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Effectiveness of Aquaflor (50% florfenicol) administered in feed to control mortality associated with Streptococcus iniae in tilapia at a commercial tilapia production facility

The efficacy of Aquaflor (florfenicol; FFC) to control mortality caused by Streptococcus iniae in tilapia was evaluated under field conditions. The trial was initiated following presumptive diagnosis of S. iniae infection in a mixed group of fingerling (mean, 4.5 g) Nile Tilapia Oreochromis niloticus and a hybrid of Nile Tilapia×Blue Tilapia O. aureus . Diagnoses included mortality in source tank; examination of clinical signs and presence or absence of gram-positive cocci in brain, and collection of samples for microbiological review and disease confirmation of 60 moribund fish. Following presumptive diagnosis, tilapia (83/tank) were randomly transferred to each of 20 test tanks receiving the same water as the source tank (test tank water was not reused). Tilapia were offered either nonmedicated control feed or FFC-medicated feed (FFC at 15 mg/kg body weight/d; 10 tanks per regimen) for 10 consecutive days followed by a 14-d observation period during which only the nonmedicated control feed was offered. Streptococcus iniae was presumptively identified during pretreatment necropsy and confirmed by polymerase chain reaction assay; S. iniae was confirmed in samples taken during the dosing period but was not detected during the postdosing period. The FFC disk diffusion zone of inhibition ranged from 29 to 32 mm, while the minimum inhibitory concentration of FFC ranged from 2 to 4 μg/mL for the S. iniae isolates collected. Survival of tilapia assigned to the FFC-dose group was significantly greater at 14 d posttreatment than that of the nonmedicated controls. The odds of tilapia assigned to the FFC-dose group surviving to the end of the postdosing period were 1.34 times the odds of survival of tilapia assigned to the nonmedicated control group. There were no clinically apparent adverse effects associated with the administration of FFC-medicated feed in this study.

North American Journal of Aquaculture

A portable freshwater closed-system fish egg incubation system

To identify fish eggs collected in the field to species, a portable closed‐system fish egg incubation system was designed and used to incubate and hatch the eggs in the laboratory. The system is portable, small in scale (2.54 × 1.52 × 2.03 m), and affordable, with the approximate cost of the system being US$8,300 (2012). The main tank is 678 L and holds a battery of up to 21 (egg) incubation jars. The system includes three independent water pumping systems to (1) provide aerated water to hatching jars, (2) filter and sterilize incubation water, and (3) provide temperature‐controlled water in the main tank bath and the incubation jars. The system was successfully used to incubate freshwater fish eggs to raise resulting larvae to the post‐yolk‐sac stage for three seasons (spring 2012, spring 2013, and fall 2013) over two consecutive years, at two different locations, enabling us to identify fish eggs to species by providing identifiable fish larvae from incubated fish eggs.

North American Journal of Aquaculture

Efficacy of iodine for disinfection of Lake Sturgeon eggs from the St. Lawrence River, New York

Optimal fish husbandry to reduce the risk of disease is particularly important when using wild fish as the source for gametes. The propagation and reestablishment of Lake Sturgeon Acipenser fulvescens in New York waters to become a viable self-sustaining population is considered a high priority by managers. While standard hatchery egg disinfection practices have been used to prevent the transmission of diseases, data on the bacterial loads present on egg surfaces following iodine disinfection is lacking. Our study investigated the bacteria present on the outer surface of Lake Sturgeon eggs and the effectiveness of an iodine disinfection treatment in eliminating bacteria that could pose a threat to egg survival and cause hatchery disease outbreaks. During the springs of 2011–2013, 12 to 41 different species of bacteria were recovered from the outer egg surfaces prior to an iodine treatment; Aeromonas , Pseudomonas , Shewanella , and Chryseobacterium were the most common genera identified. Cohort eggs treated using the standard protocol of a single treatment of 50 mg/L iodine for 30 min resulted in an average of 57.8% reduction in bacterial CFU/g. While this is a significant reduction, bacteria were not completely eliminated and hatchery managers should be aware that pathogens could remain on Lake Sturgeon eggs following the standard iodine disinfection treatment.

