Preservation of Black Tiger Shrimp Penaeus monodon Spermatophores by Chilled Storage

Abstract

Chilled storage of spermatozoa in fish has been extensively investigated for many years, but limited research was focused on crustacean species. Chilled storage of spermatophores of black tiger shrimp Penaeus monodon is needed to generate consistent and reliable supply of spermatozoa for subsequent use. The objective of this study was to develop a protocol for the chilled storage of black tiger shrimp spermatophores and to evaluate bacterial propagation during chilled storage of spermatophores. In the first experiment, spermatophores were selected and preserved using four different extenders, namely mineral oil, Ringer's solution, phosphate buffer and 0.85% sodium chloride, and stored at low temperature (2-4 C) for 42 d without antibiotic supplementation. Results showed that mineral oil was the best extender for chilled storage of spermatophores, since the highest percentage of viable sperm (58.3 f 2.9%) was observed with this extender at the end of experiment (day 42). Bacillus sp., Staphylococcus sp., and Pseudomom aeruginosa were identified as the predominant bacteria occurring during chilled storage, and the total bacteria count gradually increased during the experiment. In the second experiment, spermatophores were preserved in the mineral oil with four concentrations of the antibiotic, penicillin-streptomycin (O.l%, 1%,2%, and 3%). There was no significant difference (P> 0.05) in the percentage of viable sperm among treatments with 0.1%, 1 %, 2%, and 3% antibiotics. The total count of Bacillus sp., Staphylococcus sp., and f? aeruginosa in the antibiotic treated groups significantly decreased (P < 0.05) to undetectable levels by day 14 of the experiment. Fertility studies from artificial insemination indicated that l? monodon spermatophores preserved with mineral oil for 7-8 d at 2-4 C were capable of fertilizing eggs with hatching rates similar to the controls. This study suggests that chilled storage of spermatophores is a feasible approach for the management and spawning of black tiger prawn broodstock or other invertebrate species that produce spermatophores.

Growth performance, survival and maturation ofLitopenaeus vannamei(Boone) in an inland CRS with no water reposition

“…In both the trials, spawned eggs were lysed after 6 h, and only with unused fresh seawater was it possible to obtain shrimp nauplii (with an apparent hatching rate of 91.7–92.9%). A similar hatching rate was obtained by Babu et al (2001) and Nimrat et al (2005). Van Wyk and Scarpe (1999) found that the ratio of the divalent ions (Ca +2 and Mg +2 ) to the monovalent ions (Na+ and K+) is critical for shrimp reared at a low salinity.…”

“…The next day at 08:00 hours, females were returned to their original tanks (Peixoto et al 2005; Gandy et al 2007). The spawning rate and apparent hatching rate were evaluated using the procedure described by Huang, Jiang, Lin, Zhou and Le (2008) and Nimrat, Sangnawaiuj and Vuthiphandchai (2005) respectively. Random samples from the spawning tank were used to determin the quality and total egg numbers (Babu, Kitto, Ravi & Marian 2001; Peixoto et al 2005).…”

Assessment of viability of sperm cells of Litopenaeus vannamei on cryopreservation

“…In fact, simultaneous handling of many samples makes it difficult the execution of each step in optimal time conditions, generating a range of results in cryopreservation studies. Finally, emphasis should be given to the fact the limited success obtained in many species of penaeidae reproduction in captivity could be related with reproductive problems in the males, since most studies with shrimps describe females, a gap to be fulfilled with the development of studies concerning spermatophore production and sperm quality (Nimrat et al 2005) as well as of biotechnologies of assessment for sperm quality. In conclusion, there was no difference observed between the two cryoprotectants tested, DMSO and EG on results for cell survival, Cell viability decreased as storage time in liquid nitrogen increased and flow cytometry and eosine-nigrosine stain techniques were highly correlated, indicating that both could be used with high probability for confidence in the analyses of sperm cells viability.…”

Evaluating the impact of moulting and chilled storage of spermatophores on the integrity of plasma membrane, acrosome and DNA of black tiger prawn ( Penaeus monodon ) spermatozoa

“…Calcium free saline is the most commonly used prawn sperm extender for sperm cryopreservation (Bart, Choosuk, & Thakur, ; Vuthiphandchai, Nimrat, Kotcharat, & Bart, ) or chilled sperm storage (Bray & Lawrence, ; Morales‐Ueno, Paniagua‐Chávez, Martínez‐Ortega, Castillo‐Juárez, & Alfaro‐Montoya, ) as the depletion of calcium typically prevents sperm AR in penaeid species. Alternative sperm extenders that have been reported include artificial crustacean haemolymph (Nimrat, Sangnawakij, & Vuthiphandchai, ; Talbot, Poolsanguan, Poolsanguan, & Al‐Hajj, ), natural seawater (Bray & Lawrence, ), phosphate buffer (Nimrat et al, ) and mineral oil (Nimrat et al, ). Within the spermatophore, there is a large volume of glutinous fluid covering the sperm mass, possibly providing extra protection for the embedded spermatozoa (Sasikala & Subramoniam, ); consequently, it may be prudent to examine whether sperm quality is affected by breaching the spermatophore wall, in which case the spermatozoa are either chilled preserved as a free sperm suspension or retained within the intact spermatophore.…”

“…Within the spermatophore, there is a large volume of glutinous fluid covering the sperm mass, possibly providing extra protection for the embedded spermatozoa (Sasikala & Subramoniam, ); consequently, it may be prudent to examine whether sperm quality is affected by breaching the spermatophore wall, in which case the spermatozoa are either chilled preserved as a free sperm suspension or retained within the intact spermatophore. With respect to prawn sperm storage time, a prolonged period of sperm storage, regardless of through cryopreservation or chilled storage, has been proven to have a negative impact on the sperm quality as assessed by fertility rate (Nimrat et al, ; Vuthiphandchai et al, ).…”