Browsing by Author "Hough, Denise"
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- ItemComparison of two CYP17 isoforms : implications for cortisol production in the South African Merino(Stellenbosch : Stellenbosch University, 2012-03) Hough, Denise; Swart, Pieter; Cloete, Schalk W. P.; Stellenbosch University. Faculty of Science. Dept. of Biochemistry.ENGLISH ABSTRACT: This study describes: • the comparison of the enzymatic activities of the two ovine cytochrome P450 17 - hydroxylase/17,20-lyase (CYP17) isoforms expressed in non-steroidogenic COS-1 cells. The Km and Vmax values for the metabolism of pregnenolone and progesterone were determined, while time-dependent metabolism of pregnenolone, 17-hydroxypregenolone, progesterone and 17-hydroxyprogesterone was also reported. The cloning and sequencing of ovine cytochrome b5 is reported and was co-expressed with CYP17. The results showed that the wild type 1 (WT1) isoform of ovine CYP17 produce more cortisol precursors than the wild type 2 (WT2) isoform; • the analysis of the frequency distribution of the CYP17 genotypes within a South African Merino population, which were divergently selected for (H-line) or against (L-line) the ability of a ewe to rear multiple offspring per birthing opportunity. It was observed that the CYP17 frequency distribution was the same within the H- and L-line, with 78.3 % heterozygous WT1/WT2 and 21.7 % homozygous WT1/WT1. No homozygous WT2/WT2 individuals were identified; • the development of a UPLC-MS/MS method for the separation and quantification of all thirteen adrenal steroids that are produced in the adrenal gland; • the relative contribution of the CYP17 genotypes in the total steroidogenic output in adult adrenocortical cells from the adrenal glands of H- and L-line sheep, with particular emphasis on cortisol production. The adrenocortical cells from the H-line sheep showed a marked higher cortisol production than the L-line, while adrenocortical cells from homozygous WT1/WT1 sheep also produced more cortisol than heterozygous WT1/WT2 sheep; • the blood cortisol responses upon the stimulation of the HPA axis by insulin induced hypoglycaemia of the H- and L-line sheep with known CYP17 genotypes. It was observed that the CYP17 genotype and selection line are important factors affecting the cortisol responses of sheep, where L-line heterozygous WT1/WT2 sheep showed the lowest cortisol response and glucose recovery; • the association of the CYP17 genotype with behavioural responses of H- and L-line sheep to flock isolation stress, as well as the association of the CYP17 genotype with ewe reproduction and lamb output. While reproduction seemed to be unaffected by the CYP17 genotype, the behavioural stress responses of sheep to flock isolation correlated with the CYP17 genotype, where the heterozygous WT1/WT2 genotype was associated with a wilder nature.
- ItemThe effect of extensive human presence at an early age on stress responses and reactivity of juvenile ostriches towards humans(MDPI, 2018-07) Muvhali, Pfunzo T.; Bonato, Maud; Engelbrecht, Anel; Malecki, Irek A.; Hough, Denise; Robinson, Jane E.; Evans, Neil P.; Cloete, Schalk W. P.The effect of extensive human presence and regular gentle handling performed at an early age (0–3 months old) on stress responses and reactivity of juvenile ostriches towards humans was investigated. A total of 416 ostrich chicks over two years were exposed to one of three treatments for three months after hatching; namely, Human Presence 1 (HP1, N = 144): extensive/prolonged human presence with physical contact (touch, stroking), gentle human voice, and visual stimuli; Human Presence 2 (HP2, N = 136): extensive/prolonged human presence without physical contact, but with gentle human voice and visual stimuli; and the Standard treatment (S, N = 136): human presence limited to routine feed and water supply as a control. At 7.5 months of age, the plasma heterophil/lymphocyte (H/L) ratio was measured before and 72 h after feather harvesting and feather clipping to determine acute stress responses, while chronic stress was measured by quantification of corticosterone (CORT) concentrations in the floss feathers of the birds. Birds’ behavioural response towards a familiar or an unfamiliar handler was evaluated at 12 months using docility and fear tests, and through behavioural observations conducted on random days between the ages of 8–13 months. Willingness to approach, and to allow touch interactions, aggressiveness, and exhibition of sexual display towards the handler, was recorded. No difference in the H/L ratios before and after feather harvesting and clipping was observed in HP1 birds, whereas H/L ratios showed a significant increase 72 h post feather harvesting and clipping in HP2 and S birds (p < 0.05). Birds from the S treatment exhibited a significantly (p < 0.05) higher feather CORT concentration compared with HP1 birds, while HP2 birds had intermediate responses. Birds’ reactivity towards humans and temperament as evaluated using behavioural observations, docility, and fear tests was not affected by treatment (p > 0.05). However, HP1 and HP2 birds were more inclined (p < 0.05) to approach a familiar rather than an unfamiliar handler during the behavioural observations, indicating an ability to distinguish between a familiar and an unfamiliar handler. Overall, the results indicate that early gentle human interactions with ostrich chicks can be beneficial in reducing physiological stress sensitivity later in life and facilitate the ability of ostriches to distinguish between familiar and unfamiliar handlers.
