Enhancing performance of berseem clover genotypes with better harvesting management through farmers' participatory research at smallholder farms in Punjab.

A field study was conducted on smallholder farmer fields between 2012 to 2014 to evaluate the performance of cv. Agaitti Berseem-2002, against local landraces exchanged between farmers (LBF1) or available from local markets (LBM1). The effects of genotype and harvesting regimen on forage production, quality and seed production were evaluated. Significant differences (P < 0.05) among genotypes and cutting treatments were recorded for forage and seed yields, and forage quality across all research sites in both years. Maximum cumulative fresh forage (89.7 t/ha) and dry matter (DM; 13.4 t/ha) yields were obtained with Agaitti Berseem-2002 when harvesting occurred five times over the season. However, maximum seed yield (1048 kg/ha) with higher 1000-seed weight (3.63 g) were obtained if forage was only harvested three times and the crop then left for seed set. Agaitti Berseem-2002 also produced forage with the higher crude protein content (27%), DM digestibility (69%), digestible organic matter (DM basis; 65%) and metabolizable energy content (10%) compared to the local landraces (LBF1 and LBM1). Therefore, the harvesting regimen for greatest economic return which produced optimum fresh and DM forage yields of highest nutritive values and maximum seed yield, were comprised of taking three forage cuts (at 65, 110 and 150 days after sowing) prior to seed harvest.

Molecular characterization of the level of sialic acids N-acetylneuraminic acid, N-glycolylneuraminic acid, and ketodeoxynonulosonic acid in porcine milk during lactation.

Sialic acids (Sia) are key monosaccharide constituents of sialylated glycoproteins (Sia-GP), human sialylated milk oligosaccharide (Sia-MOS), and gangliosides. Human milk sialylated glycoconjugates (Sia-GC) are bioactive compounds known to act as prebiotics and promote neurodevelopment, immune function, and gut maturation in newborns. Only limited data are available on the Sia content of porcine milk. The objective of this study was to quantitatively determine the total level of Sia N-acetylneuraminic acid (Neu5Ac), N-glycolylneuraminic acid (Neu5Gc), and ketodeoxynonulosonic acid (KDN) in porcine milk and to compare these levels in gilt and sow milk during lactation. Milk from 8 gilts and 22 sows was collected at 3 stages of lactation (colostrum, transition, and mature milk). Standard and experimental samples were derivatized using 1,2-diamino-4,5-methylenedioxy-benzene and analyzed by ultra-high-performance liquid chromatography using a fluorescence detector. The following new findings are reported: (1) Gilt and sow milk contained significant levels of total Sia, with the highest concentration in colostrum (1,238.5 mg/L), followed by transition milk (778.3 mg/L) and mature milk (347.2 mg/L); (2) during lactation, the majority of Sia was conjugated to Sia-GP (41-46%), followed by Sia-MOS (31-42%) and a smaller proportion in gangliosides (12-28%); (3) Neu5Ac was the major form of Sia (93-96%), followed by Neu5Gc (3-6%) and then KDN (1-2%), irrespective of milk fraction or stage of lactation; (4) the concentration of Sia in Sia-GP and Sia-MOS showed a significant decline during lactation, but the level of ganglioside Sia remained relatively constant; (5) mature gilt milk contained a significantly higher concentration of Sia-GP than sow milk. The high concentration of total Sia in porcine milk suggests that Sia-GC are important nutrients that contribute to the optimization of neurodevelopment, immune function, and growth and development in piglets. These findings provide an important rationale for the inclusion of Sia-GC in pig milk replacers to mimic porcine milk composition for the optimal growth and development of piglets.

A review of recent developments in buffalo reproduction - a review.

