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Temperature influence on the development and loss of seawater tolerance in two fast-growing strains of Atlantic salmon

Development of hypo-osmoregulatory ability, gill Na+,K +-ATPase activity, condition factor and growth in Atlantic salmon during parr-smolt transformation was studied in a 2??3 factorial design with three temperatures (12.0, 8.9??C and ambient, 2.4-11.9??C, mean: 6.0??C) and two farmed strains of smolts (Mowi and AquaGen). The development of hypo-osmoregulatory ability and gill Na+,K+-ATPase activity were significantly influenced by freshwater temperature. In smolts raised at 12.0??C, maximum gill Na+,K+-ATPase activity was reached in late April, compared with late May and mid-June in the 8.9??C and ambient groups, respectively. In all groups, peak gill Na+,K +-ATPase activity was seen 350 degree days (d??C) after the onset of the smolt-related increase in enzyme activity (30 March) The period of high enzyme activity (>90% of maximum) lasted approximately 250 d??C. No distinct peak level in gill Na+,K+-ATPase activity was seen in the AquaGen strain at ambient temperature. Elevated temperatures also accelerated the loss of hypo-osmoregulatory capacity. In all groups, gill Na+,K+-ATPase activity reached pre-smolt levels approximately 500 d??C after the calculated peak level. Growth rate in freshwater was influenced by strain, temperature and their interaction, with the Mowi strain showing a higher growth rate than the AquaGen strain at 8.9??C and ambient temperatures. Following transfer to seawater, a higher growth rate was recorded in smolts from the Mowi strain than the AquaGen strain from the ambient temperature regime. Temperature influences the development and loss of smolt characteristics in both strains, and has long-term effects on post-smolt performance in seawater. ?? 2004 Elsevier B.V. All rights reserved.

Aquaculture

Umbilical scarring in hatchling American alligators

Umbilical scarring is the presence of excess scar tissue deposited between abdominal dermal layers at the site of yolk sac absorption in hatchling American alligators (Alligator mississippiensis). The presence of this dermal condition plays a key evaluatory role in the overall quality and subsequent value for various commercial leather products. Despite the prevalent nature of this condition, currently the industry has no standardized protocols for its quantification. The objectives of this study were to examine the relationship between hatchling weight and age and incidence of umbilical scarring and to develop a quantifiable and reproducible technique to measure this dermal condition in hatchling American alligators. Thirty eggs from each of nine clutches were incubated in two separate incubators at different facilities and hatchling umbilical scarring was measured at 2 and 10 days of age using digital calipers. Umbilical area was calculated by multiplying umbilical length times umbilical width. There was a significant effect of both age and clutch on umbilical area (overall decline of 64%) by 10 days post-hatch. However, only five of the nine clutches utilized expressed a noticeable decline in the size of this dermal condition (range 67-74%). We had hypothesized that larger hatchlings would have larger umbilical areas and a slower rate of improvement in this condition during the first few days post-hatch. The differences in umbilical area and percent decline with age across clutches, however, were not associated with differences in initial hatchling weights. Within clutches and time periods, hatchling weight had no significant effect on the size and/or rate of decline of this condition. ?? 2004 Published by Elsevier B.V.

Aquaculture

Efficacy of hydrogen peroxide to control saprolegniasis on channel catfish (Ictalurus punctatus) eggs

