Nano-scale applications in aquaculture: Opportunities for improved production and disease control.

Aquaculture is the fastest growing food-production sector and is vital to food security, habitat restoration and endangered species conservation. One of the continued challenges to the industry is our ability to manage aquatic disease agents that can rapidly decimate operations and are a constant threat to sustainability. Such threats also evolve as microbes acquire resistance and/or new pathogens emerge. The advent of nanotechnology has transformed our approach to fisheries disease management with advances in water disinfection, food conversion, fish health and management systems. In this review, several nano-enabled technology successes will be discussed as they relate to the challenges associated with disease management in the aquaculture sector, with a particular focus on fishes. Future perspectives on how nanotechnology can offer functional approaches for improving disinfection and innovating at the practical space of early warning systems will be discussed. Finally, the importance of "safety by design" approaches to the development of novel commercial nano-enabled products will be emphasized.

Test-Retest Reliability of a Food Frequency Questionnaire to Assess Seafood Intake Dynamics for High-End Consumers in Coastal Gulf of Mexico Communities.

Background: Estimates for fish and shellfish intake are used to inform communities and healthcare systems about potential health risks and benefits for individuals, communities, and vulnerable populations. A dietary assessment instrument was designed for use in populations of high-end consumers of seafood to examine intake of finfish, shrimp, oysters, and blue crab in coastal communities across the Gulf of Mexico. Objective: To validate the reliability of a novel food frequency questionnaire (FFQ) for seafood intake. Design: Test-retest reliability of the FFQ, which included a species-specific photographic portion guide, was evaluated by the inperson administration and readministration of the instrument with each participant by the same interviewer. Responses from coastal and noncoastal participants were compared to discern FFQ reliability in heterogeneous samples. Participants/setting. A convenience sample of 27 coastal participants from Cedar Key, Steinhatchee, and Apalachicola, Florida, reported data for 101 household members; and 15 noncoastal participants from Gainesville, Florida, reported for 42 household members. Analysis. Repeated measures from the FFQ were evaluated using correlation concordance for continuous variables (age, weight, and height) and kappa coefficient for categorical variables (type, amount, and frequency of seafood consumed). Results: Concordance correlation coefficient (1.00) and kappa coefficient (r = 0.73 to 1.00) for yearly and seasonal seafood consumption indicated substantial to almost perfect reproducibility, i.e., participants provided responses that were reproducible. Test-retest agreement was highest for coastal participants who consumed more seafood, as compared to occasional, noncoastal consumers, based on the intergroup comparison of kappa coefficients for yearly and seasonal seafood consumption (r = 0.69 to 0.99). Conclusions: The seafood FFQ instrument evaluated in this study, included as a supplement to this report, used in tandem with a photographic portion guide, provides a utilitarian tool for assessing fish, shrimp, oyster, and blue crab intake dynamics in adult and youth populations drawn from coastal communities.

Probabilistic risk assessment for high-end consumers of seafood on the northeastern Gulf coast.

The Deepwater Horizon oil spill (April 20, 2010) caused concern regarding Gulf seafood safety. Communities were skeptical of governmental risk assessments because they did not take into account the higher consumption of seafood along coastal areas. The objective of this study was to perform a probabilistic risk assessment based on the consumption rates of high-end consumers of Gulf seafood. We utilized seafood consumption data from five communities across the northeastern Gulf of Mexico. This study collected finfish, shrimp, blue crab, and oysters from these communities and analyzed their tissues for polynuclear aromatic hydrocarbons (PAHs). A probabilistic risk assessment was performed using population-specific seafood consumption rates and body weights for commercial fishers, recreational fishers, and a Filipino-American community. For non-cancer effects, 95th percentile hazard quotients for these targeted populations ranged between 1.84E-04 to 5.39E-03 for individual seafood types. The 95th percentile hazard indices for total seafood consumption ranged from 3.45E-03 to 8.41E-03. Based on total seafood consumption, highest hazard indices were modeled for the Filipino-American community followed by commercial and recreational fishers. Despite higher consumption rates, hazard indices for the high-end consumers targeted in this study were two to three orders of magnitude below the regulatory limit of 1.