The buffalo is an important livestock resource in several countries of South Asia and the Mediterranean regions. However, reproductive efficiency is compromised due to known problems of biological and management origins, such as lack of animal selection and poor nutrition. Under optimal conditions puberty is attained at 15 to 18 months in river buffalo, 21 to 24 months in swamp buffalo and is influenced by genotype, nutrition, management and climate. However, under field conditions these values deteriorate up to a significant extant. To improve reproductive efficiency, several protocols of oestrus and ovulation synchronization have been adopted from their use in commercial cattle production. These protocols yield encouraging pregnancy rates of (30% to 50%), which are comparable to those achieved in buffaloes bred at natural oestrus. The use of sexed semen in buffalo heifers also showed promising pregnancy rates (50%) when compared with conventional non-sexed semen. Assisted reproductive technologies have been transferred and adapted to buffalo but the efficiency of these technologies are low. However, these latest technologies offer the opportunity to accelerate the genetic gain in the buffalo industry after improving the technology and reducing its cost. Most buffaloes are kept under the small holder farming system in developing countries. Hence, future research should focus on simple, adoptable and impact- oriented approaches which identify the factors determining low fertility and oestrus behaviour in this species. Furthermore, role of kisspeptin needs to be explored in buffalo.

Effect of pre-weaning feeding regimens on post-weaning growth performance of Sahiwal calves.

The objective of the study was to assess the post-weaning growth response of Sahiwal calves reared on four different pre-weaning dietary regimens. The four diets were: (a) whole cow's milk, starter ration (SR; CP = 20%, total digestible nutrients (TDN) = 72%) and Berseem hay (H; Egyptian clover; CP = 21%, TDN = 63%); (b) whole cow's milk + H; (c) milk replacer (MR; reconstituted to supplier specification; Sprayfo®) + SR + H; and (d) MR + H. The protein and fat percentages of reconstituted MR were 2.22 and 1.84, respectively. Milk or MR were fed at the rate of 10% of the calves' body weight (BW) until 56 days of age, and then withdrawn gradually until weaned completely by 84 days of age. The average initial BW of calves in groups A, B, C and D were 56.3 ± 1.0, 47.5 ± 1.0, 40.4 ± 1.0 and 30.3 ± 1.0 kg, respectively. Initially, there were 12 calves in each group with six of each sex; however, one male calf died from each of groups B and C and were not replaced. During the post-weaning period, 13 to 24 weeks, the calves were fed a single total mixed ration ad libitum based on maize, canola meal, wheat straw and molasses containing 16% CP and 70% TDN. Daily feed intake and weekly BW gains were recorded. The data were analyzed by MIXED model analysis procedures using the statistical program SAS. The intake of calves as percent of their BW, feed conversion ratio and cost per kg of BW gain were not different (P > 0.05) across treatments. The daily gain at 24 weeks of age for the pre-weaning treatments A, B, C and D were 746 ± 33, 660 ± 33, 654 ± 33 and 527 ± 33 g/day and the final liveweights of calves were 119 ± 4.2, 102 ± 4.2, 95 ± 4.2 and 75 ± 4.2 kg, respectively. Gains were influenced significantly (P < 0.05) by pre-weaning treatments. The calves fed MR and H only during the pre-weaning period were unable to catch up post weaning with calves on other dietary treatments. The calves fed whole milk from birth at the rate of 10% of liveweight together with concentrates had higher weaning weight and superior growth rate post weaning as well. Thus, pre-weaning feeding was important for higher weaning weights and superior growth rates post weaning.

Evaluation of commercially available assays for the measurement of equine insulin.

Determining circulating equine insulin concentrations is becoming increasingly important in equine clinical practice and research. Most available assays are optimized for human medicine, but there is strong equine cross-reactivity because of the highly conserved nature of insulin. To identify an accurate and reliable assay for equine insulin, 6 commercial immunoassays were evaluated for precision, accuracy, and specificity. Only 1 assay initially reached the requisite standard: Mercodia Equine Insulin Enzyme-linked Immunosorbent assay (ELISA). Plasma matrix interferences were identified when the provided assay buffer was used with the Siemens Count-a-Coat Insulin radioimmunoassay (RIA) but not when charcoal-stripped equine plasma was used as the diluent. This modified RIA and the Mercodia Equine Insulin ELISA were evaluated further by directly examining accuracy by comparing their results for 18 equine plasma samples with values obtained using liquid chromatography and high-resolution/high-accuracy mass spectrometry (LC-MS). Compared with LC-MS measurements, the modified Siemens Insulin RIA rendered a moderate Lin's concordance coefficient (ρ(c)) of 0.41, whereas the Mercodia Equine Insulin ELISA rendered a very poor ρ(c) of 0.06. This suggests that the Siemens Insulin RIA is appropriate to use for routine evaluations when LC-MS is not available.

