Effects of Wash Protocol and Contamination Level on Concentrations of Cortisol and Dehydroepiandrosterone (DHEA) in Swine Hair

The effect of washing procedure and contamination level on the concentrations of cortisol and dehydroepiandrosterone (DHEA) in swine hair was explored over two studies. Hair shaved from finisher pigs (n = 8) and sows (n = 8, cortisol study 1 only) was split into two treatments (two hair samples/pig) to receive either three isopropanol or methanol washes, and two paired subsamples of hair were contaminated with feces and urine, mildly or severely. Samples were further subdivided and received one, three, or five methanol washes. Hormone concentrations were quantified from the hair and wash solvent, and the ratio of hormones in the solvent to that in the hair calculated. When grouping sow and grower hair together for analysis, hair cortisol concentrations were 13% greater after three isopropanol washes compared to methanol (22.84 ± 3.12 vs. 19.77 ± 2.64 pg/mg, respectively). When analyzing sow and grower hair separately, sow hair cortisol concentrations were 20% higher following three isopropanol washes compared to methanol washes (22.06 ± 5.21 vs. 27.72 ± 5.65 pg/mg), with no differences in grower pig hair concentrations. The solvent cortisol concentrations did not differ with wash solvent. No differences were seen for DHEA. Contamination level did not influence hormone concentrations. Hair cortisol concentrations were 24% higher after one wash compared to five washes (11.98 ± 1.47 vs. 9.05 ± 0.92 pg/mg,), whereas the solvent cortisol concentrations were 80% and 84% higher after one wash compared to three and five washes, respectively (21.09 ± 4.04 vs. 4.21 ± 1.62 vs. 3.36 ± 1.32 pg/mg). The solvent–hair cortisol ratio was 65% and 73% higher following one wash compared to three and five washes (1.36 ± 0.80 vs. 0.47 ± 0.12 vs. 0.37 ± 0.14). Hair DHEA concentrations were 39% higher after one wash compared to five washes (42.39 ± 6.87 vs. 26.02 ± 5.69 pg/mg). The solvent DHEA concentrations, and the solvent–hair ratio for DHEA were 94% and 98% and 92% and 98% higher going from one wash to three and five washes, respectively (solvent: 5.07 ± 0.26 vs. 0.28 ± 0.12 vs. 0.12 ± 0.09 pg/mg and solvent–hair ratio: 0.13 ± 0.006 vs. 0.010 ± 0.004 vs. 0.003 ± 0.002). Following three methanol washes, the non-contaminated hair had 46% and 48% higher hair (17.47 ± 1.12 vs. 9.35 ± 0.80 vs. 9.05 ± 1.06 pg/mg) and a 76% and 72% higher solvent (16.31 ± 8.07 vs. 3.92 ± 0.50 vs. 4.50 ± 2.31 pg/mg) cortisol concentration compared to mild and severely contaminated hair, respectively. Wash solvent influences cortisol concentrations in swine hair, but not DHEA. Contaminated swine hair should be avoided in analyses when possible.

Download Full-text

304 Effects of Supplementing Phix-up® on Productive, Physiological, and Rumen Responses of Feedlot Cattle Consuming a Corn-based Finishing Diet

Journal of Animal Science ◽

10.1093/jas/skab235.298 ◽

2021 ◽

Vol 99 (Supplement_3) ◽

pp. 162-163

Author(s):

Eduardo Colombo ◽

Reinaldo F Cooke ◽

Kelsey Harvey ◽

Jacob Wiegand ◽

Ana Clara Araujo ◽

...

Keyword(s):

