William J. Aspden

Reproductive responses of cattle to GnRH agonists

The response in cattle to treatment with gonadotrophin releasing hormone (GnRH) agonist includes downregulation of GnRH receptors on gonadotrophe cells, desensitisation of the anterior pituitary gland to endogenous GnRH, and the abolition of pulsatile release of LH. In bulls, a tonic pattern of LH release is associated with increased secretion of testosterone, which persists for the duration of treatment with GnRH agonist. The mechanism for this response in bulls has not been elucidated, but clearly pulsatile release of LH is not required to stimulate the synthesis of steroidogenic enzymes that sustain elevated secretion of testosterone. In heifers, desensitisation to endogenous GnRH prevents the occurrence of the pre-ovulatory surge release of LH, thus blocking ovulation. The latter provided the opportunity to evaluate the potential of a GnRH agonist bioimplant to control fertility in heifers under extensive management. Bioimplants that contained graded amounts of GnRH agonist prevented pregnancies in heifers for periods of 3 to 12 months. Zebu crossbred heifers treated with GnRH agonist from 14 to 23 months of age failed to conceive, but showed normal conception patterns when introduced into mating herds at around 26 months of age. After treatment with GnRH agonist for 4 to 6 weeks, ovarian follicular growth in heifers is restricted to relatively small (2-4 mm) antral follicles. Suppressed follicular growth in heifers treated long-term with GnRH agonist is due to a lack of gonadotrophin support, rather than a direct action of agonist at the ovaries. This was demonstrated by the ability to induce apparently normal follicular growth and ovulation by acute treatment with FSH for 4 days, followed by an injection of LH, in heifers that had been exposed to GnRH agonist for around 6 months, and which had only small (2-4 mm) antral follicles at the start of FSH treatment. GnRH agonist bioimplants have been incorporated into new multiple ovulation and embryo transfer protocols that allow control of the time of ovulation subsequent to superstimulation of ovarian follicular growth with FSH. In these protocols, the endogenous surge release of LH is blocked by treatment with agonist and ovulation is timed by injection of exogenous LH, allowing fixed-time AI. It can be concluded from recent studies that GnRH agonist bioimplants have considerable potential for both pro-fertility and anti-fertility applications in cattle. It is likely that commercial bioimplants will be available within the next 3 to 5 years.

Sustained testicular atrophy in bulls actively immunized against GnRH: potential to control carcase characteristics

The objectives were to determine whether active immunization against gonadotrophin releasing hormone (GnRH) induced a long-term suppression of testicular function in bulls, and to ascertain the effects of immunization against GnRH on carcase and meat quality characteristics. In experiment 1, 6-month-old Zebu bulls were assigned to: control (n=25), no treatment; immunized (n=31), immunized against GnRH at 0 and 4 months (anti-GnRH(2)), with a sub-set of bulls (n=17) immunized again at 10 months (anti-GnRH(3)). After the second immunization, testicular growth ceased for 2 months in 14/31 (45%) bulls and for at least 6 months in 17/31 (55%) bulls. Among the latter bulls (anti-GnRH(3)) the testes did not grow for >1 year after the third immunization in 5/17 (30%) bulls. In experiment 2, 22-month-old Zebu bulls were assigned to: control (n=14), no treatment; immunized (n=17), immunized against GnRH at 0, 2 and 4 weeks. The testes decreased (P<0.05) in size for 2 months after immunization in 11/17 (65%) bulls and then re-initiated growth, whilst in 6/17 (35%) bulls the testes continued to decrease in size for 4 months and did not re-initiate growth for 1 year. At slaughter, the latter immunocastrated bulls had carcase and meat quality characteristics the same as contemporary bulls that had been castrated before puberty. The findings demonstrated that active immunization against GnRH can induce a long-term suppression of testicular function in a proportion of bulls. Also, when bulls are immunocastrated after puberty, carcase and meat quality traits change from those typical of entire bulls to traits that are characteristic of long-term castrated bulls.

Use of GnRH agonist implants for long-term suppression of fertility in extensively managed heifers and cows.