The efficacy of hydrogen peroxide to control mortality associated with saprolegniasis in channel catfish ( Ictalurus punctatus ) eggs was evaluated at the Lost Valley State Fish Hatchery (Warsaw, MO). Two efficacy trials were conducted. In Trial 1, channel catfish eggs in their natural gelatinous matrix were treated with hydrogen peroxide at 0, 500, and 750 mg l(-1). Channel catfish eggs in Trial 2 had the gelatinous matrix removed before treatment with hydrogen peroxide at 0 and 500 mg l(-1). Each treatment regimen was tested in triplicate and each egg jar contained similar to 17,400 eggs. Hydrogen peroxide was administered as a 15-min flow-through treatment applied once daily for a total of six applications. Control jars were similarly treated with culture water. Samples of exposure water were collected during each treatment and analyzed to verify actual treatment concentrations. Hydrogen peroxide treatment efficacy was assessed by comparing the percent egg hatch in the treatment group to the untreated control group in each trial. Mean percent hatch in Trial I was 44% (control), 54% (500 mg l(-1)), and 69% (750 mg l(-1)). Hydrogen peroxide treatment at either 500 or 750 mg l(-1) significantly (P<0.01) increased the percent hatch compared to the untreated control group. In Trial 2, hydrogen peroxide treatment at 500 mg l(-1) significantly (P<0.01) increased the percent egg hatch (67%) relative to the untreated controls (57%). Hydrogen peroxide treatment reduced egg mortality and increased the percent hatch of channel catfish eggs regardless of whether eggs were incubated in the gelatinous matrix or without the matrix in comparison to the untreated control.

Missouri

Effects of phase-feeding dietary phosphorus on survival, growth, and processing characteristics of rainbow trout Oncorhynchus mykiss

A factorial experiment involving eight diets and three feeding periods was conducted to determine the minimal level of dietary phosphorus required to maintain survival, growth, and processing characteristics of post-juvenile rainbow trout. Trout were reared to an average size of 200, 300, or 400 g using a commercial feed (1.20% P), then allotted by triplicate groups of nine fish to one of seven experimental diets containing logarithmic increments of dietary phosphorus (0.15%%, 0.21%, 0.30%, 0.42%, 0.60%, 0.85%, and 1.20% P) or a commercial trout feed (1.20% P). At an average weight of 550 g, fish were transported to a commercial processing plant, mechanically filleted, and evaluated for quality. Fish survival and weight gain increased quadratically with increased dietary phosphorus for fish started on treatment at 200 and 300 g, but were similar among all fish started at 400 g. Phosphorus retention decreased with increasing dietary phosphorus level, from approximately 88% in groups fed diets containing 0.21% phosphorus to between 23% and 32% in groups fed diets containing 0.85% phosphorus. Calculated phosphorus losses increased as dietary phosphorus levels increased, from a low of approximately 0.4 g phosphorus kg-1 fish weight gain to between 9.5 and 13 g phosphorus kg-1 fish weight gain at the highest dietary phosphorus level. Dietary phosphorus did not affect carcass moisture, protein, lipid, or ash, but carcass phosphorus increased with increased dietary phosphorus among fish started on treatment at 200 and 300 g. There were no differences among any treatment group in carcass dressing or finishing percentage, or visual or textural appeal. The results indicate that available phosphorus levels can be reduced in rainbow trout diets to 0.60% at 200 g, to 0.30% at 300 g, or to 0.15% at 400 g live weight without loss in production or product quality in fish harvested at 550 g. Using these phase-feeding strategies would reduce the amount of phosphorus fed to the fish over the production cycle by 25%, and the amount lost to the environment by 12.5% for fish starting at 300 or 400 g. ?? 2004 Elsevier B.V. All rights reserved.

Aquaculture

Disparate effects of constant and annually-cycling daylength and water temperature on reproductive maturation of striped bass (Morone saxatilis)

