Reproductive toxicity of DDT in the Japanese medaka fish model: Revisiting the impacts of DDT+ on female reproductive health.

The organochlorine pesticide dichlorodiphenyltrichloroethane (DDT) is an endocrine-disrupting compound (EDC) that has been banned by most countries for decades. However, it continues to be detected in nearly all humans and wildlife due to its biological and environmental persistence. The ovarian dysgenesis syndrome hypothesis speculates that exposure to EDCs during sensitive developmental windows such as early gonadal differentiation lead to reproductive disorders later in life. Yet, mechanisms by which DDT affects developing gonads remain unclear due to the inherent challenge of getting developmental exposure data from adults presenting with reproductive disease. The Japanese medaka (Oryzias latipes) is a valuable fish model for sex-specific toxicological studies due to its chromosomal sex determination, external embryonic development, short generation time, and extensively mapped genome. It is well documented that medaka exposed to DDT and its metabolites and byproducts (herein referred to as DDT+) at different developmental time points experience permanent alterations in gonadal morphology, reproductive success, and molecular and hormonal signaling. However, the overwhelming majority of studies focus primarily on functional and morphological outcomes in males and females and have rarely investigated long-term transcriptional or molecular effects. This review summarizes previous experimental findings and the state of our knowledge concerning toxic effects DDT + on reproductive development, fertility, and health in the valuable medaka model. It also identifies gaps in knowledge, emphasizing a need for more focus on molecular mechanisms of ovarian endocrine disruption using enhanced molecular tools that have become increasingly available over the past few decades. Furthermore, DDT forms a myriad of over 45 metabolites and transformation products in biota and the environment, very few of which have been evaluated for environmental abundance or health effects. This reinforces the demand for high throughput and economical in vivo models for predictive toxicology screening, and the Japanese medaka is uniquely positioned to meet this need.

Visual, spectral, and microchemical quantification of crystalline anomalies in otoliths of wild and cultured delta smelt.

Developmental abnormalities in otoliths can impact growth and survival in teleost fishes. Here, we quantified the frequency and severity of developmental anomalies in otoliths of delta smelt (Hypomesus transpacificus), a critically endangered estuarine fish that is endemic to the San Francisco Estuary. Left-right asymmetry and anomalous crystalline polymorphs (i.e., vaterite) were quantified and compared between wild and cultured populations using digital image analysis. Visual estimates of vaterite were validated using X-ray diffraction, Raman spectroscopy, laser ablation ICPMS, and electron probe microanalysis. Results indicated that cultured delta smelt were 80 times more likely to contain a vateritic otolith and 18 times more likely to contain relatively large (≥ 15%) amounts of vaterite. Similarly, cultured fish exhibited 30% greater asymmetry than wild fish. These results indicate that cultured delta smelt exhibit a significantly higher frequency of vestibular abnormalities which are known to reduce fitness and survival. Such hatchery effects on otolith development could have important implications for captive culture practices and the supplementation of wild fish populations with cultured individuals.

Temperature and salinity preferences of endangered Delta Smelt (Hypomesus transpacificus, Actinopterygii, Osmeridae).

Temperature and salinity often define the distributions of aquatic organisms. This is at least partially true for Delta Smelt, an imperiled species endemic to the upper San Francisco Estuary. While much is known about the tolerances and distribution of Delta Smelt in relation to these parameters, little is known regarding the temperature and salinity preferences of the species. Therefore, the temperature and salinity preferences of sub-adult Delta Smelt were investigated across a wide range of thermal (8-28 °C) and salinity (0-23 ppt) conditions. Replicates of ten fish were allowed to swim between two circular chambers with different temperature or salinity, and the distribution of fish between the chambers was recorded. We found that Delta Smelt showed no temperature preference below 15 °C, a modest aversion to the warmer tank from 15 to 28 °C, and a strong aversion to the warmer tank with elevated mortality at temperatures above 28 °C. Delta Smelt also preferred lower salinities, and this preference became more pronounced as salinity increased toward 23 ppt. These results indicate that Delta Smelt can tolerate high temperatures and salinities for a short time, and that their preferences for lower temperature and salinity strengthens as these variables increase.