The ability of gonadotrophin releasing hormone (GnRH) agonist implants to suppress ovarian activity and prevent pregnancies, long-term, was examined in heifers and cows maintained under extensive management. At three cattle stations, heifers (2-year-old) and older cows (3- to 16-year-old) were assigned to a control group that received no treatment, or were treated with high-dose (12 mg, Station A) or low-dose (8 mg, Station B and Station C) GnRH agonist implants. The respective numbers of control and GnRH agonist-treated animals (heifers + cows) at each station were: Station A, 20 and 99; Station B, 19 and 89; Station C, 20 and 76. Animals were maintained with 4% bulls and monitored for pregnancy at 2-monthly intervals for approximately 12 months. Pregnancy rates for control heifers and control cows ranged from 60-90% and 80-100%, respectively, depending on the study site. The respective number of animals (heifers + cows) treated with GnRH agonist that conceived, and days to first conception, were: Station A, 9 (9%) and 336 +/- 3 days; Station B, 8 (10%) and 244 +/- 13 days; Station C, 20 (26%) and 231 +/- 3 days. Treatment with high-dose GnRH agonist prevented pregnancies for longer (approximately 300 days) than treatment with low-dose GnRH agonist (approximately 200 days). In the majority of heifers and cows treated with GnRH agonist, ovarian follicular growth was restricted to early antral follicles (2-4mm). The findings indicate that GnRH agonist implants have considerable potential as a practical technology to suppress ovarian activity and control reproduction in female cattle maintained in extensive rangelands environments. The technology also has broader applications in diverse cattle production systems.

Differential responses in anterior pituitary luteinizing hormone (LH) content and LHβ- and α-subunit MRNA, and plasma concentrations of LH and testosterone, in bulls treated with the LH-releasing hormone agonist deslorelin

Anterior pituitary gland contents of LH and LH beta- and alpha-subunit mRNAs, and circulating concentrations of LH and testosterone, were determined in bulls treated with the LH-releasing hormone (LHRH) agonist deslorelin. Brahman (Bos indicus) bulls (14-month-old) were allocated to two groups and received the following: Control (n = 5), no treatment; Deslorelin (n = 4), four deslorelin implants (approximately 200 micrograms total deslorelin/day) for 36 d. Plasma concentrations of LH were higher in bulls treated with deslorelin on Day 1, had returned to typical levels by Day 8, and did not differ for control bulls and bulls treated with deslorelin from Day 8 to Day 29. Pituitary content of LH on Day 36 was reduced (P < 0.001) in bulls treated with deslorelin (33 +/- 4 ng/mg) compared with control bulls (553 +/- 142 ng/mg). Relative pituitary content of LH beta-subunit mRNA was also reduced on Day 36 in bulls treated with deslorelin (Control, 0.65 +/- 0.10; Deslorelin, 0.22 +/- 0.04; P = 0.003). However, alpha-subunit mRNA relative content did not differ (Control, 0.73 +/- 0.15; Deslorelin, 1.06 +/- 0.12; P > 0.05). Plasma concentrations of testosterone were increased over the period of the experiment in the bulls treated with deslorelin compared with control bulls. This is the first demonstration of reduced pituitary content of LH beta-subunit mRNA and LH, and unaltered content of alpha-subunit mRNA, in bulls treated with LHRH agonist. This was associated with apparently typical plasma concentrations of LH and elevated plasma testosterone. The anterior pituitary in bulls treated with LHRH agonist, therefore, undergoes classical desensitization and downregulation, but plasma LH and testosterone are not suppressed.

Changes in Testicular Steroidogenic Acute Regulatory (STAR) Protein, Steroidogenic Enzymes and Testicular Morphology Associated with Increased Testosterone Secretion in Bulls Receiving The Luteinizing Hormone Releasing Hormone Agonist Deslorelin