Adult striped bass ( Morone saxatilis ) were exposed to various combinations of constant or anually-cycling daylength and water temperature. Constant conditions (15 h days, 18 °C) were those normally experienced at spawning and cycling conditions simulated natural changes at Chesapeake Bay latitude. Females exposed to constant long (15 h) days and cycling water temperature (TEMPERATURE group) had blood plasma levels of sex steroids (testosterone [T] and estradiol-17β [E 2 ]) and vitellogenin (Vg), and profiles of oocyte growth, that were nearly identical to those of females held under a natural photothermal cycle (CONTROL group). Several fish from these two groups were induced to spawn fertile eggs. Females constantly exposed to warm water (18 °C), with or without a natural photoperiod cycle (PHOTOPERIOD and STATIC groups, respectively), had diminished circulating levels of gonadal steroid hormones and Vg, impaired deposition of yolk granules in their ooplasm, and decreased oocyte growth, and they underwent premature ovarian atresia. Males exposed to cycling water temperature (CONTROL and TEMPERATURE groups) spermiated synchronously during the natural breeding season, at which time they also had had high plasma androgen (T and 11-ketotestosterone [11-KT]) levels. The timing of spermiation was highly asynchronous among males in groups of fish held constantly at 18 °C (STATIC and PHOTOPERIOD groups) and this asynchrony was associated with diminished plasma androgen levels. Termination of spermiation by males exposed to cycling water temperature coincided with a sharp decline in levels of plasma androgens about a month after water temperature rose above 18 °C. In contrast, most males held constantly at 18 °C sustained intermediate levels of plasma androgens and spermiated until the end of the study in late July. The annual cycle of water temperature clearly plays a prominent role in the initiation, maintenance, and termination of the striped bass reproductive cycle. In females, a decrease in water temperature below values experienced at spawning appears to be required for vitellogenesis and oocyte growth to proceed normally. Constant exposure of males to spawning temperature disrupts synchronous spermiation but also delays testicular regression, which may be useful for spawning fish after the natural reproductive season.

Aquaculture

Na+/K+/2Cl- cotransporter and CFTR gill expression after seawater transfer in smolts (0+) of different Atlantic salmon (Salmo salar) families

Smoltification involves morphological and physiological changes in the gills that prepare anadromous salmonids to osmoregulate efficiently in seawater. In a previous study, we found that different families of Atlantic salmon (Salmo salar) smolts vary in their ability to osmoregulate when abruptly transferred to cold seawater and that these differences are correlated with gill Na+/K+ ATPase activity. Here we extend these findings to test whether other key transport proteins, namely Na+/K+/2Cl- contransporter (NKCC) and the Cl- channel or cystic fibrosis transmembrane conductance regulator (CFTR), play a significant role in osmoregulatory differences between families. To facilitate molecular analysis of NKCC, we first isolated a gill cDNA containing the complete coding region (1147 aa) of an isoform previously reported as a partial sequence. Phylogenetic analysis showed that this isoform is most closely related to isoforms of the NKCC1a subfamily found in European eel and Mozambique tilapia. In a second step, we quantified NKCC protein abundance as well as mRNA expression levels for NKCC1a and two CFTR isoforms (CFTRI and CFTRII) in 0+ smolts from three families prior to and following seawater transfer. The family with the lowest salinity tolerance also showed significant increases in gill NKCC1a mRNA after seawater transfer. Taken together with our previous study, these data indicate that family differences in expression of transport proteins are in part related to salinity tolerance, although the best indicator of osmoregulatory performance between families may be gill Na+/K+ ATPase activity and CFTR I mRNA levels, rather than Na+/K+ ATPase and NKCC1a mRNA levels or NKCC protein abundance. ?? 2007 Elsevier B.V. All rights reserved.

Aquaculture

Molecular mechanisms of continuous light inhibition of Atlantic salmon parr-smolt transformation

Atlantic salmon (Salmo salar) rely on changes in photoperiod for the synchronization of the developmental events constituting the parr-smolt transformation. In the absence of photoperiod cues, parr-smolt transformation is incomplete, and such 'pseudo-smolts' normally fail to adapt to seawater. The present study addresses the endocrine and molecular mechanisms controlling the development of hypo-osmoregulatory ability and how artificial photoperiod can disrupt these changes. Juvenile Atlantic salmon reared under constant light (LL) from first feeding, were separated into two groups, and exposed to either LL or simulated natural photoperiod (LDN) from October, eight months prior to the expected completion of smoltification. Juveniles reared on LL grew well, but failed to show the smolt-related reduction in condition factor in spring. Gill mRNA levels of Na+, K+-ATPase (NKA) isoform ??1a decreased in LDN fish through completion of parr-smolt transformation, while levels remained unchanged in the LL group. In contrast, ??1b expression increased 6-fold in the LDN group between February and May, again with no change in the LL group. Further, Na+, K+, 2Cl- co-transporter (NKCC) showed a transient increase in expression in smolts on LDN between February and May, while no changes in mRNA levels were seen in juveniles under LL. Consequently, gill NKA activity and NKA ?? and NKCC protein abundance were significantly lower in juveniles on LL than in smolts on LDN. LL fish in spring had lower circulating levels of thyroid hormones (THs), growth hormone (GH) and cortisol. Gill GH-receptor mRNA levels, determined by quantitative PCR, were less than 50% of controls. In contrast, circulating levels of IGF-1 and gill IGF-1 receptor expression, were comparable to controls. Our findings show that continuous light prevents the completion of parr-smolt transformation at a very basic level, disrupting the natural up-regulation of key elements of the endocrine system involved in the regulation of the parr-smolt transformation, and consequently inhibiting the smoltification-related increase in expression, abundance and activity of gill ion transport proteins. ?? 2007 Elsevier B.V. All rights reserved.