Chronic exposure to high-density polyethylene microplastic through feeding alters the nutrient metabolism of juvenile yellow perch (Perca flavescens).

Microplastics are emergent contaminants threatening aquatic organisms including aquacultured fish. This study investigated the effects of high-density polyethylene (HDPE, 100 to 125 µm) on yellow perch (Perca flavescens) based on integrative evaluation including growth performance, nutritional status, nutrient metabolism, fish health, and gut microbial community. Five test diets (0, 1, 2, 4, or 8 g HDPE/100 g diet) containing 41% protein and 10.5% lipid were fed to juvenile perch (average body weight, 25.9 ± 0.2 g; n = 15) at a feeding rate of 1.5% to 2.0% body weight daily. The feeding trial was conducted in a flow-through water system for 9 wk with 3 tanks per treatment and 15 yellow perch per tank. No mortality or HDPE accumulation in the fish was found in any treatments. Weight gain and condition factor of fish were not significantly impacted by HDPE (P > 0.05). Compared to the control group, fish fed the 8% HDPE diet had significantly decreased levels of protein and ash (P < 0.05). In response to the increasing levels of HDPE exposure, the hepatosomatic index value, hepatocyte size, and liver glycogen level were increased, but lipid content was reduced in the liver tissues. Compared to the control treatment, fish fed the 8% HDPE diet had significant accumulations of total bile acids and different metabolism pathways such as bile acid biosynthesis, pyruvate metabolism, and carnitine synthesis. Significant enterocyte necrosis was documented in the foregut of fish fed the 2% or 8% HDPE diet; and significant cell sloughing was observed in the midgut and hindgut of fish fed the 8% HDPE diet. Fish fed the 2% HDPE diet harbored different microbiota communities compared to the control fish. This study demonstrates that HDPE ranging from 100 to 125 µm in feed can be evacuated by yellow perch with no impact on growth. However, dietary exposure to HDPE decreased whole fish nutrition quality, altered nutrient metabolism and the intestinal histopathology as well as microbiota community of yellow perch. The results indicate that extended exposure may pose a risk to fish health and jeopardize the nutrition quality of aquacultured end product. This hypothesis remains to be investigated further.

Reproductive strategy of Delta Smelt Hypomesus transpacificus and impacts of drought on reproductive performance.

Understanding reproductive biology and performance of fish is essential to formulate effective conservation and management programs. Here, we studied reproductive strategies of female Delta Smelt Hypomesus transpacificus, an endangered fish species in the State of California, the United States, focusing on (1) better understanding their distribution pattern during the winter and spring spawning season at very fine scale to predict their possible spawning grounds and (2) assessing impacts of a recent, severe drought on their reproductive performance. We formulated our hypotheses as follows; (1) female Delta Smelt migrate to particular locations for spawning so that mature females can be frequently found in those locations throughout the spawning season and (2) reproductive performance of individual female fish declined during the drought. To test the first hypotheses, we analyzed relationships between water quality parameters and maturity/distribution pattern of Delta Smelt. Salinity better explained the distribution pattern of Delta Smelt at subadult and adult stages compared with water temperature or turbidity. Although there are some freshwater locations where mature Delta Smelt can frequently be found during the spawning season, Delta Smelt at the final maturation stage (Stage 5: hydration) and post spawners appeared to be widespread in the area where salinity was below 1.0 during the spawning season. Therefore, Delta Smelt could theoretically spawn in any freshwater locations, with more specific spawning requirements in the wild (e.g., substrate type and depth) still unknown. Delta Smelt, which experienced dry and critically dry conditions (the 2013 and 2014 year-classes), showed smaller oocytes, and lower clutch size and gonadosomatic index compared with the fish caught in a wet year (2011 year-class) at the late vitellogenic stage (Stage 4 Late), suggesting reproductive performance was negatively affected by environmental conditions during the drought.

