Growth and Survival of Olive Barb, Puntius Sarana (Hamilton 1822) Larvae Produced with Cryopreserved Versus Fresh Sperm
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Aquaculture Research, 2016, 47, 228–233 doi:10.1111/are.12484 Growth and survival of olive barb, Puntius sarana (Hamilton 1822) larvae produced with cryopreserved versus fresh sperm Shirin Akter1, Md Mahbubul Hassan2,*, Md Nahiduzzaman3 & Mostafa Ali Reza Hossain1 1Department of Fisheries Biology and Genetics, Bangladesh Agricultural University, Mymensingh, Bangladesh 2Department of Fisheries Biology and Genetics, Hajee Mohammad Danesh Science and Technology University, Dinajpur, Bangladesh 3WorldFish, Bangladesh and South Asia, Rangpur, Bangladesh Correspondence: Dr M A R Hossain, Department of Fisheries Biology and Genetics, Bangladesh Agricultural University, Mymen- singh 2202, Bangladesh. E-mails: [email protected]; [email protected] *Present address: M M Hassan, School of Biological Sciences, Flinders University, Adelaide, Australia Abstract Introduction Despite the success in fertilization and hatching Lack of good quality fish seed is considered one of fish eggs with cryopreserved sperm, report on of the major challenges for the development of growth and survival of larvae produced from fro- aquaculture in Bangladesh, and many other zen-thawed sperm is inadequate. The study eval- Asian countries (Siriwardena 2007). Cryopreser- uates the applicability of cryopreserved sperm for vation (storage of biological materials in liquid mass seed production by comparing the growth nitrogen at À196°C) technique has the potential and survival of a popular food-fish olive barb, to overcome the bottleneck by supplying sperm Puntius sarana (Hamilton 1822) larvae produced from superior male during artificial fertilization. from cryopreserved and fresh sperm. The eggs Sperm cryopreservation is a proven technique to were artificially fertilized with cryopreserved and supply sperm for artificial fertilization in a large freshly collected sperm, and the growth and sur- number of freshwater and marine fish. In addi- vival of produced larvae from both group tion, commercial application of cryopreserved recorded up to 12 weeks. The independent sam- sperm and feasibility of gene banking has been ple t-test statistic showed the difference in investigated for many fish species (Butts, Feindel, lengths, t(718) = 0.241; P = 0.810 and weights, Neil, Kovacs, Urbanyi & Trippel 2011). In Ban- t(718) = 0.412; P = 0.680 were insignificant gladesh, research on fish sperm cryopreservation between two groups. There was also no signifi- started in 2004, and the protocols have been cant difference, t(718) = À0.758, P = 0.448 in developed for several commercial and threatened survival of larvae produced from cryopreserved fish species (Hossain 2010). The studies mostly and freshly collected sperm. The study indicates focused on selection of suitable extenders and that larvae of olive barb produced from cryopre- cryoprotectants, optimal dilution ratios of milt, served sperm are equally compatible in growth and optimal cryoprotectant concentrations, post- and survival as the larvae produced from fresh thawed motility of sperm, fertilization and hatch- sperm. Therefore, cryopreserved sperm can be ing rate (Nahiduzzaman, Hassan, Roy, Hossain, applied for artificial fertilization of P. sarana to Hossain & Tiersch 2012). supply quality seed for aquaculture. To date, only 10 fish species are studied for the growth and survival of larvae produced with Keywords: cryopreserved sperm, growth, sur- cryopreserved sperm worldwide. The growth and vival, Puntius sarana survival of offspring produced from cryopreserved 228 © 2014 John Wiley & Sons Ltd Aquaculture Research, 2016, 47, 228–233 Growth and survival of Puntius sarana larvae S Akter et al. sperm has been studied for pabda catfish, Ompok larvae need to be studied to know the compatibil- pabda (Sarder, Saha & Sarker 2013), tiete tetra, ity with fresh sperm. Brycon insignis (Viveiros, Isau, Caneppele & Leal In many fish species, cryopreservation reduces 2012), silver barb, Barbonymus gonionotus post-thawed sperm motility without affecting (Rahman, Sarder & Rouf 2009), common carp, embryogenesis and larval growth (Viveiros et al. Cyprinus carpio (Sarder, Salam & Hussain 2007), 2012). Freezing and thawing damage DNA of steelhead trout, Oncorhynchus mykiss (Hayes, sperm and reduces fertility (Watson 2000; Zilli, Rubin, Hensleigh, Reisenbichler & Wetzel 2005), Schiavone, Zonno, Storelli & Vilella 2003; Cabrita, turbot, Scophthalmus (Psetta) maxima (Chereguini, Robles, Rebordinos, Sarasquete & Herraez 2005). La Banda, Garcıa, Rasines & Fernandez 2002), Prior to start of mass seed production for any fish channel catfish, Ictalurus punctatus (Tiersch, species with cryopreserved sperm, growth and sur- Goudie & Carmichael 1994), African