Aquaculture

Differential hormonal responses of Atlantic salmon parr and smolt to increased daylength: A possible developmental basis for smolting

In order to elucidate the developmental basis for smolting, Atlantic salmon, Salmo salar, parr (< 11.5??cm) and smolts (> 12.5??cm) were exposed to natural daylength (LDN) and increased daylength (LD16:8) starting in late February and gill Na+,K+-ATPase activity and circulating hormone levels monitored from January to May. Gill Na+,K+-ATPase activity remained low and constant in both groups of parr. In smolts, gill Na+,K+-ATPase began increasing in late February in both photoperiods, but was significantly higher in the LD16:8 group from March through April. Smolts exposed to LD16:8 had dramatically elevated plasma GH within one week of increased daylength that remained high through April, whereas plasma GH of LDN smolts increased steadily beginning in late February and peaking in late April. Plasma GH levels of parr remained low in spring and did not respond to increased daylength. Plasma insulin-like growth factor I (IGF-I) levels were substantially higher in smolts than parr in January. Plasma IGF-I levels of parr increased steadily from January to May, but there was no influence of increased daylength. In smolts, plasma IGF-I of LD16:8 fish initially decreased in early March then increased in late March and April, whereas plasma IGF-I of LDN smolts increased steadily to peak levels in early April. Plasma cortisol was low in parr throughout spring and did not differ between photoperiod treatments. Plasma cortisol of LD16:8 smolts increased in early March and remained elevated through April, whereas in LDN smolts plasma cortisol did not increase until early April and peaked in late April. Plasma thyroid hormones were generally higher in smolts than in parr, but there was no clear effect of increased daylength in parr or smolts. The greater capacity of the GH/IGF-I and cortisol axes to respond to increased daylength may be a critical factor underlying smolt development. ?? 2007 Elsevier B.V. All rights reserved.

Aquaculture

Effects of somatostatin on the growth hormone-insulin-like growth factor axis and seawater adaptation of rainbow trout (Oncorhynchus mykiss)

Growth hormone (GH) has been shown to contribute to the seawater (SW) adaptability of euryhaline fish both directly and indirectly through insulin-like growth factor-1 (IGF-1). This study examined the role of somatostatin-14 (SS-14), a potent inhibitor of GH, on the GH-IGF-1 axis and seawater adaptation. Juvenile rainbow trout (Oncorhynchus mykiss) were injected intraperitoneally with SS-14 or saline and transferred to 20??ppt seawater. A slight elevation in plasma chloride levels was accompanied by significantly reduced gill Na+, K+-ATPase activity in SS-14-treated fish compared to control fish 12??h after SW transfer. Seawater increased hepatic mRNA levels of GH receptor 1 (GHR 1; 239%), GHR 2 (48%), and IGF-1 (103%) in control fish 12??h after transfer. Levels of GHR 1 (155%), GHR 2 (121%), IGF-1 (200%), IGF-1 receptor A (IGFR1A; 62%), and IGFR1B (157%) increased in the gills of control fish 12??h after transfer. SS-14 abolished or attenuated SW-induced changes in the expression of GHR, IGF-1, and IGFR mRNAs in liver and gill. These results indicate that SS-14 reduces seawater adaptability by inhibiting the GH-IGF-1 axis. ?? 2007 Elsevier B.V. All rights reserved.