- ItemThe effect of extensive human presence at an early age on stress responses and reactivity of juvenile ostriches towards humans(MDPI -, 2018) Muvhali, Pfunzo T.; Bonato, Maud; Engelbrecht, Anel; Malecki, Irek A.; Hough, Denise; Robinson, Jane E.; Evans, Neil P.; Cloete, Schalk W. P.The effect of extensive human presence and regular gentle handling performed at an early age (0–3 months old) on stress responses and reactivity of juvenile ostriches towards humans was investigated. A total of 416 ostrich chicks over two years were exposed to one of three treatments for three months after hatching; namely, Human Presence 1 (HP1, N = 144): extensive/prolonged human presence with physical contact (touch, stroking), gentle human voice, and visual stimuli; Human Presence 2 (HP2, N = 136): extensive/prolonged human presence without physical contact, but with gentle human voice and visual stimuli; and the Standard treatment (S, N = 136): human presence limited to routine feed and water supply as a control. At 7.5 months of age, the plasma heterophil/lymphocyte (H/L) ratio was measured before and 72 h after feather harvesting and feather clipping to determine acute stress responses, while chronic stress was measured by quantification of corticosterone (CORT) concentrations in the floss feathers of the birds. Birds’ behavioural response towards a familiar or an unfamiliar handler was evaluated at 12 months using docility and fear tests, and through behavioural observations conducted on random days between the ages of 8–13 months. Willingness to approach, and to allow touch interactions, aggressiveness, and exhibition of sexual display towards the handler, was recorded. No difference in the H/L ratios before and after feather harvesting and clipping was observed in HP1 birds, whereas H/L ratios showed a significant increase 72 h post feather harvesting and clipping in HP2 and S birds (p < 0.05). Birds from the S treatment exhibited a significantly (p < 0.05) higher feather CORT concentration compared with HP1 birds, while HP2 birds had intermediate responses. Birds’ reactivity towards humans and temperament as evaluated using behavioural observations, docility, and fear tests was not affected by treatment (p > 0.05). However, HP1 and HP2 birds were more inclined (p < 0.05) to approach a familiar rather than an unfamiliar handler during the behavioural observations, indicating an ability to distinguish between a familiar and an unfamiliar handler. Overall, the results indicate that early gentle human interactions with ostrich chicks can be beneficial in reducing physiological stress sensitivity later in life and facilitate the ability of ostriches to distinguish between familiar and unfamiliar handlers.
- ItemExploration of the hypothalamic-pituitary-adrenal axis to improve animal welfare by mean of genetic selection : lessons from the South African merino(MDPI, 2013) Hough, Denise; Swart, Pieter; Cloete, SchalkIt is a difficult task to improve animal production by means of genetic selection, if the environment does not allow full expression of the animal’s genetic potential. This concept may well be the future for animal welfare, because it highlights the need to incorporate traits related to production and robustness, simultaneously, to reach sustainable breeding goals. This review explores the identification of potential genetic markers for robustness within the hypothalamic-pituitary-adrenal axis (HPAA), since this axis plays a vital role in the stress response. If genetic selection for superior HPAA responses to stress is possible, then it ought to be possible to breed robust and easily managed genotypes that might be able to adapt to a wide range of environmental conditions whilst expressing a high production potential. This approach is explored in this review by means of lessons learnt from research on Merino sheep, which were divergently selected for their multiple rearing ability. These two selection lines have shown marked differences in reproduction, production and welfare, which makes this breeding programme ideal to investigate potential genetic markers of robustness. The HPAA function is explored in detail to elucidate where such genetic markers are likely to be found.