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1.
Analysis of 19 Minerals and Cortisol in Red Deer Hair in Two Different Areas of the Stelvio National Park: A Preliminary Study
Marta Montillo, Tanja Peric, 2019, original scientific article

Abstract: The aim of the study was to perform an investigation on the concentration of 19 minerals and cortisol in red deer (Cervus elaphus) hair, a matrix that is easy to collect with non-invasive and painless sampling, able to represent an integrative values of long-term substance concentrations, and able to give useful information, also when performed on dead animals, given its extreme stability over time. In the study thirty-five animals were included, coming from two different sides of a valley in the Stelvio National Park, where official water analysis had pointed out elevated concentrations of As in one of the two orographic sides. Hair cortisol concentrations were measured using a RIA(Radio Immuno Assay), while minerals were detected using ICP-MS (Inductively Coupled Plasma- Mass Spectrometry). Results showed a negative relationship between cortisol and some mineral concentrations (Li, Co, As, Cd, Cr and Tl) and significant differences in some mineral concentrations between park areas (Al, Co, Cu, Cd and Ni). As, Cr and cortisol differences approached statistical significance. This preliminary study represents a step forward in the study of wildlife allostatic load and a valid method for applications in wildlife management programs, in environmental studies and in public health programs.
Keywords: hair, deer, mineral, cortisol
Published in RUNG: 27.08.2019; Views: 3061; Downloads: 97
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2.
A NEW TOOL TO EVALUATE CORTISOL CONCENTRATIONS IN ANIMAL HAIR
Antonella Comin, Tanja Peric, Mara Colzani, Francesco De Vecchi, Cristina Bergamin, Alberto Prandi, 2018, published scientific conference contribution abstract

Abstract: The study of cortisol in hair requires the use of a highly sensitive assay. To date, hair cortisol concentrations (HCC) have been evaluated with different analytical methods as Radio Immune Assay (RIA), Enzyme Immunossay (EIA), Enzyme Linked Immunosorbent Assay (ELISA), ChemiLuminescent Immunoassay (CLIA) and High Performance Liquid Chromatography with Mass Spectrometry (HPLC/MS) or Liquid Chromatography with Tandem Mass Spectrometry (LC-MS/MS). Amplified luminescent proximity homogeneous assay (AlphaLISA) is an ELISA-like, nonradioactive technology first reported in 1994 [1]. This technology allows the quantitative detection of molecules of interest in a light-induced chemiluminescence immunoassay using a microplate without wash step. The aim of the study was to compare RIA with AlphaLISA method and examine the sensitivity in evaluating cortisol concentrations in animal hair of the Cortisol AlphaLISA kit (PerkinElmer, USA), originally suggested for the analysis of buffer and serum samples. The study has been carried out on 4 samples of calf, 4 samples of foal and 8 samples of sheep hair. After washing with isopropanol, two extracts were obtained from each hair sample. One extract has been evaluated for HCC by RIA as previously described [2,3]. The second extract has been reconstituted with the buffer provided by the Cortisol AlphaLISA Kit and HCC evaluated by the use of this commercially available kit. The preliminary results indicate that the Cortisol AlphaLISA Kit is capable to detect cortisol also in hair samples with high sensitivity, with a detection limit of 17 pg/ml. Intra- and inter-assay coefficients of variation (CVs) were 4.0% and 9.0%, respectively. Cortisol concentrations ranged between 1.31 and 16.94 pg/mg, 6.30 and 57.65 pg/mg, and 2.16 and 45.08 in calf, foal and sheep hair, respectively. The HCC obtained by Cortisol AlphaLISA Kit and RIA showed a good correlation (r=0.79, p<0.01). Considering the possibility to use a low amount of extracted sample, its no-washing procedure and the performances showed, we can conclude that the Cortisol AlphaLISA Kit can be considered an excellent tool to evaluate cortisol concentrations also in hair derived from animal species.
Keywords: hair, cortisol, analyses, method, AlphaLISA, Amplified luminescent proximity homogeneous assay
Published in RUNG: 09.04.2019; Views: 3523; Downloads: 0
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3.
Maternal and fetal hair cortisol concentrations in beef cattle at delivery
Tanja Peric, Monica Probo, Antonella Comin, Maria Cristina Veronesi, Marta Montillo, Alberto Prandi, 2018, published scientific conference contribution abstract