Aquaculture

Whole-body cortisol response of zebrafish to acute net handling stress

Zebrafish, Danio rerio, are frequently handled during husbandry and experimental procedures in the laboratory, yet little is known about the physiological responses to such stressors. We measured the whole-body cortisol levels of adult zebrafish subjected to net stress and air exposure at intervals over a 24 h period; cortisol recovered to near control levels by about 1 h post-net-stress (PNS). We then measured cortisol at frequent intervals over a 1 h period. Cortisol levels were more than 2-fold higher in net stressed fish at 3 min PNS and continued to increase peaking at 15 min PNS, when cortisol levels were 6-fold greater than the control cortisol. Mean cortisol declined from 15 to 60 min PNS, and at 60 min, net-stressed cortisol was similar to control cortisol. Because the age of fish differed between studies, we examined resting cortisol levels of fish of different ages (3, 7, 13, and 19 months). The resting cortisol values among tanks with the same age fish differed significantly but there was no clear effect of age. Our study is the first to report the response and recovery of cortisol after net handling for laboratory-reared zebrafish. ?? 2009 Elsevier B.V.

Aquaculture

Depletion of florfenicol amine, marker residue of florfenicol, from the edible fillet of tilapia (Oreochromis niloticus x O. niloticus and O. niloticus x O. aureus) following florfenicol administration in feed

Aquaflor ?? , a 50% feed premix containing the broad spectrum antibacterial agent florfenicol is available globally to control mortality associated with economically significant systemic bacterial diseases of fish. Florfenicol (FFC) is effective in controlling mortality associated with Streptococcus iniae in tilapia Oreochromis sp. when administered in medicated feed at a dose of 15 mg/kg bodyweight (BW)/d for 10 consecutive days. Our objective was to characterize the depletion of the FFC marker residue, florfenicol amine (FFA), from the edible tissue of market-weight Nile tilapia O. niloticus x O. niloticus and hybrid tilapia O. niloticus x O. aureus offered feed medicated with FFC at a nominal dose rate of 15 mg/kg BW/d for 12 days. Near market-weight tilapia were obtained from a commercial tilapia farm, distributed to 2 single pass (one for Nile tilapia and one for hybrid tilapia), flow-through systems and maintained at 27 ??C under a 15 h light:9 h dark photoperiod over a 41-d pre-dosing period. During the dosing period, tilapia were offered feed medicated with FFC at a concentration of 1.479 g/kg at 1% BW daily divided in three equal offerings. The initial 10-d dosing period was extended to 12 d because one tank did not consume > 75% of the feed offered during the first two dosing days. The total dose consumed by fish in each of the 2 tanks ranged from 147 to 167 mg/kg. Once during the pre-dose period and on days 1, 2, 4, 7, 14, 21, and 28 of the post-dose period, groups of fish were indiscriminately removed from each tank, measured for weight and length, scaled, filleted, and the skin-on fillets stored at <-70 ??C. Frozen fillets were individually homogenized, extracted, and FFA concentration was determined by high-performance liquid chromatography with UV detection. Florfenicol amine is rapidly eliminated from tilapia fillet after withdrawal from medication and depletion followed first-order kinetics with an estimated half-life of 2.32 d. The FFA tolerance limit, calculated as the 99th percentile of the potential residue level at 95% confidence, had depleted to less than the 1 ??g/g maximum residue level by 6.14 d after the dosing period.

Aquaculture

Gill Na+,K+-ATPase of Atlantic salmon smolts in freshwater is not a predictor of long-term growth in seawater

