Effects of incubation temperature on the upper thermal tolerance of the imperiled longfin smelt (Spirinchus thaleichthys).

Upper thermal limits in many fish species are limited, in part, by the heart's ability to meet increased oxygen demand during high temperatures. Cardiac plasticity induced by developmental temperatures can therefore influence thermal tolerance. Here, we determined how incubation temperatures during the embryonic stage influence cardiac performance across temperatures during the sensitive larval stage of the imperiled longfin smelt. We transposed a cardiac assay for larger fish to newly hatched larvae that were incubated at 9°C, 12°C or 15°C. We measured heart rate over increases in temperature to identify the Arrhenius breakpoint temperature (TAB), a proxy for thermal optimum and two upper thermal limit metrics: temperature when heart rate is maximized (Tpeak) and when cardiac arrhythmia occurs (TArr). Higher incubation temperatures increased TAB, Tpeak and TArr, but high individual variation in all three metrics resulted in great overlap of individuals at TAB, Tpeak and TArr across temperatures. We found that the temperatures at which 10% of individuals reached Tpeak or TArr and temperatures at which number of individuals at TAB relative to Tpeak (ΔN(TAB,Tpeak)) was maximal, correlated more closely with upper thermal limits and thermal optima inferred from previous studies, compared to the mean values of the three cardiac metrics of the present study. Higher incubation temperatures increased the 10% Tpeak and TArr thresholds but maximum ΔN(TAB,Tpeak) largely remained the same, suggesting that incubation temperatures modulate upper thermal limits but not Topt for a group of larvae. Overall, by measuring cardiac performance across temperatures, we defined upper thermal limits (10% thresholds; Tpeak, 14.4-17.5°C; TArr, 16.9-20.2°C) and optima (ΔN(TAB,Tpeak), 12.4-14.4°C) that can guide conservation strategies for longfin smelt and demonstrated the potential of this cardiac assay for informing conservation plans for the early life stages of fish.

Loss of plasticity in maturation timing after ten years of captive spawning in a delta smelt conservation hatchery.

Adaptation to captivity in spawning programs can lead to unintentional consequences, such as domestication that results in reduced fitness in the wild. The timing of sexual maturation has been shown to be a trait under domestication selection in fish hatcheries, which affects a fish's access to mating opportunities and aligning their offspring's development with favorable environmental conditions. Earlier maturing fish may be favored in hatchery settings where managers provide artificially optimal growing conditions, but early maturation may reduce fitness in the wild if, for example, there is a mismatch between timing of reproduction and availability of resources that support recruitment. We investigated patterns of maturation timing in a delta smelt (Hypomesus transpacificus) conservation hatchery by quantifying changes to the median age at maturity since the captive spawning program was initiated in 2008. Over the span of a decade, we observed a small, but significant increase in age at maturity among broodstock by 2.2 weeks. This trait had low heritability and was largely controlled by phenotypic plasticity that was dependent on the time of year fish were born. Fish that were born later in the year matured faster, potentially a carryover from selection favoring synchronous spawning in the wild. However, higher DI (domestication index) fish showed a loss of plasticity, we argue, as a result of hatchery practices that breed individuals past peak periods of female ripeness. Our findings suggest that the hatchery setting has relaxed selection pressures for fish to mature quickly at the end of the year and, consequently, has led to a loss of plasticity in age at maturity. Hatchery fish that are re-introduced in the wild may not be able to align maturation with population peaks if their maturation rates are too slow with reduced plasticity, potentially resulting in lower fitness.

Bifenthrin, a Ubiquitous Contaminant, Impairs the Development and Behavior of the Threatened Longfin Smelt during Early Life Stages.

