Milk Cortisol Research Articles

Composition and Antioxidant Status of Human Milk of Women Living in Bydgoszcz (Poland).

The aim of this study was to compare cortisol concentrations, nutritional composition, and the antioxidant status of human milk of women living in Bydgoszcz (Poland), taking into account maternal factors (fertility, area of residence, economic activity, and breastfeeding period). The basic composition of human milk was evaluated using the MIRIS HMATM analyzer. The level of cortisol was determined by the enzyme-linked immunosorbent method. In order to determine the antioxidant activity, the DPPH radical method was used. It was observed that the concentration of cortisol in human milk in the group of women living in the city center was higher compared to the milk of women living on the outskirts of the city. In the group of women breastfeeding from 3 to 5 weeks after childbirth, the concentration of cortisol in milk was higher compared to the group of women breastfeeding less than 12 months of age and compared to the group of women lactating over 12 months of age. The antioxidant status of human milk was highest in the group of professionally active women and in the group of breastfeeding women from 3 to 5 weeks after childbirth. The basic composition and the caloric value of human milk differed statistically significantly in the study groups. Based on this study, it can be concluded that the composition and antioxidant status of human milk depends on maternal factors (fertility, professional activity, area of residence, and breastfeeding period). Higher cortisol concentrations in breast milk are probably determined by the area of residence (city center and associated higher noise/sound and stress levels) and lactation period (hormonal imbalance, fatigue, and postpartum period). Milk from economically active women shows greater protection against reactive oxygen species compared to milk from inactive women, protecting against the occurrence of diseases of civilization. Milk from breastfeeding women over 12 months of age also shows protection against reactive oxygen species, despite the fact that the highest level of antioxidant status of human milk occurs in the initial period of lactation.

Assessment of mammary stress in early lactating crossbred (Alpine × Beetal) does during heat stress in conjunction with milk somatic cells and non-invasive indicators

Milk somatic cell counts (SCC) are crucial for monitoring dairy animals' mammary health and milk quality. This study was initiated to assess the mammary health of early lactating crossbred (Alpine × Beetal) does during periods of heat stress. The udder skin surface temperature (USST), total and differential milk SCC, milk composition, milk cortisol levels and phagocytic activity (PA) of milk neutrophils and macrophages were estimated in these does. Results indicated a significant (p<0.01) increase in the milk neutrophils to macrophages (N: M) ratio and milk cortisol levels with milk somatic cell score (SCS). The PA of milk neutrophils decreased significantly (p<0.01), while that of macrophages increased significantly (p<0.01) with higher SCC. On Pearson correlation, milk SCS revealed strong positive correlation (p<0.01) with N: M ratio (r=0.90), USST (r=0.98), milk cortisol (r=0.97), milk pH (r=0.83) and conductivity (r=0.92). Conversely, milk lactose exhibited a moderate negative correlation of (r= −0.48) with SCS. These findings suggest that USST infrared thermography and milk cortisol levels are promising non-invasive methods for rapidly assessing mammary stress in early lactating does during heat stress.

Impact of Maternal Health and Stress on Steroid Hormone Profiles in Human Milk: Implications for Infant Development

Steroid hormones are biologically active factors in human milk (HM) that influence the physical and mental development of infants. Critically, maternal psychosocial stress has been associated with changes in HM steroid composition. This work aimed to characterize the steroid hormone profile of HM and pasteurized donor human milk (DHM) and assess the interplay between maternal physical and psychosocial status, the HM steroid profile, and infant outcomes.A targeted ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) method was developed to quantify sixteen steroid hormones in HM samples. HM samples from mothers of term infants (N=42) and preterm infants (N=35) were collected at (i) recovery of birth weight or achievement of complete enteral nutrition, respectively, and (ii) 6 months later as well as DHM samples (N=19) from 11 donors. The physical and psychosocial status of mothers and infant neurodevelopment and temperament were assessed through structured interviews and validated questionnaires.Fourteen steroids were detected in HM/DHM samples, with cortisol, 20β-dihydrocortisol, dehydroepiandrosterone, pregnenolone, and cortisone being present in > 48% of samples. Pregnenolone, 17α-OH-progesterone, and dehydroepiandrosterone are reported for the first time in HM. Whereas milk cortisol levels were not directly related with maternal physical and psychosocial status nor with infant development, cortisone and pregnenolone correlated positively with maternal weight gain during pregnancy and were associated to maternal well-being and infant growth. The pasteurization process may have a detrimental effect on the steroid hormone levels in HM, what might influence the development of receptors.