Examining the responses of the zebrafish (Danio rerio) gastrointestinal system to the suspected obesogen diethylhexyl phthalate.

Epidemiological evidence suggests that phthalate plasticizers may act as "obesogens", which are chemicals that exacerbate obesity. The gastrointestinal (GI) system is the primary exposure route for phthalates, however, the relationship between phthalate-driven perturbations of GI system functions that can influence obesity has yet to be examined. To address this knowledge gap, we exposed Danio rerio (zebrafish) for 60 days to either (1) Control feeding (5 mg/fish/day), (2) Overfeeding (20 mg/fish/day) or (3) Overfeeding with diethyl-hexyl phthalate (DEHP) (20 mg/fish/day with 3 mg/kg DEHP). After 60 days, Overfed and Overfed + DEHP zebrafish had elevated body mass, and hepatosomatic and gonadosomatic indices. RNAseq analysis of the GI revealed enrichment of gene networks related to lipid metabolism in the Overfed + DEHP group. Many of the enriched networks were under transcriptional control of peroxisome proliferator activated receptor alpha (pparα), a known modulator of lipid metabolism, immune function, and GI function. Real-time PCR confirmed that pparα was overexpressed in the Overfed + DEHP zebrafish, further revealing a pathway by which DEHP may influence lipid metabolism via the GI. These data increase our understanding of phthalate-driven effects on GI function and lipid metabolism, identifying gut-specific gene networks that may drive phthalate-exacerbated obesity.

Validation of a photographic seafood portion guide to assess fish and shrimp intakes.

OBJECTIVE: To validate a novel photographic portion guide as a tool to estimate consumption of fish and shrimp. Application of such a validated tool can facilitate accurate individual and community seafood intake assessments and provide meaningful data relative to health benefits and hazard assessment, particularly in response to environmental contamination and disasters. DESIGN: A photographic fish and shrimp portion guide presenting a stepped range of cooked portion sizes was used by participants to estimate their typical portion sizes. Participants selected their typical portion size from the photographic guide and also from a selection of freshly cooked reference meals. Photographic portions selections were compared with plated reference portions for each participant. SETTING: Academic sensory testing laboratory in the USA. SUBJECTS: Separate groups of adults (25-64 years) contributed to fish (n 54) and shrimp (n 53) portion size comparison studies. RESULTS: In the fish study, there was no difference between photographic portion selections (6·59 (sd 2·65) oz (186·8 (sd 75·1) g)) and reference plate selections (7·04 (sd 2·63) oz (199·6 (sd 74·6) g); P=0·384). Similarly in the shrimp study, there was no difference between photographic portion selections (6·88 (sd 3·40) oz (195·0 (sd 96·4) g)) and reference plate selections (6·06 (sd 2·65) oz (171·8 (sd 75·1) g); P=0·159). Photographic portions predicted plated reference portions for both fish and shrimp based on linear regression (P<0·001). Bland-Altman plot analyses showed good agreement between the two methods, <1 oz (<28·3 g) bias, in both fish and shrimp studies. CONCLUSIONS: This validated photographic seafood portion guide provides a utilitarian tool for accurately assessing fish and shrimp intake in a community setting.

A multi-year study of hepatic biomarkers in coastal fishes from the Gulf of Mexico after the Deepwater Horizon Oil Spill.

Following the 2010 Gulf of Mexico oil spill, concerns were raised regarding exposure of fish to crude oil components, particularly polycyclic aromatic hydrocarbons (PAHs). This three year study examined hepatic enzymes in post-mitochondrial supernatant fractions from red snapper (Lutjanus campechanus) and gray triggerfish (Balistes capriscus) collected in the north central Gulf of Mexico between 2011 and 2014. Biomarker activities evaluated included benzo(a)pyrene hydroxylase (AHH), ethoxyresorufin O-deethylase (EROD), glutathione transferase (GST), and glutathione peroxidase (GPx). Mean EROD activity was higher in gray triggerfish (12.97 ± 7.15 pmol/min/mg protein [mean ± SD], n = 115) than red snapper (2.75 ± 1.92 pmol/min/mg protein, n = 194), p < 0.0001. In both species, EROD declined over time between 2011 and 2014. Declines in GST and GPx activities were also noted over this time period for both species. Gray triggerfish liver was fatty, and heptane extracts of the liver fat contained fluorescent substances with properties similar to known PAHs, however the origin of these PAHs is unknown.