The Longfin Smelt (Spirinchus thaleichthys) population in the San Franscisco Bay/Sacramento-San Joaquin Delta (Bay-Delta) has declined to ∼1% of its pre-1980s abundance and, as a result, is listed as threatened under the California Endangered Species Act. The reasons for this decline are multiple and complex, including the impacts of contaminants. Because the spawning and rearing seasons of Longfin Smelt coincide with the rainy season, during which concentrations of contaminants increase due to runoff, we hypothesized that early life stages may be particularly affected by those contaminants. Bifenthrin, a pyrethroid insecticide commonly used in agricultural and urban sectors, is of concern. Concentrations measured in the Bay-Delta have been shown to disrupt the behavior, development, and endocrine system of other fish species. The objective of the present work was to assess the impact of bifenthrin on the early developmental stages of Longfin Smelt. For this, embryos were exposed to 2, 10, 100, and 500 ng/L bifenthrin from fertilization to hatch, and larvae were exposed to 2, 10, and 100 ng/L bifenthrin from one day before to 3 days post-hatch. We assessed effects on size at hatch, yolk sac volume, locomotory behavior, and upper thermal susceptibility (via cardiac endpoints). Exposure to these environmentally relevant concentrations of bifenthrin did not significantly affect the cardiac function of larval Longfin Smelt; however, exposures altered their behavior and resulted in smaller hatchlings with reduced yolk sac volumes. This study shows that bifenthrin affects the fitness-determinant traits of Longfin Smelt early life stages and could contribute to the observed population decline.

Effects of turbidity, temperature and predation cue on the stress response of juvenile delta smelt.

The San Francisco Estuary (SFE) is one of the most degraded ecosystems in the United States, and organisms that inhabit it are exposed to a suite of environmental stressors. The delta smelt (Hypomesus transpacificus), a small semi-anadromous fish endemic to the SFE and considered an indicator species, is close to extinction in the wild. The goal of this study was to investigate how environmental alterations to the SFE, such as reductions in turbidities, higher temperatures and increased prevalence of invasive predators affect the physiology and stress response of juvenile delta smelt. Juvenile delta smelt were exposed to two temperatures (17 and 21°C) and two turbidities (1-2 and 10-11 NTU) for 2 weeks. After the first week of exposure, delta smelt were exposed to a largemouth bass (Micropterus salmoides) predator cue at the same time every day for 7 days. Fish were measured and sampled on the first (acute) and final (chronic) day of exposures to predator cues and later analyzed for whole-body cortisol, glucose, lactate, and protein. Length and mass measurements were used to calculate condition factor of fish in each treatment. Turbidity had the greatest effect on juvenile delta smelt and resulted in reduced cortisol, increased glucose and lactate, and greater condition factor. Elevated temperatures reduced available energy in delta smelt, indicated by lower glucose and total protein, whereas predator cue exposure had negligible effects on their stress response. This is the first study to show reduced cortisol in juvenile delta smelt held in turbid conditions and adds to the growing data that suggest this species performs best in moderate temperatures and turbidities. Multistressor experiments are necessary to understand the capacity of delta smelt to respond to the multivariate and dynamic changes in their natural environment, and results from this study should be considered for management-based conservation efforts.

Captive-reared Delta Smelt (Hypomesus transpacificus) exhibit high survival in natural conditions using in situ enclosures.

Conservation of endangered fishes commonly includes captive breeding, applied research, and management. Since 1996, a captive breeding program has existed for the federally threatened and California endangered Delta Smelt Hypomesus transpacificus, an osmerid fish endemic to the upper San Francisco Estuary. Although this program serves as a captive refuge population, with experimental releases being initiated to supplement the wild population, it was uncertain how individuals would survive, feed, and maintain condition outside hatchery conditions. We evaluated this and the effects of three enclosure designs (41% open, 63% open, and 63% open with partial outer mesh wrap) on growth, survival, and feeding efficacy of cultured Delta Smelt at two locations (Sacramento River near Rio Vista, CA and in Sacramento River Deepwater Ship Channel) in the wild. Enclosures exposed fish to semi-natural conditions (ambient environmental fluctuations and wild food resources) but prevented escape and predation. After four weeks, survival was high for all enclosure types (94-100%) at both locations. The change in condition and weight was variable between sites, increasing at the first location but decreasing at the second location. Gut content analysis showed that fish consumed wild zooplankton that came into the enclosures. Cumulatively, results show that captive-reared Delta Smelt can survive and forage successfully when housed in enclosures under semi-natural conditions in the wild. When comparing enclosure types, we observed no significant difference in fish weight changes (p = 0.58-0.81 across sites). The success of housing captive-reared Delta Smelt in enclosures in the wild provides preliminary evidence that these fish may be suitable to supplement the wild population in the San Francisco Estuary. Furthermore, these enclosures are a new tool to test the efficacy of habitat management actions or to acclimate fish to wild conditions as a soft release strategy for recently initiated supplementation efforts.