Gill Na + ,K + -ATPase activity is a widely used measure of osmoregulatory preparedness in salmonid smolts. The degree to which this measure may predict long term performance is uncertain. In order to assess the relationship of this enzyme to long term growth and ion homeostasis, a cohort of Atlantic salmon hatchery smolts was used in a controlled environment with no salinity perturbations. In May 2006, gill Na + ,K + -ATPase activity from 940 individually PIT tagged, Penobscot River smolts (USFWS, Green Lake National Fish Hatchery, Maine, United States) was measured immediately prior to isothermal transfer from freshwater to 32 ppt seawater. From the observed range of activities, individuals were classified as having “low”, “middle”, or “high” enzyme activity levels. Individual size (fork length and mass) was recorded on days 0, 1, 3, and 14 and monthly for four months. Growth rates over four time periods were calculated for individual fish maintained until the end of the experiment. Gill Na + ,K + -ATPase activities were also measured from a subset of sampled fish. All groups effectively osmoregulated as evidenced by minor perturbations in plasma osmolyte levels. Apart from initial weight loss on transfer, fish grew throughout the experiment, however, there were no differences (fish size, growth rate, and gill Na + ,K + -ATPase activity in seawater) among groups with initially different gill Na + ,K + -ATPase activities (prior to seawater entry). While gill Na + ,K + -ATPase activity may be predictive of performance during the acute phase of acclimation (first few days), typical variation in this enzyme, expressed in freshwater at the peak of smolting, does not appear to be predictive of long-term growth in seawater.

Aquaculture

Thresholds for short-term acid and aluminum impacts on Atlantic salmon smolts

Although the negative effects of acid and aluminum (Al) on smolt development have been known for some time, the thresholds for impact of short-term exposure of several days that may occur during episodic acidification have not been systematically examined. In order to determine the levels of acid and Al that impact juvenile Atlantic salmon, smolts and yolk-sac larvae were exposed to three pH levels (6.0, 5.7, and 5.3) and four added Al levels (0, 40, 80 and 175 μg/L total Al) for 48 h. Following this treatment, 10 smolts were sampled in freshwater and another 10 were subjected to a 24 h seawater challenge (35 ppt). Survival of yolk-sac larvae was > 96% in all acid and Al treatments. All smolts died within 48 h at pH 5.3, 175 μg L− 1 Al. There were some mortalities in freshwater at pH 5.3, 80 μg L− 1 Al and pH 5.7, 175 μg L− 1 Al, and further mortalities when these fish were transferred to seawater. Mortalities in these groups were associated with decreased plasma chloride in freshwater and higher plasma chloride in seawater, indicating that these smolts had lost seawater tolerance. Gill Na+/K+-ATPase (NKA) activity decreased at pH 5.7, 175 μg L− 1 Al in freshwater, and further decreases were observed at more moderate pH and Al exposures after transfer to seawater. Hematocrit and plasma glucose were the most sensitive physiological responses, increasing at all Al treatments at pH 5.7 and 5.3 in freshwater. There was no detectable increase in gill Al levels at pH 6.0 with added Al, whereas substantial increases in gill Al were observed in all added Al groups at pH 5.7 and 5.3. Our results demonstrate a critical interaction between acid and Al in their effects on smolts, and that episodic acidification events will negatively impact smolt survival in freshwater and after seawater entry.

Aquaculture

Impacts of multispecies parasitism on juvenile coho salmon (Oncorhynchus kisutch) in Oregon

We are studying the impacts of parasites on threatened stocks of Oregon coastal coho salmon (Oncorhynchus kisutch). In our previous studies, we have found high infections of digeneans and myxozoans in coho salmon parr from the lower main stem of West Fork Smith River (WFSR), Oregon. In contrast parr from tributaries of this river, and outmigrating smolts, harbor considerably less parasites. Thus, we have hypothesized that heavy parasite burdens in parr from this river are associated with poor overwintering survival. The objective of the current study was to ascertain the possible effects these parasites have on smolt fitness. We captured parr from the lower main stem and tributaries of WFSR and held them in the laboratory to evaluate performance endpoints of smolts with varying degrees of infection by three digeneans (Nanophyetus salmincola, Apophallus sp., and neascus) and one myxozoan (Myxobolus insidiosus). The parameters we assessed were weight, fork length, growth, swimming stamina, and gill Na + ,K + -ATPase activity. We repeated our study on the subsequent year class and with hatchery reared coho salmon experimentally infected with N. salmincola. The most significant associations between parasites and these performance or fitness endpoints were observed in the heavily infected groups from both years. We found that all parasite species, except neascus, were negatively associated with fish fitness. This was corroborated for N. salmincola causing reduced growth with our experimental infection study. Parasites were most negatively associated with growth and size, and these parameters likely influenced the secondary findings with swimming stamina and ATPase activity levels.

