Environmental contamination can negatively impact fish populations. In addition to acute toxicity leading to death, toxicants can reduce fish growth and lower reproduction. The potential for adverse population level effects of environmental contaminants are estimated to conduct risk assessments from laboratory toxicity tests that most often measure apical endpoints related to growth, survival and reproduction. The relationships between these effect endpoints are being evaluated to predict shifts in fish population demography better after exposure to environmental toxicants. Environmental contaminants can also affect fish populations indirectly by reducing prey biomass. However, estimating the magnitude of the combined effects of prey reduction and direct toxicity is difficult and rarely attempted. Here we describe a toxicity test designed to estimate the effect on Japanese medaka of both reduced food and chronic exposure to diazinon, an acetylcholinesterase inhibiting organophosphate pesticide. Fish were reared with limited food ration and/or diazinon exposure through a full life cycle to assess possible interactions between the two stressors in their effects on growth and reproduction. Diazinon exposure (10 or 20 μg/L), reduced ration (50% and 25% of ad libitum), or combinations of both lowered growth rates and reproductive output of Japanese medaka. In addition, growth and reproduction alone were modeled, and then various relationships between the two stressors (diazinon and ration) and how they relate to growth and reproduction were modeled.

The Medaka Extended One Generation Reproduction Test (MEOGRT) is a Tier 2 test within U.S. Environmental Protection Agency's (USEPA) Endocrine Disruptor Screening Program (EDSP), designed to characterize the potential adverse effects to fish of exposure to chemical that can cause disruption of the endocrine system. The MEOGRT focuses primarily on adverse effects to reproduction while collecting information regarding effects on growth, survival, and endocrine-related endpoints. However, the risk assessment process for fish, as mandated by legislation such as the Federal Insecticide, Fungicide, and Rodenticide Act (FIFRA) or the Toxic Substances Control Act (TSCA), could benefit from a more detailed assessment of effects on growth. Typically, fish growth data in support of risk assessment are obtained from full life-cycle tests or early life stage tests using the fathead minnow. As an alternative to these tests, a modified MEOGRT was conducted to assess the effects of diazinon on the various parameters measured in the MEOGRT. Diazinon is an organophosphate insecticide that is detected in the environment, and whose efficacy is a result of inhibition of the acetylcholine esterase enzyme at neuromuscular junctions and synapses of the nervous system. Diazinon (2.9, 5.2, 10.3, 19.8, and 40.2 μg/L) was tested with the MEOGRT protocol, and the lowest observable effect concentrations of 2.9 μg/L for fecundity and 5.2 μg/L for growth were determined. Additional growth measurements were added to the MEOGRT protocol to more robustly define growth rates and to determine the impact size has on reproductive performance. Fish size starting at the first measurement day (i.e. 21 days post-fertilization), and continuing through the duration of the test was reduced with exposure to 5.2 μg/L and higher, and asymptotic size predicted from growth modeling was reduced at 10.3 μg/L and higher. By simply adding non-destructive growth measurements at two additional time points, the MEOGRT provided enough data for the parameterization of growth models, which could be used to characterize the reproductive implications of growth impairment.

In response to various legislative mandates, the US Environmental Protection Agency (USEPA) formed its Endocrine Disruptor Screening Program (EDSP), which in turn, formed the basis of a tiered testing strategy to determine the potential of pesticides, commercial chemicals, and environmental contaminants to disrupt the endocrine system. The first tier of tests is intended to detect the potential for endocrine disruption mediated through estrogen, androgen, or thyroid pathways, whereas the second tier is intended to further characterize the effects on these pathways and to establish a dose-response relationship for adverse effects. One of these tier 2 tests, the Medaka Extended One Generation Reproduction Test (MEOGRT), was developed by the USEPA for the EDSP and, in collaboration with the Japanese Ministry of the Environment, for the Guidelines for the Testing of Chemicals of the Organisation for Economic Co-operation and Development (OECD). The MEOGRT protocol was iteratively modified based on knowledge gained after the successful completion of 9 tests with variations in test protocols. The present study describes both the final MEOGRT protocol that has been published by the USEPA and the OECD, and the iterations that provided valuable insights into nuances of the protocol. The various tests include exposure to 17β-estradiol, 4-t-octylphenol, o,p'- dichlorodiphenyltrichloroethane, 4-chloro-3-methylphenol, tamoxifen, 17β-trenbolone, vinclozolin, and prochloraz. Environ Toxicol Chem 2017;36:3387-3403. Published 2017 Wiley Periodicals Inc. on behalf of SETAC. This article is a US government work and, as such, is in the public domain in the United States of America.

