A time-course study of immune response in Japanese flounder Paralichthys olivaceus exposed to heavy oil

Purpose

The immunotoxicities of oil and its components on fish immunities have been investigated, but there is little literature on the recovery of the fish from the immune suppression. Therefore, the recovery of Japanese flounder Paralichthys olivaceus from an immunosuppressive effect due to heavy oil (HO) exposure was investigated in this study.

Methods

Fish were exposed to HO at a concentration of 0.385?g/L for 2?days, while control fish received no exposure. Seven fish were sampled at 0, 3, 7, and 14?days post-exposure. The respiratory rate was measured everyday as an indicator of the acute effect of HO exposure. Fish serum was collected and used for antibacterial activity assay against Edwardsiella tarda. Expression changes of respiratory and immune-related genes were evaluated by real-time PCR.

Results and discussion

The respiratory rate was significantly increased in the HO-exposed group until 4?days post-exposure. A respiratory-related gene, ??-hemoglobin, was also significantly downregulated in the spleen both at 0 and 7?days post-exposure and kidney at 3?days post-exposure in HO-exposed fish. Immunotoxicity, including suppression of antibacterial activities and downregulation of the IgM gene, was observed in HO-exposed fish until 3?days post-exposure, but not after that time. From these results, we conclude that the fish likely return to normal status around 1?week.     

Genotoxic effect of copper on salivary gland polytene chromosomes of Chironomus riparius Meigen 1804 (Diptera, Chironomidae)

The genotoxic action of copper (Cu) on the polytene chromosomes of Chironomus riparius was investigated by analysing structural and functional chromosome aberrations of fourth instars larvae hatched from eggs subject to acute (48 h) exposure with three environmentally relevant concentrations of aqueous Cu (0.005, 0.01, 0.05 mg/l). A dose dependent relationship was observed between Cu concentration and frequency of chromosomal aberrations. A significantly higher frequency of functional alterations, specifically decondensed centromeres and telomeres, and reduction in activity of Balbiani rings, was observed in treated material compared to control. A comparison of breakpoints resulting from treatment with chromium and lead from earlier studies with those Cu-induced identified a series of chromosomal weak points particularly vulnerable to trace metals. We also show that the appearance of structural and functional chromosome aberrations are more sensitive indicators of acute Cu toxicity in chironomid larvae than changes in external morphology.     

Toxicological effect of MPA-CdSe QDs exposure on zebrafish embryo and larvae

Cadmium selenium (CdSe) quantum dots (QDs) are semiconductor nanocrystals that hold wide range of applications and substantial production volumes. Due to unique composition and nanoscale properties, their potential toxicity to aquatic organisms has increasingly gained a great amount of interest. However, the impact of CdSe QDs exposure on zebrafish embryo and larvae remains almost unknown. Therefore, the lab study was performed to determine the developmental and behavioral toxicities to zebrafish under continuous exposure to low level CdSe QDs (0.05-31.25 mg L⁻¹) coated with mercaptopropionic acid (MPA). The results showed MPA-CdSe exposure from embryo to larvae stage affected overall fitness. Our findings for the first time revealed that: (1) The 120 h LC₅₀ of MPA-CdSe for zebrafish was 1.98 mg L⁻¹; (2) embryos exposed to MPA-CdSe resulted in malformations incidence and lower hatch rate; (3) abnormal vascular of FLI-1 transgenic zebrafish larvae appeared after exposure to MPA-CdSe including vascular junction, bifurcation, crossing and particle appearance; (4) larvae behavior assessment showed during MPA-CdSe exposure a rapid transition from light-to-dark elicited a similar, brief burst and a higher basal swimming rate; (5) MPA-CdSe induced embryos cell apoptosis in the head and tail region. Results of the observations provide a basic understanding of MPA-CdSe toxicity to aquatic organisms and suggest the need for additional research to identify the toxicological mechanism.     

Behavioral changes in fish exposed to phytoestrogens

We investigated the behavioral effects of exposure to waterborne phytoestrogens in male fighting fish, Betta splendens. Adult fish were exposed to a range of concentrations of genistein, equol, beta-sitosterol, and the positive control 17beta-estradiol. The following behaviors were measured: spontaneous swimming activity, latency to respond to a perceived intruder (mirror reflection), intensity of aggressive response toward a perceived intruder, probability of constructing a nest in the presence of a female, and the size of the nest constructed. We found few changes in spontaneous swimming activity, the latency to respond to the mirror, and nest size, and modest changes in the probability of constructing a nest. There were significant decreases, however, in the intensity of aggressive behavior toward the mirror following exposure to several concentrations, including environmentally relevant ones, of 17beta-estradiol, genistein, and equol. This suggests that phytoestrogen contamination has the potential to significantly affect the behavior of free-living fishes.     