Physiological Responses ◽

Latin Square ◽

Feedlot Cattle ◽

Hair Cortisol ◽

Finishing Cattle ◽

Rumen Ph ◽

Hair Samples ◽

Buffering Agent ◽

To Receive ◽

Feeding Systems

Abstract This experiment (d -14 to 118) compared rumen pH, productive, and physiological responses in finishing cattle supplemented or not with a rumen-buffering agent (pHix-up®; Timab, Dinard, France). Angus-influenced yearling cattle (58 heifers and 62 steers) were ranked by sex and body weight (BW) and allocated to 4 groups of 30 animals each (d -14). Groups were housed in 1 of 4 drylot pens equipped with GrowSafe automated feeding systems, and assigned to receive a corn-based diet containing monensin, and with the inclusion of pHix-up® at 0.00% (control) 0.25%, 0.50%, or 0.75% (d 0 to 118). Cattle BW was recorded twice monthly (d -14 to 118), and blood samples were collected on d 0, 28, 56, 84 and 112. Hair samples from the tail-switch were collected on d 0, 56 and 112. Cattle were slaughtered on d 119. Eight rumen-cannulated steers were also assigned to this experiment (d 42 to 98), with 2 steers housed with each group in a replicated 4 × 4 Latin Square design (4 periods of 14 d). Rumen pH was measured on d 7 and 14 of each period at 0800h, 1200h, 1600h and 2000h. No treatment effects were noted (P ≥ 0.13) for cattle BW gain, feed intake, and feed efficiency. Inclusion of pHix-up® linearly increased (P < 0.01) plasma Mg concentrations and tended to linearly decrease (P = 0.09) plasma haptoglobin concentrations. Cattle receiving pHix-up® had greater (P < 0.01) mean plasma cortisol concentrations compared with non-supplemented cattle. Hair cortisol concentration linearly decreased (P < 0.01) as pHix-up® inclusion increased on d 112 (treatment × day interaction; P = 0.02). In rumen-cannulated steers, pHix-up® inclusion linearly increased (P = 0.03) mean rumen pH. Collectively, pHix-up® supplementation improved rumen pH and modulated stress-related physiological responses in finishing cattle, but without improving their productive responses.

Download Full-text

Welfare assessment in dairy cows using hair cortisol as a part of monitoring protocols

Journal of Dairy Research ◽

10.1017/s0022029920000588 ◽

2020 ◽

Vol 87 (S1) ◽

pp. 72-78

Author(s):

Urška Vesel ◽

Tea Pavič ◽

Jožica Ježek ◽

Tomaž Snoj ◽

Jože Starič

Keyword(s):

Dairy Cows ◽

Environmental Changes ◽

Cortisol Concentration ◽

Hair Cortisol ◽

Welfare Assessment ◽

Assessment Protocol ◽

Potential Marker ◽

Hair Samples ◽

Welfare Quality ◽

Monitoring Protocols

AbstractWelfare of dairy cows can be assessed using welfare assessment protocols consisting of resource, management and animal-based measures. Welfare Quality® Assessment Protocol is one of the best-known protocols, which depends almost entirely on animal-based measures. To gain more objective and rapid welfare assessment, new techniques have been developed to measure welfare of animals, such as hair cortisol concentration. As cortisol is released in response to stress, it has long been used as a biomarker of stress in animals. While the precise mechanism of cortisol incorporation into hair is unknown, hair cortisol concentration seems to be a marker of long-term systemic cortisol concentration. Hair cortisol is, therefore, a potential marker of chronic stress and is not likely to be affected by acute stress. Studies on cattle show connections between hair cortisol concentration and factors such as pregnancy, parity, diseases, ectoparasites, body condition score, environmental changes, stocking density and milk yield. Hair cortisol concentration appears to be affected by time of sampling, cow age and breed, UV radiation, season, body region of sampled hair and hair colour. Its concentration also depends on sampling and analytical methods. Hair cortisol is a promising non-invasive tool to evaluate welfare of dairy cows, however, more research is needed to determine the extent of effects on its concentration and the appropriate method of sampling and analysis. Correlations between Welfare Quality® Assessment Protocol scores and pooled hair cortisol concentrations have not yet been found, and more research is needed with larger sample size, a standardized protocol of hair sampling, processing and analysis. With proper attention to detail, hair cortisol levels in pooled hair samples might come to be used as a reliable indicator of dairy animal welfare.

Download Full-text

Long-term stress in dogs is related to the human–dog relationship and personality traits

Scientific Reports ◽

10.1038/s41598-021-88201-y ◽

2021 ◽

Vol 11 (1) ◽

Author(s):

Amanda Höglin ◽

Enya Van Poucke ◽

Rebecca Katajamaa ◽

Per Jensen ◽

Elvar Theodorsson ◽

...

Keyword(s):