Covariance of Phytoplankton, Bacteria, and Zooplankton Communities Within Microcystis Blooms in San Francisco Estuary.

Microcystis blooms have occurred in upper San Francisco Estuary (USFE) since 1999, but their potential impacts on plankton communities have not been fully quantified. Five years of field data collected from stations across the freshwater reaches of the estuary were used to identify the plankton communities that covaried with Microcystis blooms, including non-photosynthetic bacteria, cyanobacteria, phytoplankton, zooplankton, and benthic genera using a suite of analyses, including microscopy, quantitative PCR (qPCR), and shotgun metagenomic analysis. Coherence between the abundance of Microcystis and members of the plankton community was determined by hierarchal cluster analysis (CLUSTER) and type 3 similarity profile analysis (SIMPROF), as well as correlation analysis. Microcystis abundance varied with many cyanobacteria and phytoplankton genera and was most closely correlated with the non-toxic cyanobacterium Merismopoedia, the green algae Monoraphidium and Chlamydomonas, and the potentially toxic cyanobacteria Pseudoanabaena, Dolichospermum, Planktothrix, Sphaerospermopsis, and Aphanizomenon. Among non-photosynthetic bacteria, the xenobiotic bacterium Phenylobacterium was the most closely correlated with Microcystis abundance. The coherence of DNA sequences for phyla across trophic levels in the plankton community also demonstrated the decrease in large zooplankton and increase in small zooplankton during blooms. The breadth of correlations between Microcystis and plankton across trophic levels suggests Microcystis influences ecosystem production through bottom-up control during blooms. Importantly, the abundance of Microcystis and other members of the plankton community varied with wet and dry conditions, indicating climate was a significant driver of trophic structure during blooms.

Gross morphology, histology, and ultrastructure of the olfactory rosette of a critically endangered indicator species, the Delta Smelt, Hypomesus transpacificus.

The Delta Smelt (Hypomesus transpacificus) is a small, semi-anadromous fish native to the San Francisco Bay-Delta Estuary and has been declared as critically endangered. Their olfactory biology, in particular, is poorly understood and a basic description of their sensory anatomy is needed to advance our understanding of the sensory ecology of species to inform conservation efforts to manage and protect them. We provide a description of the gross morphology, histological, immunohistochemical, and ultrastructural features of the olfactory rosette in this fish and discuss some of the functional implications in relation to olfactory ability. We show that Delta Smelt have a multilamellar olfactory rosette with allometric growth. Calretinin immunohistochemistry revealed a diffuse distribution of olfactory receptor neurons within the epithelium. Ciliated, microvillous and crypt neurons were clearly identified using morphological and immunohistochemical features. The olfactory neurons were supported by robust ciliated and secretory sustentacular cells. Although the sense of smell has been overlooked in Delta Smelt, we conclude that the olfactory epithelium has many characteristics of macrosmatic fish. With this study, we provide a foundation for future research into the sensory ecology of this imperiled fish.

Potential Impacts on Treated Water Quality of Recycling Dewatered Sludge Supernatant during Harmful Cyanobacterial Blooms.