Evaluate effects of the dilution medium and holding time on various motility parameters of delta smelt semen.

Sperm preservation is an efficient technique used for the recovery, conservation, and management of some endangered fish species. The present study was conducted to explore how preservation time would affect sperm and spawning performance in the endangered delta smelt (Hypomesus transpacificus). Sperm were preserved with the modified Hanks balanced salt solution at 14.7-16.9 °C. The Kruskal-Wallis test of sperm parameters using OpenCASA plugin in ImageJ software showed that sperm (n = 33♂) had significantly higher velocity and motility within the first 5 s after activation than that of other time points, while sperm had the lowest velocity and motility after 3 min post activation (P < 0.001). The findings (n = 30♂) also showed fresh sperm had higher velocity and motility than preserved sperm, while the sperm preserved for over 24 h showed a significantly low performance (P < 0.001). The nonlinear mixed effects models of fertilization results (n = 14♂ × 70♀) indicated the fresh sperm and sperm preserved for 1 h had higher fertilization rates than other preservation times (P < 0.001). The hatching rate (n = 14♂ × 70♀) also showed the fresh sperm and sperm preserved for 3 min and 1 h had higher hatching rates than other preservation times (P < 0.001). Overall, the study showed the best sperm performance in delta smelt was found within the first 5 s post activation, and the best fertilization and hatching rates were found when the sperm were fresh and preserved for 1 h. The findings of this study provide information for the first time about how long the delta smelt's sperm are motile for quality analysis, and how the preservation time can affect sperm quality, fertility, and hatching of this species for future applications.

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.

Characterizing the stress response in juvenile Delta smelt exposed to multiple stressors.

The Delta Smelt (Hypomesus transpacificus), once an abundant fish endemic to the Sacramento-San Joaquin Estuary, is now on the brink of extinction. Due to the high sensitivity of this species, knowledge of their stress response will be vital to their future survival and sustainability. Understanding the magnitude and kinetics of cortisol induction in Delta Smelt will provide valuable information when interpreting the degree of environmentally relevant stressors, such as warming and predator exposure. As little is known about the primary stress response and cortisol dynamics in Delta Smelt, the first aim of this study was to measure basal and maximal whole-body cortisol prior to and following exposure to a sublethal and significant netting stress at 17 and 21 °C. Our findings reveal that juvenile Delta Smelt held at 21 °C display an exacerbated stress response and a reduction in available energy compared to fish held at 17 °C. There was no evidence of the secondary stress response to the netting stress as whole-body glucose and lactate levels in treatment groups remained similar to basal values. The second aim of this study was to investigate the effect of a largemouth bass (Micropterus salmoides) predator cue, which was found to induce a significant increase in cortisol relative to control levels in juvenile Delta Smelt. Indices such as cortisol can be used as bioindicators of stress in the field and results from this study suggest that moderate temperatures and reduced predation are optimal release conditions during hatchery-based supplementation to minimize stress to this highly sensitive species.

Titanium dioxide nanoparticles decreases bioconcentration of azoxystrobin in zebrafish larvae leading to the alleviation of cardiotoxicity.