New York

Propagation of hybrid Devils Hole Pupfish × Ash Meadows Amargosa Pupfish

Recent censuses of Devils Hole Pupfish Cyprinodon diabolis revealed that fewer than 100 individuals currently remain in the wild. Captive propagation is among actions being considered to prevent their extinction, but no pure-strain Devils Hole Pupfish were available for broodstock. To help provide emergency information, we investigated techniques to propagate their most closely related relative, hybrid Devils Hole Pupfish C. diabolis × Ash Meadows Amargosa Pupfish C. nevadensis mionectes. We tested various temperatures and larval feeds with respect to egg production, larval survival, and growth. Larval survival and growth were similar from 24°C to 32°C and egg production peaked at static 28°C; however, reducing water temperatures to 23°C and then raising them to 28°C resulted in even higher production. Larvae fed infusoria, Rio Grande Silvery Minnow Chow (RGSM), or Zeigler larval diet (ZLD) had the highest survival (79.4, 71.6, and 73.4%, respectively), and those fed Otohime (OTO) had the lowest survival (60.8%), although OTO provided greatest (14 mm) 30-d growth. Supplementation of RGSM or ZLD with Artemia nauplii increased growth but decreased survival. Larval production was maximized by placing six spawning mops, constructed of yarn and tile, in each of four 437-L parental aquaria, stocked with 24 adult fish each (1:1 sex ratio) for 3 d, to attract adults and provide spawning substrate. A 30% water change conducted on the same day of mop placement lowered water temperature from 28°C to 23°C. Water temperature was raised back to 28°C over 48 h. After 3 d, mops were transferred to hatching aquaria that were held at 28°C and aerated until larval hatch. Although some differences likely exist in effective propagation techniques for hybrid and pure-strain Devils Hole Pupfish, these data help provide initial recommendations to aid recovery.

North American Journal of Aquaculture

Mark retention of calcein in Cisco and Bloater

Since 2012, a multi-agency initiative to restore these native forage species has been under way. Evaluating the restoration success of Cisco Coregonus artedi and Bloater C. hoyi in Lake Ontario waters requires methods to identify stocked fish. However, juvenile Cisco and Bloater are fragile; thus, mass marking techniques that reduce the handling of individual fish are required and have not previously been evaluated. In 2014–2015 we evaluated the usefulness of calcein (SE-MARK) as a marker on bony structures, including the otolith. Juvenile Bloater and Cisco (14, 100, 128 d old) were immersed in a calcein bath at 5,000 mg/L of water for 4 min to apply the chemical marker. Observations of the marking retention were evaluated 8 d following the treatment. All fish immersed in calcein had strong brilliant marks (rating scale 3) on all bony structures including scales, fin rays, jaw bones, and vertebrate. The otolith was the only hard structure that did not show a brilliant marking due to the opaque nature of the structure. Our results suggest that calcein produces a strong discernable mark on hard bony structures of Cisco and Bloater; however, long-term retention needs further study.

Lake Ontario

Design and testing of a mesocosm-scale habitat for culturing the endangered Devils Hole Pupfish

