Fertilization by refrigerator stored sperm of the Indian major carp,Labeo calbasu(Hamilton, 1822)

Abstract

This study reports on the spermatological properties, and on the development of a protocol for refrigerator storage (4°C) of Labeo calbasu (Hamilton, 1822) sperm for artificial breeding. Volume, motility, concentration and pH of the freshly collected sperm were 2.21 ± 0.53 (μL g−1 of fish weight) (mean ± SD), 95 ± 1 (%), 1.93 ± 0.44 × 109 (cells mL−1) and 7.56 ± 0.17 respectively. Sperm activation was evaluated at different osmolalities of NaCl solution, and motility ceased completely when osmolality of the extender was ≥287 mOsmol kg−1. Sperm retained motility for 24, 72 and 108 h, after refrigerator storage when sperm were undiluted, suspended in Alsever's solution and suspended in Alsever's solution containing 5% methanol respectively. Fertilization rate of fresh eggs with sperm stored at 4°C in Alsever's solution and Alsever's solution containing 5% methanol was 77% and 60% with a hatching rate of 60% and 43% respectively. The fertilization and hatching success of the stored sperm suggests potential to use refrigeration for transporting genetic material to hatcheries for artificial breeding of L. calbasu in Bangladesh.

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“…Sperm concentration might be related to gonadal development and maturation, which are also regulated by changing in climate, day food length and food supply (Piros et al 2002). The present study showed that the sperm density (3.14±0.165× 10 10 /ml) of B. gonionotus was similar to Hypophthalmicthys molitrix (Rahman et al, 2011), however, higher compared to L. calbasu (Hassan et al, 2013) and salmonid fish (Hatef et al, 2007). Sperm density of Salmo trutta abanticus (Hatipoglu and Akcay, 1954) and Salmo trutta macrostigma (Yavas et al, 2015) were 17.9×10 9 /ml and 14.53×10 9 / ml, respectively.…”

“…Yavas et al (2015) reported that motility duration of Salmo trutta macrostigma was 68.1s. Hassan et al (2013) observed that L. calbasu sperm remained motile for 138s after activated with 0.3% NaCl solution. The individual variation of sperm motility and duration depends on the age of male fish (Tekin et al, 2003) and time of season (Akcay et al, 2004).…”

“…81.8±1.7% in Salmo trutta (Hatipoglu and Akcay, 1954), 96.7% in Onchorhynchus mykiss (Sahin et al, 2013), 84.7% in Salmo truttamacrostigma (Yavas et al, 2015) and 77.6% motility in Mirror carp (Bozkurt and Secer, 2005). Hassan et al (2013), found that, motility of fresh sperm in Labeo calbasu decreased with the increasing osmolality. While motility of B. gonionotus were positively correlated with osmolality (r=0.538, P< 0.05) and sperm density (r=0.88, P< 0.01), but negatively correlated with motility duration (r= -0.53, P< 0.05).…”

“…Similarly Hassan et al (2013) used Alsever's solution and Abinawanto et al (2013) used skim milk with fish ringer solution for short term preservation of Indian major carp, Labeo calbasu and Java barb spermatozoa and reported 89% and 83% motility with these two extenders respectively during the storage period of 60 hours and 24 hours. Sahin et al (2013) found 64.4% motility from glucose based extender after 72h storage and 86.1% from glucose based extender after 24 h storage.…”

“…Based on the earlier studies conducted by different researchers (Sarder et al, 2009;Hassan et al, 2013;Abinawanto et al, 2013;Sahin et al, 2013) on sperm cryopreservation, five extenders such as Alsever's solution, glucose based extender, skim milk with fish ringer, egg-yolk citrate and urea-egg yolk (Table 1) and three dilution ratio (1:4, 1:6, and 1:9) were determined. The extenders Alsever solution (E1), glucose based extender (E2), skim milk with fish ringer (E3), egg-yolk citrate (E4) and urea egg-yolk (E5) were mixed with semen at respective dilution ratios and stored in an ice box about ~2°C, after a rapid shaking.…”