In Vitro Fertilization and Development of OPU Derived Goat and Sheep Oocytes

The recent upgrade in IVP technology seen in cattle can be adapted to embryo production in small ruminants to overcome limitations exhibited by surgical procedures on preserving the reproductive potential of donors and the efficiency of embryo production. The aim of the present study was to assess the current procedures used in cattle for the production of IVP embryos in goats and sheep based on laparoscopic-aided ovum pick-up (LOPU) supplied oocytes. Sexually matured goat and sheep donors were treated during the breeding season with FSH and subjected to laparoscopic-guided follicular puncture under general anaesthesia. The collected cumulus-oocyte complexes were matured in medium 199 and fertilized by frozen-thawed spermatozoa using Talp medium supplemented with heparin and oestrus-sheep serum. Cleaved ova were either cultured in sheep in vitro fertilization medium plus amino acids or transferred to sheep oviducts. Blastocyst rate, hatching rate and development rate up to term were used as markers of embryo function. The results obtained for goat and sheep involving 30 and 35 donors respectively (10 and 9 LOPU sessions) were 81.2% and 85.2% of oocyte collection rate; 88.3% and 98.6% oocyte incubation rate; 85.6% and 76.0% fertilization rate; 82.4% and 93.4% of cleavage rate; 50.0% and 61.5% IVP blastocyst rate; 42.1% and 45.5% blastocyst rate in oviducts; 73.0% and 66.7% embryo survival up to term, respectively. The results are comparable to those obtained in small ruminants and in bovines suggesting that requirements for embryo production and development are similar.     

Development of an enzyme-linked immunosorbent assay to detect serum antibody to chicken anemia agent

An enzyme-linked immunosorbent assay (ELISA) for the detection of antibodies to chicken anemia agent (CAA) has been developed. This test utilizes a CAA-specific mouse monoclonal antibody to selectively capture virus antigen. Chicken antibodies to CAA bind to the captured antigen and are detected with horseradish peroxidase-labeled anti-chicken immunoglobulin using a conventional indirect ELISA protocol. When 388 chicken sera from specific-pathogen-free and commercial flocks from the United Kingdom, West Germany, the United States and Australia were examined, 98.5% agreement was obtained between the results of the ELISA and the indirect immunofluorescence assay. This ELISA should have worldwide application in testing SPF and commercial chicken flocks for CAA antibodies.     

Effect of dietary energy, body condition and calf removal on pituitary gonadotropins, gonadotropin-releasing hormone (GnRH) and hypothalamic opioids in beef cows

Beef cows (n = 64) were slaughtered to evaluate effects of dietary energy and calf removal (CR) on hypothalamic and adenohypophysial endocrine characteristics. From d 190 of gestation until parturition, cows received maintenance (ME; n = 32) or low (LE; n = 32) energy diets (ME = 100%, LE = 70% NRC recommendations). After parturition, half (n = 16) of each prepartum diet group received low (LE; n = 32) or high (HE = 130% NRC; n = 32) energy diets. At 30 d postpartum, cows were slaughtered 0 or 48 hr after CR. Hypothalami [preoptic area (POA), hypothalamus (HYP), stalk-median eminence (SME)] and pituitaries were collected. Basal and K(+)-induced release of GnRH from SME, and pituitary luteinizing hormone (LH) and follicle stimulating hormone (FSH) did not differ among groups (P greater than .05). Hypophyseal LH was correlated (P less than .01) with body condition score (BCS) at parturition and slaughter (r = .36 and .47, respectively). Prepartum LE diet increased (P less than .05) met-enkephalin in POA compared to prepartum ME (.59 +/- .05 vs. .44 +/- .04 pmol/mg) regardless of postpartum diet or suckling status. Concentrations of beta-endorphin in combined HYP + POA were decreased (P less than .05) by 48 hr CR (15.1 +/- 1.1 vs. 18.1 +/- 0.7 fmol/mg).(ABSTRACT TRUNCATED AT 250 WORDS)     

An analysis of sap flow in mountain ash (Eucalyptus regnans) forests of different age

Diurnal measurements of sap velocity were made in 50-, 90-, 150- and 230-year-old mountain ash (Eucalyptus regnans F. Muell.) forests in the North Maroondah catchment (southeast Australia) over the periods January 8, 1990 to April 4, 1990 and October 29, 1990 to April 16, 1991. Over the two periods, daily mean sap velocities for the four forests, in order of increasing age, were 11.5, 11.4, 9.9 and 11.8 cm h(-1) respectively. Daily mean sap velocity did not differ significantly among the 50-, 90- and 230-year-old plots. However, in the 150-year-old trees it was significantly smaller by an average of 14%. Sap velocity varied diurnally and also between positions within individual trees and among trees both within and between stands. Despite this variability, the sampling intensity and duration were sufficient to establish that behavior was highly correlated among individuals within plots. There was a significant decline with age in the overstory sapwood conducting area of these forests. In order of increasing age, the values were 6.7, 6.1, 4.2 and 4.0 m(-2) ha(-1), respectively. When combined with daily mean sap velocity, these data allowed the calculation of overstory water use. Over the experimental period, water use of the overstory decreased with age ranging, on average, from 1.86 mm day(-1) for the 50-year-old plot to 0.81 mm day(-1) for the 230-year-old plot. Mean daily water use for the two intermediate-aged forests was 1.67 and 1.00 mm day(-1), respectively. Annual water use decreased with forest age from 679 mm for the 50-year-old stand to 296 mm for the 230-year-old stand. This difference corresponds to 3.8 x 10(3) m(3) ha(-1). The annual water use of the intermediate-aged stands was 610 and 365 mm for the 90- and 150-year-old stands, respectively.     