Embryonic development assay with Daphnia magna: application to toxicity of aniline derivatives.

An assay system using Daphnia magna embryos was applied to investigate the adverse effects of aniline derivatives. The data were compared with our previous data for chlorophenols. This new assay provides useful information to evaluate the toxicity of chemicals and the differences in sensitivity between the life stages. The effects of 15 aniline derivatives on embryonic development of D. magna embryos were determined. At the start of exposure, 2-6-h old eggs (between stages 1 and 2, round in shape, diameter approx. 400 microm), were used. In control and solvent control groups, embryonic development from an egg to a free-swimming animal proceeded completely within 3 days with more than 90% hatchability. Median effective concentrations (EC50s) to reduce the numbers hatched were determined and gross morphological abnormalities of hatched animals recorded. Anilines induced no obvious morphological abnormalities and no developmental delay although premature deaths occurred. However, they affected the number of embryos hatching in a dose-dependent manner. In addition, this embryo assay was more sensitive to aniline derivatives (except for aniline) than acute juveniles immobilization assay. Ratios of 48-h EC50 (juvenile)/3-day EC50 (embryo) for eight anilines were greater than 5.0. Particularly, the ratios of 4-methyl-, 4-ethyl- and 3-methylaniline were 77, 23 and 11, respectively. EC50s for embryos and juveniles were poorly correlated (r = 0.41). This indicated that the sensitivities of the two life stages were different to the effects of anilines. EC50s were poorly correlated (r = -0.097) with the log Kow (1-octanol/water partition coefficient). These results were compared with previous results for phenols.     

Synergistic effects caused by atrazine and terbuthylazine on chlorpyrifos toxicity to early-life stages of the zebrafish Danio rerio

This study examined the effects of three widely used pesticides that have been previously detected in aquatic systems neighbouring agricultural fields on the early-life stages of the zebrafish Danio rerio. Tests involving single exposures and binary combinations of the s-triazine herbicides (atrazine and terbuthylazine) and the organophosphate insecticide chlorpyrifos were performed. Several endpoints, such as swimming behaviour, morphological abnormalities and mortality, were studied. In addition, the inhibition of acetylcholinesterase (AChE) activity was investigated in order to evaluate the mode of action and toxicity of chlorpyrifos in the presence of these herbicides. Results indicate that both binary mixtures elicited synergistic responses on the swimming behaviour of zebrafish larvae. Moreover, although the herbicides were not effective inhibitors of the AChE on their own, a synergistic inhibition of the enzyme activity was obtained by exposure to mixtures with chlorpyrifos. We observed a correlation between impairment of swimming behaviour of the larvae and inhibition of AChE activity. This study supports previous studies concerning the risk assessment of mixtures since the toxicity may be underestimated when looking only at the single toxicants and not their mixtures.     

Effect of ammonia on the survival of Zacco platypus (Temminck and Schlegel) at each developmental stage

The survival rates were studied of fertilized eggs, larvae, fry, and adults of pale chub Zacco platypus (Temminck and Schlegel) exposed to various concentrations of ammonia. There was a decrease in hatching rate and an increase in deformed alevins at a concentration of 0.9 mg liter(-1) of ammonia. All alevins were deformed at a concentration of 1.6 mg liter(-1) of ammonia and no eggs hatched at a concentration of 3.1 mg liter(-1) of ammonia. No larvae survived for 84, 24 and 12 h at concentrations of 1.2, 2.0 and 4.7 mg liter(-1) of ammonia, respectively. Eighty percent of the fry died after 168 h and no fry survived for 24 h at concentrations of 1.6 and 2.1 mg liter(-1) of ammonia, respectively. Eighty percent of the adults died after 24 h and no adult survived for 12 h at concentrations of 1.0 and 1.9 mg liter(-1), respectively. Larger adults were the most susceptible to ammonia. The order of resistance to ammonia was eggs, fry, larvae, and adults.     