Personality Traits ◽

Big Five ◽

Cortisol Concentration ◽

Hair Cortisol ◽

Hunting Dogs ◽

Breed Group ◽

Hair Samples ◽

Stress Levels ◽

Inventory Survey

AbstractPreviously, we found that dogs belonging to the herding breed group, selected for human cooperation, synchronise their long-term stress levels with their owners. The aim of the current study was to investigate features that could influence long-term stress levels in ancient dog breeds, genetically closer to wolves, and dogs specifically selected to work independently of their owner. Twenty-four ancient breed dogs and 18 solitary hunting dogs were recruited and hair samples were obtained from both dogs and owners from which hair cortisol concentration (HCC) was analysed. Additionally, the owners completed lifestyle surveys, the Monash Dog Owner Relationship Scale (MDORS) on human–dog relationship, and both dog and owner personality questionnaires (Dog Personality questionnaire and Big Five Inventory survey). The results from the MDORS indicate that the subscale Perceived cost correlated to the dog HCC of tested breed groups: solitary hunting breeds (χ2 = 4.95, P = 0.026, β = 0.055), ancient breeds (χ2 = 2.74, P = 0.098, β = 0.027), and herding dogs included from a previous study (χ2 = 6.82, P = 0.009, β = − 0.061). The HCC of the solitary hunting dogs was also related to the owner personality traits Agreeableness (χ2 = 12.30, P < 0.001, β = − 0.060) and Openness (χ2 = 9.56, P = 0.002, β = 0.048) suggesting a more substantial influence of the owner on the solitary hunting dog’s HCC compared to the ancient breeds. No effect of owner HCC on dog HCC was found in either ancient or in solitary hunting breeds. Hence, the long-term stress synchronisation is likely to be a trait in breeds selected for human cooperation. In conclusion, dog HCC is often related to the owners’ personality, but is primarily influenced by the owner-dog relationship.

Download Full-text

Correlation analysis of cortisol concentration in hair versus concentrations in serum, saliva and urine.

10.21203/rs.2.24006/v1 ◽

2020 ◽

Author(s):

Lukasz Cieszynski ◽

Jaroslaw Jendrzejewski ◽

Piotr Wisniewski ◽

Przemyslaw Klosowski ◽

Krzysztof Sworczak

Keyword(s):

Clinical Practice ◽

Salivary Cortisol ◽

Serum Cortisol ◽

Individual Characteristics ◽

Cortisol Concentration ◽

Reference Ranges ◽

Hair Cortisol ◽

Free Cortisol ◽

Hair Samples ◽

Cortisol Levels

Abstract Background: Cortisol concentration is measured in blood, urine, and saliva samples. It has been recently proven that cortisol could be also detected in hair samples. Cortisol measurements in different samples have their own individual characteristics and clinical utility. We aimed to investigate the correlation between hair cortisol concentration and standard cortisol measurements used in clinical practice. Methods: Fifty adult volunteers with a negative history of endocrine disorders were enrolled into the study. Morning serum cortisol (MSC), evening serum cortisol (ESC), evening free salivary cortisol (EFSC), urine free cortisol (UFC) and hair cortisol were analyzed in all participants. Eventually, 41 volunteers were included into the study, whose cortisol concentration in the 1 mg overnight dexamethason suppression test (1mgONDST) were <50 nmol/l, and cortisol levels in serum, saliva and urine were within reference ranges. Hair cortisol concentration (HCC) was performed for 20 mg of hair strands of the proximal 1 cm hair segments. Results: HCC ranged from 0.3036 to 2.65 nmol/l/mg, and the average value was 0.8125±0.4834 nmol/l/mg. No significant correlations were found between HCC and MSC (rho=0.04419, p=0.7838), HCC and ESC (rho=-0.2071, p=0.1938), HCC and EFSC (rho=0.1005, p=0.532), and HCC and UFC (rho = 0.1793, p = 0.262). Conclusions : This work is another step in the discussion on the application of HCC determinations in clinical practice. Our results have showed no correlations between HCC and single point cortisol assessment in blood, saliva and urine in patients with reference cortisol levels. Keywords: Hair cortisol, Serum cortisol, Salivary cortisol, Urine cortisol, Cortisol assessment, Cortisol correlation

Download Full-text

Association between hair cortisol concentration and metabolic syndrome

Open Medicine ◽

10.1515/med-2021-0298 ◽

2021 ◽

Vol 16 (1) ◽

pp. 873-881

Author(s):

Eglė Mazgelytė ◽

Asta Mažeikienė ◽

Neringa Burokienė ◽

Rėda Matuzevičienė ◽

Aušra Linkevičiūtė ◽

...

Keyword(s):

Risk Factors ◽

Metabolic Syndrome ◽

Blood Serum ◽

Salivary Cortisol ◽

High Performance ◽

Cortisol Concentration ◽

Hair Cortisol ◽

Hair Samples ◽

Lower Social Support ◽

Cortisol Levels

Abstract Metabolic syndrome (MetS) is a highly prevalent disorder defined as a cluster of cardiometabolic risk factors including obesity, hyperglycemia, hypertension, and dyslipidemia. It is believed that excessive cortisol secretion due to psychosocial stress-induced hypothalamic-pituitary-adrenal axis activation might be involved in the pathogenesis of MetS. We sought to explore the association between MetS and psychosocial risk factors, as well as cortisol concentration measured in different biological specimens including saliva, blood serum, and hair samples. The study was conducted on a sample of 163 young and middle-aged men who were divided into groups according to the presence of MetS. Hair cortisol concentration (HCC) was determined using high performance liquid chromatography with UV detection, while blood serum and salivary cortisol levels were measured by enzyme-linked immunoassay. Lipid metabolism biomarkers were determined using routine laboratory methods. Anthropometric and lifestyle characteristics, as well as self-reported psychosocial indicators, were also examined. Significantly higher HCC and lower social support level among participants with MetS compared with individuals without MetS were found. However, no significant differences in blood serum and salivary cortisol levels were observed between men with and without MetS. In conclusion, chronically elevated cortisol concentration might be a potential contributing factor to the development of MetS.