1,1-Dichloropropene (1,1-DCP) is a contaminant present in both ground and surface waters used as sources for drinking water. Structural similarity to several compounds with known mutagenicity and carcinogenicity, and recent demonstration of mutagenicity in vitro, suggest this compound may be similarly mutagenic in vivo. A transgenic fish model, the lamda transgenic medaka, was used to evaluate the potential mutagenicity of this contaminant in vivo following sub-chronic exposure for 6 weeks. Mutant frequencies of the cII target gene (MF) increased six-fold in the livers of fish exposed to the lowest 1,1-DCP exposure concentration (0.44 mg/L, MF = 18.4 x 10(-5), and increased with each treatment, culminating in a 32-fold induction in fish from the highest 1,1-DCP treatment (16.60 mg/L, MF = 96.3 x 10(-5). Mutations recovered from treated fish showed a distinctive mutational spectrum comprised predominantly of +1 frameshift mutations, induced 166-fold above that of untreated animals. The majority of frameshifts were +1 insertions at thiamine and adenine. These results represent the first evidence of mutagenicity of 1,1-DCP in vivo, and of the highly characteristic spectrum of induced mutations dominated by +1 frameshift mutations. Based upon results from previous in vitro studies, the similar role of glutathione S-transferase (GSTT1-1) in the activation of 1,1-DCP to a mutagen in vivo is also suggested. This study further illustrates the utility of the lamda transgenic medaka as a model for identifying and characterizing potential genetic health risks associated with chemical exposures in the environment.

A physiologically based toxicokinetic (PBTK) model was devel- oped to describe dietary uptake of hydrophobic organic com- pounds by fish. The gastrointestinal (GI) tract was modeled using four compartments corresponding to the stomach, pyloric ceca, upper intestine, and lower intestine, and the lumenal volume of each compartment was allowed to change in time as a function of bulk flow down the GI tract and (for the pyloric ceca and upper intestine) nutrient uptake. The model was developed using data from rainbow trout that were fed a single meal of 60-day-old fathead minnows contaminated with (UL- 14 C) 2,2,5,5-tetrachlo- robiphenyl (( 14 C) PCB 52). Chemical partitioning coefficients for the gut contents and tissues were adjusted to account for changes in chemical affinity associated with uptake of dietary lipid. Per- meability constants for the absorbing gut segments were then fitted by modeling to measured ( 14 C) PCB 52 concentrations in gut

The disposition of [UL-(14)C]2,2',5,5'-tetrachlorobiphenyl (TCB) in rainbow trout (Oncorhynchus mykiss) was studied in acute dietary exposures using TCB-contaminated fathead minnows (Pimephales promelas). Trout were sampled at several postfeeding time points and TCB-derived radioactivity was measured in gut contents and selected tissues. Gastric evacuation was exponential with time and was 95% complete within 36 h of feeding. The ratio of activity in upper intestinal tissue to that in blood declined between 6 and 48 h, as did the lumenal contents/tissue ratio. Stomach content lipid declined between 0 and 24 h, while the lipid content of chyme remained relatively constant. These observations are consistent with liquid phase emptying of lipid and TCB to the upper intestine followed by rapid coassimilation. Tissue/blood activity ratios for the stomach, lower intestine, muscle, liver, and kidney were constant and probably represented near equilibrium conditions. The fat/blood activity ratio increased through 96 h, indicating that TCB was redistributing to fat. The lower intestinal tissue/feces activity ratio increased between 6 and 24 h and then declined rapidly. Fecal lipid content also increased between 6 and 24 h, but the amount of this increase was insufficient to explain observed changes in the distribution of TCB-derived activity. A small amount of 3-hydroxy TCB was detected in feces. Generally, however, metabolism had little or no impact on the uptake, distribution or elimination of TCB. Measured assimilation efficiencies exceeded 90% and are the highest ever reported in fish feeding studies with TCB.

A 10-day bioassay with larval chironomids (Chironomus tentans) was used to evaluate sediment samples from harbors at Michigan City, IN, St. Joseph, MI, Grand Haven, MI, and Toledo, OH for toxicity, based upon the endpoints of survival, dry weight, and growth. Larval responses in sediment samples from each harbor were compared to responses of larvae in reference sediments collected from or near each harbor. An inverse relationship between the number of survivors and mean organism dry weight or growth indicated that food was limiting in the bioassay for some samples. The confounding nature of this interaction was minimized by evaluating effects on the basis of total biomass per replicate. A site from Toledo Harbor was the most toxic. Reduced larval growth at this site indicated the likelihood for a negative populational impact. The importance of reference sediment selection was noted, as the assessment of sediment quality varied considerably for Toledo Harbor depending upon the particular reference sediment that was used for statistical comparisons.