Infant Temperament Is Associated With Milk Cortisol but Not With Maternal Childhood Trauma.

Previous studies have suggested that maternal childhood trauma (MCT) may influence infant temperament, but the underlying physiological mechanisms remain unclear. This study sought to confirm the involvement of breast milk cortisol in the link between MCT and infant temperament. The study sample included 90 mother-infant dyads recruited from the urban Polish population. MCT was assessed based on the Early Life Stress Questionnaire (ELSQ) and infant temperamental factors (surgency/extraversion, negative affectivity, and orienting/regulation) using the Infant Behavior Questionnaire-Revised at 12 months of life. Cortisol was assayed in milk samples collected at 5 months of life using the ELISA method. Based on the ELSQ median, the sample was divided into low and high MCT groups. The ANCOVA models with milk cortisol as a covariant were run to check the effect of low versus high MCT on infant temperament. We found a positive association between milk cortisol and orienting/regulation. Surprisingly, the low and high MCT groups did not significantly differ in milk cortisol. Furthermore, we found that MCT was unrelated to any infant temperamental factor. While recent literature on the association between milk cortisol and infant temperament is inconsistent, our results suggest that high orienting/regulation might be an adaptation to adverse environments such as stress. Moreover, the infant's temperament appears to be more responsive to the current exposition to maternal stress than her experience of traumatic stress.

Injection of antioxidant trace minerals/vitamins into peripartum crossbred cows improves the nutritional and immunological properties of colostrum/milk and the health of their calves under heat stress conditions.

Multimineral and vitamin injections can provide better nutrient availability at the cellular level, which is essential for mitigating transition period stress and improving the wellbeing and productivity of dairy cows. The present study was conducted to assess the colostrum quality and calf health after intramuscular injection of multi-minerals (MM) and multi-vitamins (MV) to peripartum cows during winter (THI = 58 to 66) and summer (THI = 78 to 82) months. In each season, twenty-four pregnant crossbred Karan Fries cows were grouped into four, each consisting of six cows. Group I, referred to as the Control, received solely the basal diet, without any additional supplements. Groups II, III, and IV were administered additional MM (T1), MV (T2), and a combined MM and MV (T3) along with their basal diet, starting 30days before calving and continuing for 30days after calving. Blood samples were collected from the calves, while colostrum/milk samples were obtained from the cows on days 1, 3, 7, and 15 after calving. The somatic cell counts (SCC) in the milk were determined using a cell counter. Cortisol, IgG, IGF1 and total immunoglobulins (TIG) in whey and plasma from cow colostrum/milk or calf blood samples were estimated by ELISA. Cows that calved in the summer exhibited notably reduced levels (P < 0.05) of IgG, milk, and plasma IGF1, along with lower calf body weights, in comparison to those calving in the winter season. Furthermore, the summer months saw significant increases (P < 0.05) in plasma and milk cortisol levels, as well as total somatic cell counts (SCC) in both colostrum and milk samples. Maximum beneficial effect was observed in T3 group. Results indicate that injections to peripartum cows could be an important strategy for improving colostrum quality and calf health during the summer seasons.

The association between maternal factors and milk hormone concentrations: a systematic review.

Breast milk is the gold standard for infant feeding. It is a dynamic biological fluid rich in numerous bioactive components. Emerging research suggests that these components, including hormones, may serve as signals between mother and offspring. From an evolutionary perspective, maternal hormonal signals could allow co-adaptation of maternal and offspring phenotype, with implications for their Darwinian fitness. However, a series of steps need to be considered to establish the role of a component as a signal and this systematic review focuses on one step: 'Do maternal factors influence the concentration of milk hormones?' To systematically review human studies which analyze the association between maternal factors and the concentration of hormones in breast milk. Three databases were searched for studies reporting the association of maternal factors including body mass index (BMI), weight, fat mass, age, ethnicity, smoking with hormones such as adiponectin, leptin, insulin, ghrelin, and cortisol in breast milk. Thirty-three studies were eligible for inclusion. Maternal BMI was positively associated with milk leptin (20/21 studies) and with milk insulin (4/6 studies). Maternal weight also displayed a positive correlation with milk leptin levels, and maternal diabetes status was positively associated with milk insulin concentrations. Conversely, evidence for associations between maternal fat mass, smoking, ethnicity and other maternal factors and hormone levels in breast milk was inconclusive or lacking. Current evidence is consistent with a signaling role for leptin and insulin in breast milk, however other steps need to be investigated to understand the role of these components as definitive signals. This review represents a first step in establishing the role of signaling components in human milk and highlights other issues that need to be considered going forward.

