Sperm cryopreservation of silver barb (Barbodes gonionotus): cryoprotectants, cooling rate and storage time on sperm quality

Abstract

Effectiveness and efficiency of frozen sperm on fertilization and hatching success of eggs from silver barb was examined in relation to cryoprotectants, freezing rate and storage period. Sperm was diluted in calcium-free Hank's balanced salt solution, equilibrated with dimethylsulphoxide (DMSO), propylene glycol, sucrose or methanol at 5%, 10%, 15% or 20% final concentrations, and frozen in 250-lL straws using a one-step freezing procedure (1, 5 and 8°C min À1 from 25 to À40°C). Highest post-thaw sperm motility was found from a treatment using 10% DMSO and 5°C min À1 (82.2 AE 2.1%), similar to that of 10% DMSO and 8°C min À1 (87.8 AE 3.2%). Post-thaw motility of sperm frozen at 5 or 8°C min À1 was significantly higher than 1°C min À1 . Relative sperm motility declined significantly after 10 months of cryostorage while viability did not change during a 12-month cryostorage. Average fertilization rates of sperm after 1 and 4 months of storage were 64.5 AE 4.6% and 61.3 AE 3.4%, respectively, similar to those of fresh sperm (69.6-72.3%). Hatching rates of cryopreserved sperm (45.4-51.2%) were similar to those of fresh sperm (51.8-57.8%). This study developed suitable methods for cryopreservation of silver barb sperm that can be used to facilitate hatchery operation.

Evaluation of the potential source of bacterial contamination during cryopreservation process of silver barb (Barbodes gonionotus) sperm

“…Afterwards, percentages of forward‐moving sperm from each male were separately evaluated based on subjective estimation following protocol of Vuthiphandchai et al . (). Briefly, semen from each male was placed on a microscopic slide, activated with distilled water (100 μL) and observed at 40× magnification using a light microscope (Zeiss model D‐7085; Berlin, Germany).…”

“…Silver barb, Barbodes gonionotus , is recognized as one of economically important freshwater fish of Thailand. Recently, cryopreservation of silver barb sperm was carried out by Vuthiphandchai, Wilairattanadilok, Chomphuthawach, Sooksawat and Nimrat () who focused on the effects of cryoprotectants, cooling rate and storage time on post‐thawed sperm quality.…”

Morphological and morphometric evaluation of silver barb, Barbodes gonionotus (Bleeker, 1849) sperm supplemented with antibiotics

“…Sperm motility was assessed from the forward movement, while vibrating spermatozoa were judged as immotile sperm. Percentage of forward‐moving sperm was expressed based on quantitative motility score from immotile (0%) and every 20% increment in a forward direction until reaching a maximum score (100%) that referred to highly active motile sperm (Vuthiphandchai et al., ). Evaluation of sperm motility was done within 15s post‐activation and by the same experienced personnel to eliminate the degree of variation among observers.…”

“…Calcium‐free Hank's balanced salt solution (Ca‐F HBSS) was used to dilute pooled semen (Vuthiphandchai et al., ). The extender comprised (g L −1 ) NaCl (8.89), KCl (0.44), Na 2 HPO 4 ·2H 2 O (0.13), NaHCO 3 (0.39), KH 2 PO 4 (0.07), MgSO 4 ·7H 2 O (0.22) and glucose (1.11).…”

Development of cryopreservation methods for silver barb ( Barbodes gonionotus , Bleeker) spermatozoa by manipulation of cooling rates in dry shipper

“…Silver barb, Barbodes gonionotus , is widely cultured as an economically important freshwater fish species in Southeast Asian countries, especially in Thailand. Several authors have reported successful cryopreservation protocols of silver barb sperm using a programmable controlled freezer (Routray et al, ; Sarder, Rahman, Hossain, & Samad, ; Vuthiphandchai, Wilairattanadilok, Chomphuthawach, Sooksawat, & Nimrat, ).…”