Acid-volatile sulfide (AVS) has been proposed as the primary normalization phase for the development of sediment quality criteria for certain cationic metals. This study was designed to assist in this development by providing necessary field data on the relationships among season, AVS concentrations, and zinc bioavailability and toxicity in freshwater sediments. Zinc was spiked into uncontaminated sediments collected from a local pond, creating five simultaneously extracted metal (SEM) concentrations ranging from 0.8 to 12.0 mu mol/g dry weight. The spiked sediments were transferred to 4-L plastic trays, returned to the bottom of the pond, and sampled on five dates during 1993-1994. Results revealed a pronounced increase in AVS concentration with increasing zinc concentration. Acid-volatile sulfide concentrations in zinc-spiked sediments displayed only minor seasonal variation but were lowest in surficial (0-2 cm) sediments. Acid-volatile sulfide concentrations always exceeded SEM concentrations at less than or equal to 6.0 pmol SEM/g; only at 12.0 mu mol SEM/g did SEM/AVS ratios exceed 1.0. Zinc was rarely detected in pore water at any treatment and never at concentrations which should have posed a hazard to benthic macroinvertebrates. No substantial effect on colonization of zinc-spiked sediments by benthic macroinvertebrates was observed. Only oligochaetes (Naididae) were significantly reduced in abundance at the high zinc treatment, although reductions were occasionally evident for other taxa. Lack of noteworthy pore-water zinc concentrations and lack of associated, ecologically meaningful effects were attributed to the increase in AVS levels observed with increasing SEM zinc sediment concentration. The increases in AVS theoretically resulted from a replacement of natural iron and manganese sulfides with the more stable zinc sulfide complex.

Ammonia commonly is present at detectable concentrations in overlying and interstitial water in solid-phase sediment toxicity tests. There are scenarios in which it is desirable to discern ammonia toxicity from that caused by other contaminants. The objective of this study was to verify the technical basis for defining the bioavailability of ammonia in solid-phase sediment toxicity tests based on interstitial- (pore-) water concentrations of the compound. We compared the toxicity of ammonia in spiked-sediment versus water-only exposures with the oligochaete Lumbriculus variegatus, the midge Chironomus tentans, and the amphipod Hyalella azteca. Sediment exposures were conducted in a test system that enabled the maintenance of target concentrations of ammonia in pore water, while keeping overlying water concentrations of the compound below potential effect levels. To enable direct comparison of the water-only and sediment exposures, the same test conditions (including pH) were used for each. There was good correspondence between the LC50 values of water-only tests and spiked-sediment toxicity tests for both L. variegatus and C, tentans. Hyalella azteca apparently avoided the spiked sediments and was frequently observed in the less contaminated overlying water, thus limiting the evaluation of a pore-water exposure model for the amphipod. Overall, at least for some benthic species, ammonia bioavailability and toxicity can be accurately predicted from pore-water concentrations of the compound; however, the model maybe less robust for more epibenthic organisms, such as H. azteca.

A laboratory study was conducted with Chironomus tentans to assess the significance of growth retardation of third-to fourth-instar larvae over a 10-d test period on long-term survival, adult emergence, and ovipositing success. Data were intended to provide interpretive guidance for the commonly used growth endpoint in 10-d sediment bioassays with C. tentans. Larval growth was controlled by using six feeding levels ranging from 0.2 to 5.9 mg dry weight Tetrafin® fish food per day. Mean 10-d survival was ≥88% at all feeding levels, but larval growth decreased significantly (p<0.05) with each decrease in feeding level. Cumulative successful emergence of adult C. tentans decreased significantly with decreasing larval growth. Mean times to emergence always increased with decreasing growth rates and effects were generally more pronounced for females than males. At the lowest 10-d mean growth where successful emergence occurred (0.27 mg), the times to emergence doubled relative to the times observed at the highest 10-d mean growth (1.03 mg). Ten-day larval growth retardation was strongly correlated with reduction in adult emergence success (r2 = 0.96). Growth retardations ≥64% resulted in 86–100% reductions in adult emergence. Growth retardation in the range of 35 to 50% equated with comparable percent reductions in adult emergence success. Although fewer females successfully emerged at 10-d growths of 0.37 and 0.74 mg relative to the highest 10-d growth (1.03 mg), ovipositing success of these females did not appear to be adversely affected by either their slower growth rates or their lower mean dry weights (0.62 and 0.99 mg, respectively). Growth of second generation larvae did not appear to be affected by maternal growth rate, but rather appeared to be solely correlated with their own feeding level.