Abstract: Hypothalamus-pituitary-adrenal (HPA) axis of both mother and fetus, is one of key mechanisms involved in fetal programming. Cortisol, the end-product of this axis, plays a critical role in regulating fetal and new-born metabolism and development. The aim of the study was to evaluate maternal and fetal cortisol concentrations shortly after delivery on hair samples non-invasively collected. The study was carried out on 11 pairs of beef cows and their calves (calves were 12 because of one case of twins) from the same farm and subjected to the same managerial conditions. Hair from new-born calves and from their mothers was collected on the day of birth shaving their withers to the level of the skin. The hair cortisol concentrations (HCC) were determined by RIA as previously described (Peric et al. 2013: J Dairy Sci 96:3023–3027). The results showed that fetal and maternal HCC ranged from 16.11 to 40.43 pg/mg and from 1.88 to 15.73 pg/mg respectively, with fetal and maternal mean ± SE HCC of 23.72 ± 2.21 and 6.40 ± 1.32 pg/mg respectively (p < 0.001). Fetal HCC were on average 3.5 higher than those of mothers but this ratio differed among the pairs. These differences were also found among the twin calves. These results indicate that in beef cows-and-calves pairs, although HCC at delivery were always higher in calves than in cows, the ratio between fetal and maternal HCC differs among pairs, suggesting an individuality in the fetal programming processes that may also rely on genetic and epigenetic factors.
Keywords: hair, cortisol, cattle, delivery
Published in RUNG: 09.04.2019; Views: 3597; Downloads: 0
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4.
A first survey on hair cortisol of an Alpine ibex (Capra ibex ibex) population.
Tanja Peric, Alberto Prandi, Mirco Corazzin, Antonella Comin, Monica Colitti, 2018, original scientific article

Abstract: Biometric measurements of horn, body and physiological variables of 35 Alpine ibex, Capra ibex ibex, (14 females and 21 males, age range: 2-15 years) were analysed in relation to gender and area of origin. Moreover, for the first time hair cortisol concentration was evaluated in this species. The ibexes were live-captured in spring in the Maritime Alps Natural Park. All the horn and body morphometric measurements were significantly different between genders and a statistical difference was found between horn and body data considering the area of origin of ibexes, with the exception of horn circumference and height as well of body and hock length. Among the physiological variables, female ibex showed higher heart rate and glycemia, but tended to have lower respiratory rate than male. The mean hair cortisol concentration was 22.40±1.44 pg/mg (±SE). A different distribution was described for the male hair cortisol concentrations compared to that in females. Obtained data indicate that the geographical area where Alpine ibex lives could affect the endurance of an ibex population but further investigations are needed to understand if intra- and inter-specific competition and different environment could be the basis of the HPA axis activation. Hair cortisol evaluation together with biometric values could be an interesting way to obtain information on the adaptation that the animal attempts to reach in relation to environmental factors and pursuing organism homeostasis. This feature could be important in attempting to preserve wildlife biodiversity that also depends from the state of animal welfare.
Keywords: allostatic load, Alps, ibex, biometry, hair cortisol
Published in RUNG: 06.03.2018; Views: 4528; Downloads: 0
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5.
Hair cortisol concentrations in New Zealand white rabbits subjected to surgery
Tanja Peric, Antonella Comin, Mirco Corazzin, Marta Montillo, Federico Canavese, Marco Stebel, Alberto Prandi, 2018, original scientific article