Ramadan fasting in the third trimester of pregnancy and postpartum colostrum cortisol concentrations in Morocco.

Breast milk cortisol concentration varies among and within lactating women, but the causes and consequences of variation are not yet fully understood. This study investigated whether maternal fasting from food and water for over 16 h per day during the third trimester of pregnancy was associated with increased concentrations of cortisol in colostrum during the month of Ramadan in a Moroccan sample. Participants were women who gave birth at the public maternity hospital in Rabat, Morocco throughout the month of and month following Ramadan. Participants completed interviews (N = 92), underwent anthropometric assessment (N = 73), and colostrum samples (N = 63) were collected approximately 48-72 h postpartum. Cortisol analysis was completed on a subset of colostrum samples (N = 43). Women were categorized as fasting every day of Ramadan before giving birth (N = 52/92, 57%), fasting some days (N = 29/92, 32%), and not fasting (N = 11/92, 11%). For women who fasted, average fasting duration was 19.5 days (range: 1-29 days), and average time between the last day of fasting and birth was 8.8 days (range: 0-43 days). The number of days of fasting was positively associated with cortisol concentrations in colostrum (p < .009) in a multivariate analysis controlling for maternal anthropometry, hypertension and/or diabetes, and reasons for opting out of fasting. Each day of fasting was associated with a 4.05% increase in colostrum cortisol. Results are consistent with the hypothesis that maternal fasting during the third trimester of pregnancy increases breast milk cortisol and thus alters the hormonal milieu to which the infant is exposed.

Association between maternal stress and premature milk cortisol, milk IgA, and infant health: a cohort study.

Maternal stress is pervasive in the neonatal intensive care unit (NICU). Maternal stress is associated with changes in human milk (HM) immunomodulatory agents, which may impact neonatal health. We sought to determine the association between maternal stress, HM immunoglobulin A (IgA) and cortisol, and to assess how these milk components correlate with infant immune and neurodevelopmental outcomes. We then compared how these associations persist over time. The study design involved a cohort study of exclusively breastfeeding mothers and their singleton moderately preterm (28-34 weeks) infants admitted to the NICU. We collected maternal serum, maternal saliva, and first-morning whole milk samples, and administered maternal stress questionnaires at 1 and 5 weeks postpartum. We analyzed the samples for HM IgA (using a customized immunoassay in skim milk) and for HM and salivary cortisol (using a chemiluminescent immunoassay). Infant illness was assessed using the Score for Neonatal Acute Physiology II (SNAP II) and SNAP II with Perinatal Extension (SNAPPE II), and infant neurodevelopment were assessed using the Test of Infant Motor Performance. We analyzed changes in HM IgA and cortisol over time using paired t-tests. Furthermore, we performed correlation and regression analyses after adjusting for gestational age (GA), corrected GA, and infant days of life. In our study, we enrolled 26 dyads, with a mean maternal age of 28.1 years, consisting of 69% white, 19% Black, and 8% Hispanic. Cortisol: Salivary and HM cortisol were closely associated in week 1 but not in week 5. Though mean salivary cortisol remained stable over time [2.41 ng/mL (SD 2.43) to 2.32 (SD 1.77), p = 0.17], mean HM cortisol increased [1.96 ng/mL (SD 1.93) to 5.93 ng/mL (SD 3.83), p < 0.001]. Stress measures were inversely associated with HM cortisol at week 1 but not at week 5. IgA: HM IgA decreased over time (mean = -0.14 mg/mL, SD 0.53, p < 0.0001). High maternal stress, as measured by the Parental Stressor Scale: neonatal intensive care unit (PSS:NICU), was positively associated with HM IgA at week 5 (r = 0.79, P ≤ 0.001). Higher IgA was associated with a lower (better) SNAP II score at week 1 (r = -0.74, p = 0.05). No associations were found between maternal stress, salivary cortisol, HM cortisol, or HM IgA and neurodevelopment at discharge (as assessed using the TIMP score). Furthermore, these relationships did not differ by infant sex. Maternal stress showed associations with HM cortisol and HM IgA. In turn, HM IgA was associated with lower measures of infant illness.

