Successful fertility experiments with cryopreserved spermatozoa of barramundi, Lates calcarifer (Bloch), using dimethylsulfoxide and glycerol as cryoprotectants

The fertility of cryopreserved Lates calcarifer sperm was studied to increase the availability of semen for routine fertilization of stripped eggs and to provide a tool for selective breeding. Semen diluted (1:4 v/v) and frozen (-196 degrees C) with 5% dimethylsulfoxide (DMSO) or 10% glycerol (final concentration) as cryoprotectants was used to inseminate freshly stripped ova. Frozen-thawed sperm were motile for about 4 min after being mixed with seawater. In the DMSO medium, post-thaw sperm activation was immediate after dilution with seawater, but in the glycerol medium maximum motility intensity was delayed for up to 1 min. When eggs and sperm were mixed before the addition of seawater, semen frozen with DMSO as cryoprotectant gave a mean hatch rate (84.1%) no different (P > 0.05) from that of unfrozen semen diluted with Ringer's solution (80.7%) or with DMSO (83.7%), but higher (P < 0.05) than that of semen frozen with glycerol (60.9%). Adding sperm to seawater 30 s before mixing with eggs did not improve the fertility of sperm cryopreserved with glycerol. Eggs inseminated with glycerol-cryoprotected sperm showed higher mortality during incubation than those inseminated with DMSO-cryoprotected sperm. Sperm held in liquid nitrogen for 90 days with DMSO as cryoprotectant yielded acceptable fertilization and hatching rates with semen-to-ova ratios of up to 1:100 (v/v) , and produced fish with no apparent abnormalities over a 29-day period after hatch. These results show that cryopreservation of L. calcarifer sperm is feasible and well suited to a variety of hatchery purposes.

Evaluation of the Australian branched broomrape (Orobanche ramosa) eradication program

Because weed eradication programs commonly take 10 or more years to complete, there is a need to evaluate progress toward the eradication objective. We present a simple model, based on information that is readily obtainable, that assesses conformity to the delimitation and extirpation criteria for eradication. It is applied to the program currently targeting the annual parasitic weed, branched broomrape, in South Australia. The model consists of delimitation and extirpation (E) measures plotted against each other to form an 'eradograph.' Deviations from the 'ideal' eradograph plot can inform tactical responses, e.g., increases in survey and/or control effort. Infestations progress from the active phase to the monitoring phase when no plants have been detected for at least 12 mo. They revert to the active phase upon further detection of plants. We summarize this process for the invasion as a whole in a state-and-transition model. Using this model we demonstrate that the invasion is unlikely to be delimited unless the amount of newly detected infested area decreases, on average, by at least 50% per annum. As a result of control activities implemented, on average approximately 70% (range, 44 to 86%) of active infestations progressed to the monitoring phase in the year following their detection. Simulations suggest that increasing this rate of transition will not increase E to a significant extent. The rate of reversion of infestations from the monitoring phase to the active phase decreased logarithmically with time since last detection, but it is likely that lower rates of reversion would accelerate the trend toward extirpation. Program performance with respect to the delimitation criterion has been variable; performance with respect to the extirpation criterion would be improved considerably by the development and application of cost-effective methods for eliminating branched broomrape soil seed populations.