Assessing ovulation detection performance in commercial dairy herds using progesterone concentrations from limited numbers of strategically collected milk samples.

It is important to assess ovulation detection performance in commercial dairy herds both to investigate low reproductive performance and to enable herd managers to monitor the effectiveness of their system for detecting ovulations. A method was developed to assess ovulation detection performance that uses limited numbers of strategically collected milk samples, assesses performance over the period when herd managers are making maximal effort to detect ovulations, and when assessing proportions of ovulations detected, accounts for false positive diagnoses of estrus and for cows that have not recommenced postpartum ovulatory cycles. Milk was sampled from cows not diagnosed in estrus early in the breeding program (about d 26 in year-round calving herds and d 22 in seasonal calving herds); milk samples were also collected from cows on the day of insemination. Cows with high milk progesterone concentrations were assumed to have had undetected ovulations and false positive diagnoses of estrus, respectively. The method was successfully implemented in 161 of 167 commercial dairy herds. Positive predictive values (PPV; the proportions of ovulation diagnoses where ovulation was, in fact, imminent) were generally high in both year-round and seasonal calving herds (median values were 0.96 and 0.97, respectively), but 25% of herds had PPV <0.95. Ovulation detection sensitivities (ODS) were low in most year-round calving herds, but many seasonal calving herds had high ODS values; median ODS were 0.73 and 0.94, respectively. However, in 25% of seasonal calving herds, ODS was <0.91. These findings indicate that this method for assessing ovulation detection performance can be successfully implemented in commercial dairy herds with appropriate professional support. The wide range of ODS and the absence of correlation between ODS and PPV suggest that it is possible for managers of many commercial herds in Australia to achieve increased reproductive efficiency through increases in ODS and PPV.

The influence of extracellular matrix and prolactin on global gene expression profiles of primary bovine mammary epithelial cells in vitro.

An in vitro bovine mammosphere model was characterized for use in lactational biology studies using a functional genomics approach. Primary bovine mammary epithelial cells cultured on a basement membrane, Matrigel, formed three-dimensional alveoli-like structures or mammospheres. Gene expression profiling during mammosphere formation by high-density microarray analysis indicated that mammospheres underwent similar molecular and cellular processes to developing alveoli in the mammary gland. Gene expression profiles indicated that genes involved in milk protein and fat biosynthesis were expressed, however, lactose biosynthesis may have been compromised. Investigation of factors influencing mammosphere formation revealed that extracellular matrix (ECM) was responsible for the initiation of this process and that prolactin (Prl) was necessary for high levels of milk protein expression. CSN3 (encoding kappa-casein) was the most highly expressed casein gene, followed by CSN1S1 (encoding alphaS1-casein) and CSN2 (encoding beta-casein). Eighteen Prl-responsive genes were identified, including CSN1S1, SOCS2 and CSN2, however, expression of CSN3 was not significantly increased by Prl and CSN1S2 was not expressed at detectable levels in mammospheres. A number of novel Prl responsive genes were identified, including ECM components and genes involved in differentiation and apoptosis. This mammosphere model is a useful model system for functional genomics studies of certain aspects of dairy cattle lactation.

A functional genomics approach to evaluate candidate genes located in a QTL interval for milk production traits on BTA6.

