Suppression of ovarian progesterone production in dairy cows using an implant of GnRH-agonist (deslorelin) for the purpose of evaluating progesterone metabolism

OBJECTIVE: To evaluate the potential of an implant of a GnRH-agonist (deslorelin) to create a progesterone free animal suitable for studying progesterone (P4) metabolism in intact cows by measuring blood P4 and faecal P4 metabolites. METHODS: Experiment 1: Eighteen non-lactating cycling Holstein-Friesian cows, 4 to 7 years old, were allocated to one of three groups to study plasma P4 concentrations preceding an intravaginal insert. These groups comprised: i) a deslorelin group (GnRH-agonist implanted); ii) a PGF group receiving two injections of prostaglandin (PGF2alpha) 12 days apart; and, iii) an ovariectomised (OVX) group. An intravaginal device (CIDR) was inserted into the vagina of each animal and left in place for 11 days. Plasma P4 concentrations were measured during the study period. Experiment 2: Twelve non-lactating cycling Holstein-Friesian cows, 4 to 7 years old, were allocated to two groups: i) a deslorelin group (GnRH-agonist implanted); and ii) an ovariectomised group. Plasma P4 and faecal P4 metabolites (20-oxo-pregnanes, 20alpha-OH and 20beta-OH) were monitored for a period of 5 weeks. RESULTS: Experiment 1: Average plasma P4 concentration did not differ between the three groups (1.28, 1.43 and 1.55 ng/mL for deslorelin, OVX and PGF cows, respectively, P = 0.8) during the period of supplementation. Experiment 2: There was no difference in plasma P4 (mean plasma P4 < 0.02 ng/mL, P = 0.9) and faecal P4 metabolites between deslorelin and OVX cows 2 weeks after the implantation (P = 0.7). CONCLUSIONS: These data showed that a GnRH-agonist (deslorelin) implant may be used as an alternative to ovariectomy to create a progesterone free animal suitable for studying the metabolism of administered P4.     

Changes in the concentration of mRNAs for the inhibin subunits in ovarian follicles after administration of gonadotropins to progestin treated ewes.

RNA was extracted from single or small groups of ovine ovarian follicles after treatment of ewes with FSH and/or LH. The content of mRNA for the alpha-inhibin and beta A-inhibin subunits was analyzed by hybridization with specific cDNA probes. All ewes were treated with progestin vaginal pessaries to suppress spontaneous preovulatory follicle maturation and ewes were given three intramuscular injections of gonadotropins at 8-hr intervals starting 24 hr prior to collection of ovaries. In experiment I, both Schering-FSH and NIDDK-oFSH-17 (oFSH) significantly increased alpha- and beta A-inhibin mRNA per ewe in 2-5 mm follicles and tended to increase alpha- and beta A-inhibin mRNA in large (greater than 5 mm) follicles. In experiment II, oFSH and NIDDK-oLH-25 (oLH) were administered in a 2X2 factorial arrangement. Separate administration of oFSH or oLH increased (P less than .05) the alpha-inhibin mRNA concentration in large follicles. alpha-inhibin mRNA concentration in 4-5 mm follicles was also increased by oFSH but was decreased by oLH. Concomitant treatment with oFSH and oLH did not change alpha-inhibin mRNA concentrations from those measured in oFSH treated ewes. In experiment II, beta A mRNA concentrations followed a pattern similar to that of alpha A mRNA, but the differences were not statistically significant. We conclude that, in the ewe, exogenous FSH increases the concentration of inhibin mRNA in the whole follicle. The ability of exogenous oLH to alter expression of the inhibin subunit genes may depend upon the stage of follicle maturation.     

Structural Characterisation of the Interaction between Triticum aestivum and the Dothideomycete Pathogen Stagonospora nodorum

The interaction between Stagonospora nodorum and a susceptible wheat cultivar was investigated using a range of microscopic techniques. Germination of pycnidiospores occurred approximately 3 h after making contact with the leaf surface and was followed by attempted penetration 8–12 h later. Penetration was observed through stomata and also directly through periclinal and anticlinal epidermal cell walls. Penetration down the anticlinal cell walls appeared to occur without a differentiated penetrating structure whilst structures identified as either lateral appressoria or hyphopodia were typically present when penetrating over a periclinal cell wall. Once inside the leaf, the fungus continued to grow for the next 4–5 days colonising all parts of the leaf except the vascular bundles. Only in the later phase of the infection was total host cell collapse apparent. Evidence of polyphenolic compounds was observed. The infection cycle was completed within 7 days as indicated by sporulation on the leaf surface. These results have allowed us to understand how the fungus physically interacts with the leaf and will help the overall understanding of the infection process.     