catfish, vival of larvae need to be tested. Whether cryopre- Clarias gariepinus (Van der Walt, Van der Bank served sperm exerts any deleterious effect on & Steyn 1993), tilapia, Oreochromis hornorum growth and survival of resulting P. sarana larvae (Chao, Chao, Liu & Liao 1987) and striped is not known. Therefore, the objective of the study bass, Morone saxatilis (Kerby, Bayless & Harrell was to compare the growth and survival of P. sar- 1985). ana larvae produced with cryopreserved and fresh The fish under study, olive barb, Puntius sarana sperm. (Hamilton 1822) is the largest barb native to inland waters of South-East Asia (Talwar & Jhin- Materials and methods gran 1991). The olive barb is a popular food-fish having high market demand in Bangladesh and in Collection and rearing of larvae other South Asian countries (Jena, Das, Das & Mondal 2007; Hossain, Nahiduzzaman & Saha The experiment was carried out during July to 2010) enlisted as critically endangered (CR) in November, 2012 in backyard hatchery of the Fac- Bangladesh (IUCN-Bangladesh 2000). Along with ulty of Fisheries, Bangladesh Agricultural Univer- its conservation in natural habitat through estab- sity. Two groups of larvae were produced by lishing fish sanctuary and stock enhancement pro- fertilizing olive barb eggs with freshly ejaculated gramme, the inclusion of the fish in carp sperm and frozen thawed sperm following the polyculture system would contribute to the diversi- protocol of Nahiduzzaman et al. (2011). After fication of aquaculture system and conservation of absorption of yolk sac, the larvae were trans- the species (Hossain et al. 2010). As the fish is ferred to three rectangular glass aquaria rarely available in the wild, there has been limited (120 9 50 9 35 cm) for each group. The initial scope of collecting fish for fry production in the density (number/L water) of the larvae was twenty hatcheries. If the stored sperm can be made avail- five. Continuous aeration was supplied in the rear- able on demand for research or commercial fry ing tank, and water exchange took place once a production, this would certainly contribute to both week during the rearing period. The temperature conservation and aquaculture enhancement of the throughout the study period was 24–31°C. The species. Sperm cryopreservation protocol has larvae were fed with finely minced hard-boiled already been developed for P. sarana to assist basic chicken egg-yolk six times a day for first 3 days. research and time/location independent supply of During 3 days post-hatching period, larvae were sperm for artificial fertilization (Nahiduzzaman, fed to apparent satiation four times a day with live Hassan, Khanam, Mamun, Hossain & Tiersch food (microalgae, zooplankton and crushed tubifi- 2011). Although the frozen-thawed sperm of the cid worm). After 4 weeks, live food was partially species is proven useful to fertilize eggs, the lower replaced with the pelleted starter feed (Paragon hatching rate of the thawed sperm made them Feeds Limited, Bangladesh) until the end of the incompatible with fresh sperm. In most of the study period. studies, fertilization and hatching is presumed to The growth and survival of larvae was studied be ultimate criterion of assessment of a cryopreser- out up to 12 weeks rearing period. Ten larvae vation protocol. However, prior to use preserved were randomly sampled once in a week with three sperm for seed production and restocking por- replicates. The total length (mm) and wet weight gramme, the growth performance and survival of (g) were taken for growth study according to © 2014 John Wiley & Sons Ltd, Aquaculture Research, 47, 228–233 229 Growth and survival of Puntius sarana larvae S Akter et al. Aquaculture Research, 2016, 47, 228–233 Memisß, Ercan, Cßelikkale, Timur and Zarkua SPSS version 20.0 (IBM Corporation, Armonk, (2009). The lengths of the fish were measured NY, USA). using Vernier calipers (Mitutoyo 500-196-20, Foshan, China) to the nearest 0.05 mm, and body Results weights were measured to an accuracy of 1 mg using digital scale (UW1020H, Kyoto, Japan). The Growth of larvae dead larvae were counted and removed from the tank every day, and the survival rate were calcu- The gain in length (mm) and weight (mg) of lar- lated by counting all dead larvae over the rearing vae were considered to measure the growth, and period. independent sample t-test performed to compare between larvae of two groups. The test statistic showed that the differences for both the lengths, t Calculation of growth and survival (718) = 0.241; P = 0.810 and weights, t(718) = Daily instantaneous length specific growth rates 0.412; P = 0.680 were insignificant between the were calculated by fitting the exponential model: larvae produced with cryopreserved sperm and gt Lt = L0e , Where, Lt = total length (mm) at time t, fresh sperm. The exponential equation fitting the