Energy-Related Hormones in Raw and Retail Cow’s Milk and Possible Risk for Consumers

Abstract The study aimed to determine the concentration of energy-related hormones in cow’s milk and to consider them from a public health perspective. Fourteen Holstein cows were subjected to milk and blood sampling on the following days in lactation (DIL): 10, 30, 60, 90, 150, 180, 210, 250 and 280 to determine milk hormones, fat and protein content and blood biochemical parameters. For the same purpose, bulk-tank milk was sampled and samples of retail m ilk with 1.5% (CM1.5) and 3.2% (CM3.2) fat was purchased. Milk insulin-like growth factor-1 (IGF-1) values were significantly lower at 90, 150, 180, 210 and 250 and significantly higher at 10, 30 and 60 DIL than lactation average (LA). Milk insulin concentrations were significantly lower at 30, 60 and 90 and higher at 210, 250 and 280 DIL than LA. Free thyroxine (fT4) level in the milk was higher at 250 DIL, while milk free triiodothyronine (fT3) concentrations were lower at 30, 60, 90 and 280 DIL, and significantly higher at 10 and 180 DIL than respective LA. Milk cortisol levels were lower at 60 and 280 DIL than LA. All measured milk hormones were significantly lower in CM1.5 compared to CM3.2, bulk-tank milk and LA. An exception was the LA of IGF-1, which was significantly lower than the IGF-1 content in CM1.5. Blood biochemical parameters fluctuated evenly during lactation and were within the reference range. Hormone concentrations in cow’s milk fluctuate during lactation, giving milk an important role in the context of public health.

Stress indicators in dairy cows adapting to virtual fencing.

Virtual fencing (VF) enables livestock grazing without physical fences by conditioning animals to a virtual boundary delimited with an audio tone (AT) and an electric pulse (EP). The present study followed the adaptation process of lactating dairy cows to a VF system with changing virtual boundaries and investigated its impact on animal welfare. Twenty cows were divided into stratified groups (2× VF; 2× electric fencing, EF) of five individuals. Each group grazed half-days in a separate EF paddock of comparable size during 3d of acclimation (P0), followed by 21, 14, 14, and 7d of experimental treatment (P1 to P4). At the start of the trial, all cows were equipped with an IceQube pedometer (Peacock Technology Ltd, Stirling, UK) and a VF collar (Nofence AS, Batnfjordsøra, Norway). During P0, cows were accustomed to their first paddock with a deactivated virtual boundary and wearing the sensors. In P1 to P4, an active virtual boundary for the VF groups, and a second EF for the EF groups was set up parallel to an outer EF within their paddock. Throughout the trial, the sensors continuously tracked cow positions and activity behavior at 15-min intervals. From P1 onwards, the VF collars additionally recorded each AT and EP per cow with a georeferenced time stamp. During P0 to P4, daily feed intake, body weight, and milk yield were recorded in the barn. A total of 26 milk samples were collected per cow to determine milk cortisol levels. Behavioral observations were conducted for 2h on day 23 to record agonistic behaviors, vocalizations, and excretions. The total number of stimuli per cow ranged from 37 to 225 ATs (mean ± SD: 1.9 ± 3.3 per day) and 3 to 11 EPs (mean ± SD: 0.1 ± 0.7 per day) throughout the trial. The maximum number of EPs per day was 8 for an individual cow and occurred once on D1. Mean EP/AT decreased by 55% during the first three half-days of grazing and with each paddock change from 0.2 EP/AT in week 1 to 0.03, 0.02, and 0 EP/AT in weeks 4, 6, and 8, respectively. Linear and generalized mixed effects models revealed that milk yield and cortisol, feed intake, body weight, and activity and lying behavior did not significantly differ between VF and EF groups. A higher number of agonistic behaviors were observed in the VF groups when the VF system was activated. However, due to the short observation periods only few contacts were observed in total. Overall, all cows adapted to the VF system without evidence of lasting adverse effects on animal welfare.

PSVI-16 Conditioning Dairy Cows to a Virtual Fencing System Is Compatible with Animal Welfare