Tracking and quantification of single-walled carbon nanotubes in fish using near infrared fluorescence.

Detection of SWCNTs in complex matrices presents a unique challenge as common techniques lack spatial resolution and specificity. Near infrared fluorescence (NIRF) has emerged as a valuable tool for detecting and quantifying SWCNTs in environmental samples by exploiting their innate fluorescent properties. The objective of this study was to optimize NIRF-based imaging and quantitation methods for tracking and quantifying SWCNTs in an aquatic vertebrate model in conjunction with assessing toxicological end points. Fathead minnows (Pimephales promelas) were exposed by single gavage to SWCNTs and their distribution was tracked using a custom NIRF imaging system for 7 days. No overt toxicity was observed in any of the SWCNT treated fish; however, histopathology observations from gastrointestinal (GI) tissue revealed edema within the submucosa and altered mucous cell morphology. NIRF images showed strong SWCNT-derived fluorescence signals in whole fish and excised intestinal tissues. Fluorescence was not detected in other tissues examined, indicating that no appreciable intestinal absorption occurred. SWCNTs were quantified in intestinal tissues using a NIRF spectroscopic method revealing values that were consistent with the pattern of fluorescence observed with NIRF imaging. Results of this work demonstrate the utility of NIRF imaging as a valuable tool for examining uptake and distribution of SWCNTs in aquatic vertebrates.

Chronic nanoparticulate silver exposure results in tissue accumulation and transcriptomic changes in zebrafish.

Increasing utilization of metallic nanomaterials in recent years implies an increasing rate of release to the environment, with potentially serious adverse effects on environmentally important species. Previously, we demonstrated that exposure to nanoparticulate silver for 24-48 h results in dramatic alterations in global gene expression patterns and increased tissue burdens in zebrafish gills. The present study reports outcomes associated with chronic exposure to nanoparticulate silver in zebrafish. Adult female Danio rerio were exposed to 5, 15, 25, or 50 µg/L nanoparticulate silver in a time course up to 28 days. A soluble silver treatment (5 µg/L) was also included. Results indicate that use of flow-through systems for chronic nanometal studies is a viable concept; measured concentrations of approximately 60% of nominal values over the course of the 28-day exposure were observed. Dissolution of nanoparticulate silver was measured twice weekly throughout the exposure ranging between 0.5 and 1.0 µg/L, and was relatively consistent between nanoparticulate silver tanks, with no differences between treatments. Gill samples from the 28-day time point were analyzed for global gene expression patterns and histopathology. Tissue accumulation in both gill and eviscerated carcass was dose-dependent, and remained elevated 4 days after the silver was removed. Microarray analysis also revealed a dose-dependent response pattern, with the largest number of genes affected in the 50 µg/L AgNP exposure. Pathway analysis of affected genes identified a number of GO terms that were significantly over-represented in the high AgNP dataset. These terms are associated with DNA damage repair, cellular restructuring, and developmental processes.

Oxytetracycline pharmacokinetics in rainbow trout during and after an orally administered medicated feed regimen.