aptive propagation of desert spring fishes, whether for conservation or research, is often difficult, given the unique and often challenging environments these fish utilize in nature. High temperatures, low dissolved oxygen, minimal water flow, and highly variable lighting are some conditions a researcher might need to recreate to simulate their natural environments. Here we describe a mesocosm-scale habitat created to maintain hybrid Devils Hole × Ash Meadows Amargosa Pupfish ( Cyprinodon diabolis × C. nevadensis mionectes ) under conditions similar to those found in Devils Hole, Nevada. This 13,000-L system utilized flow control and natural processes to maintain these conditions rather than utilizing complex and expensive automation. We designed a rotating solar collector to control natural sunlight, a biological reactor to consume oxygen while buffering water quality, and a reverse-daylight photosynthesis sump system to stabilize nighttime pH and swings in dissolved oxygen levels. This system successfully controlled many desired parameters and helped inform development of a larger, more permanent desert fish conservation facility at the U.S. Fish and Wildlife Service’s Ash Meadows National Wildlife Refuge, Nevada. For others who need to raise fish from unique habitats, many components of the scalable and modular design of this system can be adapted at reasonable cost.

North American Journal of Aquaculture

Enhancing hatch rate and survival in laboratory-reared hybrid Devils Hole Pupfish through application of antibiotics to eggs and larvae

We evaluated the effectiveness of four antibiotics in enhancing the hatch rate, larval survival, and adult survival of hybrid Devils Hole Pupfish Cyprinodon diabolis (hybridized with Ash Meadows Amargosa Pupfish C. nevadensis mionectes ). Cephalexin (CEX; concentration = 6.6 mg/L of water), chloramphenicol (CAM; 50 mg/L), erythromycin (ERY; 12.5 mg/L), and trimethoprim sulfamethoxazole (TMP-SMX; 25 mg/L) were applied as a constant bath either to incubating eggs or to larvae that hatched from untreated eggs. Hatch rate was roughly doubled by incubation in the presence of CAM (68% hatch) and TMP-SMX (66%) relative to the control (28%). Cephalexin and ERY conferred no benefit upon the hatch rate. Among fry that hatched from treated eggs, there was no increase in 15-d larval survival. However, fish that hatched from eggs treated with CAM, ERY, and TMP-SMX demonstrated enhanced survival at 360 d (51.2, 38.4, and 43.6%, respectively) and at 540 d (22.6, 6.8, and 20.2%, respectively); the untreated control had no survivors to those time points. All groups of eggs treated with antibiotics showed reductions in bacterial colony-forming units (CFUs) at 24 h posttreatment. At 120 h posttreatment, CEX-treated eggs had CFU counts similar to those of the control, whereas the TMP-SMX-treated eggs had the lowest CFU counts. Eggs treated with CAM and ERY had similar CFU counts, which were significantly reduced from the control counts. Larvae that were treated with CAM and TMP-SMX within 12 h posthatch showed enhanced 15-d survival (74% and 72%, respectively) in comparison with the control (56%). For pupfish rearing efforts in which antibiotic use is appropriate, CAM and TMP-SMX appear to provide the greatest benefit, particularly when applied to incubating eggs rather than to hatched larvae.

North American Journal of Aquaculture

An accurate method for measuring triploidy of larval fish spawns

A standard flow cytometric protocol was developed for estimating triploid induction in batches of larval fish. Polyploid induction treatments are not guaranteed to be 100% efficient, thus the ability to quantify the proportion of triploid larvae generated by a particular treatment helps managers to stock high-percentage spawns and researchers to select treatments for efficient triploid induction. At 3 d posthatch, individual Grass Carp Ctenopharyngodon idella were mechanically dissociated into single-cell suspensions; nuclear DNA was stained with propidium iodide then analyzed by flow cytometry. Following ploidy identification of individuals, aliquots of diploid and triploid cell suspensions were mixed to generate 15 levels (0–100%) of known triploidy ( n = 10). Using either 20 or 50 larvae per level, the observed triploid percentages were lower than the known, actual values. Using nonlinear regression analyses, quadratic equations solved for triploid proportions in mixed samples and corresponding estimation reference plots allowed for predicting triploidy. Thus, an accurate prediction of the proportion of triploids in a spawn can be made by following a standard larval processing and analysis protocol with either 20 or 50 larvae from a single spawn, coupled with applying the quadratic equations or reference plots to observed flow cytometry results. Due to the universality of triploid DNA content being 1.5 times the diploid level and because triploid fish consist of fewer cells than diploids, this method should be applicable to other produced triploid fish species, and it may be adapted for use with bivalves or other species where batch analysis is appropriate.