Abstract Virtual fencing (VF) enables livestock grazing without physical fences by conditioning animals to an audio tone (AT) followed by an electric pulse (EP) that is up to 25 times weaker than conventional electric fencing (EF). However, the operating principle of a virtual fencing system raises concerns about animal welfare. Therefore, the present study investigated the learning process of dairy cows in a virtual fencing environment and its effects on animal behavior and welfare. Twenty lactating cows were divided into four groups of five individuals each (2x VF; 2x EF), balanced by lactation stage (203 ± 62 days) and age (3.7 ± 1.5 lactations). All cows were accustomed to daily grazing with EF but were naïve to VF. Each group grazed in a separate paddock for 3 days of acclimation (P0) and 21, 14, 14, and 7 days of experimental treatment (P1-4). All paddocks were electrically fenced, and of similar vegetation structure and botanical composition. During P1-P4, a virtual boundary was activated within each paddock of the VF groups, and a second electric fence was set up within each paddock of the EF groups to maintain comparable paddock sizes. All cows were fitted with a VF collar (Nofence AS, Batnfjordsøra, Norway) and an IceQube pedometer (Peacock Technology Ltd, Stirling, UK). The sensors continuously tracked cow positions, the daily numbers of AT and EP, and activity behavior, respectively. Throughout P0-P4, daily milk yield, feed intake, and body weight were recorded. A total of 26 milk samples were collected per cow to determine milk cortisol concentrations. Behavioral observations were conducted for 2 h on 23 days to record agonistic behaviors, vocalizations, and excretions. During P1-P4, each cow received an average of 1.9 ± 3.3 AT and 0.1 ± 0.7 EP per day. The total number of stimuli ranged from 37 to 225 AT and 3 to 11 EP per cow throughout the experimental period, indicating individual variations in learning (P &amp;lt; 0.003). Within the first three days after virtual fence activation, most cows learned the concept of the VF system. The conditioning of the cows was successful at herd level after three repetitions at a new virtual boundary, reflected by a decreasing ratio of mean EP/AT from 20 to 3, 2, and 0 % with each paddock change. Milk yield and cortisol, feed intake, body weight, and cow activity did not differ between VF and EF groups. Overall, there were a mean of 11.2 vocalizations and 5.9 displacements more per cow in the VF groups (P &amp;lt; 0.05), but similar across P0-P4. Our results suggest that learning the virtual fencing system did not lead to lasting adverse effects on cow welfare.

PENGARUH STRES PSIKOLOGIS TERHADAP KADAR KORTISOL ASI PADA IBU MENYUSUI DI KABUPATEN SELUMA

&lt;p&gt;Breast milk is the gold standard in infant nutrition, which is the best food for babies compared to formula milk. Apart from containing the necessary nutrients, breast milk also contains various bioactive compounds that can affect the growth and development of the baby. Among these bioactive compounds there is also a glucocorticoid (GC) content, such as cortisol. It is known that cortisol levels are affected by the level of stress experienced by mothers during pregnancy and after childbirth. Purpose: this study aims to analyze the effect of maternal stress on breast milk cortisol levels in breastfeeding mothers in Seluma District. &lt;strong&gt;Method&lt;/strong&gt; : : This research is an analytic survey, cross sectional study, conducted in Seluma Regency. A total of 77 rbreastfeeding mothers of children aged 0-24 months were taken as respondents by random sampling. Breast milk samples were taken to examine breast milk cortisol levels and were measured using the ELISA method. interviews using a questionnaire were conducted to collect data on maternal characteristics, while the stress level data were obtained by conducting interviews using the DASS 42 questionnaire. The data obtained was then processed and analyzed using univariate and bivariate analysis. &lt;strong&gt;Results&lt;/strong&gt;: The results of measuring psychological stress showed that 35.1% of mothers experienced stress, while the results of measuring breast milk cortisol levels showed that 50% of mothers had high cortisol levels. The results of the Mann Whitney test found that there was a significant difference in the average breast milk cortisol levels in mothers who experienced psychological stress and those who did not (201.65 ± 97.82 μ/mL vs 150.32 ± 81.80 μ/mL, p=0.028) . &lt;strong&gt;Conclusion&lt;/strong&gt;: psychological stress affects breast milk cortisol levels in breastfeeding mothers in Seluma Regency. Education related to handling stress in breastfeeding mothers needs to be done so that it does not hinder the breastfeeding process which can have an impact on the health of mothers and children.&lt;/p&gt;&lt;p&gt; &lt;/p&gt;&lt;p&gt;&lt;strong&gt;&lt;em&gt;Keywords:&lt;/em&gt;&lt;/strong&gt;&lt;em&gt; Breastfeeding mothers, breast milk cortisol level, psychological stress&lt;/em&gt;&lt;/p&gt;

Predictors of milk cortisol in North American women.

