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Assessing consumption of bioactive micro-particles by filter-feeding Asian carp

Silver carp Hypophthalmichthys molitrix (SVC) and bighead carp H. nobilis (BHC) have impacted waters in the US since their escape. Current chemical controls for aquatic nuisance species are non-selective. Development of a bioactive micro-particle that exploits filter-feeding habits of SVC or BHC could result in a new control tool. It is not fully understood if SVC or BHC will consume bioactive micro-particles. Two discrete trials were performed to: 1) evaluate if SVC and BHC consume the candidate micro-particle formulation; 2) determine what size they consume; 3) establish methods to evaluate consumption of filter-feeders for future experiments. Both SVC and BHC were exposed to small (50-100 μm) and large (150-200 μm) micro-particles in two 24-h trials. Particles in water were counted electronically and manually (microscopy). Particles on gill rakers were counted manually and intestinal tracts inspected for the presence of micro-particles. In Trial 1, both manual and electronic count data confirmed reductions of both size particles; SVC appeared to remove more small particles than large; more BHC consumed particles; SVC had fewer overall particles in their gill rakers than BHC. In Trial 2, electronic counts confirmed reductions of both size particles; both SVC and BHC consumed particles, yet more SVC consumed micro-particles compared to BHC. Of the fish that ate micro-particles, SVC consumed more than BHC. It is recommended to use multiple metrics to assess consumption of candidate micro-particles by filter-feeders when attempting to distinguish differential particle consumption. This study has implications for developing micro-particles for species-specific delivery of bioactive controls to help fisheries, provides some methods for further experiments with bioactive micro-particles, and may also have applications in aquaculture.

Journal of Aquaculture Research & Development

Identifying sources and year classes contributing to invasive grass carp in the Laurentian Great Lakes

Relative contributions of aquaculture-origin and naturally-reproduced grass carp ( Ctenopharyngodon idella ) in the Laurentian Great Lakes have been unknown. We assessed occurrence and distribution of aquaculture-origin and wild grass carp in the Great Lakes using ploidy and otolith stable oxygen isotope (δ 18 O) data. We inferred natal river and dispersal from natal location for wild grass carp using otolith microchemistry and estimated ages of wild and aquaculture-origin fish to infer years in which natural reproduction and introductions occurred. Otolith δ 18 O indicated that the Great Lakes contain a mixture of wild grass carp and both diploid and triploid, aquaculture-origin grass carp. Eighty-eight percent of wild fish (n = 49 of 56) were caught in the Lake Erie basin. Otolith microchemistry indicated that most wild grass carp likely originated in the Sandusky or Maumee rivers where spawning has previously been confirmed, but results suggested recruitment from at least one other Great Lakes tributary may have occurred. Three fish showed evidence of movement between their inferred natal river in western Lake Erie and capture locations in other lakes in the Great Lakes basin. Age estimates indicated that multiple year classes of wild grass carp are present in the Lake Erie basin, recruitment to adulthood has occurred, and introductions of aquaculture-origin fish have happened over multiple years. Knowledge of sources contributing to grass carp in the Great Lakes basin will be useful for informing efforts to prevent further introductions and spread and to develop strategies to contain and control natural recruitment.

Michigan, New York, Ohio, Ontario

First record of the non-indigenous parasitic copepod Neoergasilus japonicus (Harada, 1950) in the Lake Ontario Watershed: Oneida Lake, New York

Four specimens of the Asiatic parasitic copepod Neoergasilus japonicus (Harada, 1930) were collected from Oneida Lake, New York in September 2018; one specimen was from a white sucker Catostomus commersonii , another from a green sunfish Lepomis cyanellus , and two from a bluegill Lepomis macrochirus . The four adult female specimens were found attached to the base of the gills of their respective hosts along with other ergasilid species. The average total length of the adult female N. japonicus specimens we found was 0.609 mm. These detections represent the first known occurrence of this non-native species in the state of New York, extends the easternmost distribution of this parasite over 400 miles, and now includes the Lake Ontario watershed for the first time. It is commonly believed that the international aquaculture industry and aquarium fish trade are the most likely vectors of dispersal for N. japonicus . Monitoring the spread of non-indigenous aquatic species is an important step towards the development of management plans and mitigation efforts with regards to the anthropogenic causes of dispersal, and fish parasites are no exception.