North American Journal of Aquaculture

Bioenergetics estimate of the effects of stocking density on hatchery production of smallmouth bass fingerlings

Production of and consumption by hatchery-reared fingerling (age-0) smallmouth bass Micropterus dolomieu at various simulated stocking densities were estimated with a bioenergetics model. Fish growth rates and pond water temperatures during the 1996 growing season at two hatcheries in Oklahoma were used in the model. Fish growth and simulated consumption and production differed greatly between the two hatcheries, probably because of differences in pond fertilization and mortality rates. Our results suggest that appropriate stocking density depends largely on prey availability as affected by pond fertilization and on fingerling mortality rates. The bioenergetics model provided a useful tool for estimating production at various stocking density rates. However, verification of physiological parameters for age-0 fish of hatchery-reared species is needed.

Oklahoma

Effect of dietary ingredient substition on dorsal fin erosion of steelhead

A feeding trial was conducted to determine the effect of supplementing a diet based on hydrolyzed fish meal with either 6% chitin (as ground krill shell), 6% squid meal, or 200 mg carotenoid pigment (canthaxanthin) per kilogram on dorsal fin erosion in juvenile steelhead Oncorhynchus mykiss. Three hundred fry in triplicate tanks were fed one of the four test diets, and 300 fry in a single tank were fed a commercial trout starter diet. The feeding trial was conducted for 15 weeks at a water temperature of 15°C. Growth was significantly higher ( P < 0.05) for fish fed the diet containing chitin than for those fed other feeds. Dorsal fin index (100 × mean dorsal fin height/total fish length) was similar among fish consuming the hydrolyzed fish meal diets but lower for fish consuming the commercial formulation. Results suggest that removal of bones from fish meal may influence dorsal fin condition in steelhead, perhaps through altered dietary mineral composition or form.

North American Journal of Aquaculture

Control of predacious flatworms Macrostomum sp. in culturing juvenile freshwater mussels

Flatworms of the genus Macrostomum are voracious predators on newly metamorphosed juvenile freshwater mussels (Unionidae), which require a fish host to transform mussel larvae into free-living juveniles. Toxicity tests were performed with formalin (paracide-F, 37% formaldehyde) to determine the appropriate levels of treatment for eradicating these flatworms from host fish tanks without adversely affecting the culture of juvenile mussels. Results indicate that a 1-h shock treatment of 250 mg/L formalin or a 3-d continuous exposure to 20 mg/L of formalin kills adult Macrostomum but not fish. Observations indicate that a single treatment is insufficient to kill Macrostomum eggs, so a second treatment after 3 d is necessary to kill newly hatched flatworms. Newly metamorphosed freshwater mussels exposed to similar shock and continuous treatments of formalin were also killed. Thus, all host fish introduced for the purpose of mussel production should be quarantined and treated prophylactically to avoid the infestation of mussel culture systems with predacious flatworms.

North American Journal of Aquaculture

Probabalistic projections for recovery of the endangered Cui-ui

The cui-ui Chamistes cujus , a lake sucker found in only Pyramid Lake, Nevada, is listed as endangered under the U.S. Endangered Species Act, and a recovery plan has been formulated in an effort to save it. We used a population simulation model that incorporated environmental stochasticity in water availability to project the persistence of cui-ui over 200 years, Based on this approach, recovery of the species in terms of persistence was evaluated for the existing hydrological situation and for situations involving water supplementation in various amounts. Our calculations indicate that immediate (as of spring 1991) acquisition of 70,000 acre-feet per year of supplemental water to raise levels in Pyramid Lake would have assured recovery and persistence over 200 years with a probability of 0.95. Postponement of acquisition for 3 years followed by incremental increases of 3,000 or 10,000 acre-feet per year would require, respectively, 83,000 and 120,000 acre-feet of annual supplemental water to assure persistence. The approach is applicable to a wide range of species and conditions.