Cyanobacterial blooms and the associated release of cyanotoxins pose problems for many conventional water treatment plants due to their limited removal by typical unit operations. In this study, a conventional water treatment process consisting of coagulation, flocculation, sedimentation, filtration, and sludge dewatering was assessed in lab-scale experiments to measure the removal of microcystin-LR and Microcystis aeruginosa cells using liquid chromatography with mass spectrometer (LC-MS) and a hemacytometer, respectively. The overall goal was to determine the effect of recycling cyanotoxin-laden dewatered sludge supernatant on treated water quality. The lab-scale experimental system was able to maintain the effluent water quality below relevant the United States Environmental Protection Agency (US EPA) and World Health Organisation (WHO) standards for every parameter analyzed at influent concentrations of M. aeruginosa above 106 cells/mL. However, substantial increases of 0.171 NTU (Nephelometric Turbidity Unit), 7 × 104 cells/L, and 0.26 µg/L in turbidity, cyanobacteria cell counts, and microcystin-LR concentration were observed at the time of dewatered supernatant injection. Microcystin-LR concentrations of 1.55 µg/L and 0.25 µg/L were still observed in the dewatering process over 24 and 48 h, respectively, after the initial addition of M.aeruginosa cells, suggesting the possibility that a single cyanobacterial bloom may affect the filtered water quality long after the bloom has dissipated when sludge supernatant recycling is practiced.

The health and condition responses of Delta Smelt to fasting: A time series experiment.

There is an extensive literature establishing, validating, and quantifying a wide range of responses of fishes to fasting. Our study complements this work by comparing fed and unfed treatments of hatchery-raised Delta Smelt (Hypomesus transpacificus)-an imperiled fish that is endemic to the San Francisco Estuary and its tributaries in California, USA-across a diverse suite of endpoints over a two-month time series. The experiment was conducted at 15.9°C, and individuals were sampled at 12 time points as starvation became increasingly severe. We found that hepatosomatic index and condition factor were relatively sensitive to starvation, becoming significantly depressed at Day 4 and 7, respectively. Histological analysis of liver showed elevated cytoplasmic inclusion bodies at Day 7, followed by increased glycogen depletion, single cell necrosis, and hydropic vacuolar degeneration at Day 14, 21, and 28, respectively. Of four antioxidants measured, glutathione decreased at Day 4, superoxide dismutase increased at Day 14, catalase increased at Day 56, and glutathione peroxidase was not affected by starvation. The net result was a ~2-fold increase in lipid peroxidation (malondialdehyde) in fasted fish that was highly inconsistent through time. RNA to DNA ratio and triglycerides in muscle were relatively insensitive to starvation, only consistently decreasing with fasting after mortality began increasing in the 'No Feeding' treatment, at Day 21. Together, these results suggest that Delta Smelt mobilize hepatic energy stores far more rapidly than lipids in muscle when subjected to fasting, leading to rapid atrophy of liver and the development of cytoplasmic inclusion bodies-possibly autophagosomes-in hepatocytes.

Histopathological assessment of seven year-classes of Delta Smelt.

The Delta Smelt (Hypomesus transpacificus) is an imperiled, annual fish endemic to the Sacramento-San Joaquin Delta and San Francisco Estuary. This study examined the severity and prevalence of liver and gill lesions of juvenile through adult Delta Smelt from 2011 through 2017 collected from five regions throughout its habitat (n = 1,053). The first and last years of the study were wet, but bracketed an extreme drought in CA (2012-2016), during which the Delta Smelt population reached historical lows. Overall, the three most common lesions were gill ionocyte hyperplasia, liver lipidosis, and gill aneurysm. Individuals with higher fork lengths exhibited increased gill and liver lesion score (summations of the severity scores), suggesting that Delta Smelt accumulate lesions through their lives, and that larger individuals were more tolerant of liver and gill lesions. Liver lesion score showed significant regional differences, while salinity was a better predictor of gill lesions than region, with lower gill lesion scores associated with higher salinities. Largely consistent with previously reported histopathology patterns, Delta Smelt collected from the Confluence and Suisun Marsh had the lowest liver lesion score, while Delta Smelt collected from Cache Slough and Suisun Bay had the highest lesion scores, and Suisun Marsh had the lowest glycogen depletion, suggesting heterogeneous levels of exposure to environmental stressors across regions. Gill and liver lesion score also varied significantly with year-class. The highest gill lesion score occurred in the 2015/16 year-class, and the lowest occurred in the 2017/18 year-class, a 2.8-fold difference. Controlling for size and regional effects, individuals with comparatively high liver lesion scores were prevalent in the population until the 2014/15 year-class. In the two subsequent year-classes, Delta Smelt livers were in the best condition, coinciding with peak drought conditions and record low abundances.