Chronic dietary exposure to pyrolytic and petrogenic mixtures of PAHs causes physiological disruption in zebrafish - part I: Survival and growth

The release of polycyclic aromatic hydrocarbons (PAHs) into the environment has increased very substantially over the last decades leading to high concentrations in sediments of contaminated areas. To evaluate the consequences of long-term chronic exposure to PAHs, zebrafish were exposed, from their first meal at 5 days post fertilisation until they became reproducing adults, to diets spiked with three PAH fractions at three environmentally relevant concentrations with the medium concentration being in the range of 4.6–6.7 μg g^?1 for total quantified PAHs including the 16 US-EPA indicator PAHs and alkylated derivatives. The fractions used were representative of PAHs of pyrolytic (PY) origin or of two different oils of differing compositions, a heavy fuel (HO) and a light crude oil (LO). Fish growth was inhibited by all PAH fractions and the effects were sex specific: as determined with 9-month-old adults, exposure to the highest PY inhibited growth of females; exposure to the highest HO and LO inhibited growth of males; also, the highest HO dramatically reduced survival. Morphological analysis indicated a disruption of jaw growth in larvae and malformations in adults. Intestinal and pancreatic enzyme activities were abnormal in 2-month-old exposed fish. These effects may contribute to poor growth. Finally, our results indicate that PAH mixtures of different compositions, representative of situations encountered in the wild, can promote lethal and sublethal effects which are likely to be detrimental for fish recruitment.     

Embryonic exposure to low concentration of bisphenol A affects the development of Oryzias melastigma larvae

The prevalence of bisphenol A (BPA) in the environment has attracted increasing attention because of the toxicity of this manmade pollutant. However, the toxicity related to cardiac development remains largely unknown. In the present paper, we investigated the cardiac toxicity of BPA using marine medaka (Oryzias melastigma) embryos. At 2?days postfertilization (dpf), the embryos were continuously exposed to a low concentration of BPA (200?μg/L) for the whole embryonic stage. Heart rate and sinus venosus (SV)-bulbus arteriosus (BA) distance were measured under microscopy. The mRNA expression levels of genes were quantified by SYBR real-time RT-PCR, and hematoxylin and eosin (H&E) staining was used to examine the histology of fish larvae hearts. Neither the heart rate nor the SV-BA distance of the embryos was affected by BPA exposure. However, the mRNA expression levels of Na(+)-K(+)-ATPase, BMP4, COX-1, FGF8, GATA4, and NKX2.5 were all downregulated at the critical developmental stages (6 and 10?dpf). Interestingly, the mRNA expression levels of COX-2 and LERP were significantly upregulated at 10?dpf. The mRNA expressions of inflammation-related genes (TNFα, IL1β, SOD, and CCL11) were all significantly upregulated after exposure. Moreover, we found that both the body length and the body width decreased in the larvae after embryonic exposure to BPA. The distributed foci of inflammation were observed in the juveniles after 2?weeks' depuration. Exposure to BPA at embryonic stages could alter the expression of heart development-related genes and inflammation-related genes of O. melastigma. The larvae hatched from exposed embryos showed the foci of inflammation in the heart ventricles and the decrease of the body length and width.     

Chronic dietary exposure to pyrolytic and petrogenic mixtures of PAHs causes physiological disruption in zebrafish—part II: behavior

In the last 10 years, behavior assessment has been developed as an indicator of neurotoxicity and an integrated indicator of physiological disruption. Polycyclic aromatic hydrocarbon (PAH) release into the environment has increased in recent decades resulting in high concentrations of these compounds in the sediment of contaminated areas. We evaluated the behavioral consequences of long-term chronic exposure to PAHs, by exposing zebrafish to diets spiked with three PAH fractions at environmentally relevant concentrations. Fish were exposed to these chemicals from their first meal (5 days postfertilization) until they became reproducing adults (at 6 months old). The fractions used were representative of PAHs of pyrolytic (PY) origin and of two oils differing in composition (a heavy fuel oil (HO) and a light crude oil (LO)). Several tests were carried out to evaluate circadian spontaneous swimming activity, responses to a challenge (photomotor response), exploratory tendencies, and anxiety levels. We found that dietary PAH exposure was associated with greater mobility, lower levels of exploratory activity, and higher levels of anxiety, particularly in fish exposed to the HO fraction and, to a lesser extent, the LO fraction. Finally, our results indicate that PAH mixtures of different compositions, representative of situations encountered in the wild, can induce behavioral disruptions resulting in poorer fish performance.     

