Shrimp Penaeus monodon spermatophore collection and cryopreservation | |
| Author | Sudharma Choosuk |
| Call Number | AIT Thesis no.AQ-03-16 |
| Subject(s) | Shrimps -- Spermatozoa Shrimps -- Germplasm resources -- Cryopreservation Penaeus monodon -- Germplasm resources --Cryopreservation |
| Note | A thesis submitted in partial fulfillment of the requirements for the degree of Master of Science, School of Environment, Resources and Development |
| Publisher | Asian Institute of Technology |
| Series Statement | Thesis ; no. AQ-03-16 |
| Abstract | Shrimp Penaeus monodon spermatophore method from wild caught male brood was developed. The mean weight (grams) of spematophore, sperm count (x106 sperm cells) and viability(%) in the first of stripping was 0.2290 ± 0.0241,151.04 ± 18.09 and 99.67 ± 0.13, respectively. In the second time of stripping from the same male was 0.2400 ± 0.0163, 161.18 ± 12.94 and 99.68 ±0.20, respectively. There were no significant differences among weight, number of sperm and viability (P<0.05) between first and second time of stripping. Fertilization rate of artificial insemination with stripping spermatophores and natural matting were 90.50 ± 0.60 and 90.60 ± 0.60, respectively and were not significant different (P<0.05). Hatching rate of artificial insemination with stripping spermatophores and natural matting were 88.80 ± 0.57 and 88.30 ± 0.54, respectively and were not different. Penaeus monodon spermatophores could be collected by stripping method. There is no need to kill male brood and reuse in 15 days interval. This study also provides the preliminary data on cryopreservation of spermatophore for this specie as well as fertilization success. Spermatophores were also frozen to - 196 °C using a variety of concentrations and cryoprotectants. Sperm viability was assessed using the staining method with 0.5% eosin and 10% nigrosin. Spermatophores were put in the cryovial with 0.5 ml solution of known concentration, 5% DMSO, 10% DMSO, 5% MeOH, 10 % MeOH, 5% EG and 10% EG. After 30 minutes equilibration time these spermatophores were frozen using controlled rate freezer. The freezing rate that used in this study was 15 °C.min-1 from 25 °C to - 10 °C and - 10 °C to -80 °C at 2 °C.min-1 then plunged into liquid nitrogen (-196 °C). Frozen spermatophores were thawed in 30 °C water for 2 minutes. The highest sperm survival (79.6%) was obtain with spermatophores frozen in 5% DMSO. Seeding and holding time did not affect viability of sperm used 5% DMSO as cryoprotectants. Frozen - thawed spermatophores used for artificial insemination resulted in high fertilization rate, (82.0 ± 1.0 %) was lower significantly than fresh spermatophores (90.50%). Hatching rate of frozen-thawed spermatophores (87.0 ± 1.0 % ) was not significant different between fresh and frozen-thawed spermatophore. |
| Year | 2003 |
| Corresponding Series Added Entry | Asian Institute of Technology. Thesis ; no. AQ-03-16 |
| Type | Thesis |
| School | School of Environment, Resources, and Development (SERD) |
| Department | Department of Food, Agriculture and Natural Resources (Former title: Department of Food Agriculture, and BioResources (DFAB)) |
| Academic Program/FoS | Aquaculture and Aquatic Resources Management (AQ) |
| Chairperson(s) | Bart, Amrit; |
| Examination Committee(s) | Amararatne Amakupitiyage ;Mair, Graham C. ;Verapong Vuthiphandchai ;Yi, Yang; |
| Scholarship Donor(s) | Asian Institute of Technology; |
| Degree | Thesis (M.Sc.) - Asian Institute of Technology, 2003 |