clinical history to provide early interventions to increase breastfeeding duration and exclusivity.

Human Milk Sodium and Potassium as Markers of Mastitis in Mothers of Preterm Infants.

Introduction: This prospective longitudinal study examined changes in milk sodium concentration (Na) and sodium:potassium ratio (Na:K), microbiological culture, milk production, and breast health in relation to mastitis after preterm birth. Methods: We studied women who gave birth at 29-34 weeks of gestation in a tertiary obstetric hospital in Perth, Western Australia. Milk samples, 24-hour milk production, and breast health data were collected every second day to day 10 postpartum, then every third day until infant discharge from the neonatal unit. Milk Na and K were measured at point of care (POC) using handheld ion selective meters, and Na:K calculated. Cultures were performed on postnatal days 8, 13, and every 6 days thereafter. For episodes of mastitis, milk was cultured at onset, and Na and Na:K measured daily until resolution. Women were followed up at 4 and 8 weeks postpartum. Results: In a sample of 44 women, 4 mastitis cases were detected in 3 women during their infants' neonatal stay; all had elevated milk Na and Na:K that resolved within 48 hours; 2/4 experienced reduced milk production and 1/4 had heavy growth of Staphylococcus epidermidis. A further 2 mastitis cases were reported in 39 women followed up to 8 weeks postpartum. Four women had elevated milk Na and Na:K without clinical signs of mastitis; three also had reduced milk production. Conclusions: POC testing of milk Na and/or Na:K may offer a useful indicator of breast health. Mastitis may cause an acute reduction in milk production regardless of the presence of culture-positive infection.

Association of pre-treatment somatic cell counts with bacteriological cure following diagnosis of intramammary infection

Antibiotic administration is crucial to ensure the health and productivity of dairy cattle. Mastitis is a disease that is typically a result of an intramammary infection (IMI), and antibiotic regimens are implemented to aid in curing IMI. Diagnosis is usually by detection of elevated milk somatic cell counts (SCC) and/or presence of culturable pathogens in the milk. Antibiotic treatment success is associated with the SCC at the time of treatment, though this correlation is still poorly understood. The objective of this project was to evaluate pre-treatment SCC and its association with IMI cure incidence following a standard antibiotic treatment. We hypothesized that pre-treatment SCC would be significantly lower in cases where the IMI ultimately cured compared to cases where the IMI failed to cure. Milk samples were collected aseptically from lactating cow quarters experiencing clinical or subclinical mastitis (n = 52). Clinical mastitis was diagnosed by a trained milking technician and subclinical mastitis was diagnosed at the quarter level as a SCC > 200,000 cells/mL and presence of bacterical growth in milk at time of treatment. After collection of the day 0 (D0) milk samples, the SCC was enumerated, and the milk sample cultured. Intramammary antibiotic therapy Cetftiofur hydrochloride (Spectramast® LC) was administered once/day for 5 days. Post-treatment samples were collected 14 d (D14) and 28 d (D28) later. A bacteriological cure was confirmed when both the D14 and D28 samples were free of culturable pathogens. The overall cure rate was 46.2%. Interestingly, the cure rates of antibiotic therapy decreased as pre-treatment SCC increased. Quarters that experienced bacteriological cure demonstrated a lower pre-treatment SCC (507,041 cells/mL ± 127.86 SEM, P = 0.01) compared to cows that did not cure, which had high pre-treatment SCC (1,640,392 cells/mL ± 333.28 SEM). Quarters that failed to cure had higher SCC values 28 days post-treatment in comparison to quarters that cured (P < 0.001). Future studies should investigate whether we can develop unique SCC-dependent mastitis treatment protocols which increase mastitis cure rates and enhance overall mammary health.