The pharmacokinetic-pharmacodynamic predictor of antimicrobial activity for tetracyclines is reported to be the area under the concentration-time curve at steady state (AUC(ss)) divided by the minimal inhibitory concentration of the targeted pathogen. Here, we estimate AUC(ss) values for oxytetracycline (OTC) in serum of rainbow trout Oncorhynchus mykiss by using a destructive sampling study design. Seventy-two rainbow trout were fed OTC-medicated feed at 74.7 +/- 1.5 mg/kg (mean +/- SD) body weight (BW) by oral gavage for 10 consecutive days. Serum was collected from nine fish at 1, 3, 6, 8, 10, 12, 15, and 22 d after dosing began. Serum OTC concentrations were measured by high-performance liquid chromatography with a 0.01-microg/mL limit of detection. The average OTC AUC(ss) was 29.2 microg x h/mL and was estimated using nonlinear mixed-effects modeling and bootstrap resampling techniques. The elimination half-life was estimated as 85.0 h, and the fraction of steady state achieved was estimated as 0.85. The calculated AUC(ss) (24.8 microg x h/mL) following 10 d of oral dosing with 75 mg OTC/kg BW was less than the estimated AUC(ss). Results suggest that the pharmacokinetics of OTC exposure, including the AUC(ss), is better evaluated by using multiday dosimetry than by using a standard single-dose protocol.

Poultry litter-induced endocrine disruption in fathead minnow, sheepshead minnow, and mummichog laboratory exposures.

Animal feeding operations in the United States produce more than 500 million tons of manure annually. Disposal of poultry waste via application as fertilizer results in substantial runoff of poultry litter-associated contaminants (PLAC). Of particular concern are sex steroids, 17ß-estradiol, estrone and testosterone, responsible for sex differentiation and development of reproductive structures. In a series of laboratory assays, mature male and mixed-sex larval/juvenile fish were continuously exposed to environmentally relevant PLAC solutions. Effects on gonads were assessed histologically, and vitellogenin (VTG) induction was measured as a gauge of estrogenicity. Twenty-one-day exposures to laboratory-generated PLAC solutions routinely induced VTG in mature male Pimephales promelas. Vitellogenesis in Fundulus heteroclitus only occurred at the highest tested PLAC concentration, and Cyprinodon variegatus were unresponsive at any tested concentration. All species produced considerable VTG in response to a 17ß-estradiol-positive control. A pronounced feminization was seen in P. promelas when exposed to PLAC as larvae but not when exposed as juveniles. Runoff from a poultry litter-amended field cropped under standard agronomic practices induced significant VTG in male P. promelas. Results indicate that environmentally relevant PLAC concentrations exhibit endocrine activity sufficient to induce VTG production in male fish and possibly affect sex ratios in resident fish populations.

17alpha-ethinylestradiol alters reproductive behaviors, circulating hormones, and sexual morphology in male fathead minnows (Pimephales promelas).

Ecologically relevant indicators of endocrine disruption in fish must be linked with measures of reproductive success. The ability of male fathead minnows (Pimephales promelas) to compete for, maintain, and defend a spawning substrate is paramount to reproductive success. The present study quantified alterations in male fathead minnow reproductive behaviors after exposure to environmentally relevant concentrations (0, 10, 20, or 40 ng/L) of 17alpha-ethinylestradiol (EE2) for 21 d. A video-based behavioral quantification system examined changes in male-male competitive behaviors (chasing and head-butting) and ability of males to maintain spawning substrates (nibbling and scrubbing). Behaviors analyzed included time under the spawning substrate, frequency of substrate cleaning, and conspecific aggression. Plasma hormone levels (11-ketotestosterone [11-KT], testosterone, and estradiol [E2]), vitellogenin (VTG), secondary male characteristics (tubercle count and dorsal nape pad rank), gonadosomatic index (GSI), and gonad histology also were evaluated. Exposure to 40 ng/L of EE2 decreased the ability of exposed males to compete with control males for spawning substrates (p = 0.09). Furthermore, exposed males displayed reduced frequency of substrate cleaning activities as well as chasing male competitors (p < or = 0.05). 11-Ketotestosterone, testosterone, and E2 were lower, and VTG was notably higher, in EE2-exposed males compared with control males (p < or = 0.03). 17alpha-Ethinylestradiol exposure in males also was associated with reductions in tubercles; lower GSI, gonadal maturity ranks, and number of resorbed tubercles; and presence of an ovipositor (p < or = 0.001). These data reveal alterations in male reproductive behavior that coincide with decreased hormone levels and secondary sex characteristics. Behavioral endpoints to discern potential ecological consequences in fish exposed to low concentrations of endocrine-disrupting chemicals may provide sensitive and functional indices of effect.

