This study aimed to develop a cryopreservation protocol for sperm of the mud spiny lobster, Panulirus polyphagus. Sperm of P. polyphagus were successfully cryopreserved using a protocol with cooling periods of 15 min per temperature, 25, 20, 16, 4, 2, −4, −20, −80, and −150°C, followed by immediate storage in liquid nitrogen (at −196°C). The efficacy of the cryopreserved protocol was determined by assessing the viability of sperm. The optimal thawing temperature for cryopreservation of sperm was 26°C for 30 s, with a viability rate of 76.09% ± 7.81. At room temperature, −20 and −80°C, 10% glycine provided the highest percentage of sperm viability at 91.87 ± 2.03% (5 min at room temperature), 91.31 ± 2.65% (6 h at − 20°C) and 75.88 ± 10.81% (6 h at − 80°C). In conclusion, we developed a protocol (Protocol I) for the successful cryopreservation of P. polyphagus sperm using Ca-F saline as an extender and 10% glycine as a cryoprotectant. Statement of relevance: This article is suitable with Aquaculture Journal because the findings will give a big contribution in mud spiny lobster aquaculture. Mud spiny lobster seedling especially P. polyphagus is getting less landing every year, new approach needed in ensuring enough supplies of P. polyphagus to be available. The content in this article would help developing the research innovation for the hatchery production of P. polyphagus to increase.
Many crustacean species including mud crab, genus Scylla are incapable of natural maturation under captivity, thus putting high pressure on the wild catch. Therefore, to increase the availability of mature broodstocks in captivity, this study determined the effect of Eyestalk Ablation (EA) and Methyl Farnesoate (MF) administration on ovarian maturation stages of the orange mud crab, Scylla olivacea. The study was conducted using a control group (T1) consisting of 95% alcohol (widely used as a chemical solvent), and three treatment groups consisting of: 5 ml/g oral administrated MF in the diet (T2), EA (T3) and a combination of both treatments of MF and EA (T4). The highest percentage of ovarian maturation Stage 4 was found in the T4 treatment (20.8%) compared to the other treatments which were T1 (0%), T2 (8.33%) and T3 (12.5%). Ovarian development of the treated groups (T2, T3, and T4) was significantly different compared to the control group (T1) (p < 0.05). However, there was no significant difference observed in the mean Gonadosomatic Index (GSI) in orally administrated MF (T2) compared to the EA group (T3) (p > 0.05), but it was significantly different when compared to the combination group of MF and EA (T4) (p < 0.05). It can be concluded that the combination method of oral administration of MF through diet and EA (T4) is the best technique for producing mature ovaries.
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