Widening the Lens on UV Filters: Newfound Ubiquity of Triazine UV Filters in Breast Milk from South China and Implications for Augmented “Cocktail” Exposure in Infants

Our previous work demonstrated triazine UV filters (TA-UVFs) as an emerging class of abundant, ubiquitous pollutants in the indoor environment; however, information about human exposure to these novel chemicals remains unknown. In this study, 17 TA-UVFs as well as 9 benzotriazole (BTR)-UVFs and 6 benzophenone (BP)-UVFs were included in a comprehensive, dedicated screening in breast milk from South China. In addition to some previously unrecognized BTR- and BP-UVFs, 11 of the 17 TA-UVFs were detected for the first time in breast milk, with ethylhexyl triazone (EHT) and bis-ethylhexyloxyphenol methoxyphenyl triazine (BEMT) as the predominant, omnipresent pollutants in humans. The median total concentration of TA-UVFs reached 1350 pg/mL, which is nearly half that of BTR-UVFs (3080 pg/mL) or BP-UVFs (3220 pg/mL). More daily use of personal care products (≥5 types) significantly elevated breast milk levels for all three classes of UVFs (p < 0.05) in the studied population. The overall daily intake of UVFs for infants via breast-feeding will substantially increase by 28% under the median exposure scenario if TA-UVFs are included in an integrative assessment. The current breast milk levels of TA-UVFs indicate a demand for additional monitoring and actions to reduce maternal and infant exposure to these emerging contaminants.

Got milk? Maternal immune activation during the mid-lactational period affects nutritional milk quality and adolescent offspring sensory processing in male and female rats.

Previous studies have underscored the importance of breastfeeding and parental care on offspring development and behavior. However, their contribution as dynamic variables in animal models of early life stress are often overlooked. In the present study, we investigated how lipopolysaccharide (LPS)-induced maternal immune activation (MIA) on postnatal day (P)10 affects maternal care, milk, and offspring development. MIA was associated with elevated milk corticosterone concentrations on P10, which recovered by P11. In contrast, both milk triglyceride and percent creamatocrit values demonstrated a prolonged decrease following inflammatory challenge. Adolescent MIA offspring were heavier, which is often suggestive of poor early life nutrition. While MIA did not decrease maternal care quality, there was a significant compensatory increase in maternal licking and grooming the day following inflammatory challenge. However, this did not protect against disrupted neonatal huddling or later-life alterations in sensorimotor gating, conditioned fear, mechanical allodynia, or reductions in hippocampal parvalbumin expression in MIA offspring. MIA-associated changes in brain and behavior were likely driven by differences in milk nutritional values and not by direct exposure to LPS or inflammatory molecules as neither LPS binding protein nor interleukin-6 milk levels differed between groups. These findings reflected comparable microbiome and transcriptomic patterns at the genome-wide level. Animal models of early life stress can impact both parents and their offspring. One mechanism that can mediate the effects of such stressors is changes to maternal lactation quality which our data show can confer multifaceted and compounding effects on offspring physiology and behavior.

Food antigen consumption and disease activity affect food-specific IgG4 levels in patients with eosinophilic esophagitis (EoE).

High levels of serum food-specific IgG4 (sIgG4) have been reported in patients with EoE. The objective of this study was to examine whether serum sIgG4 levels to foods and aeroallergens are higher in EoE patients than allergic controls and to investigate the association between sIgG4 and EoE clinical characteristics. This was a case-control study nested in a prospective EoE Cohort. EoE cases were defined per consensus guidelines, and controls were individuals with symptoms who were confirmed to be EoE-negative on upper endoscopy. Demographic and clinical information was prospectively collected. Serum IgE and sIgG4 were measured to foods and aeroallergens by ImmunoCAP. Mean levels of sIgG4 were compared between cases and controls, and logistic regression models were used to examine predictors of elevated milk sIgG4 levels. The analysis included 123 individuals (EoE n=93, control n=30) with a similar distribution of allergic disease between EoE patients and controls (86% vs. 93%; p=.30). EoE patients had significantly higher sIgG4 levels to all allergens evaluated, with the exception of birch (p=.24). Milk sIgG4 levels were independently associated with milk consumption (OR 4.95; p=.01) and the presence of sIgE to milk (OR 4.23; p=.008). Serum sIgG4 levels to food and aeroallergen proteins were higher in patients with EoE than non-EoE controls, and higher levels of milk sIgG4 were independently associated with milk consumption and the presence of sIgE to milk proteins. Whether sIgG4 plays a pathogenic role in EoE or could be used as an EoE biomarker remains unknown and warrants further study.