New York

Comparative hydraulics of two fishery research circular tanks and recommendations for control of experimental bias

Hydraulic characteristics inside two research circular tanks (1.5-m and 1.2-m diameter) with the same volume of water were studied to understand how they might affect experimental bias by influencing the behavior and development of juvenile fish. Water velocities inside each tank were documented extensively and flow behavior studied. Surface inflow to the 1.5-m tank created a highly turbulent and aerated surface, and produced unevenly distributed velocities within the tank. A low-flow velocity, or "dead" zone, persisted just upstream of the surface inflow. A single submerged nozzle in the 1.2-m tank created uniform flow and did not cause undue turbulence or introduce air. Flow behavior in the 1.5-m tank is believed to have negatively affected the feeding behavior and physiological development of a group of juvenile fall chinook salmon, Oncorhynchus tshawytscha. A new inflow nozzle design provided comparable flow behavior regardless of tank size and water depth. Maintaining similar hydraulic conditions inside tanks used for various biological purposes, including fish research, would minimize experimental bias caused by differences in flow behavior. Other sources of experimental bias are discussed and recommendations given for reporting and control of experimental conditions in fishery research tank experiments.

Journal of Applied Aquaculture

Tests of rhodamine WT dye for toxicity to oysters and fish

Because of the toxicity to oyster larvae and eggs of rhodamine B dye in concentrations greater than 1 mg/l in earlier tests, there was a concern that rhodamine WT, a similar tracer dye, would have a detrimental effect on marine life being developed under the aquaculture program of the Lummi Indian Tribe near Bellingham, Wash. Tests showed that 48-hour exposures at 24° C of 11,000 oyster eggs per liter and 6,000 12-day-old larvae per liter, in sea water with concentrations of rhodamine WT ranging from 1 μg/l to 10 mg/l, resulted in development of the eggs to normal straight-hinge larvae and no abnormalities in the larvae development. Tests made on the smolt of both silver salmon and Donaldson trout, with the fish held for 17.5 hours in a tankfull of sea water with a dye concentration of 10 mg/l at 22°C showed no mortalities or respiratory problems. With the concentration increased to 375 mg/l, and the time extended an additional 3.2 hours, still no mortalities or abnormalities were noted. The fish remained healthy in dye-free water when last checked a month after the test.

Journal of Research of the U.S. Geological Survey

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

Isothermal recombinant polymerase amplification and CRIPSR (CAS12A) assay detection of Renibacterium salmoninarum as an example for wildlife pathogen detection in environmental DNA samples

Improving rapid detection methods for pathogens is important for research as we collectively aim to improve the health of ecosystems globally. In the northern hemisphere, the success of salmon ( Oncorhynchus spp.) populations is vitally important to the larger marine, aquatic, and terrestrial ecosystems they inhabit. This has led to managers cultivating salmon in hatcheries and aquaculture to bolster their populations, but young salmon face many challenges, including diseases such as bacterial kidney disease (BKD). Early detection of the BKD causative agent, Renibacterium salmoninarum , is useful for managers to avoid outbreaks in hatcheries and aquaculture stocks to enable rapid treatment with targeted antibiotics. Isothermal amplification and CRIPSR-Cas12a systems may enable sensitive, relatively rapid, detection of target DNA molecules from environmental samples compared to quantitative PCR (qPCR) and culture methods. We used these technologies to develop a sensitive and specific rapid assay to detect R. salmoninarum from water samples using isothermal recombinase polymerase amplification (RPA) and an AsCas12a RNA-guided nuclease detection. The assay was specific to R. salmoninarum (0/10 co-occurring or closely related bacteria detected) and sensitive to 0.0128 pg/µL of DNA (approximately 20–40 copies/µL) within 10 min of Cas activity. This assay successfully detected R. salmoninarum environmental DNA in 14/20 water samples from hatcheries with known quantification for the pathogen via previous qPCR (70% of qPCR-positive samples). The RPA-CRISPR/AsCas12a assay had a limit of detection (LOD) of >10 copies/µL in the hatchery water samples and stochastic detection below 10 copies/µL, similar to but slightly higher than the qPCR assay. This LOD enables 37 C isothermal detection, potentially in the field, of biologically relevant levels of R. salmoninarum in water. Further research is needed to develop easy-to-use, cost-effective, sensitive RPA/CRISPR-AsCas12a assays for rapidly detecting low concentrations of wildlife pathogens in environmental samples.

Journal of Wildlife Diseases

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