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1.
High‐throughput Cryopreservation of Sperm from Sex‐reversed Southern Flounder,Paralichthys lethostigma 
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下载免费PDF全文 E Hu Rafael Cuevas‐Uribe Huiping Yang Robin Sanderson Adriane O. Gill Harry Daniels Terrence R. Tiersch 《Journal of the World Aquaculture Society》2016,47(4):555-565
The Southern flounder, Paralichthys lethostigma, is a valuable aquaculture fish with established markets in the USA. All‐female production in this species is an important technology for aquaculture because the females usually have body sizes twice those of males at the same age, and sex‐reversed males (genotypic XX neomales) are used for all‐female production by crossing with genetically normal females. However, sperm volume from the neomales is usually small (<0.5 mL) and limits their application for all‐female fish production. Cryopreservation of sperm from these sex‐reversed neomales will provide access on demand with increased efficiency to extend the application of neomales. The goal of this study was to develop a protocol for cryopreservation of sperm from the Southern flounder by using an automated high‐throughput processing system. The objectives were to: (1) determine the effect of osmolality on activation of sperm motility; (2) evaluate the effect of extender solutions on sperm motility capacity; (3) evaluate the acute toxicity of cryoprotectants (dimethyl sulfoxide [DMSO], propylene glycol, and polyethylene glycol) on sperm motility, and (4) estimate the effect of cooling rate on sperm cryopreservation and post‐thaw fertilization. Sperm motility was activated when osmolality was 400 mOsmol/kg or higher. Of the three extender buffers tested, HEPES4‐(2‐hydroxyethyl)‐1‐piperazineethanesulfonic acid (HEPES) at 300 mOsmol/kg resulted in better protection for sperm motility than did Hanks' balanced salt solution and Mounib solution at 300 mOsmol/kg during 7 d of refrigerated storage. After 30 min equilibration with the cryoprotectant of 15% DMSO, sperm motility was 24 ± 21% (fresh sperm motility without any cryoprotectants was 42%). After cooling at a rate of 20 C/min, post‐thaw sperm motility was 8 ± 5% and fertilization was 63 ± 40% evaluated at the 32–64 cell stage (5 × 105 sperm per egg). Overall, a protocol was developed for sperm cryopreservation in the Southern flounder with high‐throughput processing, which provides a tool to preserve the valuable genetic resources from neomale flounders, and enables germplasm repository development for the Southern flounder. 相似文献
2.
Steven G. Hall James Finney R. Paul Lang Terrence R. Tiersch 《Aquacultural Engineering》2002,26(4):277-289
A control system was designed to raise and maintain water temperatures within 0.03-ha earthen ponds to a range conducive for spawning (24–30 °C) channel catfish Ictalurus punctatus. Heating was done during February 2001 to April 2001, when temperatures would have otherwise prohibited spawning (<24 °C). Temperature was increased from 10 °C (ambient) by 2 °C per day, and maintained at 27 °C, by the addition of geothermally warmed water (36 °C). The control system substantially increased the controllability and precision of heating ponds compared to manual operation. Systems were designed to control sets of four ponds. In designing this control system, consideration of biological constraints was essential. Reproduction in channel catfish is most strongly influenced by temperature. Because cold fronts are common during the winter and early spring (January–March), it was essential to ensure that pond temperatures did not fall below the range for spawning. Constraints on the heating rate and temperature variability to maintain fish health and stimulate spawning behavior were considered. Components of the control system included temperature measurement devices (type-T thermocouples), a central electronic control unit, electronic switches and electrically actuated ball valves. In response to the temperature sensed by each thermocouple, the controller sent a message to close or open the valve. When the valve was opened, warm water was added to the pond to increase the average pond temperature. Hardware and algorithm design and initial system testing were the major components of this project. The final design incorporated information on relevant biological parameters and safety features including peak pond temperature, independent aeration and water pressure control mechanisms. Initial results indicate successful control of this biological system, and ongoing studies suggest similar mechanisms may be used for additional control objectives. In particular, this system could be used to vary pond temperatures to study biological responses and to cool ponds by addition of well water during summer months. 相似文献
3.
Abstract— Channel catfish Icralurus punctatus are commonly spawned for research purposes by pairing of a hormonally treated female with a male in flow-through aquaria. A technique that allows hormonal induction of ovulation in females without pairing would accelerate genetic improvement and production of hybrid catfish. Over a 3-yr period (1994, 1995, and 1996) we conducted a series of trials to demonstrate the potential for artificial spawning in recirculating systems, and in 1996 we included trials with grouped females in addition to male-female pairs. Females were induced to spawn with injection of synthetic leuteinizing hormone-releasing hormone, and those that ovulated were stripped and the eggs were artificially fertilized. During 1994 and 1995, all fish were spawned by pairing, and in 1996, half of the females were spawned by pairing and half were grouped in tanks without males. Spawning success (percent of females that produced eggs), latency (time between injection and ovulation), and percent fertilization were observed for the paired and grouped trials. Spawning success was 36% in 1994 (N= 36). 22% in 1995 (N= 54). 41% in 1996 (N= 27). and 58% for grouped females (N= 26). The latency period was 113 ± 69 h in 1994, 109 ± 57 h in 1995, 44 ± 8 h in 1996, and 50 ± 9 h for grouped females. Percent fertilization was 16 ± 26% for eggs stripped in 1994, 72 ± 25% in 1995, 43 ± 20% in 1996, and 16 ± 37% for grouped females. In 1995, water quality problems were associated with high mortality of females (24 of 44 females; 4 of 44 males). The metabolic demands of final oocyte maturation in combination with methemoglobinemia caused by high nitrite levels could account for the increased vulnerability of females. These trials indicate that with adequate biofiltration, artificial spawning is possible in recirculating systems and with females grouped rather than paired. Further research on hormone dosage and timing of egg stripping will increase the utility of grouped spawning of channel catfish. 相似文献
4.