Interactions between titanium dioxide nanoparticles (n-TiO2) and pollutants in the aquatic environment may alter the bioavailability of pollutants, and thus altering their toxicity and fate. In order to investigate the bioconcentration of azoxystrobin (AZ) and its mechanism of cardiotoxicity in the presence of n-TiO2, the experiment was divided into control, n-TiO2 (100 µg/L), AZ (40, 200 and 1000 µg/L) and AZ (40, 200, 1000 µg/L) + n-TiO2 groups, and the zebrafish embryos were exposed to the exposure solution until 72 h post-fertilization. Results suggested the presence of n-TiO2 notably reduced the accumulation of AZ in larvae compared with exposure to AZ alone, thereby significantly decreasing AZ-induced cardiotoxicity, including heart rate changes, pericardium edema, venous thrombosis, increased sinus venosus and bulbus arteriosus distance and changes in cardiac-related gene expression. Further studies showed that AZ + n-TiO2 together restrained total-ATPase and Ca2+-ATPase activities, while the activity of Na+K+-ATPase increased at first and then decreased. Furthermore, there were significant changes in the expressions of oxidative phosphorylation and calcium channel-related genes, suggesting mitochondrial dysfunction may be the potential mechanism of cardiotoxicity induced by AZ and n-TiO2. This study supplies a new perspective for the joint action of AZ and environmental coexisting pollutants and provides a basis for ecological risk management of pesticides.

Distinct vulnerability to oxidative stress determines the ammonia sensitivity of crayfish (Procambarus clarkii) at different developmental stages.

Red swamp crayfish (Procambarus clarkii) has increasingly become a high-value freshwater product in China. During the intensive cultivation, excessive ammonia exposure is an important lethal factor of crayfish. We investigated the toxic effects and mechanisms of ammonia on crayfish at two different developmental stages. A preliminary ammonia stress test showed a 96-h LC50 of 135.10 mg/L and 299.61 mg/L for Stage_1 crayfish (8.47 ± 1.68 g) and Stage_2 crayfish (18.33 ± 2.41 g), respectively. During a prolonged ammonia exposure (up to 96 h), serum acid phosphatase and alkaline phosphatase showed a time-dependent decrease. Histological assessment indicated the degree of hepatopancreatic injury, which was mainly characterized as tubule lumen dilatation, degenerated tubule, vacuolization and dissolved hepatic epithelial cell, increased with exposure time. Enhanced malondialdehyde level and reduced antioxidant capacity of hepatopancreas were also observed. The mRNA expression and activity of catalase and superoxide dismutase showed an initial up-regulation within 24 h, and then gradually down-regulated with the exposure time. In the post-treatment recovery period, the Stage_2 crayfish exerted a stronger antioxidant and detoxification capacity than that of the Stage_1 crayfish, and thus quickly recovered from the ammonia exposure. Our findings provide a further understanding of the adverse effects of ammonia stress and suggest guidelines for water quality management during crayfish farming.

Growth, osmoregulation and ionoregulation of longfin smelt (Spirinchus thaleichthys) yolk-sac larvae at different salinities.

Longfin smelt (Spirinchus thaleichthys) is a threatened anadromous fish species that spawns in freshwater to moderately brackish (i.e. 5-10 ppt) reaches of the upper San Francisco Estuary and has declined to ~1% of its pre-1980s abundances. Despite 50+ years of population monitoring, the efficacy of 10+ years of conservation efforts for longfin smelt remain uncertain due to a limited understanding of how the species responds to environmental variation, such as salinity. For example, high mortality during larval stages has prevented culture efforts from closing the life cycle in captivity. Here, we investigated the effects of salinity on longfin smelt yolk-sac larvae. Newly hatched larvae from four single-pair crosses were acutely transferred to and reared at salinities of 0.4, 5, 10, 20 or 32 ppt. We compared whole-body water and sodium ion (Na+) content, notochord length and yolk-sac volume at 12, 24, 48, 72, and 96 hours post-transfer for each salinity treatment. We found that larvae maintained osmotic and ionic balance at 0.4-10 ppt, whereas salinities ˃10 ppt resulted in decreased water and increased whole-body Na+ content. We also found that larvae grew largest and survived the longest when reared at 5 and 10 ppt, respectively, and that yolk resorption stalled at 0.4 ppt. Finally, there were significant but small interclutch variations in responses to different salinities, with clutch accounting for <8% of the variance in our statistical models. Overall, our results indicate that longfin smelt yolk-sac larvae likely perform best at moderately brackish conditions, thus yielding a mechanism that explains their distribution in field surveys and providing key information for future conservation efforts.