North American Journal of Aquaculture

Use of hydrogen peroxide during incubation of landlocked fall Chinook salmon eggs in vertical-flow incubators

Six different hydrogen peroxide treatment regimes were evaluated in a series of three trials with landlocked fall Chinook salmon Oncorhynchus tshawytscha eggs incubated in vertical-flow incubators. Six daily 15-min hydrogen peroxide treatment regimes (1,000 mg/L; 1,000 mg/L with a decrease to 500 mg/L during estimated blastopore formation; 2,000 mg/L; 2,000 mg/L with a decrease to 500 mg/L during estimated blastopore formation; 2,500 mg/L; and 2,500 mg/L with a decrease to 500 mg/L during estimated blastopore formation) were compared with daily 15-min treatments of 1,667 mg/L of formalin. Mortality at egg eye-up and fry hatch and from eye-up to hatch was significantly greater in eggs receiving the 2,500-mg/L hydrogen peroxide treatments throughout incubation and in those receiving 2,500 mg/L hydrogen peroxide with a decrease to 500 mg/L during blastopore formation than in either of the 1,000-mg/L hydrogen peroxide treatment regimes or the formalin-treated eggs in the first trial. No significant differences in mortality were observed among any of the treatments in the subsequent two trials with maximum hydrogen peroxide concentrations of 2,000 mg/L. Fungal infestations were observed primarily in the incubation trays treated at either of the 1,000-mg/L hydrogen peroxide regimens, as well as in those trays whose treatment concentrations were dropped to 500 mg/L during blastopore formation. Infestations were not observed in any of the formalin-treated trays. If minor fungal infestation is acceptable, then daily hydrogen peroxide treatments of 1,000 mg/L for 15 min would probably provide adequate fungal control compared with formalin usage.

South Dakota

Dietary uptake of polychlorinated biphenyls (PCBs) by rainbow trout

The presence of detectable levels of polychlorinated biphenyls (PCBs) in commercially produced fish feed has raised a concern about the degree of biomagnification of these contaminants in hatchery-reared trout. Our objectives were to (1) define the relationship between concentrations of PCBs in fish feed and in fish tissue and (2) estimate the relative contributions of feed and hatchery supply water to PCB concentrations in fish. We conducted a 6-month feeding trial with fingerling rainbow trout Oncorhynchus mykiss fed commercial diets with four concentrations of PCBs: a low-PCB diet (69 ng/g); a typical commercial diet (126 ng/g); and the typical diet spiked with PCBs at two levels (220 and 280 ng/g). The concentrations of PCBs in fillets after 1 month were commensurate with those in the feeds and remained relatively stable for the next 5 months; mean PCB concentrations in fillets ranged from 54 to 94 ng/g. Low levels of PCBs were detected in the hatchery supply water. We used the concentrations of PCBs in the feeds, absorption rates of PCBs, and two different rates of PCB depuration to estimate the potential uptake of PCBs from supply water. When we used a low depuration rate (half-life = 219 d), the computed body burdens of PCBs could be entirely attributed to the feeds. When a high depuration rate (half-life = 66 d) was used, some uptake of PCBs from the supply water was likely, but most of the total body burden originated from the feeds. We concluded that rainbow trout fed a diet with 126 ng/g PCBs would have a PCB concentration of about 60 ng/g in their fillets, which is high enough to warrant issuance of a consumption advisory (no more than one meal of fish per week) under a protocol adopted by some Great Lakes states.