The Effect of Herbicide Formulations and Herbicide-Adjuvant Mixtures on Aquatic Food Web Species of the Sacramento-San Joaquin Delta, California, USA.

Herbicides have recently been designated as one of the most frequently detected chemical types in aquatic systems. We evaluated the effect of the herbicide formulations Clearcast® (active ingredient imazamox) and Galleon® SC (active ingredient penoxsulam), in conjunction with the adjuvant Agri-dex®, on the diatom Thalassiosira pseudonana and the copepod Eurytemora affinis and whether their application in invasive aquatic weed control has acutely negative effects on important delta food web species. Herbicide-adjuvant mixtures were more toxic than herbicide formulations alone. For the diatom, 50% inhibition concentration (IC50) were 84.4 mg/L for Clearcast, >100 mg/L for Galleon SC, and 38.5 mg/L for Agri-dex. In the herbicide mixture diatom tests, IC50s decreased to 2.8 mg/L for Clearcast + 64 mg/L Agri-dex and to 6.6 mg/L for Galleon SC + 41 mg/L Agri-dex. In the copepod tests, 50% effect concentrations (EC50s) were 45.4 mg/L for Agri-dex, 31.2 mg/L for Galleon SC, and >100 mg/L for Clearcast. When tested together, EC50s were reduced to 24.1 for Galleon SC and 67.6 mg/L for Clearcast in the presence of 50 mg/L Agri-dex. Environmentally relevant herbicide-adjuvant mixture ratios were at the no-observed-effect level. Mixture interactions between herbicides and adjuvants indicate the potential for increased toxicity in herbicide formulations and tank mixes, especially in consideration of the unlisted, proprietary ingredients which are included in herbicide formulations, making predicting nontarget effects challenging. Environ Toxicol Chem 2020;39:1375-1381. © 2020 SETAC.

Analysis of Covalently Bound Microcystins in Sediments and Clam Tissue in the Sacramento-San Joaquin River Delta, California, USA.

Harmful cyanobacterial blooms compromise human and environmental health, mainly due to the cyanotoxins they often produce. Microcystins (MCs) are the most commonly measured group of cyanotoxins and are hepatotoxic, neurotoxic, and cytotoxic. Due to MCs ability to covalently bind to proteins, quantification in complex matrices is difficult. To analyze bound and unbound MCs, analytical methods were optimized for analysis in sediment and clam tissues. A clean up step was incorporated to remove lipids, improving percent yield. This method was then applied to sediment and clam samples collected from the Sacramento-San Joaquin River Delta (Delta) in the spring and fall of 2017. Water samples were also tested for intracellular and extracellular MCs. These analyses were used to quantify the partitioning of MCs among sediment, clams, and water, and to examine whether MCs persist during non-summer months. Toxin analysis revealed that multiple sediment samples collected in the Delta were positive for MCs, with a majority of the positive samples from sites in the San Joaquin River, even while water samples from the same location were below detection limit. These data highlight the importance of analyzing MCs in complex matrices to accurately evaluate environmental risk.

Toxicity of herbicides to cyanobacteria and phytoplankton species of the San Francisco Estuary and Sacramento-San Joaquin River Delta, California, USA.