Captive Rearing of Longfin Smelt Spirinchus thaleichthys: First Attempt of Weaning Cultured Juveniles to Dry Feed.

The rapid decline of longfin smelt Spirinchus thaleichthys, a threatened euryhaline forage fish in California, is a serious concern for scientists and resource managers. To recover and conserve this species, a captive culture program was initiated, focusing on the collection, captive rearing and breeding of wild broodstock, and the rearing of their offspring. Although progress has been made in the collection of broodstock and the production and culturing of larvae, no studies have evaluated the rearing of juvenile life stages in captivity. The present study examines methodological considerations for culturing F1 juvenile longfin smelt, specifically, the first efforts toward weaning juveniles to a dry commercial pellet feed. Cultured juvenile longfin smelt were fed live Artemia only or co-fed Artemia and dry feed for 62 days, and the effects of feed type on juvenile survival, growth, body condition, and fatty acid profiles were examined. No significant differences were observed between feeding treatments, despite an 80% reduction in Artemia in the co-feeding treatment. Furthermore, examination of fish stomach contents at the end of the trial confirmed the transition to dry feed. This is the first study to indicate successful feeding by longfin smelt on dry commercial pellets, and suggests that juvenile longfin smelt can be fully weaned onto dry feeds. Results of this study are critical for closing the lifecycle of longfin smelt in captivity and developing a successful conservation culture program for this imperiled species.

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.

The presence of polystyrene nanoplastics enhances the MCLR uptake in zebrafish leading to the exacerbation of oxidative liver damage.

The accumulation of diminutive plastic waste in the environment, including microplastics and nanoplastics, has threatened the health of multiple species. Nanoplastics can adsorb the pollutants from the immediate environment, and may be used as carriers for pollutants to enter organisms and bring serious ecological risk. To evaluate the toxic effects of microcystin-LR (MCLR) on the liver of adult zebrafish (Danio rerio) in the presence of 70 nm polystyrene nanoplastics (PSNPs), zebrafish were exposed to MCLR alone (0, 0.9, 4.5 and 22.5 µg/L) and a mixture of MCLR + PSNPs (100 µg/L) for three months. The results indicated that groups with combined exposure to MCLR and PSNPs further enhanced the accumulation of MCLR in the liver when compared to groups only exposed to MCLR. Cellular swelling, fat vacuolation, and cytoarchitectonic damage were observed in zebrafish livers after exposure to MCLR, and the presence of PSNPs exacerbated these adverse effects. The results of biochemical tests showed the combined effect of MCLR + PSNPs enhanced MCLR-induced hepatotoxicity, which could be attributed to the altered levels of reactive oxygen species, malondialdehyde and glutathione, and activities of catalase. The expression of genes related to antioxidant responses (p38a, p38b, ERK2, ERK3, Nrf2, HO-1, cat1, sod1, gax, JINK1, and gstr1) was further performed to study the mechanisms of MCLR combined with PSNPs aggravated oxidative stress of zebrafish. The results showed that PSNPs could improve the bioavailability of MCLR in the zebrafish liver by acting as a carrier and accelerate MCLR-induced oxidative stress by regulating the levels of corresponding enzymes and genes.

Experimental validation of otolith-based age and growth reconstructions across multiple life stages of a critically endangered estuarine fish.

