AQUACULTURE RESEARCH

Use of Cryopreserved Sperm of Grass Carp () for Seed Production at the Hatchery Level of Bangladesh-A Need for Development of Germplasm Repositories
Hossian S, Reza MF, Rahman MM, Mariom , Alam MJ, Razzak MA, Mollah MFA, Tiersch TR and Sarder MRI
Grass carp (), a freshwater nonnative fish species, is a potential aquaculture candidate in Bangladesh. The seed of the species is produced in the hatcheries by hypophysation, but the quality of seedstock of grass carp is deteriorated due to inbreeding, negative selection, and interspecific introgression among fishes. To increase the availability of quality seed and best genetic traits of grass carp, this study dealt with finding suitable conditions of sperm cryopreservation protocols and evaluated the effectiveness of cryopreserved sperm through breeding trials of . A broodstock population was developed from fingerlings imported from China by the Bangladesh Department of Fisheries. Sperm was collected from hormone-induced mature males, with an estimated concentration of 2.4 ± 0.3 × 10 cells/mL and a pH of 8.3 ± 0.2. Sperm motility was evaluated in different concentrations of NaCl solution. The highest motility (96% ± 1%) and longest motility duration (22 ± 1 min) were achieved at 0.4% of NaCl (128 mOsmol/kg). The toxicity of DMSO and methanol at concentrations of 5%, 10%, and 15% was evaluated for 5-40 min using Alsever's solution and egg yolk citrate. The highest motility was achieved during 5 and 10 min of incubation with cryoprotectants at 5% and 10%. Alsever's solution with 10% DMSO at 1:9 dilution with sperm produced the highest equilibration motility (93% ± 2%) and when cooled at 10°C/min yielded the highest postthaw motility (85% ± 3%). Fertilization of 24% ± 3% to 51% ± 2% and hatching of 18% ± 2% to 41% ± 2% were achieved by using cryopreserved sperm in six selected private hatcheries. The fertilization rate for fresh sperm sourced from hatchery-reared males was 64% ± 5% to 85% ± 3%, and the hatching rate ranged from 53% ± 6% to 74% ± 5%. Thus, the cryopreservation protocol of sperm was found to be feasible for fry production at commercial hatcheries, but further research is needed to improve the fertilization and hatching rates.
Growth, morphometrics, and nutrient content of farmed eastern oysters, (Gmelin), in New Hampshire, USA
Grizzle RE, Ward KM, Peter CR, Cantwell M, Katz D and Sullivan J
When harvested, oysters represent a removal from the ecosystem of nutrients such as nitrogen (N) and carbon (C). A number of factors potentially affect nutrient content, but a quantitative understanding across the geographic range of the eastern oysters is lacking. The present study was designed to quantify the relationships among various metrics of farmed eastern oysters near its northern geographic range focusing on nutrient content. Hatchery-reared oysters were deployed in polyethylene bags at six sites, and were measured on multiple occasions from 2010-2012. A quadratic polynomial fit to the combined datasets for shell height indicated that on average a 'cocktail' size oyster (63 mm shell height) would be reached after 2 yr, and 'regular' size (76 mm) would require 3 yr. There were significant differences in growth rates and oyster nutrient content among the sites; means for %N in soft tissue ranged from 6.9 to 8.6, and 0.07 to 0.18 in shell. Percent N in soft tissue and shell were highest at two sites at the mouths of rivers with elevated dissolved inorganic N concentrations in the water. Grand means (all sites, seasons and years combined) of soft tissue N and C for regular size oysters were 7.3% and 38.5%, respectively; and for shell N and C were 0.13% and 12.0%, respectively. Our study extends the range of data on nutrient content of the eastern oyster to northern New England, and indicates that oyster size, seasonality, and nutrient concentration in ambient water potentially affect %N and %C content of oysters.
Vitrification of Sperm from Marine Fishes: Effect on Motility and Membrane Integrity
Cuevas-Uribe R, Chesney EJ, Daly J and Tiersch TR
Our goal was to develop a standardized approach for sperm vitrification of marine fishes that can be applied generally in aquatic species. The objectives were to: 1) estimate acute toxicity of cryoprotectants over a range of concentrations; 2) evaluate the properties of vitrification solutions (VS); 3) evaluate different thawing solutions, and 4) evaluate sperm quality after thawing by examination of motility and membrane integrity. Sperm were collected from red snapper (), spotted seatrout (), and red drum (). A total of 29 combinations of cryoprotectants were evaluated for toxicity and glass formation. Samples were loaded onto 10-µL polystyrene loops and plunged into liquid nitrogen. There was a significant difference ( < 0.05) in post-thaw motility among VS and among species when using the same VS. The sperm in VS of 15% DMSO + 15% ethylene glycol + 10% glycerol + 1% X-1000™ + 1% Z-1000™ had an average post-thaw motility of 58% and membrane integrity of 19% for spotted seatrout, 38% and 9% for red snapper, and 30% and 19% for red drum. Adaptations by marine fish to high osmotic pressures could explain the survival in the high cryoprotectant concentrations. Vitrification offers an alternative to conventional cryopreservation.