Abstract: The aim of this study was to assess hair cortisol concentrations in New Zealand white rabbits (Oryctolagus cuniculus) that were subjected to relocation and surgery to evaluate HPA-axis activity; in addition, we used this marker of cortisol secretion to evaluate the allostatic load of animals undergoing surgery. After a period of acclimatisation, which lasted 40 days from their arrival at the enclosure, 19 rabbits were subjected to T1–T12 dorsal arthrodesis (RS), 19 were sham-operated (SS), and 19 were non-operated (CON). Hair samples were collected at the time of arrival (ST1) at the animal facility, and seven other sets of hair samples were collected at 40-day intervals from the same area of skin for a period of 240 days as re-shaved hair (anagen phase): immediately before surgery (ST2) and after the surgery (ST3, ST4, ST5, ST6, ST7, and ST8). The transition from the rabbitry to the animal breeding facility led to a significant increase in cortisol concentration (ST2) in all of the groups. At ST3, the RS group presented higher cortisol concentrations than those of the SS group and the CON group. At ST4, the experimental groups showed similar values that remained constant until ST8. The results show that the management of rabbits undergoing surgery should be evaluated very carefully, and hair cortisol concentrations may provide a means of avoiding the dangerous cumulative effects of additional stressors close to surgery.
Keywords: animal welfare, cortisol, hair, HPA axis, rabbit, surgery
Published in RUNG: 09.02.2018; Views: 4178; Downloads: 0
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6.
HAIR CORTISOL LEVEL IN THE GOLDEN JACKAL (CANIS AUREUS MOREOTICUS I. GEOFFROY SAINT HILAIRE, 1835) IN RELATION TO THE SAMPLE CONSERVATION, SEX AND ECOLOGICAL CHARACTERISTICS.
Marta Montillo, Stefano Pesaro, Tanja Peric, Veronica Maran, Stefano Filacorda, 2017, published scientific conference contribution abstract

Abstract: Hair cortisol analysis is a complementary method for monitoring HPA axis activity, capturing systemic cortisol exposure over longer periods of time. We have tested the hair cortisol concentration on 16 specimens of Golden jackal (Canis aureus moreoticus): 5 from frozen animals, 10 from stuffed animals and 1 taken from injured animal during recovery in an Animal Care Center; the samples came from Italy (Friuli Venezia Giulia and Veneto Region) and Croatia (Istria and Dalmatian region). We have studied the hair cortisol concentration by RIA method in respect to conservation method, sex and areas of origin, with non parametric tests and mixed model. The hair cortisol concentration (values expressed in pg/mg) from the frozen animals (mean=3.38 SD=3.13, n=5) was not different in respect to the stuffed samples (mean=2.81 SD=2.72, n=9), also the sex has not showed any statistical effect (males mean=1.96 SD=0.63 n=9; females mean=4.56 SD=3.99 n=6). The hair cortisol concentration in respect to the sites of origin of samples, dispersal and core areas was not different (dispersal areas: mean=1.56 SD=0.77 n=5, core areas: mean=3.72 SD=3.14 n=10). The use of mixed model to detect the effect of areas (core and dispersal areas) and interaction between sex and areas has showed the presence of significative higher hair concentrations of cortisol in core areas for the females (mean=6.27 SD=3.84 n=4)in respect to the males (mean=2.02 SD=0.52 n=6). The concentration of hair cortisol of sick and injured animal recovered in the Animal Care Center was higher (> of 12 pg/mg) in respect to the other samples. Hair sample can be used to develop a clearer understanding of the interrelationship of health and physiology, mobility and social behaviour.
Keywords: hair, Canis aureus, cortisol
Published in RUNG: 19.07.2017; Views: 4985; Downloads: 0
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7.
Cortisol and DHEA concentrations in the hair of dairy cows managed indoor or on pasture
Tanja Peric, Mirco Corazzin, Alberto Romanzin, Stefano Bovolenta, Alberto Prandi, M. Montillo, Antonella Comin, 2017, original scientific article

Abstract: This study was conducted in order to assess the changes of cortisol and dehydroepiandrosterone (DHEA) concentrations in hair of Italian Simmental dairy cows managed indoor in tie-stall barns or grazing on high mountain pasture. In experiment I, animals were kept in tie-stalls throughout the year (n = 27). In experiment II, animals remained on mountain pastures during summer (from 20th June to 20th September) and then kept indoors in tie-stalls for the rest of the year (n = 18). At the end of July, following vertical transhumance, dairy cows were moved to a pasture located at a higher altitude. In the two studies, hair samples, obtained using clippers from the animal's forehead, were collected on the 20th June (P1), 20th July (P2), 20th August (P3), 20th September (P4) and 20th October (P5) and analysed for cortisol and DHEA. In experiment I, DHEA concentration was unaffected by sampling time (P > 0.05), whereas, cortisol/DHEA ratio was similar between P1 and P2 (P > 0.05) and then increased during the experimental period reaching the highest value at P5 (P < 0.05). In experiment II, dairy cows had higher concentration of DHEA during grazing compared to cows kept indoors (P < 0.05), conversely cortisol/DHEA ratio decreased from P1 to P2 (P < 0.05), remained constant from P2 to P3 (P > 0.05) and then increased from P3 to P4 (P < 0.05), the highest value was recorded at P5. In conclusion, cortisol and DHEA concentrations and their ratio measured in the hair of dairy cows can be useful to evaluate the allostatic load in animals managed indoor in tie-stall barn or grazing on mountain pasture.
Keywords: Dairy cow, Mountain pasture, Hair, Cortisol, DHEA
Published in RUNG: 28.05.2017; Views: 4834; Downloads: 0
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8.
Hair cortisol and testosterone concentrations and semen production of Bos taurus bulls
Giuseppe Stradaioli, Tanja Peric, Marta Montillo, Antonella Comin, Mirco Corazzin, Maria Cristina Veronesi, Alberto Prandi, 2017, original scientific article