The potential genetic and economic advantage of marker-assisted selection for enhanced production in dairy cattle has provided an impetus to conduct numerous genome scans in order to identify associations between DNA markers and future productive potential. One area of focus has been a quantitative trait locus on bovine chromosome 6 (BTA6) found to be associated with milk yield, milk protein and fat percentage, which has been subsequently fine-mapped to six positional candidate genes. Subsequent investigations have yet to resolve which of the potential positional candidate genes is responsible for the observed associations with productive performance. In this study, we analysed candidate gene expression and the effects of gene knockdown on expression of beta- and kappa-casein mRNA in a small interfering RNA transfected bovine in vitro mammosphere model. From our expression studies in vivo, we observed that four of the six candidates (ABCG2, SPP1, PKD2 and LAP3) exhibited differential expression in bovine mammary tissue over the lactation cycle, but in vitro functional studies indicate that inhibition of only one gene, SPP1, had a significant impact on milk protein gene expression. These data suggest that the gene product of SPP1 (also known as osteopontin) has a significant role in the modulation of milk protein gene expression. While these findings do not exclude other positional candidates from influencing lactation, they support the hypothesis that the gene product of SPP1 is a significant lactational regulatory molecule.

Ghrelin secretion is more closely aligned to energy balance than with feeding behaviour in the grower pig.

Secretory characteristics of the ghrelin profile for the pig are still unknown. Our objective was to clarify the mechanisms that influence ghrelin secretion during differing feeding patterns. Pigs were initially fed a commercial pelleted diet offered ad libitum and blood samples collected for 24 h at intervals of 1 h. The pigs were then entrained for 17 days to a twice daily interval feeding regimen (0900-1000 and 1600-1700 h) and blood samples were collected for 12 h (0800-2000 h). This was followed by a similar interval feeding and blood sampling regimen with the 0900-1000 h feeding period being replaced by a sham feed where pigs were shown their usual feed but none offered. During the ad libitum feeding regimen, there was no preprandial rise or postprandial fall in circulating plasma total ghrelin concentration, which remained constant throughout the sampling period. In addition, no preprandial rise or postprandial fall in ghrelin concentrations was observed when pigs were fed either twice or once daily; however, plasma ghrelin concentration rose gradually over the 12-h sampling period during the twice daily feeding regimen and increased further when pigs were fed once per day. This increase in ghrelin levels coincided with an increase in plasma GH and non-esterified fatty acid concentrations and was not associated with either plasma glucose or insulin concentrations. These results suggest that circulating total plasma ghrelin concentrations in the pig appear to be influenced by chronic changes in energy balance rather than the feeding pattern per se.

Interaction between oestrogen and oxytocin on hypothalamic-pituitary-adrenal axis activity.

In addition to its role in reproduction, oxytocin has central actions modulating behavioural and hypothalamic-pituitary-adrenal (HPA) axis responses during late pregnancy and lactation. The hypothesis that ovarian hormones modulate the effects of oxytocin on HPA axis activity was studied in 7-day ovariectomised rats receiving oestradiol with or without progesterone replacement and intracerebroventricular (i.c.v) minipump infusion of oxytocin (100 ng/h). In an initial experiment, i.c.v. oxytocin had no effect on basal or restraint-stimulated plasma adrenocorticotrophic hormone (ACTH) and corticosterone concentrations or hypothalamic corticotrophin-releasing factor (CRF) mRNA expression with low oestradiol replacement alone but it had a stimulatory effect in the presence of low oestradiol and progesterone. To investigate further whether oestradiol modulates central actions of oxytocin, rats received low dioestrous (low), pro-oestrous (medium) or pregnancy (high) oestradiol replacement levels, yielding plasma concentrations of < 5, 17.3 +/- 4.5 and 258 +/- 32 pg/ml, respectively, with or without i.c.v. oxytocin. Oestradiol caused dose-dependent increases in basal plasma ACTH and corticosterone concentrations but decreased the ACTH response to restraint stress. In parallel to the changes in basal plasma ACTH, high oestrogen increased basal CRF hnRNA, CRF mRNA in the paraventricular nucleus and pro-opiomelanocortin (POMC) mRNA in the pituitary gland, while decreasing restraint stress-stimulated levels. Intracerebroventricular administration of oxytocin reduced basal and stress-stimulated plasma ACTH, hypothalamic CRF hnRNA (30 min), CRF mRNA and pituitary POMC mRNA (4 h) levels parallel to the increases induced by elevating plasma oestradiol. The present study demonstrates the converse effects of oestradiol on basal and restraint stress-stimulated basal HPA axis activity, and that the ability of central oxytocin to inhibit HPA axis activity depends on the levels of circulating oestradiol.

