Effects of different nutritional regimens on the productivity of Australian Cashmere goats and the partitioning of nutrients between cashmere and hair growth

The influence of energy or protein supplementation or energy restriction on cashmere growth was studied in 35 highly productive cashmere wether goats. The goats were shorn on 3 December and randomly allocated to 3 levels of energy intake: M, goats fed to maintain liveweight; 0.8M, goats fed to lose 5 kg liveweight from December to April and then fed ad libitum; and >M, goats fed to gain liveweight. Nested within >M were ADLIB (goats offered feed ad libitum), and 1.25M and l.5M (goats fed M plus 25 or 50% of the difference in mean intake between M and ADLIB). The metabolisable energy requirement to maintain liveweight was 250 kJ kg-0.75 day-1 but to maintain body condition (l.25M) it was 3 12 kJ kg-0.75 day-1. Goats fed 0.8M had a mean intake of 0.68M and lost 26 g day-1 liveweight until April, but when fed ad libitum consumed 2.15M in June and grew rapidly in late autumn and winter at 93 g day-1. Goats fed ADLIB consumed 2.30M in February and gained 87 g day-1 from December to February, but intake declined to 1.61 M in June and they gained 20 g day-1 from April to June. Cashmere growth and fibre diameters of fleeces shorn on 17 June of goats fed >M (221g, 17.69 pm) were significantly greater (P< 0.02) than those of goats fed 0.8M (146 g, 16.67 ¦m), with levels of M-fed goats being intermediate. Within >M, there were no significant differences in cashmere growth. Protein supplementation within M (27 or 54 g day -1 formaldehyde- treated casein) resulted in 40% more wool growth in sheep (P<0.001), but no increase in cashmere or hair growth in goats. Goats fed ADLIB had significantly reduced cashmere yields (P < 0.05) and grew more hair (P<0.05) than did goats in other treatments. About 4 weeks after energy supplementation, fibre diameter of previously energy-deprived goats increased (P< 0.01). Midside patches indicated that energy-deprived goats, which lost liveweight, diverted nutrients preferentially to cashmere growth, while goats fed ADLIB partitioned nutrients towards hair growth. To maximise cashmere growth, supplementary energy should be supplied to avoid liveweight loss from December to April. Goats that had small (1-2 kg) liveweight gains and maintained body condition achieved near maximal levels of cashmere growth.

Functional analysis of the sheep Wilson disease protein (sATP7B) in CHO cells

In this study we investigated the function of the sheep orthologue of ATP7B (sATP7B), the protein affected in the human copper toxicosis disorder Wilson disease. Two forms of sATP7B are found in the sheep, a ‘normal’ form and one with an alternate N terminus, both of which were expressed in CHO-K1 cells. Cells expressing either form of sATP7B were more resistant to copper than the parental CHO-K1 cells. Subcellular localisation studies showed that both forms of sATP7B were similarly located in the trans-Golgi network (TGN). When the extracellular copper concentration was increased, each form of sATP7B redistributed to a punctate, vesicular compartment that extended throughout the cytoplasm. Both forms of sATP7B recycled to the perinuclear location within one hour when the cells were subsequently incubated in basal medium. After treatment of cells with bafilomycin A1 sATP7B accumulated in cytoplasmic vesicles, implying that ATP7B continuously recycles via the endocytic pathway. These results suggest that both forms of sATP7B are functional copper-transport proteins and that the intracellular location and trafficking of the sheep protein within the cell also appears normal.

Correction of the copper transport defect of Menkes patient fibroblasts by expression of two forms of the sheep Wilson ATPase

The Wilson disease (WD) protein (ATP7B) is a copper-transporting P-type ATPase that is responsible for the efflux of hepatic copper into the bile, a process that is essential for copper homeostasis in mammals. Compared with other mammals, sheep have a variant copper phenotype and do not efficiently excrete copper via the bile, often resulting in excessive copper accumulation in the liver. To investigate the function of sheep ATP7B and its potential role in the copper-accumulation phenotype, cDNAs encoding the two forms of ovine ATP7B were transfected into immortalised fibroblast cell lines derived from a Menkes disease patient and a normal control. Both forms of ATP7B were able to correct the copper-retention phenotype of the Menkes cell line, demonstrating each to be functional copper-transporting molecules and suggesting that the accumulation of copper in the sheep liver is not due to a defect in the copper transport function of either form of sATP7B.

