William H. Gingerich

Safety of Aquaflor (Florfenicol, 50% Type A Medicated Article), Administered in Feed to Channel Catfish, Ictalurus punctatus

Aquaflor, a feed premix containing the broad spectrum antibacterial agent florfenicol (50% w/w), is being developed for use to control enteric septicemia (ESC) in channel catfish Ictalurus punctatus caused by the gram-negative enterobacterium Edwardsiella ictaluri. The recommended dose of Aquaflor to control ESC is 10 mg/kg body weight (BW)/day for 10 days. The study objective was to determine the safety of Aquaflor administered in feed to channel catfish at doses of 0 (control), 10, 30, and 50 mg/kg BW/day for 20 consecutive days. Parameters evaluated included daily mortality, behavioral (appetite, distribution, flight/fright response), and water chemistry observations, initial and terminal weight measurements, and gross and microscopic pathology. Medicated feed consumption was 67—86% of target with group mean doses of 8.5 mg/kg BW/day, 24.6 mg/kg BW/day, and 34.9 mg/kg BW/day. There were no mortalities or clinically observable changes noted at any of the dose levels tested. Aquaflor-related changes were limited to the food consumption and histopathology data. Although Aquaflor-related decreased feed consumption was noted in the 30 and 50 mg/kg BW/day groups, there were no differences in fish growth among the treatment groups. Aquaflor-related histopathology findings were limited to a histomorphologically evident dose-dependent decrease in hematopoietic/lymphopoietic tissue in the anterior kidneys, posterior kidneys, and spleens of channel catfish.

Liquid chromatographic determination of florfenicol in the plasma of multiple species of fish.

A simple method was developed for determining florfenicol concentration in a small volume (250 micro l) of plasma from five phylogenetically diverse species of freshwater fish. Florfenicol was isolated from the plasma matrix through C(18) solid-phase extraction and quantified by reversed-phase high-performance liquid chromatography with UV detection. The accuracy (84-104%), precision (%RSD<or=8), and sensitivity (quantitation limit <30 ng/ml) of the method indicate its usefulness for conducting pharmacokinetic studies on a variety of freshwater fish.

Efficacy of Hydrogen Peroxide for Treating Saprolegniasis in Channel Catfish

Abstract Hatchery-reared fish and their eggs are commonly afflicted with saprolegniasis, a fungal disease that can cause significant losses in production. Fish culturists need safe and effective fungicides to minimize losses and meet production demands. The efficacy of hydrogen peroxide was evaluated for preventing or controlling mortality associated with saprolegniasis in channel catfish Ictalurus punctatus. Saprolegniasis was systematically induced in channel catfish so various therapies could be evaluated in a controlled laboratory environment. Both prophylactic and therapeutic hydrogen peroxide bath treatments of 50, 100, and 150 μL/L for 1 h were administered every other day for seven total treatments. All untreated positive control fish died of saprolegniasis during the prophylactic and therapeutic tests. Hydrogen peroxide treatments of 150 μL/L were harmful (relative to lower concentrations) to test fish and resulted in 73–95% mortality. Mortality was attributed to a combination of abrasion, temper...

A simple analytical procedure to replace HPLC for monitoring treatment concentrations of chloramine-T on fish culture facilities

Abstract Concentrations of chloramine-T must be monitored during experimental treatments of fish when studying the effectiveness of the drug for controlling bacterial gill disease. A surrogate analytical method for analysis of chloramine-T to replace the existing high-performance liquid chromatography (HPLC) method is described. A surrogate method was needed because the existing HPLC method is expensive, requires a specialist to use, and is not generally available at fish hatcheries. Criteria for selection of a replacement method included ease of use, analysis time, cost, safety, sensitivity, accuracy, and precision. The most promising approach was to use the determination of chlorine concentrations as an indicator of chloramine-T. Of the currently available methods for analysis of chlorine, the DPD ( N , N -diethyl- p -phenylenediamine) colorimetric method best fit the established criteria. The surrogate method was evaluated under a variety of water quality conditions. Regression analysis of all DPD colorimetric analyses with the HPLC values produced a linear model ( Y =0.9602 X +0.1259) with an r 2 value of 0.9960. The average accuracy (percent recovery) of the DPD method relative to the HPLC method for the combined set of water quality data was 101.5%. The surrogate method was also evaluated with chloramine-T solutions that contained various concentrations of fish feed or selected densities of rainbow trout. When samples were analyzed within 2 h, the results of the surrogate method were consistent with those of the HPLC method. When samples with high concentrations of organic material were allowed to age more than 2 h before being analyzed, the DPD method seemed to be susceptible to interference, possibly from the development of other chloramine compounds. However, even after aging samples 6 h, the accuracy of the surrogate DPD method relative to the HPLC method was within the range of 80–120%. Based on the data comparing the two methods, the U.S. Food and Drug Administration has concluded that the DPD colorimetric method is appropriate to use to measure chloramine-T in water during pivotal efficacy trials designed to support the approval of chloramine-T for use in fish culture.

