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* old and bolonia study programme


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17-ß-Estradiol and Testosterone concentrations in claws of puppies up to 60 days of age.
Jasmine Fusi, Tanja Peric, Cristina Bergamin, Maria Cristina Veronesi, 2018, published scientific conference contribution abstract

Abstract: The current 30% of canine perinatal mortality (Tonnessen et al., 2012) claims the need to deepen the knowledge about this phase, characterized by long-term hormonal and metabolic changes, that could benefit of long time-frame methods of investigation, reducing the quantity of samplings and thus respecting animal welfare (Veronesi et al., 2015). Indeed, apart from the well-known invasiveness of blood samplings, the necessary repeated collection of feces, urine and saliva, providing punctual information, could be stressful for the newborns and their mothers with negative effects on health status and maternal behavior. Given that sexual steroid hormones were reported to have an influence on health outcomes and development already from the perinatal period (Frey et al., 2017), in this work 17-β-Estradiol (E2) and Testosterone (T) concentrations were assessed from the claws of dogs up to 60 days of age, to identify possible endogenous biomarkers. Ten male and 10 female puppies, viable and healthy, born by elective cesarean section, were enrolled. Samplings were performed by trimming claws at birth, and the regrowth at 30 and 60 days of age after the breeder or the owner signed an informed consent. Then, E2 and T concentrations were analyzed by RIA (Veronesi et al., 2015) and a possible effect of gender evaluated by ANOVA. All the hormonal claws concentrations showed a significant (p<0.001) drop from birth to 30 and 60 days of age, while no significant changes were observed between 30 and at 60 days of age (Table 1). No influence of the newborns’ gender was found. Because of the higher levels of E2 and T observed at birth and at 30 days of age, it could be supposed that a source of production could be the placental and maternal compartments. However, the direct involvement of the fetus itself could not be excluded, given the reported accumulation from the nail capillary bed of those hormones (de Berker et al., 2007) and the production of sexual steroid hormones by feline fetal gonads (Braun and Jewgenow, 2017).
Found in: osebi
Keywords: Dog, Puppy, Claws, 17-ß-Estradiol, Testosterone.
Published: 09.04.2019; Views: 691; Downloads: 0
.pdf Fulltext (876,50 KB)

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.
Found in: osebi
Keywords: hair, cortisol, analyses, method, AlphaLISA, Amplified luminescent proximity homogeneous assay
Published: 09.04.2019; Views: 794; Downloads: 0
.pdf Fulltext (4,05 MB)

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.
Found in: osebi
Published: 09.04.2019; Views: 642; Downloads: 0
.pdf Fulltext (4,05 MB)

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.
Found in: osebi
Keywords: fattening pigs, hair, steroid hormones, allostatic load
Published: 12.06.2019; Views: 928; Downloads: 0
.pdf Fulltext (277,52 KB)

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