Epigenetic states of genes controlling immune responsiveness in bovine chronic mastitis

Abstract Mastitis is a common disease in dairy cows, causing substantial economic losses. The leading cause of elevated milk somatic cell count (SC), which is the best indicator for detecting mastitis, is the invasion of pathogens. A major pathogen responsible for bovine mastitis is Staphylococcus aureus, a member of the coagulase-positive staphylococci. Some strains of coagulase-negative staphylococci can also be a cause of clinical or subclinical mastitis. Our study used bisulfite sequencing PCR (BSP) to detect the methylation status of nine candidate genes (CCL2, HCK, F11R, CD8A, PDIA3, LGMN, HSPA1A, IL18 and NFKBIA). We investigated the mechanisms associated with overexpression of these genes, in the mammary gland secretory tissue of cows diagnosed with mastitis and infected with coagulase-positive or coagulase-negative staphylococci. The results showed no changes at the DNA methylation level between the mastitis (CoPS and CoNS) and control groups (H), except for in the HCK region, where the observed differences between the CoPS and H groups were statistically significant. The low methylation level of the CpG sequence seems not to correspond to the previously observed increased activity of these genes, suggesting that mechanisms other than DNA methylation may control mRNA expression at the analyzed loci.

Effect of different dietary regimens at dry-off on performance, metabolism, and immune system in dairy cows

Concentrate withdrawal and feed restriction are commonly used to reduce milk production and to facilitate dry-off, but may impair immune function in dairy cows. We investigated the effect of feed rations providing different amounts of nutrients in combination with feed restriction on performance, endocrine, and metabolic responses, as well as on leukocyte function before and after abrupt dry-off. Forty-three cows were studied from d 12 before until d 6 after dry-off (56 d before scheduled calving). Cows were fed experimental concentrates rich in crude protein (nitrogenic, n = 14), glucogenic precursors (glucogenic, n = 14), or lipids (lipogenic, n = 15). On d 3 before dry-off, total feed allowance was restricted to 50% in half of the animals of each dietary group, whereas feed allowance remained unchanged in the other animals. Performance parameters (milk yield, milk composition, and dry matter intake) were recorded, and daily blood and milk samples were taken and analyzed for various metabolic and endocrine parameters. Additionally, activity and mRNA abundance of several genes in leukocytes were measured at selected time points before and after feed restriction and dry-off, respectively. Feed restriction immediately resulted in a negative energy balance and decreased milk production. Concomitantly, concentrations of nonesterified fatty acids increased, whereas insulin, insulin-like growth factor-1, and glucagon decreased. After dry-off, energy balance turned positive and plasma nonesterified fatty acids decreased. Plasma glucose, insulin, and insulin-like growth factor-1 concentrations increased in all groups after dry-off. Glucose, insulin, and glucagon concentrations in plasma were higher in nonrestricted compared with restricted animals after dry-off. The experimental concentrate types marginally affected the investigated metabolic and endocrine factors, with the exception of elevated milk and plasma urea concentrations in cows fed the nitrogenic concentrate. Chemotactic and phagocytic activity of leukocytes were not affected by diets, feed restriction, or dry-off. Likewise, blood leukocyte mRNA abundance encoding for tumor necrosis factor α (TNF), heat shock protein family A (HSP70), and the glucose transporters (GLUT) 1 and 3 remained unchanged throughout the study period. Overall, the short-term negative energy balance induced by feed restriction was temporarily accompanied by metabolic adaptations, but did not alter the studied factors related to the immune system. Metabolic and endocrine adaptations supporting milk synthesis were continued during the first days after dry-off despite cessation of milking. Thus, the abrupt dry-off resulted in a short-term increase of glucose and triglyceride concentrations, with a delayed endocrine response to re-establish nutrient homeostasis in blood.

