Computer simulations to determine the efficacy of different genome resource banking strategies for maintaining genetic diversity.

Genome resource banks (GRBs) and assisted reproductive techniques are increasingly recognized as useful tools for the management and conservation of biodiversity, including endangered species. Cryotechnology permits long-term storage of valuable genetic material. Although, the actual application to endangered species management requires technical knowledge about sperm freezing and thawing, a systematic understanding of the quantitative impacts of various germ plasm storage and use scenarios is also mandatory. In this study, various GRB strategies were analyzed using the historical data from three managed populations of endangered species with varied pedigrees (Eld's deer, Przewalski's horse, and Sumatran tiger). The following types of sperm banks were assessed: (1) a "Wild Bank" consisting of sperm (i.e., genes) from 5 to 10 males unrelated to the managed population and to each other; and (2) a "Best Male" bank containing sperm from only the most genetically valuable males alive in the ex situ population at the time the bank was established. These different bank types were then used to evaluate the effectiveness of different bank usage frequencies. The efficiency of each scenario was assessed by examining the level of inbreeding and gene diversity in the population. Overall, a sperm usage frequency of five times per year was determined to be the most efficient and "wild banks" were highly successful at enhancing genetic diversity. The value of a GRB established from the ex situ population depends on how closely related the banked males are to future generations. A GRB will have significantly less benefit when banked males also produce many successful offspring, or when donors are already genetically over-represented in the population at the time of establishing the GRB.

Sperm capacitation in vitro in the eld's deer.

Sperm capacitation was examined in the endangered Eld's deer (Cervus eldi thamin). Sperm motility and viability (percentage of sperm cells with intact membranes) were assessed in vitro over time after attempting to induce capacitation in TALP alone and TALP supplemented with calcium (10 mM CaCl2), dibutyryl cAMP (1 mM dbcAMP), or fetal calf serum (20% FCS). Sperm aliquots were evaluated at 0, 3, 6, 9, and 12 h for motility, viability, and ability to acrosome react after exposure to calcium ionophore (A23187, CI; 10 microM) or lysophosphatidylcholine (LC; 100 microg/mL). Fresh sperm aliquots in TALP + 10 mM CaCl2 exposed to CI had fewer (P < 0.05) intact acrosomes than the TALP control (TALP alone) or dbcAMP and FCS treatments after 9 h. Mean (+/- SEM) percentage of intact acrosomes of spermatozoa incubated in medium with increased CaCl2 declined (P < 0.05) from 80.2 +/- 2.6% (0 h) to 49.7 +/- 7.3% after prolonged incubation (9 h). The proportion of capacitated fresh spermatozoa was not influenced by LC treatment. Capacitation was not induced (P > 0.05) by any of the presumptive sperm capacitators after freeze-thawing. Likewise, neither CI nor LC induced the acrosome reaction (AR) in these spermatozoa, suggesting that the freeze-thawing process may have caused membrane damage. Results revealed that the supplementation of medium with CaCl2 evokes capacitation in some spermatozoa. However, Eld's deer spermatozoa appear remarkably resistant to conventional stimulators of capacitation and the AR.

Characterization of northern pintail (Anas acuta) ejaculate and the effect of sperm preservation on fertility.

Northern pintail duck semen and sperm traits were characterized, and the fertility of cold-stored spermatozoa was investigated using artificial insemination. Excellent quality ejaculates containing high proportions of motile spermatozoa were collected from drakes within 20 s by a massage technique. Semen was collected in Beltsville poultry semen extender, pooled and cold-stored (4 degrees C) for 0, 24, 48 or 72 h. Hens were inseminated with 100 microl twice a week, and eggs were assessed for fertilization and hatch success. Fertilization success was similar (P > 0.05) for semen cold-stored for 0 (51.6%), 24 (51.5%), 48 (41.1%) and 72 h (22.3%; P > 0.05). Similar (P > 0.05) percentages of fertilized eggs hatched to live offspring (73.1, 71.4, 87.0 and 80.0%, respectively). Fresh semen was also equilibrated with 1 or 4% dimethylsulphoxide or glycerol, and cryopreserved at the following rates: (1) approximately 60 degrees C min(-1) (in liquid nitrogen [LN(2)] vapour) for 10 min; (2) 1 degrees C min(-1) to -20 degrees C, LN(2) vapour for 10 min; and (3) 1 degrees C min(-1) to -35 degrees C, all followed by immersion in LN(2). After thawing for 30 s at 37 degrees C or 20 min at 4 degrees C, sperm motility and viability were assessed. The highest numbers of motile spermatozoa were recovered after slow-fast freezing (2) and thawing at 0 degrees C (P < 0.05), but survival was inadequate to allow artificial insemination. Nonetheless, cold storage provides an effective means of short-term storage with no loss of fertility in this waterfowl species.