The herbicides glyphosate, imazamox and fluridone are herbicides, with low toxicity towards fish and invertebrates, which are applied to waterways to control invasive aquatic weeds. However, the effects of these herbicides on natural isolates of phytoplankton and cyanobacteria are unknown. Three species of microalgae found in the San Francisco Estuary (SFE)/Sacramento-San Joaquin River Delta (Delta) (Microcystis aeruginosa, Chlamydomonas debaryana, and Thalassiosira pseudonana) were exposed to the three herbicides at a range of concentrations in 96-well plates for 5-8 days. All three algal species were the most sensitive to fluridone, with IC50 of 46.9, 21, and 109 µg L-1 for M. aeruginosa, T. pseudonana and C. debaryana, respectively. Imazamox inhibited M. aeruginosa and T. pseudonana growth at 3.6 × 104 µg L-1 or higher, and inhibited C. debaryana growth at 1.0 × 105 µg L-1 or higher. Glyphosate inhibited growth in all species at ca. 7.0 × 104 µg L-1 or higher. Fluridone was the only herbicide that inhibited the microalgae at environmentally relevant concentrations in this study and susceptibility to the herbicide depended on the species. Thus, the application of fluridone may affect cyanobacteria and phytoplankton community composition in water bodies where it is applied.

Toxic effects of fluridone on early developmental stages of Japanese Medaka (Oryzias latipes).

The herbicide fluridone is intensively applied to control invasive aquatic plants globally, including in the Sacramento and San Joaquin Delta (the Delta), California, USA. Our previous study revealed that the adult stage of Delta Smelt showed acute and sub-lethal adverse effects following 6 h of exposure to environmentally relevant concentrations of fluridone. To further investigate mechanisms of toxicity of fluridone and to assess its toxicity to early life stages of fish, we performed additional exposures using the fish model Japanese Medaka (Oryzias latipes). Male and female Medaka embryos were exposed to concentrations of fluridone for 14 d and showed reduced hatching success in a dose dependent manner. The half maximal effective concentration for the hatching success was 2.3 mg L-1. In addition, male and female Medaka larvae were acute exposed to fluridone for 6 h to assess their swimming behavior and gene expression patterns. Fish exposed to fluridone at 4.2 mg L-1 or higher became lethargic and showed abnormal swimming behavior. The response to the stimuli was significantly impaired by fluridone at 21 mg L-1 and above in males, and at 104 mg L-1 in females. Transcriptome analysis identified a total of 799 genes that were significantly differentially expressed, comprising 555 up-regulated and 244 down-regulated genes in males exposed to 21 mg L-1 of fluridone. The gene set enrichment analysis indicated a number of biological processes altered by fluridone. Among the genes involved in those biological processes, the expression of the genes, acetylcholinesterase, retinoic acid receptor, insulin receptor substrate, glutathione reductase, and glutathione S transferase, exhibited dose- and sex-dependent responses to fluridone. The study indicated that fluridone exposure led to detrimental toxic effects at early developmental stages of fish, by disturbing the biological processes of growth and development, and the nervous system, inducing oxidative stress and endocrine disruption.

Hydrodynamic Modeling Coupled with Long-term Field Data Provide Evidence for Suppression of Phytoplankton by Invasive Clams and Freshwater Exports in the San Francisco Estuary.

The San Francisco Estuary (California, USA) had abundant pelagic fish in the late 1960s, but has few pelagic fish today. A primary cause for this decline in fish is thought to be a trophic cascade, triggered by declining phytoplankton. Here, we describe the changes in pelagic community structure of the San Francisco Estuary. Then, we examine whether changes in hydrodynamics due to freshwater exports, which increased exponentially beginning in 1967, in addition to the 1986 invasion by the clam Potamocorbula amurensis, explain the phytoplankton loss. Hydrodynamic variables were reconstructed back to 1956 using statistical models fit to, and cross-validated against, output from a hydrodynamic model. Then, we regressed mean summer/fall chlorophyll a-the season with the largest phytoplankton decline-against the reconstructed hydrodynamic variables and the presence/absence of P. amurensis for 1969-2014. The regression model, which explained 78% of the interannual variation in chlorophyll a, was then used to quantify the influence of P. amurensis and exports on chlorophyll a. Based on monitoring data, chlorophyll a declined 22-fold from 1969-2014, zooplankton declined 32-fold from 1972-2014, and pelagic fish declined 92-fold from 1968-2014. Averaged over 1990-2014, the chlorophyll a model ascribed an 88% decline in chlorophyll a to P. amurensis, a 74% decline to exports (at minimum), and a 97% decline to the combined influence of P. amurensis and exports (at minimum). Thus, the decline in pelagic productivity in the San Francisco Estuary has occurred largely due to the combined impacts of the P. amurensis invasion and increased freshwater exports.

