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.     

Determination of bacterial N portions in feces and differently collected ileum chymus from swine]

In faeces and ileal digesta samples of 31 intact (INT) as well as 73 surgically differently prepared pigs bacterial fractionations and 2.6-diaminopimelic acid (DAP) estimations were carried out in order to calculate the bacterial N proportion in faeces N and digesta N after feeding various diets. Because of a high individual variability and the analytical variation width of the DAP/N-ratios no distinct influences of the fed diets could be found. The average DAP/N-values in the faeces (0.224) of INT pigs ranged in the same magnitude as in the digesta (0.0272) of ileorectostomized (IRA) pigs with open colon descendens (IRAo), where a digesta backflow is possible. Distinct lower DAP/N-ratios (0.0125 resp. 0.0043), however, were found in the digesta of pigs with ileo-caecal cannulae (IZB) or IRA pigs with closed colon (IRAg). On the base of various premises (N of the "bacterial fraction" C is only bacterial N; the DAP found in fraction A originates from intestinal bacteria adhering to feed particles) conducted calculations of the bacterial N proportions (in per cent of total N) led to the following data: Faeces of INT pigs: 43.0 ... 68.2 vs. 69.6 ... 89.0; digesta of operated pigs (except protein free diet) IRAo: 22.3 ... 57.0 vs. 46.2 ... 73.8; IZB: 17.0 ... 35.7 vs. 25.2 ... 53.6; IRAg (only 3 pigs): 23.6 vs. 24.2. The proportion of bacterial N in the digesta N of protein free fed IRAo pigs was 22.0 vs. 22.6%.