Lactational performance of Quackenbush Swiss line 5 mice.

We evaluated 2 strains of mice for their utility in the investigation of nutritional and molecular regulatory mechanisms of lactation. The lactational performance and milk composition were characterized for an inbred mouse strain, inbred Quackenbush Swiss line 5 (QSi5) selected persistently for fecundity, and a nonselected strain, CBA. The milk yield assessed by changes in BW in response to suckling of sustainable litter sizes for each strain was 3-fold greater (P < 0.001) in QSi5 mice than the CBA strain. The QSi5 mice also produced milk more efficiently (P < 0.001) than CBA mice, despite having the same quantity of mammary tissue per unit of BW. Milk composition did not vary between strains or by stage of lactation, with the exception of lactose concentration, which was greater (P = 0.003) in QSi5 mice. Expression of epsilon-casein was > or = 10-fold greater, and alpha(S1)-casein was > or = 3-fold greater, during mid and late lactation compared with early lactation in both strains, whereas kappa-casein underwent an apparent alteration in posttranslational modifications in both strains from early to mid lactation. Changes in casein composition coincided with an increased susceptibility to proteolytic degradation; hence milk from early lactation may be more readily degraded to facilitate digestion in the neonate. The greater milk synthetic capacity of QSi5 mice over the lactation cycle provides a useful model for studies of nutritional and molecular regulation of lactation.

The effect of active immunization against adrenocorticotropic hormone on cortisol, beta-endorphin, vocalization, and growth in pigs.

Because the poor growth performance of intensively housed pigs is associated with increased circulating glucocorticoid concentrations, we investigated the effects of glucocorticoid suppression by inducing a humoral immune response to ACTH on physiological and production variables in growing pigs. Grower pigs (28.6 +/- 0.9 kg) were immunized with amino acids 1 through 24 of ACTH conjugated to ovalbumin and suspended in diethylaminoethyl (DEAE) dextran-adjuvant or adjuvant alone (control) on d 1, 28, and 56. The ACTH-specific antibody titers generated suppressed increases in cortisol concentrations on d 63 in response to an acute stressor (P = 0.002; control = 71 +/- 8.2 ng/mL; ACTH-immune = 43 +/- 4.9 ng/mL) without altering basal concentrations. Plasma beta-endorphin concentrations were also increased (P < 0.001) on d 63 (control = 18 +/- 2.1 ng/mL; ACTH-immune = 63 +/- 7.3 ng/mL), presumably because of a release from negative feedback on the expression of proopiomelanocortin in pituitary corticotropes. Immunization against ACTH did not alter ADG (P = 0.120; control = 1,077 +/- 25; ACTH-immune = 1,143 +/- 25 g) or ADFI (P = 0.64; control = 2,719 +/- 42; ACTH-immune = 2,749 +/- 42 g) and did not modify behavior (P = 0.681) assessed by measuring vocalization in response to acute restraint. In summary, suppression of stress-induced cortisol responses through ACTH immunization increased beta-endorphin concentrations, but it did not modify ADG, ADFI, or restraint vocalization score in growing pigs.

Effects of genetic merit and varying dietary protein degradability on lactating dairy cows.