Characterization of ketolactia in dairy cows during early lactation

Fourier transform infrared spectroscopy (FTIR) allows for the determination of milk acetone (mACE) and β-hydroxybutyrate (mBHB) concentrations, providing a potential herd monitoring tool for hyperketolactia, defined as elevated milk ketone bodies. The study aim was to characterize mACE and mBHB concentration dynamics during early lactation in Polish Holstein-Friesian cows. Milk samples (n = 3,867,390) were collected within 6 to 60 days in milk (DIM) over a 4-yr period (April 1, 2013 to March 31, 2017) from approximately 21,300 dairy herds (average 38.7 cows/herd). Fixed effects of parity, DIM, and their interaction on mACE and mBHB concentrations were determined using a mixed model with a herd-year-season fixed effect and random cow effect. Published hyperketolactic mACE (≥0.15 mmol/L) and mBHB (≥0.10 mmol/L) threshold concentrations were used to classify study milk samples into ketolactia groups of normal (mACE <0.15 mmol/L and mBHB <0.10 mmol/L) and hyperketolactic (HYKL; either mACE ≥0.15 mmol/L or mBHB ≥0.10 mmol/L). Additionally, HYKL samples were categorized into subpopulations as having elevated mBHB and mACE (HYKLACEBHB, mACE ≥0.15 mmol/L and mBHB ≥0.10 mmol/L), only elevated mBHB (HYKLBHB; mACE <0.15 mmol/L and mBHB ≥0.10 mmol/L), or only elevated mACE (HYKLACE; mACE ≥0.15 mmol/L and mBHB <0.10 mmol/L). Effects of parity, DIM, ketolactia group or subpopulation, and their interactions on mACE and mBHB concentrations were also determined using the mixed model that included ketolactia group or subpopulation as an independent variable. Across all data, mACE and mBHB concentrations were influenced by effects of parity, DIM, and their interaction as well as parity, DIM, ketolactia group or subpopulation, and their interactions. For all samples, mACE and mBHB concentrations decreased with increasing DIM, with mACE concentration declining more rapidly compared with mBHB. In the data set, 68% and 32% of all samples were defined as normal or HYKL, respectively. Among HYKL samples, mACE was elevated soon after calving and declined over time. In contrast, mBHB started lower after calving and increased reaching peak concentrations around 30 DIM, and then decreased. Within HYKL samples, 50.8, 41.3, and 7.9% were categorized as HYKLACEBHB, HYKLBHB, and HYKLACE respectively. Between 6 and 21 DIM, 11.3% of HYKL were classified as HYKLACE. Primiparous cows had greater (14.8%) HYKLACE samples in this time period. In conclusion, this study has characterized mACE and mBHB concentrations during early lactation and determined effects of parity, DIM, and their interaction. Using published criteria interpreting mACE and mBHB concentrations, it was intriguing to identify a unique population of samples having elevated mACE without mBHB in early lactation, especially in primiparous cows. Further research is needed to determine if this sample population represents an unhealthy metabolic status that adversely affects cow health and performance.

Hypernatremic Dehydration Risk Factors in Newborns: Prospective Case-Controlled Study.

The prevalence of hypernatremic dehydration (HND) has increased in recent years most likely due to insufficient intake of breast milk as the most important factor. This study aimed to investigate risk factors of HND. In this study, 47 neonates whom were diagnosed to have HND were included in the study group and 96 healty neonates whom were included in the control group. While demographic data of the patients were recorded, mothers were asked to fill out the sociodemographic/ psychosocial data form, Edinburgh Postpartum Depression Scale(EPDS) and STAI I and II State and Trait Anxiety Scale. Breast milk sodium concentrations were studied from mothers of all infants. The relationship between the development of neonatal HND and risk factors affecting this condition were evaluated. Being the first-born baby of the family was found to be a significant risk factor for HND. Breast milk sodium concentration was 25.8±7.9 mmol/L in the HND group which was significantly higher than the control group. Median depression score was similar in both groups and there was no statistical difference in terms of groups. The anxiety score was higher in the control group compared to the study group. There was no difference in terms of other sociodemographic / psychosocial data of mothers. Primiparity or insufficient breastfeeding may result in elevated breast milk sodium levels and related neonatal HND. Breastfeeding support should principally target primiparous women to improve breastfeeding outcomes like as especially HND.