BACKGROUND: The application of otolith-based tools to inform the management and conservation of fishes first requires taxon- and stage-specific validation. The Delta Smelt (Hypomesus transpacificus), a critically endangered estuarine fish that is endemic to the upper San Francisco Estuary (SFE), California, United States, serves as a key indicator species in the SFE; thus, understanding this species' vital rates and population dynamics is valuable for assessing the overall health of the estuary. Otolith-based tools have been developed and applied across multiple life stages of Delta Smelt to reconstruct age structure, growth, phenology, and migration. However, key methodological assumptions have yet to be validated, thus limiting confidence in otolith-derived metrics that are important for informing major water management decisions in the SFE. METHODS: Using known-age cultured Delta Smelt and multiple independent otolith analysts, we examined otolith formation, otolith-somatic proportionality, aging accuracy and precision, left-right symmetry, and the effects of image magnification for larval, juvenile, and adult Delta Smelt. RESULTS: Overall, otolith size varied linearly with fish size (from 10-60 mm), explaining 99% of the variation in fish length, despite a unique slope for larvae < 10 mm. Otolith-somatic proportionality was similar among wild and cultured specimens. Aging precision among independent analysts was 98% and aging accuracy relative to known ages was 96%, with age estimates exhibiting negligible differences among left and right otoliths. Though error generally increased with age, percent error decreased from 0-30 days-post-hatch, with precision remaining relatively high (≥ 95%) thereafter. Increased magnification (400×) further improved aging accuracy for the oldest, slowest-growing individuals. Together, these results indicate that otolith-based techniques provide reliable age and growth reconstructions for larval, juvenile, and adult Delta Smelt. Such experimental assessments across multiple developmental stages are key steps toward assessing confidence in otolith-derived metrics that are often used to assess the dynamics of wild fish populations.

Microcystin-Induced Immunotoxicity in Fishes: A Scoping Review.

Cyanobacteria (blue-green algae) have been present on Earth for over 2 billion years, and can produce a variety of bioactive molecules, such as cyanotoxins. Microcystins (MCs), the most frequently detected cyanotoxins, pose a threat to the aquatic environment and to human health. The classic toxic mechanism of MCs is the inhibition of the protein phosphatases 1 and 2A (PP1 and PP2A). Immunity is known as one of the most important physiological functions in the neuroendocrine-immune network to prevent infections and maintain internal homoeostasis in fish. The present review aimed to summarize existing papers, elaborate on the MC-induced immunotoxicity in fish, and put forward some suggestions for future research. The immunomodulatory effects of MCs in fish depend on the exposure concentrations, doses, time, and routes of exposure. Previous field and laboratory studies provided strong evidence of the associations between MC-induced immunotoxicity and fish death. In our review, we summarized that the immunotoxicity of MCs is primarily characterized by the inhibition of PP1 and PP2A, oxidative stress, immune cell damage, and inflammation, as well as apoptosis. The advances in fish immunoreaction upon encountering MCs will benefit the monitoring and prediction of fish health, helping to achieve an ecotoxicological goal and to ensure the sustainability of species. Future studies concerning MC-induced immunotoxicity should focus on adaptive immunity, the hormesis phenomenon and the synergistic effects of aquatic microbial pathogens.

Exposure to permethrin or chlorpyrifos causes differential dose- and time-dependent behavioral effects at early larval stages of an endangered teleost species.