J. Michael Christensen Terrence R. Tiersch 《Journal of the World Aquaculture Society》1996,27(3):340-346
Abstract.— Refrigerated storage of sperm is useful for genetic study and artificial breeding of fishes. Due to the potential loss of donor males, storage is important in species such as channel catfish Icralurus punctatus from which sperm cannot be stripped. This study addresses short-term storage (at 4 C) of channel catfish sperm by evaluation of storage methods employed for other species and for cryopreservation of channel catfish sperm. The objectives were to evaluate: 1) storage of intact testes and storage of sperm suspended in an extender solution; 2) use of various storage containers with and without supplemental oxygen; 3) use of extender solution with and without the addition of an antibiotic/antimycotic cocktail; 4) use of extender solution with and without the addition of methanol; and 5) use of extender solution with and without the addition of glucose and methanol. Sperm suspended in extender solution retained motility significantly longer (9 d) than did sperm in intact testis (2 d). Sperm stored in Zip-loc® plastic bags inflated with pure oxygen retained motility significantly longer (12 d) than did sperm stored in Zip-loc® plastic bags without supplemental oxygen (7 d), or sperm stored in plastic beakers (8 d) or test tubes (8 d) without supplemental oxygen. Sperm stored with the addition of antibiotic/antimycotic cocktail or methanol retained motility significantly longer (10–12 d) than did sperm stored without additives (6–8 d). Sperm stored in extender solution without glucose retained motility significantly longer (19–21 d) than did sperm stored in extender with glucose (13–16 d). Motility was retained for as long as 21 d in sperm stored in extender solution with 5% methanol and without glucose. In each experiment, loss of motility was associated with bacterial growth. 相似文献
5.
Evaluation of Commercial‐scale Approaches for Cryopreservation of White Crappie,Pomoxis annularis,Sperm 下载免费PDF全文
下载免费PDF全文 Charlie M. Culpepper III Amy M. Guitreau Shay Allred Terrence R. Tiersch Peter J. Allen 《Journal of the World Aquaculture Society》2018,49(4):725-734
Crappie, Pomoxis spp., are popular game fish throughout North America and are produced by public and private hatcheries. However, production is limited by a lack of information on tank culture and induced spawning methods. Development of techniques for storage of sperm and in vitro fertilization would increase flexibility in spawning. Therefore, techniques for sperm cryopreservation were examined in white crappie, Pomoxis annularis. Sperm from adult wild white crappie were used to evaluate sperm extender, cryoprotectant agent and concentration, and cooling technique based on post‐thaw sperm motility. Percent egg fertilization was also compared between sperm stored in the two best cryopreservation protocols and two different osmotic activator solutions. Sperm were cryopreserved using treatment combinations of two extenders (350 mOsmol/kg Hanks' balanced salt solution [HBSS] and 350 mOsmol/kg Ca2+free HBSS) and two cryoprotectants (dimethyl sulfoxide [DMSO] and methanol) at concentrations of 5, 10, and 15% that were cooled at four different rates: 5, 10, 20, and 40 C/min. Post‐thaw sperm motility and fertilization rates indicated white crappie sperm can be cryopreserved using either extender, cryoprotectants of either 5% DMSO or 10% methanol, and cooling at 40 C/min. A follow‐up experiment demonstrated sperm in suspensions on ice retained viability after overnight transport. 相似文献
6.
Production of inbred larvae through self‐fertilization using oocytes and cryopreserved sperm from the same individuals after sex reversal in eastern oyster Crassostrea virginica 下载免费PDF全文
下载免费PDF全文 The eastern oyster Crassostrea virginica can change sex which makes self‐fertilization possible if sperm can be cryopreserved. In this study, small (~1 year old) and large (~2–3 years old) oysters were biopsied for sperm collection. Survival of the biopsied oysters after 1 year was 50% for small oysters and 17% for large oysters. Oocytes were collected from sex‐reversed females, and self‐fertilized with cryopreserved sperm. Of the 24 cryopreserved samples, 14 individuals had ≤1% fertility when crossed with oocytes from unrelated females, indicating that the cryopreserved sperm had reduced fertility. The other 10 individuals had a fertility of 39 ± 25% when crossed with oocytes from unrelated females (non‐selfing), but showed a significantly lower success of self‐fertilization (12 ± 16%) (P = 0.008), while aliquots of the same oocytes had a fertilization of 83 ± 11% when crossing with fresh sperm. Larvae were produced at day 3 in the self‐fertilized families (12–94% of the fertilized oocytes), and survived to eyed‐larvae stage at days 11–14. Genotyping with 9 microsatellite markers confirmed that the larvae resulted from self‐fertilization in four families. This study demonstrated the feasibility of creating self‐fertilized inbred lines of oysters by use of non‐lethal sperm collection and cryopreservation. 相似文献
7.