Responses of the hypothalamopituitary adrenal axis and the sympathoadrenal system to isolation/restraint stress in sheep of different adiposity

There is evidence that levels of adipose tissue can influence responses of the hypothalamopituitary-adrenal (HPA) axis to stress in humans and rats but this has not been explored in sheep. Also, little is known about the sympathoadrenal responses to stress in individuals with relatively different levels of adipose tissue. We tested the hypothesis that the stress-induced activation of the HPA axis and sympathoadrenal system is lower in ovariectomized ewes with low levels of body fat (lean) than ovariectomized ewes with high levels of body fat (fat). Ewes underwent dietary manipulation for 3 months to yield a group of lean ewes (n = 7) with a mean (±SEM) live weight of 39.1 ± 0.9 kg and body fat of 8.9 ± 0.6% and fat ewes (n = 7) with a mean (±SEM) live weight of 69.0 ± 1.8 kg and body fat of 31.7 ± 3.4%. Fat ewes also had higher circulating concentrations of leptin than lean ewes. Blood samples were collected every 15 min over 8 h when no stress was imposed (control day) and on a separate day when 4 h of isolation/restraint was imposed after 4 h of pretreatment sampling (stress day). Plasma concentrations of adrenocorticotropic hormone (ACTH), cortisol, epinephrine and norepinephrine did not change significantly over the control day and did not differ between lean and fat ewes. Stress did not affect plasma leptin levels. All stress hormones increased significantly during isolation/restraint stress. The ACTH, cortisol and epinephrine responses were greater in fat ewes than lean ewes but norepinephrine responses were similar. Our results suggest that relative levels of adipose tissue influence the stress-induced activity of the hypothalamopituitary-adrenal axis and some aspects of the sympathoadrenal system with fat animals having higher responses than lean animals.

Isolation and restraint stress results in differential activation of corticontrophin-releasing hormone and arginine vasopressin neurones in sheep

This study investigated sex differences in the stress-induced activation of neurons containing corticotrophin-releasing hormone (CRH), arginine vasopressin (AVP) and enkephalin in the paraventricular nucleus (PVN) of gonadectomized male and female sheep. Groups (n=3) of both sexes were either subjected to 90 min isolation and restraint stress (stress group) or were not stressed. Blood samples were taken every 10 min for 90 min prior to and after stress to monitor cortisol levels in plasma. Brains were harvested after 90 min of stress. Stress caused elevation of plasma cortisol levels to a similar extent in both sexes. Double-labeling immunohistochemistry for Fos and either CRH, AVP or enkephalin was undertaken to quantify the numbers of neurons staining for CRH, AVP and enkephalin that also immunostained for Fos. Stress increased Fos immunostaining in all cell types. There was a greater proportion of CRH than AVP neurons activated in stressed animals. There were no sex differences in the activation of CRH and AVP neurons although females had a greater proportion of enkephalin cells staining for Fos than males in both control and stressed animals. There were no differences between control and stressed animals in the proportion of cells co-staining for CRH and AVP. We conclude that isolation and restraint stress activates neurons producing CRH, AVP and enkephalin in sheep and that CRH may play a greater role than AVP in regulating adrenocorticotrophic hormone secretion in response to this stressor in sheep. Finally, isolation and restraint stress does not influence co-localization of CRH and AVP in sheep.

Projections to the preoptic area from the paraventricular nucleus, acuate nucleus and the bed nucleus of the stria terminalis are unlikely to be involved in stress-induced suppression of GnRH secretion in sheep