Milk Phospholipid Profiling Among Japanese Women with Differing Docosahexaenoic Acid Levels

Objectives: To characterize the milk phospholipids (PLs) profile using phosphorus-31 nuclear magnetic resonance (31P-NMR) and to investigate the effect of elevated milk docosahexaenoic acid (DHA) levels on PLs profiles in Japanese mothers. Methods: Milk samples from eligible patients with high and low DHA from a former cross-sectional study (n = 20; n = 10 for each group) were included. Fifteen milk PLs were analyzed using 31P-NMR, and the profiles were compared group-wise using Mann–Whitney U-test. The P value of &lt;0.05 was considered statistically significant. Results: The median DHA content in milk was 1.13% and 0.29% for the high and low milk DHA groups, respectively. Twelve PLs, excluding lysophosphatidylserine, cardiolipin, and phosphatidylglycerol, were detected in all participants with 100% positive results. The median concentrations and proportions of total PLs, sphingophospholipids, and glycerophospholipids were comparable between groups. The proportions of choline-containing glycerophospholipid were significantly higher in the high milk DHA group than that in the low milk DHA group (24.09% [median, interquartile range: 23.08%–26.38%] and 21.41% [20.74%–22.84%], P = 0.019). Although the proportions of phosphatidylinositol were significantly lower in the high milk DHA group than that in the low milk DHA group (6.62% [5.75%–6.72%] versus 7.63% [7.11%–8.16%], P = 0.002), while that of phosphatidylcholine (21.90% [18.51%–23.22%] versus 19.78% [18.17%–20.26%], P = 0.059) and alkyl-acyl phosphatidylcholine (0.60% [0.40%–0.74%] versus 0.33% [0.14%–0.51%], P = 0.059) were higher in the former than that in the latter. Conclusions: Our results were comparable to that of the previous literature. Large variations in the milk DHA might affect the composition of choline-containing glycerophospholipids in Japanese mothers. However, possible confounders were not excluded in the study populations.

Preweaning nutrient supply alters serum metabolomics profiles related to protein and energy metabolism and hepatic function in Holstein heifer calves

Lifting the preweaning milk restriction in dairy calves has been causally associated with beneficial effects on growth and future lactation performance. However, the biological mechanisms linking early-life nutrient supply and future performance remain insufficiently understood. Thus, the objective of this study was to characterize growth and the metabolic profiles of calves fed a restricted (RES) and an elevated (ELE) milk supply preweaning. A total of 86 female Holstein Friesian calves were blocked in pairs by maternal parity and received identical colostrum supply within block. Treatments randomized within block consisted of a milk replacer (MR; 24% crude protein, 18% crude fat, and 45% lactose) supplied at either 5.41 Mcal of ME in 8 L of MR/d (ELE) or 2.71 Mcal of ME in 4 L of MR/d (RES) from d 2 after birth until they were stepped down by 50% during wk 7 and fully weaned at wk 8. All calves had ad libitum access to pelleted calf starter (17.3% crude protein, 24.4% neutral detergent fiber, 2.0% crude fat, and 18.2% starch), chopped wheat straw, and water. At 2 and 49 d of age, blood samples were taken for metabolomics analysis. The ELE group by design consumed more milk replacer, resulting in a lower starter intake and a greater body weight and average daily gain. The ELE calves consumed 20.7% more ME and 9.7% more crude protein. However, efficiency of growth was not different between groups. Metabolomic profiling using 908 identified metabolites served to characterize treatment-dependent biochemical differences. Principal component analysis revealed clearly distinct metabolic profiles at 49 d of age in response to preweaning milk supply. Changes in energy (fatty acid metabolism and tricarboxylic acid metabolites), protein (free AA, dipeptides, and urea cycle), and liver metabolism (bile acid and heme metabolism) were the main effects associated with the dietary differences. The ELE group consumed proportionately more glucogenic nutrients via milk replacer, whereas the RES group consumed proportionately more ketogenic nutrients from the digestion of the calf starter, comprising a larger portion of total intake. Associated with the higher growth rate of the ELE group, hepatic changes were expressed as differences in bile acid and heme metabolism. Furthermore, energy metabolism differences were noted in fatty acid and AA metabolism and the urea cycle. The metabolic profile differences between the ELE and RES groups reflect the broad differences in nutrient intake and diet composition and might point to which metabolic processes are responsible for greater dairy performance for cows fed a greater milk supply preweaning.