Human milk content varies across mother-child dyads, environments, and populations. Among the hormones in milk is cortisol, a glucocorticoid; its impact on the breastfeeding child is unknown. Milk cortisol may constitute a signal to the child's developing physiology which can shape characteristics (e.g., growth, temperament) to prevailing environmental conditions. This exploratory study evaluated the maternal, breastfeeding, and infant characteristics associated with milk cortisol. We evaluated archived milk specimens for cortisol using enzyme immunoassay and employed an information-theoretic approach to assess associations between milk cortisol and participant characteristics with linear regression modeling. Because we employed secondary data, information for some variables likely to impact milk cortisol variation (e.g., time of day, socioeconomic status, maternal or infant body mass index, milk energy density) was unavailable. Participants were 48 lactating mothers from upstate New York, aged 21-40 years. Milk cortisol ranged from 0.098 to 1.007 μg/dL. Child age ranged from 1 to 26 months. In linear regression employing best fit modeling criteria, milk cortisol increased with child age (B: 0.069; p: .000; a 7.1% increase in milk cortisol for each month of child age), while child symptoms of illness (B: -0.398; p: .057; a 33% decrease) and consumption of complementary foods (B: -.525; p: .020; a 41% decrease) were associated with lower milk cortisol. We speculate that increasing milk cortisol with child age plays a role in signaling development (e.g., as increasing independence increases risk for injury and other negative health outcomes), independent of the maternal stressors we could capture.

Determinants of maternal breast milk cortisol increase: Examining dispositional and situational factors

BackgroundBreast milk is a rich nutritional source, containing numerous proteins, carbohydrates, and hormones that impact long-term offspring development. Strikingly, predictors and correlates of breast milk composition remain largely unknown. Building on a previously discovered increase in breast milk cortisol concentration from 2 to 12 weeks postpartum, we investigated potential predictors of maternal breast milk cortisol in the first three months post-delivery by examining a suite of maternal dispositional (e.g., attachment, adverse childhood experiences or ACEs) and situational factors (e.g., partner support, self-efficacy). MethodsData from 73 mothers were collected prenatally, at birth, and 2-, 6- and 12 weeks postpartum. The analyses, which sought to predict postnatal changes in breast milk cortisol, included a pool of theoretically-sound constructs (Table 1) and an exploratory data-driven approach. We fit models differing in complexity as preregistered: 1) Random Forest models, capable of modeling interactions and non-linear relationships, and 2) Bayesian linear models, allowing to model change over time while less prone to overfitting. ResultsOverall, we found that no single variable had strong predictive value beyond the known predictors of cortisol, such as time since awakening and time of collection. However, results from both models suggest that ACEs carry information that warrants future investigations, pointing towards a negative relationship with cortisol concentration in breast milk, albeit with a minimal effect size. ConclusionUsing sophisticated models, we found that early life stress may play a role in physiological stress markers in breast milk in the first three months postpartum, with potential implications for offspring development.

Assessment of Stress Levels in Lactating Cattle: Analyzing Cortisol Residues in Commercial Milk Products in Relation to the Temperature-Humidity Index.

Chronic stress in the dairy cattle industry has negative impacts on animal health, productivity, and welfare. It has been confirmed that cortisol transfers to milk and resists the high temperature during milk processing. This study evaluated the relationship between the milk cortisol concentration (MCC) in commercial milk products and the temperature-humidity index (THI) at the time of milk production. Eleven commercially produced pasteurized and sterilized milk products, purchased in Chuncheon, Korea, with production dates ranging from July to October 2021 were analyzed. The MCC was extracted using diethyl ether and analyzed using an enzyme immunoassay. The average THI values based on microclimate data provided by the Korea Meteorological Administration were 77 ± 0.8, 75 ± 1.4, 69 ± 1.4, and 58 ± 1.8, in July, August, September, and October, respectively. The average MCC levels were 211.9 ± 95.1, 173.5 ± 63.8, 109.6 ± 53.2, and 106.7 ± 33.7 pg/mL in July, August, September, and October, respectively. The MCC in July was higher than in August, September, and October (p < 0.05), while it was lower in September and October than in August (p < 0.05). Significant variations in the MCC were observed in commercial milk products across the four production months (p < 0.05), except for two milk products. Overall, monitoring the cortisol residue in commercial dairy milk products can be an alternative indicator of stress in dairy cattle of farms.

Associations between milk cortisol and activity of the immune system of milk.