Effects of temperature on the elimination of benzocaine and acetylated benzocaine residues from the edible fillet of rainbow trout (Oncorhynchus mykiss)

The effect of temperature (7 degrees C and 16 degrees C) on the extent of accumulation and the elimination of benzocaine (BNZ) and its metabolite, acetylated benzocaine (AcBNZ), in the fillet tissue of rainbow trout was investigated. Residues were measured after bath exposure to an anesthetizing concentration of benzocaine (30 mg/l for 5 min) followed by a maintenance concentration (15 mg/l for 30 min). Immediately after exposure, the BNZ concentration in fillet tissue was approximately 27 micrograms/g at both temperatures; AcBNZ was 0.3 microgram/g at 7 degrees C and 0.6 microgram/g at 16 degrees C. The rates for elimination (alpha and beta) of BNZ and AcBNZ were not significantly different between the two temperatures. Terminal half-lives of elimination for BNZ were 1.62 h at 7 degrees C and 1.63 h at 16 degrees C; half-lives for AcBNZ were 2.36 h at 7 degrees C and 2.77 h at 16 degrees C.

Short-Duration Electrical Immobilization of Lake Trout

Abstract Chemical anesthetics induce stress responses, and most leave residues in fish tissues that require a certain withdrawal time before the animal can be released into the environment. Therefore, alternatives are needed in cases when fish must be released immediately, for example, during egg-collecting operations or after implanting elastomer tags. To evaluate pulsed direct current as an alternative method of immobilization, individual lake trout Salvelinus namaycush were electrically immobilized using various pulsed-DC voltage gradients and shock durations. Duration of opercular recovery and narcosis were measured for individual fish. Fish were euthanized 24 h after electrical immobilization and processed for lateral radiograph analysis and assessment of perivertebral hemorrhaging by dissection. Survival of lake trout after electrical immobilization at 0.6 V/cm for 30 or 40 s or 0.8 V/cm for 5 or 15 s was monitored for 81 or 84 d after immobilization. Mean narcosis duration increased with voltage gr...

Pharmacokinetics of benzocaine in rainbow trout (Oncorhynchus mykiss) after intraarterial dosing

Abstract The pharmacokinetics of benzocaine were analyzed in rainbow trout ( Oncorhynchus mykiss ) after intraarterial bolus administration of benzocaine at nominal concentrations of 6 and 9 mg/kg. Distributive phase pharmacokinetic parameters were difficult to estimate. Benzocaine concentrations at 2 min after dosing were highly variable and decreased rapidly within the first 10 min. Benzocaine concentrations were near or below the quantitation limit 90 min after dosing. A three-compartment pharmacokinetic model best described benzocaine concentrations in plasma. The apparent volume of distribution at steady state and total body clearance increased with respect to dose. Since the mean residence time was similar at both dosages, the differences in the apparent volume of distribution at steady state and total body clearance may have been the result of data variability or the degree of anaesthesia rather than dose dependency. The model parameters indicated that distribution of benzocaine outside of the plasma was limited. Despite the initial rapid distribution and elimination of benzocaine from plasma, terminal phase benzocaine elimination was relatively slow (terminal elimination phase half-lives of 89 and 109 min).