Kinetics of hepatic phase I and II biotransformation reactions in eight finfish species.

Hepatic microsomes and cytosols of channel catfish (Ictalurus punctatus), rainbow trout (Oncorhynchus mykiss), Atlantic salmon (Salmo salar), red tilapia (Oreochromis sp.), largemouth bass (Micropterussalmoides), striped bass (Morone saxatilis), hybrid striped bass (M. saxatilis x M. crysops), and bluegill (Lepomis macrochuris) (n=8) were used to study the kinetics of phase I (ECOD, EROD, PROD, BROD) and phase II (UDP-glucuronosyltransferase (UDPGT)-, sulfotransferase (ST)- and glutathione-s-transferase (GST)-mediated) reactions. The best catalytic efficiency for ECOD and GST activities was performed by channel catfish, Atlantic salmon, rainbow trout and tilapia. The highest EROD catalytic efficiency was for Atlantic salmon. None of the species had either PROD or BROD activities. Rainbow trout had very similar UDPGT catalytic efficiency to tilapia, channel catfish, Atlantic salmon, largemouth bass and bluegill. Sulfotransferase conjugation had no significant differences among the species. In summary, tilapia, channel catfish, Atlantic salmon and rainbow trout had the best biotransforming capabilities; striped bass, hybrid striped bass and bluegill were low metabolizers and largemouth bass shared some capabilities with both groups.

Epidemiologic analysis of factors associated with local disappearances of native ranid frogs in Arizona.

We examined factors that may independently or synergistically contribute to amphibian population declines. We used epidemiologic case-control methodology to sample and analyze a large database developed and maintained by the Arizona Game and Fish Department that describes historical and currently known ranid frog localities in Arizona, U.S.A. Sites with historical documentation of target ranid species (n= 324) were evaluated to identify locations where frogs had disappeared during the study period (case sites) and locations where frog populations persisted (control sites). Between 1986 and 2003, 117 (36%) of the 324 sites became case sites, of which 105 were used in the analyses. An equal number of control sites were sampled to control for the effects of time. Risk factors, or predictor variables, were defined from environmental data summarized during site surveys and geographic information system data layers. We evaluated risk factors with univariate and multifactorial logistic-regression analyses to derive odds ratios (OR). Odds for local population disappearance were significantly related to 4 factors in the multifactorial model. Disappearance of frog populations increased with increasing elevation (OR = 2.7 for every 500 m, p < 0.01). Sites where disappearances occurred were 4.3 times more likely to have other nearby sites that also experienced disappearances (OR = 4.3, p < 0.01), whereas the odds of disappearance were 6.7 times less (OR = 0.15, p < 0.01) when there was a source population nearby. Sites with disappearances were 2.6 times more likely to have introduced crayfish than were control sites (OR = 2.6, p= 0.04). The identification of factors associated with frog disappearances increases understanding of declines occurring in natural populations and aids in conservation efforts to reestablish and protect native ranids by identifying and prioritizing implicated threats.

LOCALIZATION OF OXYTETRACYCLINE IN CHLAMYDOMONAS REINHARDTII (CHLOROPHYCEAE)(1).

Oxytetracycline (OTC) is an important antimicrobial used in aquaculture. However, residues of OTC have been isolated from nontarget aquatic organisms, sediments, and water located near aquaculture facilities. Identifying OTC in plant material is particularly difficult due to interference from pigments and polyphenol substances but is important especially for algae since they are a primary food source for fish in early life stages. In this study, we describe the effect of OTC (0.1, 1, 10, 25, 50, 100 µg · mL(-1) ) on cell growth, and the localization of OTC (0, 1, 25, 100 µg · mL(-1) ) in vacuoles of Chlamydomonas reinhardtii P. A. Dang. (wildtype, ATCC 18798). We also present a method for semiquantifying OTC in living cells using fluorescent microscopy and Adobe Photoshop. We exposed algal cells to OTC and sampled after 2 or 7 d exposure. On day 7, OTC significantly inhibited algal growth at 1, 10, 25, 50, and 100 µg · mL(-1) . When viewed with fluorescent microscopy, cells exposed to the 25 and 100 µg · mL(-1) contained yellow fluorescent areas, ≤1 µm in diameter that were easily discernable against the red fluorescence of the intracellular chl. The fluorescent areas corresponded to small spherical vacuoles (i.e., polyphosphate bodies that contain calcium and magnesium complexed with polyphosphate) seen in the cells by LM. Since OTC has a high affinity for divalent cations, we suggest that OTC is localized in these vacuoles.