Eighty two multiparous Holstein cows were blocked by genetic merit (high vs. low) and assigned to one of two treatments [high rumen-undegradable protein (RUP): rumen-degradable protein (RDP) vs. low RUP: RDP] from d 21 before to d 150 after calving to study the effects of these treatments on production and reproductive performance. Diets were isonitrogenous (dry cow 10.5% crude protein; lactating cow 19.3%), isoenergetic (dry cow 10.0 MJ of metabolizable energy (ME); lactating cow 10.9 MJ of ME) and fed as total mixed rations. Feeding more RUP significantly increased dry matter intake and milk yield, reduced body tissue mobilization, and lowered concentrations of serum nonesterified fatty acids (NEFA) and plasma urea. Expression of estrus at first ovulation was improved, first service conception rate was higher, and calving to conception interval was shorter for the high RUP group. Cows of high genetic merit produced more milk, mobilized more body tissue, and had higher concentrations of plasma growth hormone. The dry matter intake and concentrations of blood metabolites did not significantly differ with genetic merit. Expression of estrus at first ovulation was significantly lower for cows of high genetic merit. Serum NEFA concentrations were significantly higher, and estrus was not observed at first ovulation for cows of higher genetic merit fed the low RUP diet. The interaction between dietary RUP and genetic merit was not significant for other measures of performance or fertility. Feeding a low RUP: high RDP diet had negative effects on some aspects of production and reproductive performance. The effects of diet on NEFA concentrations and estrus display were greater in cows of high genetic merit, indicating that potential interactions should be evaluated in future reproductive studies involving protein and fertility.

The effects of fibroblast growth factors 1 and 2 on fibre growth of wool follicles in culture.

The effects of fibroblast growth factor-1 (FGF-1) and FGF-2 on fibre growth and follicle function were examined using a previously described procedure to culture wool follicles. Because the FGFs bind to glycosaminoglycan components of the extra-cellular matrix, we also investigated interactions between FGF-1 and FGF-2 with heparin on wool fibre growth. Individual follicles were microdissected from Merino sheepskin and transferred to culture. Follicles increased in length for 6 days, and in all groups, no significant differences in follicle length were observed. Increase in follicle length was associated with fibre growth, follicles maintained normal anagen morphology and incorporated [3H]thymidine into the bulb and outer root sheath cells. Follicles in all treatments continued to produce fibre keratins, as detected by immunohistochemistry. However, the patterns of fibre and cytoskeletal proteins incorporating [35S]methionine in control and treated follicles were significantly different. We found a considerable decrease in the intermediate filament keratins or low sulphur proteins in follicles cultured in the presence of FGF-1 and FGF-2 compared to controls. The majority of proteins detected in these samples were acidic high sulphur proteins. These studies provide evidence for a specific role for the fibroblast growth factors in the regulation of fibre differentiation.

Effects of epidermal growth factor and transforming growth factor alpha on the function of wool follicles in culture.

The development of a procedure to culture wool follicles from Merino sheep in serum-free conditions has enabled us to investigate the actions of epidermal growth factor (EGF) and transforming growth factor alpha (TGF alpha) on follicle function, including fibre growth. Follicles grown in the absence of growth factors maintained their anagen morphology for 6 days as determined by light microscopy. During this time they incorporated [3H]thymidine into the DNA of the bulb matrix and outer root sheath (ORS) cells and produced fibre keratins as detected by immunohistochemistry. In the presence of EGF and TGF alpha, fibre production ceased after 4 days, as it does following the administration of EGF in vivo. Cessation of fibre growth was not accompanied by regression of the follicle bulb which occurs in vivo. Follicle length growth did not differ significantly from controls and cells in the bulb continued to proliferate. Usually, the structure of the dermal papillae resembled that in control follicles, which was also in marked contrast to changes reported in vivo. In EGF- and TGF alpha-treated follicles, [3H]thymidine continued to be incorporated into DNA of the ORS and bulb after fibre growth ceased. Although wool keratin synthesis ceased, cytokeratins of the epidermis and ORS continued to be produced in the bulb as detected by immunochemistry. These bulb cells were also positive for the periodic acid-Schiff (PAS) reaction indicating the presence of glycogen, a normal component of ORS cells. The observations that cell proliferation continued in the bulb, that glycogen was present and that soft keratins were expressed in these cells suggest that the bulb cell population was induced to differentiate into an ORS phenotype by EGF and TGF alpha.

Characterization and distribution of epidermal growth factor receptors in the skin and wool follicles of the sheep fetus during development.