Establishment of an agroforestry research network in Victoria,Australia (1983–85)

From 1983 to 1985, a network of agroforestry research trials was established at six localities throughout Victoria. This paper describes experimental designs and establishment procedures used at the trial sites. The overall aim of the research program is to provide definitive information on the costs and benefits of a wide range of agroforestry combinations. Accordingly, trials were established at the following locations to test a total of 93 tree species grown in combination with specific types of agriculture:

1. Carngham (near Ballarat) — sheep grazing andPinus radiata D. Don (radiata pine);
2. Hamilton (several sites) — sheep grazing and mixed tree species;
3. Kyabram — irrigated pastures andEucalyptus grandis W. Hill ex Maiden (flooded gum);
4. Rutherglen — cereal cropping and shelterbelts;
5. Myrtleford — irrigatedMentha spp. (mint) and two clones ofPopulus spp. (poplar); and
6. Neerim South (near Warragul) — cattle grazing and mixed tree species.

Conventional and systematic experimental designs have been used in setting up these trials, which in total constitute the most comprehensive agroforestry research in Australia. This paper describes these trials in detail, and stresses the importance of their continued expert management, so that the full benefits of the work can be realised for rural Victoria.     

Nutrient and heavy metals in decaying harvest residue needles on drained blanket peat forests

Harvest residue decomposition can significantly contribute to nutrient and heavy metal exports to receiving water courses. This study monitors the nutrient and heavy metal dynamics in decaying Sitka spruce and lodgepole pine harvest residue needles on Atlantic blanket peat forests in the west of Ireland. Using the litterbag method, harvest residue was placed both within and between furrows in two uncut forest and two clear-cut sites. On the clear-cut sites, the litterbags were positioned outside the harvest residue piles (i.e. brash windrows). Over the 2-year monitoring period, the needles decomposed slower at the clear-cut sites than the uncut forest sites, with mass losses of 46–55 and 58–77 %, respectively. Approximately 20 % less phosphorous (P) was released from the decaying needles at the clear-cut sites, while nitrogen (N) was released only at the uncut sites. Tree species was a significant factor contributing to nutrient and heavy metal release and accumulation patterns, with higher concentrations of aluminium (Al), nickel (Ni), cadmium (Cd) and zinc (Zn) in the decaying spruce needles than in pine. Conversely, the spruce needles showed accelerated depletion of calcium (Ca) and magnesium (Mg) relative to the pine. The harvest residue needle positioning (inside furrow/between furrows) and the site soil characteristics contributed significantly to Al transformations in spruce needles and iron (Fe) in both spruce and pine needles, with more accumulation occurring inside the furrows where Al and Fe contents of the peat were high. Manganese (Mn) was released from the needles in three of the four sites with a total release of over 90 % within 2 years. In the remaining site, where the Mn content of the peat was high, an accumulation of Mn in the needles was observed. The decomposition of needles on blanket peat catchments may be a significant source of P to receiving water courses, owing to their fast release of P, but not a likely source for N export.     

Environmental influences on histological changes in the reproductive cycle of four New South Wales sea urchins

The suitability of a sea urchin population for commercial harvesting and culturing depends on the timing and duration of the spawning season. Pilot studies have shown that the New South Wales echinoids produce mature gonads for several months.Investigations were made into the influence of those exogenous factors which could be modified under aquacultural conditions.Temperature and food appeared to affect gonad (roe) development, while depth and density did not. The relationship between macro and micro changes in the gonads was established and methods of estimating stages of gametogensis were compared.     

Aspartic Proteinase Members Secreted by the Ruminant Placenta: Specificity of Three Radioimmunoassay Systems for the Measurement of Pregnancy-associated Glycoproteins

Pregnancy‐associated glycoproteins (PAGs) isolated from the placenta of various ruminant species are enzymatically inactive members of the aspartic proteinase family. The measurement of these proteins in the maternal blood can be a good indicator of the presence of a live embryo. As certain aspartic proteinases are present in biological fluids in physiological and pathological conditions at various concentrations, it was necessary to determine the specificity of three radioimmunoassay (RIA) systems currently used for the detection of PAG molecules. Commercially available members of the aspartic proteinase family like pepsinogen, pepsin, chymosin, rennet, cathepsin D and renin were tested in a wide concentration range (10 ng/ml – 1 mg/ml). Pepsinogen cross‐reacted in RIA 1, RIA 2 and RIA 3 over 1 mg/ml, 50 μg/ml and 500 μg/ml concentrations, respectively. In the presence of pepsin, cross‐reaction was observed in RIA 1, RIA 2 and RIA 3 over 1 mg/ml, 500 μg/ml and 1 mg/ml concentrations, respectively. Chymosin and rennet could cross‐react in RIA 2 and RIA 3, while renin and cathepsin D did not decrease the binding of the tracer to antisera more, than that of the minimal detection limit. As the plasma/serum concentrations of the examined aspartic proteinases reported in the literature were outside the concentration range where cross‐reaction was observed, it can be concluded that these RIA systems were specific for the detection of PAGs in biological fluids.