Mycobacteria as environmental portent in Chesapeake Bay fish species.

Infection with environmental mycobacteria is increasing among many Chesapeake Bay fish species. Prevalence in juvenile Atlantic menhaden differed between tributaries and ranged from 2% to 57%. Mycobacterial infection may be a syndromic sentinel of altered environmental conditions that threaten aquatic animal health.

Sex differences in neuronal morphology in the killifish hypothalamus.

This study examined the neuroarchitecture of the male and female killifish (Fundulus heteroclitus) hypothalamus to evaluate whether sexual dimorphism of this brain region exists in fishes as it does in mammals and other vertebrates. The rostral medulla, a brain region distinct from the hypothalamic-pituitary-gonadal axis, was also examined to determine if any observed differences were region-specific. With the use of Golgi-Cox impregnation, five dendritic characteristics were measured from neurons of both the hypothalamus and medulla including: spine density, number of branch points, dendrite length, surface area and volume. Dendritic spines are associated with excitatory synapses, and changes in density are associated with a variety of normal and pathological changes. Consistent with mammalian studies, we found that adult female killifish have 25% greater dendritic spine densities in the hypothalamus than male killifish (densities of 0.34+/-0.06 microm-1 and 0.25+/-0.08 microm-1, respectively). By contrast, no statistically significant difference between males and females was detected in spine densities in the rostral medulla. This finding supports the conclusion that hypothalamic sexual dimorphism is conserved in killifish.

A video-based movement analysis system to quantify behavioral stress responses of fish.

Behavioral alterations can be measured as endpoints for sublethal toxicity, and serve as a tool for environmental risk assessment and analysis of toxicological impact. Numerous technical and biological factors have made sublethal effects on fish behavior difficult to quantify. In order to investigate stress- and contaminant-induced behavioral alterations, a video analysis system was designed by our laboratory. With this system up to 12 fish may be individually housed in 20 L exposure arenas and automatically videotaped at multiple and discrete intervals during an experimental period. Analog video data can then digitized, converted into x,y coordinates, and finally transformed into relevant behavioral endpoints using software designed for tracking fish movement combined with specific algorithms. These endpoints include velocity, total distance traveled, angular change, percent movement, space utilization, and fractal dimension (path complexity). Data from fish exposed to a reference toxicant, MS222, and simulation experiments, are presented to exemplify alterations in fish behavior associated with exposure, and accuracy and precision, respectively. The system provides flexibility to analyze any observed movement behavior, is remotely controlled, and can be transportable. These movement analyses can be used to identify characteristic behavioral responses to a variety of environmentally-relevant stressors, and assist in risk assessment and the development of more sensitive lowest observable effect level and no observable effect level for sentinel species.

Acoustical stress and hearing sensitivity in fishes: does the linear threshold shift hypothesis hold water?