We have determined the binding affinity and capacity and relative distribution of epidermal growth factor (EGF) receptors in the skin of the Merino sheep fetus before and during the development of the wool follicle population. Autoradiography of tissue sections incubated with [125I]EGF revealed that label was confined predominantly to the epidermis and dermoepidermal junction before follicle formation, at 30 and 55 d of gestation. During follicle initiation (Days 60 to 65), receptor activity was distributed over the epidermis, including the epidermal aggregations of primordia at the dermoepidermal junction. However, receptor concentrations, as revealed by grain counts of autoradiographs, were reduced in these regions when compared with 55-d skin. The receptor distribution over the epidermis and its derivatives did not alter during subsequent follicle development, although the intensity of labeling increased as the follicles matured. Specific receptor binding was not observed above background levels in the dermis and dermal papillae during all stages of follicle development. At follicle maturation, EGF receptors were widely distributed over the cells of the epidermis and the epidermal derivatives of the cutaneous appendages but were particularly localized in the sebaceous glands and outer root sheath (see also Wynn et al. 1989). EGF immunoreactive material has also been found at these sites (du Cros et al. 1992), suggesting an autocrine role for EGF in the regulation of cell function. It is likely that the differentiation-promoting activities of EGF may predominate over those of growth, because the receptor-bearing cells were not members of rapidly proliferating populations.

Growth of wool follicles in culture.

A procedure for the culture of isolated wool follicles from Merino sheep is described. Follicles were microdissected from midside skin samples of 2-yr-old wethers and transferred, individually, to 24-well tissue culture plates. When maintained in supplemented Williams' E medium containing 5 to 10% fetal bovine serum (FBS), insulin, hydrocortisone, and a trace element mixture, fibre growth rates of 40 to 80 microns/day were observed. Follicles maintained their morphologic integrity for up to 7 days, incorporated [methyl-3H]thymidine into DNA and [35S]methionine into intermediate-filament keratins of the growing fiber. Insulin and hydrocortisone stimulated fiber growth at concentrations of 10 micrograms/ml and 50 ng/ml, respectively, but higher doses were inhibitory. The growth of fibers in response to hydrocortisone and the changes in follicle morphology was similar to those induced in skin after systemic administration of cortisol in vivo. A positive interaction between hydrocortisone and trace elements for follicle survival and hydrocortisone, insulin, and FBS for fiber growth was also found. The successful culture of Merino sheep follicles provides a model with which to study the direct influence of endocrine, nutritional and local factors on wool keratin synthesis independently of systemic shifts in the animals' metabolism.

The development of corticosteroid binding globulin-like activity in fetal sheep blood.

Parturition in sheep is initiated by the fetus and is preceded by a rise in fetal cortisol and corticosteroid-binding globulin (CBG) late in gestation. In this study plasma cortisol and CBG concentrations were measured in fetal and maternal circulation from 40 days gestation to early post-partum. The fetal cortisol profile was shown to be triphasic in nature; being high in both the first and last trimester but low in the middle period of gestation. In the last trimester, total cortisol increased steadily, reaching it's highest level just prior to parturition (145 days gestation), before falling to maternal levels over the first 10 days post-partum. The changes seen in CBG concentrations throughout gestation and post-partum mirrored the triphasic nature seen in cortisol levels. CBG was significantly higher at 40, 56 and 140 days gestation than at mid-gestation (77 and 90 days). However, at 145 days gestation there was a significant fall in CBG levels. CBG levels were higher at 1 day post-partum when compared to 145 days gestation, the former rapidly falling to maternal levels over the subsequent 9 days. The maximum binding capacity at 40, 56, 70 and 90 days gestation exceeds the total serum cortisol concentration. However at 140 and 145 days gestation and 1 day post-partum the total serum cortisol exceeds the Bmax. The highest cortisol:Bmax ratio is seen at 145 days gestation due to the fall of CBG binding capacity at this time.(ABSTRACT TRUNCATED AT 250 WORDS)