North American Journal of Aquaculture

Use of prepositioned grid electrofishers for the collection of robust redhorse broodstock

We investigated the potential of prepositioned grid electrofishers as a means of collecting broodstock for the rare robust redhorse Moxostoma robustum. We found that combined with visual observation, this technique allowed for the efficient capture of fish in breeding condition. We were able to harvest eggs in the field and bring only fertilized eggs into the hatchery. There was no need to induce spawning hormonally. Although their use is limited by water depth and clarity, prepositioned grid electrofishers used in conjunction with visual observation warrants further consideration as an effective tool for the collection of reproductively active broodstock for conservation purposes. ?? Copyright by the American Fisheries Society 2005.

North American Journal of Aquaculture

Survival and passage of ingested New Zealand mudsnails through the intestinal tract of rainbow trout

We conducted laboratory trials to determine the transit time and survival of New Zealand mudsnails Potamopyrgus antipodarum in the gastrointestinal tract of rainbow trout Oncorhynchus mykiss. To assess the rate of snail passage, we force-fed groups of fish a known quantity of snails and then held them in tanks. At selected intervals we removed individual fish from the test tanks and recorded the number of snails, their condition (live or dead), and their location in the gastrointestinal tract (stomach, anterior intestine, and posterior intestine). Feces were removed from tanks and examined for live snails. We repeated evaluations of passage rate and snail survival to determine the effects of varying the number of snails ingested, fish size, snail size, and feeding a commercial diet to fish after snail ingestion. We plotted and modeled gut evacuation using a stochastic model for ordinal data to consider each test variable. Snail passage rates were faster in fish that were fed smaller snails. Surprisingly, fish fed snails and then administered rations of commercial fish feed retained the snails longer in their stomach than did fish that were not administered fish feeds after being fed snails. Increased retention time of snails in the stomach decreased the probability of snail survival when voided in fecal material. Snails that passed through the gastrointestinal tract within 12–24 h of ingestion were often recovered live in fecal samples. However, no live snails were recovered from the posterior intestine or fecal material collected 24 h after ingestion. Using our results we propose potential management options that could reduce the risks of introducing live snails into new locations when stocking fish from infested hatcheries.

Idaho

A minimally invasive method for extraction of sturgeon oocytes

Fishery biologists, hatchery personnel, and caviar fishers routinely extract oocytes from sturgeon (Acipenseridae) to determine the stage of maturation by checking egg quality. Typically, oocytes are removed either by inserting a catheter into the oviduct or by making an incision in the body cavity. Both methods can be time-consuming and stressful to the fish. We describe a device to collect mature oocytes from sturgeons quickly and effectively with minimal stress on the fish. The device is made by creating a needle from stainless steel tubing and connecting it to a syringe with polyvinyl chloride tubing. The device is filled with saline solution or water, the needle is inserted into the abdominal wall, and eggs are extracted from the fish. Using this device, an oocyte sample can be collected in less than 30 s. Such sampling leaves a minute wound that heals quickly and does not require suturing. The extractor device can easily be used in the field or hatchery, reduces fish handling time, and minimizes stress.

North American Journal of Aquaculture

An evaluation of selective feeding by three age-groups of the rainbow mussel Villosa iris

A tri-algal diet was fed to three age-groups of the rainbow mussel Villosa iris: ages 2-3 d, 50-53 d, and 3-6 years. Changes in the relative abundance of each algal species were determined in 5-h feeding trials from feeding chambers and by gut content analyses. All age-groups rejected Scenedesmus quadricauda and preferentially selected Nannochloropsis oculata and Selenastrum capricornutum, principally on the basis of size. Changes in the relative abundance of algae in feeding chambers did not differ significantly among age-groups. Observed differences in the ingested quantities of the similar-sized N. oculata and S. capricornutum were attributed to other particle-related characteristics. Results indicate that the rainbow mussel can be fed similar-sized algae at ali ages in captive propagation facilities. When developing a suitable algal diet for rearing juvenile mussels, one probably need not investigate different species at each stage of development if the algae used are in the 2.8-8.5-??m size range.

North American Journal of Aquaculture