Biodiversity of cyanobacteria and other aquatic microorganisms across a freshwater to brackish water gradient determined by shotgun metagenomic sequencing analysis in the San Francisco Estuary, USA.

Blooms of Microcystis and other harmful cyanobacteria can degrade water quality by producing cyanotoxins or other toxic compounds. The goals of this study were (1) to facilitate understanding of community structure for various aquatic microorganisms in brackish water and freshwater regions with emphasis on cyanobacteria, and (2) to test a hypothesis that Microcystis genotypes that tolerate higher salinity were blooming in brackish water environments during the severe drought, 2014. Shotgun metagenomic analysis revealed that cyanobacteria dominated the brackish water region while bacteria dominated the freshwater region. A group of cyanobacteria (e.g., Aphanizomenon, Microcystis, Planktothrix, Pseudanabaena), bacteria (e.g., Bacillus, Porphyrobacter), and diatoms (Phaeodactylum and Thalassiosira) were abundant in the brackish water region. In contrast, Hassallia (cyanobacteria) and green algae (Nannochloropsis, Chlamydomonas, and Volvox) were abundant in the landward freshwater region. Station variation was also apparent. One landward sampling station located downstream of an urbanized area differed substantially from the other stations in terms of both water chemistry and community structure, with a higher percentage of arthropods, green algae, and eukaryotes. Screening of the Microcystis internal transcribed spacer region revealed six representative genotypes, and two of which were successfully quantified using qPCR (Genotypes I and VI). Both genotypes occurred predominantly in the freshwater region, so the data from this study did not support the hypothesis that salinity tolerant Microcystis genotypes bloomed in the brackish water region in 2014.

Sub-lethal effects of herbicides penoxsulam, imazamox, fluridone and glyphosate on Delta Smelt (Hypomesus transpacificus).

Concerns regarding non-target toxicity of new herbicides used to control invasive aquatic weeds in the San Francisco Estuary led us to compare sub-lethal toxicity of four herbicides (penoxsulam, imazamox, fluridone, and glyphosate) on an endangered fish species Delta Smelt (Hypomesus transpacificus). We measured 17ß-estradiol (E2) and glutathione (GSH) concentrations in liver, and acetylcholinesterase (AChE) activity in brain of female and male fish after 6 h of exposure to each of the four herbicides. Our results indicate that fluridone and glyphosate disrupted the E2 concentration and decreased glutathione concentration in liver, whereas penoxsulam, imazamox, and fluridone inhibited brain AChE activity. E2 concentrations were significantly increased in female and male fish exposed to 0.21 µM of fluridone and in male fish exposed to 0.46, 4.2, and 5300 µM of glyphosate. GSH concentrations decreased in males exposed to fluridone at 2.8 µM and higher, and glyphosate at 4.2 µM. AChE activity was significantly inhibited in both sexes exposed to penoxsulam, imazamox, and fluridone, and more pronounced inhibition was observed in females. The present study demonstrates the potential detrimental effects of these commonly used herbicides on Delta Smelt.

Direct and indirect effects of different types of microplastics on freshwater prey (Corbicula fluminea) and their predator (Acipenser transmontanus).