Stress compromises reproductive function and the major physiological system activated during stress is the hypothalamo-pituitary-adrenal axis. Corticotrophin-releasing hormone and arginine vasopressin (AVP), which are produced in neurones of the paraventricular nucleus (PVN), drive the hypothalamo-pituitary-adrenal axis and are also implicated in the suppression of the reproductive axis. We used retrograde tracing and Fos labelling to map the projections from the PVN to the preoptic area (POA) where most gonadotrophin releasing hormone (GnRH) neurones are found. Fluorogold (FG) injections were made into the POA of gonadectomised male and female sheep (n = 5/sex), the animals were stressed and the brains recovered for histochemistry. All animals responded to stress with an increase in the number of Fos-labelled nuclei in the PVN. Few retrogradely labelled cells of the PVN were activated by stress. Dual labelling showed that very few FG-labelled cells also stained for corticotrophin-releasing hormone, none for AVP or enkephalin. Dual labelling for FG and Fos in the bed nucleus of the stria terminalis (BNST) and the arcuate nucleus showed that no FG-labelled cells in the BNST and only few in the ARC were activated by stress. No sex differences were observed in the activation of FG-labelled cells in any of the nuclei examined. We conclude that, although cells of the PVN, BNST and/or arcuate nucleus may affect reproduction via the GnRH cells of the POA, this is unlikely to involve direct input to the POA. If cells of these regions are involved in GnRH suppression during stress, this may occur via interneuronal pathways.

Sex difference in the suppressive effect of cortisol on pulsatile secretion of luteinizing hormone in sheep

We tested the hypothesis that there are sex differences in the inhibitory effect of cortisol on pulsatile LH secretion and pituitary responsiveness to GnRH in gonadectomized sheep. In experiment 1, pulsatile LH secretion was examined in gonadectomized ewes and rams infused with either saline, a low (250 µg/kg·h) or a high (500 µg/kg·h) dose of cortisol for 30 h. In experiment 2, direct pituitary actions of cortisol were assessed by monitoring LH pulse amplitude in response to exogenous GnRH in hypothalamo-pituitary disconnected ewes and rams infused with the low dose of cortisol. In experiment 1, the mean (±SEM) plasma LH concentration was (P < 0.05) reduced significantly during cortisol infusion in both sexes, but the effect was greater in rams. In ewes, LH pulse amplitude and frequency were reduced (P < 0.05) at the high, but not the low, cortisol dose, whereas total LH output (LH pulse amplitude multiplied by frequency) was reduced (P < 0.05) at both doses. In rams, LH pulse frequency and amplitude and total LH output were (P < 0.05) reduced significantly at both cortisol doses. In experiment 2, plasma LH concentration and pulse amplitude in response to exogenous GnRH were not affected by infusion of cortisol in either sex. We conclude that gonadectomized rams are more sensitive than gonadectomized ewes to the effects of cortisol to inhibit LH secretion and that sex differences exist in the specific actions of cortisol on LH pulses. The results of experiment 2 suggest that intact hypothalamic input to the pituitary is necessary for cortisol to inhibit pituitary responsiveness to GnRH.

Critical evaluation of cashmere nutrition experiments and suggestions for the design and conduct of successful experiments

Cashmere fibre production is an order of magnitude less than fibre production of Merino sheep or Angora goats and is more difficult to measure. Based on a comparison between cashmere experiments reporting responses to nutrition and those reporting no response, 13 design and management characteristics were identified that are related to the ability of experiments to discriminate among treatments. Methods must be adopted to reduce the variance in cashmere production within treatments, by using sufficient. animals per treatment, having enough replication to provide plenty of degrees of freedom to reduce error terms in analysis, and using pre-experimental cashmere production attributes as co-variants in analysis. It is preferable to use more productive and older goats, and goats that are used to handling, and to the conditions and feed to be used. Nutrition treatments need to produce different live weight growth curves and an appropriate control is needed such as live weight maintenance. As the raw cashmere fleece is composed primarily of hair and other contaminants, careful attention is required to measure, sample and test cashmere. Cashmere growth experiments should start by midsummer and last for at least four and preferably six months. These requirements make it more difficult for many university students to plan, undertake and complete long-term cashmere nutrition experiments without considerable management support.

Polyunsaturated fats in meat from Merino, first- and second-cross sheep slaughtered as yearlings