Abstract: The experiment was aimed to evaluate the usefulness of hair cortisol concentrations (HCC) in revealing the activation of the hypothalamic-pituitary-adrenal (HPA) axis due to the passage of young bulls from genetic rearing stations to artificial insemination (AI) centre and the influence of this passage on hair testosterone concentrations (HTC). Hair samples on 33 yearling bulls were collected at the time of arrival (ST1) to the AI centre and 3 more hair samples (ST2, ST3, ST4) were collected at monthly intervals. After the end of quarantine each animal was submitted to the standard procedures for collection and production of semen. Overall HCC were significantly affected by period of collection (p<.01). In particular, HCC was higher in samples obtained after finishing the quarantine period (ST2) compared with those obtained on ST1. After ST2, the HCC decreased reaching at ST4 a value similar to that recorded at ST1. An effect of sampling time on HTC was not found (p>.05). The effect of breed was observed only for HTC that was higher in PRI than BS (p<.01). The HCC group have not influenced the semen variables (p>.05). On the contrary, the effect of breed was evident. The results of this study replicated the effects of a major environmental stressor on mean cortisol concentrations assessed in hair, and add to the growing body of evidence that HCC is an effective and simply collected marker for long-term activity of the HPA system in response to persistent environmental challenge.
Keywords: Hair, bulls, cortisol, testosterone, semen
Published in RUNG: 20.03.2017; Views: 5696; Downloads: 0
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9.
Use of hair cortisol analysis for comparing population status in wild red deer (Cervus elaphus) living in areas with different characteristics
Chiara Caslini, Antonella Comin, Tanja Peric, Alberto Prandi, Luca Pedrotti, Silvana Mattiello, 2016, original scientific article

Abstract: We tested a method to measure Hair Cortisol Concentration (HCC) in 174 red deer (Cervus elaphus) culled in the hunting season 2011/12 in four areas of Central Italian Alps, with different population densities (SPN, 11.8 deer/km2; HD-AV, 3.6 deer/km2; HD-SO, 2.1 deer/km2; HD-MO, 2.0 deer/km2) and environmental conditions. Our hypothesis was that higher population densities, associated with more difficult environmental conditions, may result in higher allostatic load for these wild ungulates. No significant differences in HCC were detected between sexes (males, 4.77 ± 0.69 pg/mg; females, 5.75 ± 0.63 pg/mg) nor among age classes (calves, 6.17 ± 0.66 pg/mg; yearlings, 4.47 ± 0.83 pg/mg; adults, 5.15 ± 0.74 pg/mg; least square mean ± SE), but HCC difference between calves and yearlings was close to statistical significance (P = 0.059). HCC showed high individual variation, but on average it was higher in areas with higher deer density (SNP, 7.45 ± 1.01 pg/mg; HD-AV, 6.07 ± 0.89 pg/mg; HD-SO, 4.67 ± 1.14 pg/mg; HD-MO, 2.87 ± 1.56 pg/mg), with significant differences between HD-AV and HD-MO (P = 0.01). Carcass weight was significantly lower in SNP (46.74 ± 1.49 kg) than in HD-MO (62.71 ± 4.01 kg), HD-SO (61.73 ± 2.9 kg) and HD-AV (62.07 ± 2.04 kg) (P < 0.001). These results seem to confirm our hypothesis that allostatic load is higher in areas with higher density and harder environmental conditions. We suggest that the methodology used in this study to measure HCC provides good information on long-term HPA axis activity and allostatic load and constitutes a highly promising, reliable and non-invasive method in wildlife management for assessing HPA axis activity over extended time periods.
Keywords: Red deer, Cervus elaphus, Hair, Cortisol, Allostatic load
Published in RUNG: 29.09.2016; Views: 4471; Downloads: 0
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10.
EFFECT OF DIET ON HAIR CORTISOL AND DHEA CONCENTRATIONS IN MOUSE
Antonella Comin, Marta Montillo, Tanja Peric, Silvia Gazzin, Mirco Corazzin, Alberto Prandi, published scientific conference contribution abstract