Oregon

Depletion of eugenol residues from the skin-on fillet tissue of rainbow trout exposed to 14C-labeled eugenol

The U.S. is lagging in access to an approved immediate-release sedative, i.e. a compound that can be safely and effectively used to sedate fish and has no withdrawal period. AQUI-S® 20E (10% active ingredient, eugenol) is under investigation as an immediate-release sedative for freshwater finfish. Because of its investigational status, data are needed to characterize the depletion, distribution, and identity of AQUI-S® 20E residues in fillet tissue. Rainbow trout ( Oncorhynchus mykiss ) were exposed to uniformly ring labeled 14 C-eugenol at a nominal concentration of 10 mg/L for 60 min in 18 °C water. Fish (n = 6) were sampled immediately after the exposure (0 min) then at 30, 60, 120, and 240 min. Eugenol concentrations and characterization of 14 C residues in the fillet tissue were determined by high pressure liquid chromatography and flow-through liquid scintillation counting techniques. Total 14 C-residue burdens in fillet tissue were determined by tissue oxidation and static liquid scintillation counting techniques. Maximum eugenol and 14 C-eugenol equivalent residue concentrations in the fillet tissue were measured immediately after the exposure (44.5 and 38.8 μg/g, respectively). Eugenol was the primary 14 C-residue (> 90% of all 14 C-residues) in extracts from fillet tissue taken from fish sampled immediately after the exposure (0 min) and from fish sampled at 30 and 60 min after the exposure. The depletion of 14 C-eugenol residues from the fillet tissue was rapid (t 1/2 = 26.25 min) after transferring the exposed fish to fresh flowing water.

Aquaculture

Experimental infection of six North American fish species with the North Carolina strain of spring Viremia of Carp Virus

Spring viremia of carp virus (SVCV) is a rhabdoviral pathogen associated with disease outbreaks in cultured and wild fish worldwide. Common carp ( Cyprinus carpio carp ), and koi ( C. carpio koi ) suffer the highest mortalities from SVCV infections, while other cyprinid fish species have varying susceptibility. Although salmonid fish typically are considered refractory to infection by SVCV, there have been a few reports suggesting infection has occurred in rainbow trout ( Oncorhynchus mykiss ). There have been no reports of Percid fish being infected with SVCV. Since the first North American outbreak of SVCV at a North Carolina koi farm in 2002 there have been eight subsequent detections or outbreaks of SVCV among fish species from the families of Cyprinidae and Centrarchidae within the US and Canada. Thus, this exotic virus is considered a potential threat to native and cultured fish populations in North America. We performed multiple experimental challenges with fish species from three families ( Salmonidae , Cyprinidae , and Percidae ) to identify the potential risk associated with SVCV exposure of resident fish populations in North America. Three salmonid species, rainbow and steelhead trout ( Oncorhynchus mykiss ), Chinook salmon ( O. tshawytscha ), and sockeye salmon ( O. nerka ), were challenged by immersion or injection with the North Carolina SVCV isolate. Two cyprinid species, koi and fathead minnow ( Pimephales promelas ) and one percid species, yellow perch ( Perca flavescens ) were also challenged. Koi were highly susceptible to SVCV up to 11 months of age and fathead minnows had chronic disease expression with moderate mortality (29%). SVCV also induced moderate mortalities (33%) in yellow perch fry. Virus challenged salmonid fish had cumulative percent mortalities ranging from 0 to 100%, with sockeye salmon fry being the most vulnerable. A sub-sample of mortalities and survivors were screened for virus by plaque assay and reverse transcription polymerase chain reaction. In general, all mortalities tested positive for SVCV with high viral titers while survivors had variable persistence of SVCV with overall lower virus titers. Our SVCV challenges of multiple North American fish species suggested that host age is a key factor in determining disease outcome. Other factors, such as fish broodstock, virus strain, water temperature, and rearing conditions in association with the intrinsic level of species susceptibility may also impact infection dynamics. This is the first report of SVCV infecting a species (yellow perch) in the family Percidae and that sockeye salmon fry can suffer similarly high mortalities as the primary SVCV host species.