Integrated disease management of ascochyta blight in pulse crops

Ascochyta blight causes significant yield loss in pulse crops worldwide. Integrated disease management is essential to take advantage of cultivars with partial resistance to this disease. The most effective practices, established by decades of research, use a combination of disease-free seed, destruction or avoidance of inoculum sources, manipulation of sowing dates, seed and foliar fungicides, and cultivars with improved resistance. An understanding of the pathosystems and the inter-relationship between host, pathogen and the environment is essential to be able to make correct decisions for disease control without compromising the agronomic or economic ideal. For individual pathosystems, some components of the integrated management principles may need to be given greater consideration than others. For instance, destruction of infested residue may be incompatible with no or minimum tillage practices, or rotation intervals may need to be extended in environments that slow the speed of residue decomposition. For ascochyta-susceptible chickpeas the use of disease-free seed, or seed treatments, is crucial as seed-borne infection is highly effective as primary inoculum and epidemics develop rapidly from foci in favourable conditions. Implemented fungicide strategies differ according to cultivar resistance and the control efficacy of fungicides, and the effectiveness of genetic resistance varies according to seasonal conditions. Studies are being undertaken to develop advanced decision support tools to assist growers in making more informed decisions regarding fungicide and agronomic practices for disease control.     

催乳素对内蒙古绒山羊毛囊体外培养影响的研究

【目的】 研究催乳素（PRL）对内蒙古绒山羊初级毛囊和次级毛囊体外生长及形态变化的影响。【方法】 机械法结合切割法分离内蒙古绒山羊的初级毛囊和次级毛囊,在初级毛囊培养液中分别添加0、5、10、50、100 ng/mL催乳素进行体外培养,每组24根,共培养5 d,每天在显微镜下观察其形态并拍照,统计其生长长度、生长速度和存活率,筛选出最适催乳素处理浓度。然后将初级毛囊与次级毛囊分别分为初级毛囊对照组（PF-K）、初级毛囊试验组（PF-PRL）、次级毛囊对照组（SF-K）、次级毛囊试验组（SF-PRL）,每组24根,对照组用基础培养液培养,试验组在基础培养液中添加最适浓度的催乳素,培养5 d,每天观察毛囊的形态并拍照,同时测量各组毛囊的生长长度。【结果】 10 ng/mL催乳素组毛囊的平均日生长长度均极显著高于其他浓度组（P<0.01）,最终生长长度和存活率均最高,因此,后续试验选择10 ng/mL催乳素处理毛囊。试验组和对照组初/次级毛囊的毛干与根鞘部位同时伸长,随着培养时间的增加均出现不同程度的弯曲。PF-PRL、SF-PRL组毛囊在2~5 d的总长度分别极显著高于PF-K、SF-K组（P<0.01）。PF-K组除第1天与第0天差异不显著外,1~5 d毛囊的总长度依次显著增加（P<0.05）;PF-PRL组0~5 d毛囊的总长度依次显著增加（P<0.05）。SF-K组毛囊第5天的总长度显著高于0~4 d （P<0.05）;SF-PRL组第4、5天毛囊的总长度均显著高于0~3 d （P<0.05）,第3天毛囊的总长度显著高于0~2 d （P<0.05）。PF-PRL、SF-PRL组毛囊在2~5 d的平均日生长长度分别极显著高于PF-K、SF-K组（P<0.01）。【结论】 10 ng/mL催乳素是体外促进毛囊生长的最适浓度,10 ng/mL催乳素对体外培养的内蒙古绒山羊的初级毛囊和次级毛囊均有极显著的促生长作用。     

Purification of fusaproliferin from cultures of Fusarium subglutinans by preparative high-performance liquid chromatography

Thirty-six Fusarium strains were grown on cracked yellow corn and evaluated for optimum fusaproliferin production, with Fusarium subglutinans E-1583 producing the highest levels (1600 microg/g). Three solvent systems were tested for extracting fusaproliferin from the cultures of F. subglutinans E-1583. Methanol gave the highest fusaproliferin recovery, followed by methanol/1% aqueous NaCl (55:45, v/v) and acetonitrile/methanol/H(2)O (16:3:1, v/v/v). Hexane partitioning was effective in removing many impurities from the crude fusaproliferin extracts prior to the liquid chromatography step. Fusaproliferin samples were further purified by high-performance liquid chromatography (HPLC) with a C18 preparatory column using a mobile phase of acetonitrile/H(2)O (80:20, v/v). The purity of the fusaproliferin was verified by analytical HPLC, GC/MS, (1)H NMR spectroscopy, and electrospray ionization (ESI) MS. The isolated fusaproliferin was shown to be free of impurities and can be used as a standard for routine analysis. Fusaproliferin was shown to be temperature-sensitive when samples were stored at room temperature (20-24 degrees C) for more than several days. After 30 days at 4 degrees C, approximately 8% of the fusaproliferin had been transformed to deacetyl-fusaproliferin; however, samples stored at -20 degrees C for 1 year contained only trace amounts of the deacetylated form.     