Simulation modeling of high-throughput cryopreservation of aquatic germplasm: a case study of blue catfish sperm processing
Hu E, Liao TW and Tiersch TR
Emerging commercial-level technology for aquatic sperm cryopreservation has not been modeled by computer simulation. Commercially available software (ARENA, Rockwell Automation, Inc. Milwaukee, WI) was applied to simulate high-throughput sperm cryopreservation of blue catfish () based on existing processing capabilities. The goal was to develop a simulation model suitable for production planning and decision making. The objectives were to: 1) predict the maximum output for 8-hr workday; 2) analyze the bottlenecks within the process, and 3) estimate operational costs when run for daily maximum output. High-throughput cryopreservation was divided into six major steps modeled with time, resources and logic structures. The modeled production processed 18 fish and produced 1164 ± 33 (mean ± SD) 0.5-ml straws containing one billion cryopreserved sperm. Two such production lines could support all hybrid catfish production in the US and 15 such lines could support the entire channel catfish industry if it were to adopt artificial spawning techniques. Evaluations were made to improve efficiency, such as increasing scale, optimizing resources, and eliminating underutilized equipment. This model can serve as a template for other aquatic species and assist decision making in industrial application of aquatic germplasm in aquaculture, stock enhancement, conservation, and biomedical model fishes.
Environmental effects of a marine fish farm of gilthead seabream () in the NW Mediterranean Sea on water column and sediment
Morata T, Falco S, Gadea I, Sospedra J and Rodilla M
This study examined the effects of organic enrichment on water column, sediments and macrofauna caused by a fish farm in the Mediterranean Sea. Samples were collected on four sampling campaigns over a one-year cycle. Significant differences were found in the water column in dissolved oxygen, dissolved inorganic nitrogen, phosphate and total phosphorus concentrations between the fish farm and the control. The increase in the dissolved inorganic nitrogen and phosphate concentrations at the fish farm modified the stoichiometric ratios between nutrients, with silicate acting as limiting nutrient at the fish farm 11% more than at the control. Nevertheless, chlorophyll concentration in the water column was higher at the control station, probably due to the fouling of the underwater fish farm structures. Significant differences were found in sediment concentrations of organic matter, total phosphorus and redox potential between the fish farm and the control. The Canonical Correlation Analysis indicated that organic matter, total phosphorus, redox potential and% of gravels accounted for 68.9% of the total variance in the species data. Changes were observed in macrofauna, with a decrease in number of species and up to a nine-fold increase in abundance with respect to the control.
Gene expression kinetics of the yellow head virus in experimentally infected
Cedano-Thomas Y, De La Rosa-Vélez J, Bonami JR and Vargas-Albores F
The yellow head virus (YHV) has been reported to be one of most pathogenic viruses for cultivated shrimp; however, serious problems have only been reported in farms in south and southeastern Asian. Recently, a YHV strain was detected in cultivated in Mexican farms that lacked virus-associated mortalities or epizooties, and the animals were apparently healthy. The identity of the virus was confirmed by sequencing replicative and structural protein-encoding regions and comparing with homologous virus sequences. Phylogenic relationships and genetic distances were also determined and, although some differences were observed, an influence on virulence was uncertain. In addition, the expression levels of several transcripts (3CL, POL, GP64 and GP116) were evaluated by quantitative real-time polymerase chain reaction during an experimental infection. Although the transcript showed varying kinetics, viral genes were expressed in infected , demonstrating the replicative capability of this YHV strain.
Yellow head-like viruses affecting the penaeid aquaculture industry: a review
Munro J and Owens L
This review focuses on relevant scientific information regarding the current knowledge of the yellow head complex viruses, yellow head virus and gill-associated virus. The yellow head complex viruses have been problematic within the aquaculture industry for over 10 years and still retain their research topicality. Presently, there are numerous research papers from different journals covering the identification, disease expression and spread, pathogenesis, detection, morphology, genomic sequence and protein profiles of the yellow head complex viruses. Indeed, there has been no extensive review to compare these studies, and as a corollary, to assess flaws in contemporary research and knowledge. Additionally, the yellow head complex viruses rank within the top four prawn viruses with respect to disease impact and economic loss. This review collectively reports on all the findings and current methods of research and aims to identify weak areas of research where conclusions have been unjustifiably drawn and furthermore to elucidate areas that have a gap of knowledge.