Download Full-text

Increased Scalp Hair Cortisol Concentrations in Obese Children

The Journal of Clinical Endocrinology & Metabolism ◽

10.1210/jc.2013-2924 ◽

2014 ◽

Vol 99 (1) ◽

pp. 285-290 ◽

Cited By ~ 62

Author(s):

Margriet A. B. Veldhorst ◽

Gerard Noppe ◽

Mieke H. T. M. Jongejan ◽

Chantine B. M. Kok ◽

Selma Mekic ◽

...

Keyword(s):

Body Weight ◽

Waist Circumference ◽

Scalp Hair ◽

Blood Lipid ◽

Normal Weight ◽

Cortisol Concentration ◽

Hair Cortisol ◽

Obese Children ◽

Hair Samples

Context: Pathologically increased cortisol exposure induces obesity, but it is not known whether relatively high cortisol within the physiological range is related to childhood obesity. Objective: The aim of the study was to compare hair cortisol concentrations between obese and normal-weight children. Design: We performed an observational case-control study. Participants: Twenty obese children (body mass index-SD score [BMI-SDS] > 2.3) and 20 age- and sex-matched normal-weight children (BMI-SDS < 1.1) aged 8–12 years were recruited. Main Outcome Measures: Scalp hair samples from the posterior vertex were collected, and hair cortisol concentrations were measured using ELISA. Body weight, height, and waist circumference were measured. From the obese children, additional data on blood pressure and blood lipid concentrations were collected. Results: In both groups, five boys and 15 girls were included; their mean age was 10.8 ± 1.3 vs 10.8 ± 1.2 years (obese vs normal weight; not significant). Body weight, BMI, BMI-SDS, and waist circumference were higher in the obese children compared with the normal-weight children (69.8 ± 17.2 vs 35.5 ± 7.2 kg; 29.6 ± 4.9 vs 16.4 ± 1.6 kg/m2; 3.4 ± 0.5 vs −0.2 ± 0.8 SDS; 94 ± 13 vs 62 ± 6 cm; P < .001 all). Hair cortisol concentration was higher in obese than normal-weight children (median [interquartile range], 25 [17, 32] vs 17 [13, 21] pg/mg; P < .05). Conclusions: Hair cortisol concentration, a measure for long-term cortisol exposure, was higher in obese children than normal-weight children. This suggests long-term activation of the hypothalamus-pituitary-adrenal axis in obese children and may provide a novel target for treatment of obesity in children.

Download Full-text

Unraveling the Complexity to Observe Associations Between Welfare Indicators and Hair Cortisol Concentration in Dairy Calves

Frontiers in Animal Science ◽

10.3389/fanim.2021.793558 ◽

2021 ◽

Vol 2 ◽

Author(s):

Lena-Mari Tamminen ◽

Linda J. Keeling ◽

Anna Svensson ◽

Laurie Briot ◽

Ulf Emanuelson

Keyword(s):

Coping Styles ◽

Generalized Additive Models ◽

Cortisol Concentration ◽

Additive Models ◽

Dairy Calves ◽

Regularized Regression ◽

Hair Cortisol ◽

Welfare Indicators ◽

Hair Samples ◽

Cortisol Levels

Using levels of the stress hormone cortisol as an indicator for welfare is a common, but debated practice. In this observational study, hair cortisol concentration (HCC) of samples from 196 dairy calves from 7 to 302 days of age collected from 12 Swedish farms was determined using a commercially available ELISA. An assessment of animal welfare, assessed using animal-based indicators, was performed on the day of sampling. First, methodological factors with the potential to impact HCC and the effect of age were analyzed using generalized additive models. This revealed a significant peak in hair cortisol in young calves (around 50 days of age) and an association between fecal contamination of hair samples and the level of cortisol extracted. Second, associations between welfare indicators and HCC were explored using cluster analysis and regularized regression. The results show a complex pattern, possibly related to different coping styles of the calves, and indicators of poor welfare were associated with both increased and decreased hair cortisol levels. High cortisol levels were associated with potential indicators of competition, while low cortisol levels were associated with the signs of poor health or a poor environment. When running the regularized regression analysis without the contaminated hair samples and with the contaminated samples (including a contamination score), the results did not change, indicating that it may be possible to use a contamination score to correct for contamination.

Download Full-text