Mammals exposed to loud aerial sounds exhibit temporary threshold shifts (TTS) that are linearly related to increases of sound pressure above baseline hearing levels. It was unknown if this relationship held true for aquatic ectotherms such as fishes. To test this linear threshold shift hypothesis (LINTS) in fishes, we examined the effects of increased ambient sound on hearing of two species differing in hearing capabilities: goldfish (Carassius auratus; a hearing specialist) and tilapia (Oreochromis niloticus; a hearing generalist). Fish were exposed to 1-28 days of either quiet (110 dB re 1 microPa) or continuous white noise. First, we examined the effect of noise sound pressure level (SPL; 130, 140, 160 or 170 dB re 1 microPa) on goldfish hearing thresholds after 24 h of noise exposure. Second, in a long-term experiment using 170 dB re 1 microPa white noise, we continuously exposed goldfish and tilapia for either 7 or 21-28 days. In both experiments, we measured alterations in hearing capabilities (using auditory brainstem responses) of noise-exposed fish. While tilapia exposed to noise for 28 days showed little or no hearing loss, goldfish exhibited considerable threshold shifts that reached an asymptote of up to 25 dB after only 24 h of exposure. There was a positive linear relationship between noise-induced TTS and the sound pressure difference between the noise and the baseline hearing thresholds in goldfish but not in tilapia. A similar relationship was found for published noise-induced threshold shifts in birds and mammals, but the slope of the linear relationship was greater in these groups than for fish. The linear threshold shift relationship provides insights into differential susceptibility of hearing specialist and generalist fishes to noise-induced hearing loss for a given SPL and provides a framework for future research on noise-induced threshold shifts in fishes and other animals.

Morphometry of hepatic neoplasms and altered foci in the mummichog, Fundulus heteroclitus.

The goal of this study was to intensively sample a small number of livers from a population of mummichog exposed to PAH-contaminated sediments and evaluate them for lesion pathology, distribution, shape, and volume, and the number of histological sections needed to adequately describe the extent of various lesions. Volumetric data for each lesion type from each step section was derived from digitized section images. The total number of hepatic alterations ranged from 10-125 per fish. Alterations included: eosinophilic, basophilic, and clear cell foci; hepatocellular carcinomas; hemangiopericytomas; and cholangiomas. Lesion volumes ranged from 0.00012-64 mm3 and represented 0.21%-67% of total liver volume. There was a tendency for the lesions to be more dorsal-ventrally compressed than spherical or ropelike when observed from longitudinal sections. Periodic subsampling of the data indicated that. on average, 6 evenly spaced, longitudinal histological sections were required to accurately estimate lesion volume and extent in our model population. These data provide a formulation for histological sampling techniques and methodological support for piscine and other cancer study models that observe lesion volume changes over time. Further, this study fosters the development of early quantitative endpoints. rather than using a large number of animals and waiting for tumor progression or death to occur.

Altered thyroid status in lake trout (Salvelinus namaycush) exposed to co-planar 3,3',4,4',5-pentachlorobiphenyl.

Recent studies indicate that co-planar 3,3',4,4',5-pentachlorobiphenyl (PCB) congeners or their metabolites may disrupt thyroid function in fishes. Although co-planar PCB have been detected at microgram per kilogram levels in fish from contaminated areas, few studies have examined mechanisms whereby, co-planar PCBs may alter thyroid function in fish. We treated immature lake trout by intraperitoneal (i.p.)-injection or dietary gavage with vehicle containing 0, 0.7, 1.2, 25 or 40 microg 3,3',4,4',5-pentachlorobiphenyl (PCB 126) per kgBW. Blood and tissue samples were collected at various times up to 61 weeks following exposure. The treatments produced sustained dose-dependent elevations of tissue (PCB 126) concentrations. Thyroid epithelial cell height (TECH), plasma thyroxine (T4) and 3,3',5-triiodo-l-thyronine (T3) concentrations, hepatic 5'-monodeiodinase, hepatic glucuronidation of T4 and T3, as well as plasma T4 kinetics and fish growth were analyzed. Exposure to the highest doses of PCB 126 caused increased TECH, plasma T4 dynamics and T4-glucuronidation (T4-G). PCB 126 did not affect 5'-monodeiodinase and T3-glucuronidation (T3-G) and there were no effects on fish growth or condition. Because T3 status and growth were unaffected, the thyroid system was able to compensate for the alterations caused by the PCB 126 exposure. It is clear that concentrations of co-planar PCBs similar to those found in predatory fish from contaminated areas in the Great Lakes are capable of enhancing metabolism of T4. These changes may be of significance when T4 requirements are high for other reasons (e.g. periods of rapid growth, warm temperatures, metamorphosis, and parr-smolt transformation).