巴美肉羊发情期外周血E2和P4浓度变化规律与排卵数关系研究

绵羊在发情周期中各生殖激素浓度变化与卵泡发育、成熟和排卵有着密切的关系。为了研究巴美肉羊血清生殖激素的动态变化及其与排卵数关系,试验采用电化学法,测定了12只成年母羊发情期血清中2种类固醇激素(E₂和P₄)的浓度水平,分析其动态变化规律,并用SAS 9.0的方差分析程序分析激素浓度与排卵数的关系。结果表明,两种激素在排单卵组和排双卵组绵羊间变化规律不同,E₂在排单卵组表现为先下降后升高的变化趋势,在排双卵组表现为持续下降趋势;P₄在排单卵组表现为持续上升的趋势,在排双卵组为先上升后下降的变化趋势。排单卵和排双卵组绵羊在各时间点的E₂和P₄激素浓度差异均不显著(P＞0.05)。     

The Expanding Role of Recombinant Gonadotropins in Assisted Reproduction

Using recombinant gonadotropins for assisted reproduction of domestic species is still in its infancy. Yet, the purity, potency and pathogen-free nature of recombinant gonadotropins make them attractive alternatives to tissue-derived gonadotropic agents. In this study, the authors summarize the work to date using recombinant gonadotropins to enhance the - fertility of domestic animals and they discussed their recent studies examining the biopotency of single chain analogues of human gonadotropins. In these studies, single chain analogues of follicle stimulating hormone (Fcα), chorionic gonadotropin (CGβα) or a gonadotropin construct with dual activity (FcCGβα) were administered to sheep pre-treated with antisera directed against GnRH. Ovulation was induced 3 days after analogue administration using hCG (1000 IU, iv). Although Fcα or CGβα alone induced only modest oestradiol production during the pre-hCG period, serum concentrations of oestradiol were markedly increased (p < 0.05) 3 days after administration of FcCGβα or the Fcα + CGβα combination. Final ovarian weight was significantly increased (p < 0.05) in animals receiving Fcα, Fcα + CGβα or FcCGβα. Collectively, these observations demonstrate that the single chain analogues of the human gonadotropins are active in sheep.     

Survival rates for a declining population of bottlenose dolphins in Doubtful Sound,New Zealand: an information theoretic approach to assessing the role of human impacts

• 1. The bottlenose dolphins of Doubtful Sound, New Zealand are a declining population at the southern limit of the species' range, exposed to impacts from tourism and habitat modification. Patterns in apparent annual survival were analysed from photographic resightings of naturally marked adults (1990 to 2008) and calves within the first year of life (1994 to 2008) using capture‐recapture models.
• 2. The most parsimonious model for adults provided a time‐invariant, sex‐invariant estimate of survival (?_{a(1990–2008)}=0.9374; 95% CI: 0.9170–0.9530), marginally lower than prior estimates for wild bottlenose dolphins.
• 3. The most parsimonious model for calves indicated a significant time‐variant decline in survival from an estimate similar to other populations (?_{c(1994–2001)}=0.8621; 95% CI: 0.6851–0.9473) to a current estimate that is, to our knowledge, the lowest recorded for free‐ranging bottlenose dolphins (?_{c(2002–2008)}=0.3750; 95% CI: 0.2080–0.5782).
• 4. Information theoretic evidence ratios suggested that observed patterns in calf survival were 22 times more likely to be explained by a decline coincident with the opening of a second tailrace tunnel for a hydroelectric power station than by a decline in any other year or across multiple years.
• 5. Projections using an age‐structured stochastic population model indicated that the current level of calf survival was unsustainable (population decline: 100% of model runs; population extinction: 41.5% of model runs) and was a key factor in the observed population decline in Doubtful Sound.

Copyright © 2008 John Wiley & Sons, Ltd.     

Enabling natural resource managers to self-assess their adaptive capacity

We describe the development of a low cost, repeatable self-assessment process enabling community-based natural resource management (CBNRM) groups to set priorities for building their capacity to adopt sustainable farming practices and adapt to global change. Regional measures of adaptive capacity derived from rural livelihoods analysis were populated with secondary data and used to communicate the multiple dimensions of adaptive capacity to groups of landowners. This conceptual framework was then used to derive locally relevant measures of adaptive capacity via focus groups drawn from pre-existing networks of land managers. The key issue discussed at the workshop was what constrained or enabled private land managers to effectively manage natural resources. This self-assessment process was designed to support dialogue between CBNRM groups, industry and governments to prioritise collective action for building adaptive capacity. The approach was piloted with CBNRM groups across New South Wales, Australia.