Abstract: Obesity and MetS (Metabolic Syndrome) are both linked to persistent long-term hormonal and metabolic changes. In most of the studies, cortisol (C) and dehydroepiandrosterone (DHEA) concentrations have been measured in obese and normal-weight subjects, obtaining heterogeneous results. Plasma, saliva and urine, matrices that represent timepoint or short-term steroids exposure, were used for these studies. The aim was to study C, DHEA and C/DHEA ratio of mice pups in the hair, matrix capable of providing cumulative hormonal exposure. Sixty C57Bl/6 mice pups (30 males and 30 females) were housed in a temperature-controlled environment (22±2°C) and on a 12h light/dark schedule, under ad-libitum access to food and water for 16 weeks (welfare: Italian Law Decree 116-92 and EC Directive 86-609-EEC). Control and experimental diets were offered immediately after weaning (3 weeks old pups), for 16 weeks. Four experimental checkpoints were established (T1: 4 weeks, T2: 8 weeks, T3: 12 weeks and T4: 16 weeks of diet). T3 and T4 showed the hormonal concentrations of pubertal animals. Twentyseven pups (13 females and 14 males) were randomly group-housed in cages (6 for T1, 5 for T2, 6 for T3, 10 for T4) and assigned to control diet (CTRL: D12328, Research Diets, New Brunswick, NJ). 33 pups (17 females and 16 males) were randomly group-housed in cages (5 for T1, 6 for T2 and T3, 16 for T4) and assigned to the HFHC diet (HFHC: D12331, Research Diets, New Brunswick, NJ - plus 42g/L fructose/sucrose in drinking water). At each experimental checkpoint, all the animals of one cage for each experimental group were suppressed. Hair strands were carefully cut with scissors as close as possible to the skin from the back of the mice, paying attention to not to wound the animals. Hair was stored in an envelope at RT in a dry room until use. C and DHEA hair concentrations was measured by a solid-phase microtiter RIA assay (Peric et al., 2016 adapted in the mouse). Only at 8 weeks the HFHC group showed significantly higher C concentrations than the CTRL group (1.56±0.06 vs 1.92±0.130 pg/mg; P<0.05). DHEA concentrations were significantly reduced in the HFHC group than the CTRL group at 4 (114.64±13.93 vs 69.08±5.33 pg/mg; P<0.05), 8 (71.67±7.08 vs 50.38±4.74 pg/mg; P<0.05), 12 (73.27±8.29 vs 41.59±1.60 pg/mg; P<0.01) and 16 weeks (65.26±3.35 vs 55.05±1.84 pg/mg; P<0.05). The C/DHEA ratio was significantly increased in the HFHC than the CTRL group, at 8 (0.023±0.002 vs 0.040±0.005; P<0.01), 12 (0.024±0.003 vs 0.036±0.004; P<0.05) and 16 weeks (0.019±0.001 vs 0.025±0.002; P<0.01). The gender effect was not significant. Taking into account the lag time required for the hair emersion from the skin (about 1 week), the significant stimulation of the C at 8 weeks in the HFHC group refers to its chronic elevation from 5 to 7 weeks of treatment when the animals were not pubertal. Conversely, DHEA shows a chronic reduction in obese mice leading to consider an independent adrenal regulation of C and DHEA, both stimulated by ACTH. Research was supported by the Project CBM (MIUR DM60643). Peric et al. 2016 J Appl Anim Welf Sci 18:1-8.
Keywords: DIET, HAIR, CORTISOL, DHEA, MOUSE
Published in RUNG: 21.07.2016; Views: 5116; Downloads: 0
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