Aquaculture

Successful mitigation of viral disease based on a delayed exposure rearing strategy at a large-scale steelhead trout conservation hatchery

In 2009, the largest steelhead trout conservation hatchery in the state of Idaho, Dworshak National Fish Hatchery (NFH), lost over 50% of the juvenile steelhead trout ( Oncorhynchus mykiss ) population being reared for release. The causative agent of this high mortality was the viral pathogen infectious hematopoietic necrosis virus (IHNV). This was neither the first nor the worst epidemic of IHNV to occur at the hatchery, but it was the worst in over a decade. Genetic analysis of IHNV isolates taken from juveniles suffering epidemic IHN disease in 2009 revealed that the virus was of the M group of IHNV viruses, known to have high virulence for trout. The water supply for steelhead trout rearing at Dworshak NFH is untreated water taken directly from the Clearwater River. Further genetic analysis of IHNV isolates from adults spawned in 2009 indicated that adult steelhead trout in the river (in the hatchery water supply) were the most probable transmission source for the epidemic IHN disease in the juvenile fish. Previously, Dworshak NFH had been able to gain access to reservoir water from behind the Dworshak Dam for nursery egg incubation and the earliest stage of fry rearing, which nearly eliminated incidence of IHN disease in that stage of rearing. Additionally, the nearby Clearwater State Fish Hatchery (SFH), which operates entirely with reservoir water, has never had a case of IHN disease in juvenile steelhead trout. Therefore, staff at Dworshak NFH sought and obtained access to a limited supply of reservoir water for the first few months of outdoor rearing of juvenile steelhead trout, beginning in 2010. This strategy delayed the exposure of juvenile steelhead trout to river water for several months. The effects of this program change were: drastic reduction in IHN disease in juvenile steelhead trout; interruption in the transmission of highly virulent M group IHNV from adult steelhead trout; no interruption in the transmission of low virulent U group IHNV from adult Chinook salmon; and a shift of IHNV types in adult fish spawned at Dworshak NFH in subsequent years from M to U group viruses. While juvenile steelhead trout may still be infected via exposure to IHNV in river water, the disruption of virulent M group IHNV has been successful in dramatically reducing IHN disease in steelhead trout every year since 2010.

Idaho

Pulsed flow-through auto-feeding beaker systems for the laboratory culture of juvenile freshwater mussels

Newly metamorphosed freshwater mussels are small and delicate, so that captive laboratory culture presents challenges for handling; for maintenance of suitable microhabitat, water quality, and food; and for avoidance of competitors and predators. To address these challenges, a new pulsed flow-through auto-feeding beaker system was developed for culturing juvenile mussels. In this system, groups of mussels were maintained in 300- to 1000-mL beakers with a thin layer of sand substrate. The water in the beakers was static except for pulses that were delivered every 1 or 2 h and that displaced about half of the water in each beaker per water cycle. A peristaltic pump delivered food to multiple mixing cells where the water was automatically mixed with food just before the water delivery. In testing this approach, newly metamorphosed mussels of 4 species were cultured in the system for 84 to 357 d. The sand and beakers were replaced weekly. Survival was high (>85% at day 84) for Lampsilis siliquoidea and Villosa iris , but relatively lower for Anodonta californiensis (29% at day 155) and Margaritifera falcata (23% at day 357). Growth rate ranged among the 4 species from 27 to 60 μm/d, with the slowest rate for M. falcata and fastest for A. californiensis. Overall, the new pulsed flow-through auto-feeding beaker system improved survival and growth of juvenile mussels versus other methods previously tested. Additionally, a simplified system for the water and food delivery was developed with a single mixing cell. The use of both systems indicate that they are suitable for laboratory experiments and for captive culture of juvenile mussels.

Aquaculture