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1.
Cortisol, DHEA, and Sexual Steroid Concentrations in Fattening Pigs’ Hair
Cristina Bergamin, Antonella Comin, Mirco Corazzin, Massimo Faustini, Tanja Peric, Annalisa Scollo, Flaviana Gottardo, M. Montillo, Alberto Prandi, 2019, original scientific article

Abstract: The aim of this study was to analyze the feasibility and reliability of using hair as a matrix to determine the dehydroepiandrosterone (DHEA) and sexual steroid concentrations and the cortisol/DHEA ratio in fattening pigs. The results could be also used to plan future research to identify threshold values in order to set up strategies to control the allostatic load and increase the resilience of fattening pigs before slaughter. The study was conducted on 107 commercial crossbred rearing pigs. The hair samples were taken by shaving at the age of 36 weeks, and concentrations of the hormones were measured using a solid-phase microtiter radioimmunoassay. Females had significantly higher cortisol levels (p < 0.01), significantly lower DHEA concentrations (p < 0.05) and significantly higher cortisol/DHEA ratios (p < 0.01) than barrows. Progesterone was significantly higher in gilts than in barrows (p < 0.01). Testosterone and 17β-estradiol were significantly higher in barrows than in gilts (p < 0.05). If future research can produce threshold values for the different markers examined, the evaluation of animals under subclinical stress conditions will be possible.
Keywords: fattening pigs, hair, steroid hormones, allostatic load
Published in RUNG: 12.06.2019; Views: 3542; Downloads: 0
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2.
ASSESSMENT OF CORTISOL AND DHEA CONCENTRATIONS IN THE GRIFFON VULTURE (Gyps fulvus) FEATHERS TO EVALUATE ITS HEALTH CONDITION
Gian Nicola Frongia, Tanja Peric, Marco Muzzeddu, Fiammetta Berlinguer, Giovanni Leoni, Valentina Satta, Cristina Bergamin, Marcello Franchini, Antonella Comin, Salvatore Naitana, Alberto Prandi, 2018, published scientific conference contribution abstract

Abstract: During their life, birds face several challenges which can threaten the stability of physiological functions. These situations typically trigger a cascade of endocrine secretions involving the hypothalamic–pituitary–adrenal (HPA) axis, and resulting in the release of both cortisol and corticosterone also in birds. The use of a non-invasive approach to collect samples of biological material derived from natural populations represents a greatly combination for an improvement of knowledge avoiding handling animals. Steroid concentrations in feathers are mainly acquired from the capillary around the follicle during the long period of feather grown reflecting their bloodstream concentrations. Accordingly, a chronic HPA axis activation could be assessed using molted feathers cortisol concentrations. Another hormone, dehydroepiandrosterone (DHEA), is secreted by adrenals and it could be assessed in molted feathers. This study was aimed to evaluate the feasibility of using feathers cortisol and DHEA concentrations to provide a retrospective assessment of the activity of the HPA axis in griffon vulture acquired for health monitoring purposes. For this, we introduced two groups of animals with a known health condition: one group composed by griffons that have experienced a long-term physiologically compromised period (PC group; n=8) and the other group composed by griffons physiologically not compromised (CTRL group; n=9). The feathers cortisol and DHEA analyses were carried out using RIA [1,2] adapted for feathers. The results revealed different feather hormone concentrations between the two groups of animals. The feathers cortisol had a median value higher in the PC group than in the CTRL group, with cortisol concentrations showing a central value 1.6 times higher in those animals that suffered a long-term physiological impairment. The PC group showed also (Kruskal- Wallis test) higher feather concentrations of DHEA (P=0.01) than the CTRL group. Pearson’s correlation coefficients were used to examine the associations between cortisol and DHEA, showing no correlation between cortisol and DHEA feather concentrations in the PC group (r=0.18, P=0.34) and a moderate positive correlation in the CTRL group (r=0.51, P=0.011). In conclusion, our study reveals that molted feathers can be an interesting way to evaluate the physiological status of wild animals by using a non-invasive approach. Our analyses reveal that in addition to cortisol also DHEA could be evaluated to better understand the relationships between these hormones and to determine the resilience condition of wild species.
Keywords: CORTISOL, DHEA, GRIFFON VULTURE, Gyps fulvus, FEATHER
Published in RUNG: 09.04.2019; Views: 3017; Downloads: 0
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3.
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: 3307; Downloads: 0
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4.
Hair dehydroepiandrosterone in newborn beef calves from birth up to 10 months of age
Monica Probo, Tanja Peric, Massimo Faustini, Alberto Prandi, Antonella Comin, 2018, published scientific conference contribution abstract