Rafael Cuevas‐Uribe Edward J Chesney Jonathan Daly Terrence R Tiersch 《Aquaculture Research》2015,46(7):1770-1784
Our goal was to develop a standardized approach for sperm vitrification of marine fish that can be applied generally in aquatic species. The objectives were to: (i) estimate acute toxicity of cryoprotectants over a range of concentrations; (ii) evaluate the properties of vitrification solutions (VS); (iii) evaluate different thawing solutions and (iv) evaluate sperm quality after thawing by examination of motility and membrane integrity. Sperm were collected from red snapper (Lutjanus campechanus), spotted seatrout (Cynoscion nebulosus) and red drum (Sciaenops ocellatus). 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 (P < 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 higher osmotic pressures could explain the survival in the high cryoprotectant concentrations. Vitrification offers an alternative to conventional cryopreservation. 相似文献
8.
Schwarz K Huang SW German JB Tiersch B Hartmann J Frankel EN 《Journal of agricultural and food chemistry》2000,48(10):4874-4882
The activity of alpha-tocopherol, Trolox, propyl gallate, gallic acid, methyl carnosoate, and carnosic acid was studied in two oil-in-water (o/w) emulsions, in two water-in-oil (w/o) emulsions, and in bulk oil with and without added emulsifiers. All antioxidants had either moderate or higher activity in bulk oil than in the emulsions. In most emulsions, the most polar antioxidants, propyl gallate and gallic acid, exhibited either prooxidant activity or no antioxidant activity. Methyl carnosoate was the most active antioxidant in w/o emulsions but was less active than Trolox in o/w emulsions. alpha-Tocopherol was less active in bulk oil than in emulsions, but its activity in bulk oil was markedly enhanced by the addition of o/w emulsifiers. Partitioning of antioxidants, hydrogen bonding, interphase transport, surface accessibility, and interaction of emulsifier with antioxidants are considered to be important parameters that determine antioxidant activity in lipid-containing systems. 相似文献
9.
Abstract.— This study was designed to simulate conditions encountered routinely during refrigerated storage of channel catfish sperm. Sperm samples were stored at 4 C in non-sterile and sterile Hanks' balanced salt solution (HBSS). Non-sterile HBSS was prepared with distilled water stored for 2 wk in a plastic carboy prior to use. Observations were made on the frequency and abundance of bacteria in samples, and on changes in sperm motility and quality. Sperm samples stored in non-sterile HBSS had a complete loss of motility within 72 h. Samples maintained in sterile HBSS showed an initial decrease in motility between 48 and 72 h, and a complete loss of motility within 10 d. Quality of the sperm in each buffer decreased as motility decreased; morphologic changes and reduced motility of sperm were coincident with increased bacterial numbers. Bacteria were cultured on tryptic soy agar and Pseudomonas F agar (PFA) by spread-plating 10–μL aliquots from each sample onto bacteriologic media and incubating for 5 d. The dominant bacteria observed were members of the genus Pseudomonas , representing 67% of the total bacteria identified. The dominant pseudomonad ( Pseudomonas sp.) cultured from sperm samples stored in sterile buffer produced caseinase, lecithinase, and was β-hemolytic, whereas the dominant bacteria ( P. putida ) cultured from samples stored in the non-sterile buffers did not. Highly motile pseudomonads, present in two samples stored in sterile buffer, colonized below the surface of the PFA media at 4 C. The attributes of the bacterial contaminants that likely contributed to the decrease in sperm quality were production of extracellular enzymes, consumption of oxygen, and a high level of motility. Potential sources of degradative bacteria were commensal flora of channel catfish and the water used in preparing the storage buffer. 相似文献
10.
The effects of long-term feeding of cottonseed meal on growth, testis development, and sperm motility of male channel catfish Ictulurus punctatus were studied. Brood-sized male channel catfish were stocked into 0.04-ha earthen ponds in April 1992 at the rate of 120 fish per pond. The fish were fed a diet (32% protein floating catfish feed) containing either 0, 25, 375, or 52% cottonseed meal to satiation daily, except for winter months when fish were fed on days when the water temperature was 15 C or above. Fish were harvested in July 1994. Feed consumption was similar for fish regardless of dietary treatment. Feed conversion was higher for fish fed the two highest levels of cottonseed meal and weight gain was depressed in these groups. Testis weight, gonosomatic index, and sperm motility were not negatively affected by high levels of dietary cottonseed meal indicating that reproductive capacity was not diminished. Thus it wonld appepr that up to 52% cottonseed meal could be used in the diets of brood-sized channel catfish unless maximum gain is important. In a practical situation, growth rates of brood-sized channel catfish are not as important as reproductive performance. 相似文献