This study examined the level of long chain omega-3 and omega-6 polyunsaturated fats, the ratio of polyunsaturated fat to saturated fat (PUFA/SFA) and the ratio of omega-6 to omega-3 (n-6/n-3) fat in sheep grown under grazing conditions in Australia. The sheep genotypes used were Poll Dorset_growth × Border Leicester Merino (PDg × BLM), Poll Dorset_growth × Merino (PDg × M), Poll Dorsetmuscling × Merino (PDm × M), Border Leicester × Merino (BL × M) and Merino × Merino (M × M). Loin muscles (Longissimus lumborum) collected from 40 ewe and wether sheep slaughtered at 14 months of age were processed for fatty acid determination. After frozen storage, 20 g samples were minced and a 7 g homogenate was processed for muscle lipid extraction using a chloroform:methanol (2:1) procedure. There was an increase in PUFA/SFA as the proportion of Merino genetics increased in the progeny (second-cross < first-cross < Merino), but this was not shown in the n-6/n-3 ratio. The PUFA/SFA trend appeared to be associated with an increase in the level of total polyunsaturated fats, but not a decrease in the level of total saturated fats. The results demonstrate that there is a need to improve the PUFA/SFA content in first- and second-cross animals which are mainly used for meat production in Australia so as to maintain the healthy lipids in meat. Nutritional manipulation through feeding systems or selection of sires for greater heritability of omega-3 fat deposition may be suitable pathways to elevate the ratio of polyunsaturated fatty acids, and in particular omega-3.

Estradiol enables cortisol to act directly upon the pituitary to suppress pituitary responsiveness to GnRH in sheep

We have shown that cortisol infusion reduced the luteinizing hormone (LH) response to fixed hourly GnRH injections in ovariectomized ewes treated with estradiol during the non-breeding season (pituitary-clamp model). In contrast, cortisol did not affect the response to 2 hourly invariant GnRH injections in hypothalamo-pituitary disconnected ovariectomized ewes during the breeding season. To understand the differing results in these animal models and to determine if cortisol can act directly at the pituitary to suppress responsiveness to GnRH, we investigated the importance of the frequency of GnRH stimulus, the presence of estradiol and stage of the circannual breeding season. In experiment 1, during the non-breeding season, ovariectomized ewes were treated with estradiol, and pulsatile LH secretion was restored with i.v. GnRH injections either hourly or 2 hourly in the presence or absence of exogenous cortisol. Experiments 2 and 3 were conducted in hypothalamo-pituitary disconnected ovariectomized ewes in which GnRH was injected i.v. every 2 h. Experiment 2 was conducted during the non-breeding season and saline or cortisol was infused for 30 h in a cross-over design. Experiment 3 was conducted during the non-breeding and breeding seasons and saline or cortisol was infused for 30 h in the absence and presence of estradiol using a cross-over design. Samples were taken from all animals to measure plasma LH. LH pulse amplitude was reduced by cortisol in the pituitary clamp model with no difference between the hourly and 2-hourly GnRH pulse mode. In the absence of estradiol, there was no effect of cortisol on LH pulse amplitude in GnRH-replaced ovariectomized hypothalamo-pituitary disconnected ewes in either season. The LH pulse amplitude was reduced in both seasons in experiment 3 when cortisol was infused during estradiol treatment. We conclude that the ability of cortisol to reduce LH secretion does not depend upon the frequency of GnRH stimulus and that estradiol enables cortisol to act directly on the pituitary of ovariectomized hypothalamo-pituitary disconnected ewes to suppress the responsiveness to GnRH; this effect occurs in the breeding and non-breeding seasons.

Hair braiding : working the boundaries of methodology in globalisation research

This paper engages the constitutive as well as the representative role of metaphor in research. Metaphors are understood to provide possibilities for representations and conceptualisations. The use of metaphorical redescription permits us to 'use familiar words in unfamiliar ways' (Rorty, 1989, p. 18) and provides new language that deters the use of repeated ways of knowing. It invites us to see things differently and to act differently. Lakoff and Johnson (1980) argue that metaphor does more than represent: 'New metaphors have the power to construct a new reality' (p. 145). I have drawn on both these understandings of metaphor. In qualitative research, metaphorical analysis has a well-established history. Researchers analyse metaphors used by research participants and apply metaphors to participant actions and understandings (Koro-Ljungberg, 2001; Gregory & Noblit, 1998). Researchers also use metaphors to reflect or represent their methodological decision-making (Richardson, 2003; Gadamer, 1989). In this article, I have nudged the conceptual boundaries of methodology. I have argued the constructive nature of metaphor in methodological positioning and decision making. I use the writing of a doctoral thesis to argue this role of metaphor. There, the metaphor of hair braiding constructed and communicated my methodological decision-making and my researcher stance as a braider.