We examined whether environmentally relevant concentrations of different types of microplastics, with or without PCBs, directly affect freshwater prey and indirectly affect their predators. Asian clams (Corbicula fluminea) were exposed to environmentally relevant concentrations of polyethylene terephthalate (PET), polyethylene, polyvinylchloride (PVC) or polystyrene with and without polychlorinated biphenyls (PCBs) for 28 days. Their predators, white sturgeon (Acipenser transmontanus), were exposed to clams from each treatment for 28 days. In both species, we examined bioaccumulation of PCBs and effects (i.e., immunohistochemistry, histology, behavior, condition, mortality) across several levels of biological organization. PCBs were not detected in prey or predator, and thus differences in bioaccumulation of PCBs among polymers and biomagnification in predators could not be measured. One of the main objectives of this study was to test the hypothesis that bioaccumulation of PCBs would differ among polymer types. Because we could not answer this question experimentally, a bioaccumulation model was run and predicted that concentrations of PCBs in clams exposed to polyethylene and polystyrene would be greater than PET and PVC. Observed effects, although subtle, seemed to be due to microplastics rather than PCBs alone. For example, histopathology showed tubular dilation in clams exposed to microplastics with PCBs, with only mild effects in clams exposed to PCBs alone.

Impairment of antioxidant mechanisms in Japanese Medaka (Oryzias latipes) by acute exposure to aluminum.

Increasing aluminum (Al) concentrations in aquatic habitats as a result of anthropogenic acidification and industrialization is a global issue. Moreover, in extensive areas of the humid tropics and subtropics, high Al concentrations in freshwater are observed because of both naturally low pH and high Al concentrations in soil. Al increases production of reactive oxygen species and enhances oxidative damage in mammals. However, no studies have examined the effect of environmentally relevant concentrations of Al at low pH on oxidative stress in fish. This study assessed Al-induced effects on enzymatic and non-enzymatic antioxidants, lipid peroxidation, and on expression of oxidative stress-related genes at low pH using Japanese Medaka (Oryzias latipes). Larval fish were exposed to dissolved Al concentrations of 0, 1.7, 6.2 and 16.7µgL-1 for 4days at pH5.3 in soft water. Al caused a significant reduction in activity of glutathione peroxidase at 6.2 and 16.7µgL-1, and of glutathione reductase at 16.7µgL-1 in whole body homogenates. No changes were observed in the expression of the glutathione peroxidase gene, and expression levels of the glutathione reductase gene were too low to be quantitated. Even though there was an overall decrease in the activity of catalase and in the concentration of glutathione, differences were not significant compared to the control. Changes in lipid peroxidation were not found. This study showed that exposure to environmentally relevant concentrations of Al at low pH impairs antioxidant defense mechanisms of Medaka.

Effects of low levels of ultraviolet radiation on antioxidant mechanisms of Japanese Medaka (Oryzias latipes).

Extreme weather events like drought are expected to increase with climate change, which will increase exposure of freshwater fish to ultraviolet (UV) radiation. Compared to fully grown adult fish, fish in early life stages are more susceptible to UV radiation due to the lack of well-developed pigmentation. Even though several studies have described affectation of fish health after exposure to UV radiation, most of the studies have used intensities that are only found on the surface of the earth crust or at shallow depths in water bodies, and little is known about impacts of weaker UV radiation, which can be found in deep water. This study showed effects on the antioxidant system of Japanese Medaka after 7 days of exposure to very low intensities of UV radiation, levels that can be found at deep locations in lakes and rivers. Exposure to UV radiation (UVA: 360.1 ± 18.4 µW cm-2 for a dose of 21.6 ± 1.2 mJ cm-2 min-1, and UVB: 6.3 ± 0.5 µW cm-2 for a dose of 0.38 ± 0.03 mJ cm-2 min-1) caused a reduction in the catalase activity (over 50%) and enhanced oxidative damage to lipids. Results of this study showed that environmentally relevant, low levels of UV radiation affect mechanisms by which fish deal with enhanced production of reactive oxygen species and oxidative damage. The results raise concerns about early life stages of fish under scenarios of increased exposure to solar light, such as in tropic regions or during summer in temperate regions.