Both the immune system of human milk and milk cortisol have complex short- and long-term effects on child health and development. As understanding continues to grow of the independent effects of each of these components of milk, it is also important to investigate their intersection, including how milk cortisol affects the immune system of milk. We began this important endeavor through secondary analyses of archived milk specimens. Participants were 31 lactating mothers from upstate New York. We estimated milk cortisol concentrations via enzyme immunoassay. We assessed milk proinflammatory cytokine (interleukin-6, IL-6) responses to pathogenic (Salmonella) and commensal (Escherichia, Lactobacillus, Bifidobacterium) bacteria via in vitro stimulation. We estimated ordered logistic regression models to assess associations between milk cortisol and IL-6 responses to bacteria. Milk cortisol ranged from 0.098 to 1.007 μg/dL. Milk cortisol was positively associated with IL-6 responses to S. enterica (B: 4.035; 95% CI: 0.674, 7.395) and B. breve (B: 3.675; 95% CI: 0.426, 6.924); this association persisted after controlling for child age. Results were less clear for associations between milk cortisol and IL-6 responses to L. acidophilus (B: 2.318; 95% CI: -1.224, 5.859) and E. coli (B: 2.366; 95% CI: -0.960, 5.692). Complex interactions between cortisol and the immune system extend to milk. Milk cortisol was positively associated with proinflammatory responses to some bacteria in vitro. This may suggest that milk cortisol is causally upstream of protective immune activity.

Is the temperament of crossbred dairy cows related to milk cortisol and oxytocin concentrations, milk yield, and quality?

Reactive dairy cows are more susceptible to stress, and this may result in negative effects on milk yield and quality. The aims of this study were to investigate the relationships between temperament traits and concentration of milk cortisol and oxytocin, milk yield, milkability, and milk quality in Holstein-Gyr cows. Temperament traits were assessed in 76 Holstein-Gyr cows in the milking parlor (by scoring milking reactivity and recording the numbers of steps and kicks during pre-milking udder preparation and when fitting the milking cluster) and during handling in the corral (by measuring the time to enter in the squeeze chute, ET and flight speed, FS). Milk samples were collected for milk quality (% fat, % protein, % lactose, and somatic cell count, SCC), and milk cortisol and oxytocin. Milk yield, milking time, and average flow were also measured. The calmer cows during milking management (class 'low') produced milk with higher protein (p = 0.028) content and tendencies for lower fat (p = 0.056) and higher lactose (p = 0.055) contents. Regarding the hormones, the most reactive cows (class 'high') in the milking and handling corral produced milk with higher concentrations of cortisol (p<0.001) and oxytocin (p = 0.023). In addition, the temperament of the animals affected some of the productive measures evaluated. Cows with reactive temperament had lower milk flow and longer milking time than the intermediate ones and had higher fat and a tendency for lower protein percentage in milk compared to cows with intermediate temperaments. Calm and intermediate cows in the handling corral produced more milk and presented better milkability parameters, such as a shorter milking time and greater average milk flow. Our results suggest that the cows' behavioral reactivity can be related to the intensity of their response to stress during handling.

Using behavioral observations in freestalls and at milking to improve pain detection in dairy cows after lipopolysaccharide-induced clinical mastitis

This study aimed to determine the effect of lipopolysaccharide (LPS)-induced mastitis with or without nonsteroidal anti-inflammatory drug (NSAID) on dairy cows' clinical, physiological, and behavioral responses in the milking parlor and freestalls as well as the specificity (Sp) and sensitivity (Se) of behavioral responses in detecting cows with LPS-induced mastitis. Twenty-seven cows received an intramammary infusion of 25 µg of Escherichia coli LPS in 1 healthy quarter. Following LPS infusion, 14 cows received a placebo (LPS cows), and 13 cows received 3 mg/kg of body weight of ketoprofen i.m. (LPS+NSAID cows). Cow response to the challenge was monitored at regular intervals from 24 h before to 48 h postinfusion (hpi) through direct clinical observations, markers of inflammation in milk, and via point-in-time direct behavioral observations in the barn and at milking. In LPS cows, infusion induced a significant increase of plasma cortisol levels at 3 and 8 hpi, milk cortisol levels at 8 hpi, somatic cell counts from 8 to 48 hpi, IL-6 and IL-8 at 8 hpi, milk amyloid A (mAA) and haptoglobin at 8 and 24 hpi, rectal temperature at 8 hpi, and respiratory rate at 8 hpi. Their rumen motility rate decreased at 8 and 32 hpi. Compared with before the challenge, significantly more LPS cows stopped feeding/ruminating and pressed their tail between their legs at 3 and 5 hpi, increased feeding/ruminating at 24 hpi, and had the tendency to be less responsive, dropping their head, and dropping their ears at 5 hpi. At milking, compared with before challenge, significantly more LPS cows lifted their hooves at forestripping at 8 hpi. The 2 groups showed similar patterns of response for milk cortisol, somatic cell count, respiratory rate, mAA, haptoglobin, and IL-6, IL-1β, and IL-8. Compared with LPS cows, LPS+NSAID cows had significantly lower plasma cortisol levels at 3 hpi, their rectal temperature decreased at 8 hpi, their rumen motility rate increased at 8 and 32 hpi, and their heart rate increased at 32 hpi. Compared with LPS cows, a significantly larger proportion of LPS+NSAID cows were feeding/ruminating, a lower proportion had ears down at 5 hpi, and a larger proportion lied down at 24 hpi. At milking, whatever the phase of milking, for "hoof to belly," 9 out of 14 cows did not show this behavior before infusion (Sp = 64%) and 14/14 did not kick during pre-infusion milking (Sp = 100%). Regarding sensitivity, at maximum, 5 cows out of 14 (Se = 36%) displayed "hoof to belly" after infusion. For "lifting hoof," 14/14 did not show hoof-lifting before infusion (Sp = 100%) and 6/14 displayed it after infusion (Se = 43%) at forestripping only. In the freestall barn, 9 behaviors had a Sp >75% (at minimum, 10/14 did not show the behavior) whatever the time point but Se < 60% (at maximum, 8/14 displayed the behavior). Finally, "absence of feeding and ruminating" had Sp of 86% (12/14 ate/ruminated) and Se of 71% (10/14 did not eat/ruminate) at 5 hpi. This study shows that feeding/ruminating, tail position, and reactivity at forestripping could be used as behavioral indictors for early detection of mastitis-related pain in dairy cows.