Oxytetracycline depletion from skin-on fillet tissue of coho salmon fed oxytetracycline medicated feed in freshwater at temperatures less than 9°C

Abstract Oxytetracycline (OTC) is a broad spectrum antibacterial agent approved in the USA for treating certain bacterial diseases in salmonids cultured in freshwater at temperatures greater than or equal to 9°C. This study was conducted to provide the information necessary to expand the OTC label to include treatment of diseased salmonids cultured in freshwater at temperatures below 9°C. The study was designed to treat juvenile coho salmon ( Oncorhynchus kisutch ) with OTC-medicated feed and determine the depletion of OTC from the skin-on fillet tissue. Oxytetracycline depletion was evaluated in juvenile coho salmon (weight range, 13–62 g) fed OTC-medicated feed at a rate of 88.2 mg OTC/kg body weight/day for 10 days. Pairs of skin-on fillets were taken from individual fish on days 4 and 10 during the treatment phase and on days 1, 4, 8, 14, and 19 during the depletion phase. Water temperatures during the study period ranged from 4.1°C to 8.5°C. The OTC concentrations in medicated feed and skin-on fillets were determined with high-performance liquid chromatography methods. The maximum mean OTC concentration in fillet tissue was 932 ng/g, 1 day after the last treatment and decreased to 32 ng/g 19 days after the last treatment. The log-linear loss of OTC from the fillet tissue was biphasic with a terminal phase half-life of 4.9 days.

Depletion of the chloramine-T marker residue, para-toluenesulfonamide, from skin-on fillet tissue of hybrid striped bass, rainbow trout, and yellow perch

Abstract Waterborne exposure to n-sodium-n-chloro-p-toluenesulfonamide (chloramine-T) is an effective treatment for controlling fish mortalities caused by bacterial gill disease (BGD). Currently, data are being generated to gain United States Food and Drug Administration (FDA) approval for the use of chloramine-T in aquaculture. As part of the data required for an approval, depletion of the chloramine-T marker residue (para-toluenesulfonamide [p-TSA]) from the edible fillet tissue of exposed fish must be determined. Hybrid striped bass (Morone saxatilis×Morone chrysops; mean weight 357 g), rainbow trout (Oncorhynchus mykiss; mean weight 457 g), and yellow perch (Perca flavescens; mean weight 144 g) were exposed to 20 mg/l of chloramine-T for 60 min on 4 consecutive days (the most aggressive treatment expected for approved use in the United States). Groups of fish (n=15 or 19) were sampled immediately after the last treatment and periodically through 48 or 168 h after the treatment phase. Duplicate subsamples of skin-on fillet tissue from each fish were analyzed for p-TSA. Mean p-TSA concentrations in fillet tissue from fish sampled immediately after the last treatment were 142 ng/g (hybrid striped bass), 97 ng/g (rainbow trout), and 150 ng/g (yellow perch). Mean p-TSA concentrations at terminal sample times were 94 (168 h; hybrid striped bass), 74 (48 h; rainbow trout), and 35 ng/g (168 h; yellow perch). The half-lives of p-TSA in fillet tissue from fish near or at market size were 11.4 (hybrid striped bass), 4.3 (rainbow trout), and 3.2 days (yellow perch).

Safety of Oxytetracycline (Terramycin TM-100F) Administered in Feed to Hybrid Striped Bass, Walleyes, and Yellow Perch

Abstract Oxytetracycline (Terramycin TM-100F, a medicated premix containing oxytetracycline at 220 g/kg) is approved in the United States to control certain systemic bacterial diseases of salmon and catfish when fed at a rate of 55–82.5 mg per kilogram of bodyweight per day for 10 d. Although oxytetracycline may also control certain systemic bacterial infections in coolwater or scaled warmwater fish, no safety data for such species are available. Our objective was to determine the safety of oxytetracycline administered in feed at nominal doses of 0, 82.5, 248, or 413 mg · kg−1 · d−1 to yellow perch Perca flavescens and hybrid striped bass (striped bass Morone saxatilis × white bass M. chrysops) for 10 d and to walleye Sander vitreus (formerly Stizostedion vitreum) for 20 d. Yellow perch and hybrid striped bass consumed 50% to 100% of the diet, whereas walleye feed consumption was occasionally less than 50% of the diet. Feed or fecal material was present in the gastrointestinal tract of all necropsied wall...