Abstract: Although steroidogenesis in maternal, placental and fetal compartments is interdependent, the maternal and fetal hypothalamic-pituitary-adrenal (HPA) axes represent separate biological systems, with dehydroepiandrosterone (DHEA) recognized as the main fetal steroid. Fetal steroids are likely to influence development and have long-term effects on HPA function. Hair analysis represents a promising methodological approach for the non-invasive measurement of steroids, allowing for a retrospective analysis of the total exposure to steroids over time, and avoiding the influence of acute events or circadian fluctuations (Schury et al. 2017, BMC Psychiatry 17:213). Hair coat DHEA (hcD) concentrations have been investigated in cows (Peric et al. 2017, Livestock Sci 202:39–43), but no studies have been performed on newborn calves. Hair samples of 12 beef calves (7 males, 5 females) were collected by shaving at calving (T0) and monthly up to 10 months of age (T1-T10), only on the re-growth area. Hair DHEA was analyzed by RIA2. Statistical analysis revealed that hcD concentrations in calves were influenced by age, with higher levels at T1 and T2 compared to the other samples (p < 0.05). HcD levels were not influenced by newborn gender, birth weight and Apgar score. These data demonstrate that DHEA is quantifiable in the hair coat of newborn calves, and that hcD levels are influenced by the age of calves. The higher accumulation of DHEA was detected from birth to 2 months of age, suggesting that DHEA continues to be secreted by the newborn calf also beyond birth and could be involved in the events occurring during the first months of age.
Keywords: hair, dehydroepiandrosterone, cattle, calf
Published in RUNG: 09.04.2019; Views: 3070; Downloads: 0
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5.
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: 3360; Downloads: 0
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6.
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: 4291; Downloads: 0
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7.
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: 3954; Downloads: 0
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8.
Cortisol claws concentrations in dogs from birth to 60 days of age
Jasmine Fusi, Tanja Peric, Barbara Bolis, Alessandro Rota, Antonella Comin, Maria Cristina Veronesi, 2017, published scientific conference contribution abstract

Abstract: During the last fetal stage of development and in the neonatal period, the hypothalamic- pituitary- adrenal (HPA) axis secretes cortisol (C), responsible for several physiologic processes. The claws were recently proved to be a useful, non- invasive matrix for long time- frame retrospective C levels analysis also in puppies (Veronesi et al. 2015, Theriogenology 84:791–796). Because of the scarce knowledge on canine perinatology, this study was aimed to assess the C concentrations in claws of newborn puppies collected at birth, 30 and 60 days of age, and to evaluate the possible influence of age, gender and type of birth on C claws accumulation. The study was performed on 58 large purebred, normal, healthy puppies, born by spontaneous or cesarean parturition. The mean C claws concentrations significantly decrease from birth (26 ± 16.39 pg/mg) to 30 (10 ± 4.53 pg/mg) and 60 (6 ± 3.12 pg/mg) days of age (p < 0.0001); a decrease was also seen from 30 to 60 days of age (p < 0.05). No influence of newborn gender and type of birth was found. This trend of decrease is in agreement with data reported by (Veronesi et al. 2015, Theriogenology 84:791–796) in dead puppies, and suggests the important role of C around the time of birth. However the exact source of the C accumulated in claws of puppies at birth must be clarified, while the source of C accumulated at 30, and even more at 60 days, can be supposed to be addressed to the newborn itself.
Keywords: Cortisol, claws, dog
Published in RUNG: 01.09.2017; Views: 4508; Downloads: 0
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9.
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: 4613; Downloads: 0
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10.
Luteal activity and effect of dietary energy restriction on follicular development in lactating cows
Antonella Comin, Tanja Peric, M. Montillo, Alberto Cappa, Vittorio Marchi, Maria Cristina Veronesi, Alberto Prandi, 2017, original scientific article

Abstract: The aim of this research has been to evaluate the presence of anomalies in the ovarian cycle activity during postpartum and to verify whether 72-hr dietary fasting during the dominance phase, the phase before ovulation, might modify the ovarian follicle population. The presence of anomalies in ovarian cycle activity has been evaluated in 30 Italian Friesian cows starting from 20 days postpartum until 211 days of lactation. Long oestrus and brief dioestrus or scarce luteal activity have been the main anomalies found through measuring progesterone concentrations in the whey. Until 100 days of lactation, the BCS values of the problematic animals have been significantly lower than those in animals with normal ovarian activity. After 100 days of lactation, the ovarian anomalies continued to appear despite the fact that all the animals have reached comparable BCS values. Starting from the results of this trial, the effect of 72-hr dietary fasting on dominant follicles has been studied in six cows. Ultrasonography revealed that the diameter of the follicles at 71 days postpartum has been significantly lower than at 181 days. A 72-hr dietary restriction at 101 and 211 days postpartum did not affect the size of the dominant follicle. However, at 101 days postpartum, half of the animals presented follicular cysts. The effect of fasting differed if the animal has been in early postpartum or 211 days of lactation. Further researches are necessary to understand how different metabolic conditions can modify the follicular population but on the other hand the study shows the utility for farmers and field veterinarians of monitoring the resumption of the ovarian cycle postpartum through the whey progesterone concentrations.
Keywords: dairy cow, cyclic activity, follicles, fasting
Published in RUNG: 06.04.2017; Views: 4270; Downloads: 0
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