The Relationship of Preterm, Term, and Post-Term Births to Maternal Stress and Human Milk Cortisol Levels.

Introduction: Maternal stress can lead to changes in the composition of human breast milk. The present study evaluates cortisol levels in the breast milk of mothers after giving birth preterm, term, or post-term, and ascertains whether the levels are associated with maternal stress. Materials and Methods: Included in the study were mothers who gave birth vaginally after 32 weeks of gestation between January and April 2022. The breast milk was expressed with an electronic pump under the supervision of a nurse on day 7 following birth, and 2 mL samples of the milk were transferred into microtubes and stored at -80°C. Stress in the mothers was measured using the perceived stress scale developed by Cohen et al. The human breast milk cortisol levels were determined using an enzyme-linked immunoassay in a single session. Results: A total of 90 mothers, including 30 with preterm births, 38 with term births, and 22 with post-term births, were included in the study. The median stress scale score was 28 (17-50) and the median breast milk cortisol level was 0.49 ng/mL (0.1-1.96 ng/mL). A significant positive correlation was noted between the stress scale scores and breast milk cortisol levels (r = 0.56, p < 0.01). The breast milk cortisol levels and maternal stress scale scores were significantly higher in the preterm birth group than in the term birth group (p = 0.011 and p = 0.013, respectively). Conclusion: Although there is an association between maternal stress and preterm labor and milk cortisol levels, we believe that more studies are needed to establish a causal link.

Optimal Parameters to Milk Murciano-Granadina Goats in Mid and Low-Line Milking Parlours.

Recent short-term studies on Murciano-Granadina goats have established that the optimal parameters to set up the milking machines are different according to the milk pipes height. Two groups of 52 fresh goats each were employed in 2 different experiments to confirm during an entire lactation period the best combinations of system vacuum pulsation rate and pulsator ratio in low-line and mid-line milking parlours. The experiment performed in the low-line milking parlour included one group milked with 40 KPa vacuum system level, 90 puls/min pulsation rate and 60/40 pulsation and a second group milked with 38 KPa vacuum system level, 90 puls/min pulsation rate and 60/40 pulsation ratio. The experiment carried out in mid-line included one group milked with 40 KPa vacuum system level, 90 puls/min pulsation rate and 60/40 pulsation ratio and a second group milked with 40 KPa vacuum system level, 120 puls/min pulsation rate and 60/40 pulsation ratio. Variables studied included milking efficiency, milk composition, cortisol, SCC and intramammary infections, teat-end oedema after milking and vacuum dynamics during milking. Considering the results of an entire lactation period, it was confirmed that when milking in mid-line, the combination of 40 KPa system vacuum, 90 cycles/min pulsation rate and 60/40 pulsator ratio showed optimal results of the above-mentioned variables. On the other hand, the use of 40 KPa in a low-line system increased the milk cortisol values (0.34 ± 0.1 vs. 0.44 ± 0.1 ng/mL) without any other advantage. Thus, the recommendation is to use a combination of 38 KPa system vacuum, 90 cycles/min pulsation rate and 60/40 pulsator ratio to enhance animal welfare.