Testosterone secretion and the expression and relative contents of steroidogenic acute regulatory (StAR) protein and steroidogenic enzymes cholesterol side-chain cleavage cytochrome P450 (P450SCC), 3beta-hydroxysteroid dehydrogenase/delta(5)-->delta(4)-isomerase (3 beta-HSD), and (17)alpha-hydroxylase cytochrome P450/C17-20 lyase (P450(17)alpha) were determined in testicular tissues of bulls treated with a LHRH agonist. Testis morphology and spermatogenesis were also examined. In Experiment 1, bulls (30-mo-old) received no treatment (control, n = 7) or were implanted for 10 days with the LHRH agonist deslorelin (n = 7). Bulls were castrated on Day 10 and testis tissues prepared for Western and Northern blotting. At castration, bulls implanted with deslorelin had greater plasma testosterone (5-fold) and testis content of testosterone (10-fold) compared with control bulls. Relative content (per micrograms total testis protein or RNA) of StAR protein, 3beta-HSD, P450SCC, and mRNA for P450(17)alpha in bulls treated with deslorelin ranged from 3- to 6-fold that of control bulls. In Experiment 2, bulls (20-mo-old) were left untreated (control, n = 6) or implanted with deslorelin (n = 12) for 120 days. On Day 120, bulls were castrated and right testis tissues prepared for morphology. Testis volume and weight were increased (P < 0.01) in bulls treated with deslorelin compared with control bulls. Stereological analysis revealed that this increase occurred in all compartments (seminiferous epithelium, lumen and interstitium) studied, but was significant (P < 0.01) only for the seminiferous epithelium. Absolute numbers of round spermatids per testis were increased (P < 0.05) in bulls treated with deslorelin compared with control bulls. Increased testosterone secretion in bulls treated with deslorelin was associated with increased testicular StAR protein and steroidogenic enzymes. Bulls treated long-term with deslorelin had a faster rate of testis growth and increased daily sperm production at the end of the experiment.

Superstimulation of ovarian follicular growth with FSH, oocyte recovery, and embryo production from Zebu (Bos Indicus) calves: effects of treatment with a GnRH agonist or antagonist

The capacity of heifer calves of a late sexually maturing Zebu (Bos indicus) genotype to respond to superstimulation with FSH at a young age and in vitro oocyte development were examined. Some calves were treated with a GnRH agonist (deslorelin) or antagonist (cetrorelix) to determine whether altering plasma concentrations of LH would influence follicular responses to FSH and oocyte developmental competency. Brahman calves (3-mo-old; 140 +/- 3 kg) were randomly assigned to 3 groups: control (n = 10); deslorelin treatment from Day -8 to 3 (n = 10); and cetrorelix treatment from Day -3 to 2 (n = 10). All calves were stimulated with FSH from Day 0 to 2, and were ovariectomized on Day 3 to determine follicular responses to FSH and to recover oocytes for in vitro procedures. Before treatment with FSH, heifers receiving deslorelin had greater (P < 0.001) plasma LH (0.30 +/- 0.01 ng/ml) than control heifers (0.17 +/- 0.02 ng/ml), while plasma LH was reduced (P < 0.05) in heifers treated with cetrorelix (0.13 +/- 0.01 ng/ml). Control heifers had a surge release of LH during treatment with FSH, but this did not occur in heifers treated with deslorelin or cetrorelix. All heifers had large numbers of follicles > or = 2 mm (approximately 60 follicles) after superstimulation with FSH, and there were no differences (P > 0.10) between groups. Total numbers of oocytes recovered and cultured also did not differ (P > 0.05) for control heifers and heifers treated with deslorelin or cetrorelix. Fertilization and cleavage rates were similar for the 3 groups, and developmental rates to blastocysts were also similar. Zebu heifers respond well to superstimulation with FSH at a young age, and their oocytes are developmentally competent.

Spatial Subdivision and Genetic Diversity in Populations on the East and West Coasts of Australia: The Multi-Faceted Case of Nautilus pompilius (Mollusca, Cephalopoda)

The fragmented distribution of Nautilus pompilius and its biology suggest there will be significant genetic divergence and spatial subdivision between east and west Australian populations. Samples were collected from the northern Great Barrier Reef, the Coral Sea, and the Scott Reef off Western Australia. Phylogenetic trees and a minimum spanning tree were developed from these data to elucidate evolutionary relationships. These data demonstrate significant evolutionary separation of each of the three populations into strongly supported discrete clades matching geographic stratification. Within each of the discrete populations, genetic variation is evident. Strong inter-population variation is evident, with discrete geographic clades being recognized for each extant group. The distinct spatial subdivision between east and west Australian populations of Nautilus are related to geographic and physical isolation over evolutionary time, and this has important fisheries management implications. The distinct geographic patterns of genetic structuring demonstrated by these data indicate the existence of discrete eastern and western Australian management units, and as such, these Nautilus populations should be managed discretely, as each has a high conservation value containing unique genetic variation.

