Evaluation of Sperm-Activating Solutions in Atlantic Salmon Salmo salar Fertilization Tests

The use of saline solutions was tested to determine their efficacy as replacements for ovarian fluid as sperm activators and to eliminate variability encountered with the use of Ovarian fluid. We tested fertilization rate of semen from eight males on Atlantic salmon Salmo salar eggs after five sperm-activating solutions and a non-activating saline were substituted for ovarian fluid. We used solutions shown acceptable for use with other salmonid species. The six solutions tested were a non sperm-activating phosphate-buffered saline (PBS, 7.2 g/L NaCl, 1.48 g/L Na₂HPO₄, 0.43 g/L K H₂PO₄), a Tris buffer (6.99 g/L NaCl, 3.63 g/L Tris and 2.42 g/L glycine), a Borax buffer (12.2 g boric acid/L in solution 1, 76 g disodium tetraborate/L combined 100:118, then 1 L combined with 3.7 L water and 18 g NaCl), and three solutions of 9.25 g/L (125 mM) NaCl buffered to pH 6.0, 7.5, and 8.9. The latter five solutions activated sperm immediately on contact, while PBS required additional water to activate sperm. The PBS solution was the least effective (mean percent eyed eggs 37.6%) for egg fertilization. The mean percent eyed eggs for the other five saline solutions (range 78% to 86%) were not significantly different. Sperm from one male provided significantly lower egg fertilization (39.6%) when compared with the other seven males (67.2–87.4% egg fertilization). Percent egg fertilization was not related to number of live sperm cells per egg. Our results show that osmotically-balanced sperm-activation solutions, even those with a pH range from 6.0 to 8.9 provide adequate media for fertilization of Atlantic salmon eggs. Fertilization in a deactivation saline and water reactivation of sperm resulted in low egg fertilization.     

Dietary protein for young grazing sheep: Interactions with gastrointestinal parasitism

Effects of protein supplementation and of nematode control on production responses in young grazing sheep and on nematode population dynamics were assessed. Young Merino wether sheep (n = 270) were allocated to one of three supplementation (Su) treatments and one of three regimes of nematode control (drench, Dr) in a factorial design. Each of the nine treatments was replicated three times in a randomised complete block. Animals received no supplement (SuO) or were given supplements at a level equivalent to 200 g day⁻¹, fed three times a week. The supplements contained 25% lucerne meal with either 75% sunflower meal (Su1) or 75% formaldehyde-treated sunflower meal (Su2). Nematode control regimes were: not drenched unless survival of individual animals was threatened (DrO); drenched according to a strategic drenching programme (Dr1); treatment with a controlled-release albendazole capsule (Dr2). Infections with Haemonchus contortus were suppressed by use of closantel. Due to drought conditions and lack of pasture growth, lucerne hay was fed to all sheep from Week 14 onwards (350 g per head per day, fed twice a week). Supplementation reduced the need for ‘survival drenching’ in Dr0 sheep considerably. Live-weight gain was increased significantly by supplementation with Su1 or Su2 during the 36-week experiment. Undrenched animals given Su1 or Su2 tended to grow faster than unsupplemented animals in the Dr1 and Dr2 groups. Greasy wool production and fibre diameter were increased by supplementation and anthelmintic treatment. Faecal worm egg counts (FEC) in undrenched sheep were significantly lower for Su1 and Su2 sheep than for Su0 sheep. There were no effects of supplementation on FEC in Dr1 or Dr2 sheep. Worm burden was generally unaffected by supplementation, while the drenching treatments Dr1 and Dr2 reduced worm burden at Week 16 and to a lesser extent at Week 27. Trichostrongylus colubriformis was the predominant nematode species, with smaller numbers of Nematodirus spp., Trichostrongylus vitrinus, Trichostrongylus axei and Ostertagia circumcincta also present. It was concluded that supplementary feeding with protein meal substantially reduced production losses attributable to nematode infections in young grazing sheep. This appears to be due to an enhanced resilience of the host, rather than any major changes in development of protective immunity.     

Heterologous Radioimmunoassay for Llama and Alpaca Luteinizing Hormone with a Monoclonal Antibody,an Equine Standard and a Human Tracer

A radioimmunoassay for llama and alpaca LH was developed using a human I¹²⁵LH tracer from a commercial kit, equine LH diluted in human LH free serum as standard, and a monoclonal antibody (518B7) specific for LH but with low species specificity. A 60-min delay in the addition of the tracer and overnight incubation gave a sensitivity of 0.8 μg L⁻¹. The intra-assay coefficient of variation was 37% at 1 μg L⁻¹, declined to 15% at 4 pg L⁻¹ and was below 6% for concentrations up to 32 μg L⁻¹. The inter-assay coefficients of variation for 3 control samples were 20% (2.8 μg L⁻¹), 16% (7.1 μg L⁻¹) and 9.8% (19 μg L⁻¹). In an attempt to increase sensitivity, all tubes were preincubated for 4 h at room temperature before adding the tracer, and the sample volume was increased from 50 μL to 100 μL· (in the standard curve the increased volume was compensated for by human LH free serum). With this protocol, the assay sensitivity was 0.5 μg L⁻¹. The assay was validated clinically and demonstrated increased concentrations of LH after mating in llamas and alpacas. Furthermore, the assay was used to monitor LH responses to a single dose of GnRH in llamas (adult males and females at different ages).     

Salinity Tolerance of Rice (Oryza sativa L.) with Reference to Endogenous and Exogenous Abscisic Acid

Experiments conducted m a phytotron on three rice varieties of different salinity tolerance revealed an increase in the content of endogenous abscisic acid (ABA) with increasing NaCl salinity in IR20 (semi salt-tolerant), but in Pokkali (salt-tolerant) and IR28 (salt-sensitive) the increase in ABA content was marginal. Under sahnity stress, in general, 5 weekly sprayings of ABA (10^-4 mol L^-1) decreased Na and K concentrations in the shoot to the extent of 29.5 % and 3.3 %, respectively. However, ABA application significantly improved the K/Na ratio as well as the chlorophyll fluorescence decrease ratio (Rfd, indicator for potential photosynthetic activity), the number of green leaves per plant and the shoot dry weight. The response of IR20 and IR28 to ABA application was significantly better than that of Pokkali. Increasing salinity caused marked nutrient imbalances, decreased Rfd values and shoot dry weight. The results are discussed in relation to possible mechanism of salinity tolerance.