Pyrethroid and organophosphate pesticides are two of the most commonly used classes of insecticide worldwide. At sublethal concentrations, permethrin (a pyrethroid) and chlorpyrifos (an organophosphate) impact behavior in model fish species. We investigated behavioral effects of environmentally relevant concentrations of permethrin or chlorpyrifos on early larval delta smelt Hypomesus transpacificus, a Critically Endangered teleost species endemic to the San Francisco Bay Delta, California, USA. Using a photomotor behavioral assay of oscillating light and dark periods, we measured distance moved, turn angle, meander, angular velocity, rotations, thigmotaxis (time spent in the border versus center), and swim speed duration and frequency. The lowest concentrations of permethrin used in the tests (0.05 and 0.5 µg l-1) caused significant increases in distance moved at 72 and 96 h, respectively. At 48, 72, and 96 h of exposure, 5 µg l-1 of permethrin caused a hyperactive state in which the larvae significantly decreased thigmotaxis, quickly turning in short bouts of activity, characterized by significant increases in rotations and freezing events. Larvae exposed to 0.05 µg l-1 chlorpyrifos significantly increased thigmotaxis at 72 and 96 h. In response to 5 µg l-1 chlorpyrifos, larvae significantly increased velocity at 72 h exposure, and significantly increased freezing events at 96 h. Behavioral data on larval delta smelt exposed to contaminants present in their limited habitat have the potential to aid evaluations of the suitability of spawning and rearing habitats for this endangered species, thus improving conservation management strategies focused on this sensitive life stage.

Mechanisms of parental co-exposure to polystyrene nanoplastics and microcystin-LR aggravated hatching inhibition of zebrafish offspring.

The combined toxicity effects of microcystins-LR (MCLR) and polystyrene nanoplastics (PSNPs) on the hatching of F1 zebrafish (Danio rerio) embryos were investigated in this study due to the increasing concerns of both plastic pollution and eutrophication in aquatic environments. Three-month-old zebrafish were used to explore the molecular mechanisms underlying the combined effect of MCLR (0, 0.9, 4.5, and 22.5 µg/L) on egg hatching in the existence of PSNPs (100 µg/L). The results demonstrated the existence of PSNPs further increased the accumulation of MCLR in F1 embryos. The hatching rates of F1 embryos were inhibited after exposure to 22.5 µg/L MCLR, and the presence of PSNPs aggravated the hatching inhibition induced by MCLR. The decrease of hatching enzyme activity and the abnormality of spontaneous movement were observed. We examined the altered expression levels of the genes associated with the hatching enzyme (tox16, foxp1, ctslb, xpb1, klf4, cap1, bmp4, cd63, He1.2, zhe1, and prl), cholinergic system (ache and chrnα7), and muscle development (Wnt, MyoD, Myf5, Myogenin, and MRF4). The results suggested the existence of PSNPs exacerbated the hatching inhibition of F1 embryos through decreasing the activity of enzyme, interfering with the cholinergic system, and affecting the muscle development.

The joint effect of parental exposure to microcystin-LR and polystyrene nanoplastics on the growth of zebrafish offspring.

The coexistence of nanoplastics (NPs) and various pollutants in the environment has become a problem that cannot be ignored. In order to identify the microcystin-LR (MCLR) bioaccumulation and the potential impacts on the early growth of F1 zebrafish (Danio rerio) offspring in the presence of polystyrene nanoplastics (PSNPs), PSNPs and MCLR were used to expose adult zebrafish for 21days. The exposure groups divided into MCLR (0, 0.9, 4.5 and 22.5µgL-1) alone groups and PSNP (100µgL-1) and MCLR co-exposure groups. F1 embryos were collected and developed to 120 h post-fertilization (hpf) in clear water. Compared with the exposure to MCLR only, the combined exposure increased the parental transfer of MCLR to the offspring and subsequently exacerbated the growth inhibition of F1 larvae. Further research clarified that combined exposure of PSNPs and MCLR could reduce the levels of thyroxine (T4) and 3, 5, 3'-triiodothyronine (T3) by altering the expression of hypothalamus-pituitary-thyroid (HPT) axis-related genes, eventually leading to growth inhibition of F1 larvae. Our results also exhibited combined exposure of PSNPs and MCLR could change the transcription of key genes of the GH/IGF axis compared with MCLR single exposure, suggesting the GH/IGF axis was a potential target for the growth inhibition of F1 larvae in PSNPs and MCLR co-exposure groups. The present study highlights the potential risks of coexistence of MCLR and PSNPs on development of fish offspring, and the environmental risks to aquatic ecosystems.