Increased testosterone secretion in bulls treated with a luteinizing hormone releasing hormone (LHRH) agonist requires endogenous LH but not LHRH

The requirement for endogenous LHRH and LH action in the maintenance of elevated plasma concentrations of testosterone in bulls receiving the LHRH agonist deslorelin was examined. In Experiment 1, bulls were either (i) left untreated (control); (ii) implanted with deslorelin; (iii) actively immunized against LHRH; or (iv) implanted with deslorelin and immunized against LHRH. Experiment 2 was of similar design to Experiment 1, except that bulls were immunized against LH in place of LHRH. In Experiment 1, plasma LH declined in bulls immunized against LHRH, but not in the bulls immunized against LHRH and implanted with deslorelin. Also in Experiment 1, plasma testosterone declined in bulls immunized against LHRH but was elevated in bulls treated with deslorelin and bulls treated with deslorelin and immunized against LHRH. In Experiment 2, bulls immunized against LH and treated with deslorelin had plasma concentrations of testosterone similar to controls, whereas bulls treated only with deslorelin had elevated plasma testosterone. It was concluded from these experiments that endogenous LHRH action was not required for increased steroidogenic activity in bulls treated with a LHRH agonist. However, circulating LH was necessary for increased plasma testosterone in bulls implanted with deslorelin. LH is therefore involved in mediating the response of bulls to treatment with deslorelin, either by acting directly at the testes or through a permissive role that allows a direct action of deslorelin at the testes.

Mitochondrial phylogeography of the critically endangered Capricorn yellow chat (Epthianura crocea macgregori)

Abstract. The critically endangered Capricorn yellow chat (CYC) is endemic to coastal central Queensland on marine plains where it occurs in three areas, numbering <300 birds. Recent industrial expansion in the region has increased the threat to the CYC. To assist management of the subspecies, a phylogeographical evaluation of the CYC using mitochondrial DNA was undertaken. We found no genetic diversity within, nor genetic divergence between, the two areas at the northern and southern extremes of their current distribution, and only slight morphological differences. These findings suggest that the two groups of CYC represent daughter populations of an ancestral population that was affected by a genetic bottleneck in the recent past. Implications for conservation of the subspecies could be increased vulnerability to environmental change. A preliminary evaluation of the divergence between the CYC and its nearest subspecies, the widespread inland yellow chat, indicate a time to the most recent common ancestor of 215 000 years or less. This timespan overlaps two periods of glacial aridity during which xeric habitats used by yellow chats for breeding, such as semiarid and arid swamps, may have expanded, allowing colonisation of the coastal marine plains. CYCs may represent a relictual population from a previously more xeric era that has subsequently become isolated as the region became wetter following glacial maxima.

Ultrastructure of the gastro intestinal tract of healthy Japanese quail (Coturnix japonica) using light and scanning electron microscopy

The Japanese quail (Coturnix japonica) are popular both as an alternative protein source and as a model of choice for scientific research in several disciplines. There is limited published information on the histological features of the intestinal tract of Japanese quail. The only comprehensive reference is a book published in 1969. This study aims to fill that niche by providing a reference of general histological features of the Japanese quail, covering all the main sections of the intestinal tract. Both light and scanning electron microscope (SEM) images are presented. Results showed that the Japanese quail intestinal tract is very similar to that of the chicken with the exception of the luminal koilin membrane of the gizzard. Scanning electron microscopic photomicrographs show that in the Japanese quail koilin vertical rods are tightly packed together in a uniform manner making a carpet-like appearance. This differs in chicken where the conformations of vertical rods are arranged in clusters.