Lethal effects on different marine organisms, associated with sediment�Cseawater acidification deriving from CO2 leakage

CO(2) leakages during carbon capture and storage in sub-seabed geological structures could produce potential impacts on the marine environment. To study lethal effects on marine organisms attributable to CO(2) seawater acidification, a bubbling CO(2) system was designed enabling a battery of different tests to be conducted, under laboratory conditions, employing various pH treatments (8.0, 7.5, 7.0, 6.5, 6.0, and 5.5). Assays were performed of three exposure routes (seawater, whole sediment, and sediment elutriate). Individuals of the clam (Ruditapes philippinarum) and early-life stages of the gilthead seabream, Sparus aurata, were exposed for 10?days and 72?h, respectively, to acidified clean seawater. S. aurata larvae were also exposed to acidified elutriate samples, and polychaete organisms of the specie Hediste diversicolor and clams R. philippinarum were also exposed for 10?days to estuarine whole sediment. In the fish larvae elutriate test, 100?% mortality was recorded at pH?6.0, after 48?h of exposure. Similar results were obtained in the clam sediment exposure test. In the other organisms, significant mortality (p?相似文献   

The impact of sewage sludge exposure on the reproduction of the sand goby, Pomatoschistus minutus

Adult sand gobies were exposed to 0.1% sewage sludge for 19 weeks prior to the end of spawning. Exposure to sewage sludge had a significant effect on male mortality rates but no significant effects on the gonadosomatic index of males or females. There were no major effects of sludge exposure on testes androgen content or on testes release of androgens after in vitro gonadotrophin stimulation. Fecundity and the number of larvae produced were not significantly affected by the sludge exposure. There was a tendency for eggs and larvae from sludge-exposed females to have a higher mortality rate. At a population level, however, there was a major reduction in the number of eggs and larvae produced in the sludge-exposed population which reflected a failure of some females to spawn. Of the larvae produced, 60-70% from sludge-exposed parents were lighter and had a larger yolk-sac volume compared to larvae from non-exposed parents which may have indicated impaired yolk utilisation.     

Residues of lindane and its metabolites in eggs, chicks, and body tissues of hen pheasants after ingestion of lindane-14C via treated wheat seed or gelatin capsules.

Studies were carried out to investigate possible contamination of pheasants with residues of lindane used as seed dressings for the control of wireworms in cereal crops. One group of laying hen pheasants was fed 20 mg of lindane-14C in gelatin capsules while another group was fed wheat seed treated with 100 ppm of lindane-14C for 15 days. Residues in eggs were monitored for about 70 days. Residues in chicks hatched from the eggs were also measured. Concentrations of residues in muscle, liver, brain, and fatty tissues were determined at various times. 14C-labeled residues in eggs increased sharply in the capsule treated group and reached an average maximum of about 19 ppm in 8 days. This level decreased to less than 0.5 ppm in about 50 days. Residues in eggs from the pheasants fed treated seed increased gradually to an average maximum of 17 ppm 22 days after commencement of the feeding program. This level decreased to less than 0.5 ppm 50 days later. Residues in hatched chicks were significantly lower than those in the eggs. Highest concentrations of residues were found in fatty tissues which decreased to non-detectable level in 6 months. Several chlorobenzene metabolites were identified in egg yolk and chick tissues. Chlorophenolic metabolites were indicated only in chicks.     

Effects of low ambient pH on perivitelline fluid of Ambystoma maculatum (Shaw) eggs

Egg clumps of Ambystoma maculatum were collected from a pond on the south side of the Gaspereau Valley, Kings County, Nova Scotia, to determine the effects of low ambient pH levels on the perivitelline fluid pH. Egg masses were placed in aquaria filled with pond water adjusted to pH 4.0, 5.0, 6.0 and 7.0. Perivitelline fluid was withdrawn from the eggs at 1, 3, 5, 7, 24, 48, 96 and 120 h after initial immersion, and its pH measured. Initial perivitelline fluid values decreased significantly 7 to 96 h after initial immersion in treatments altered to pH 4.0 and 5.0. Rate of H(+) influx into the perivitelline fluid is much greater in eggs immersed in water at pH 4 than in water at pH 5 or 6. Median Tolerance Limit tests indicated a critical pH value between 3.5 and 4.0 for newly hatched larvae.     

Tributyltin exposure causes brain damage in Sebastiscus marmoratus

Tributyltin (TBT) is a ubiquitous marine environmental contaminant characterized primarily by its reproductive toxicity. However, the neurotoxic effect of TBT has not been extensively described, especially in fishes which have a high number of species in the marine environment. This study was conducted to investigate the neurotoxic effects of TBT at environmental levels (1, 10, and 100ngl(-1)) on female Sebastiscus marmoratus. The results showed that TBT exposure induced apoptosis in brain cells of three regions including the pallial areas of the telencephalon, the granular layer of the optic tectum, and the cerebellum. In addition, the increase of reactive oxygen species and nitric oxide levels, and the decrease of Na(+)/K(+)-ATPase activity were found in the brain. The results strongly indicated neurotoxicity of TBT to fishes. According to the regions in which apoptosis was found in the brain, TBT exposure might influence the schooling, sensory and motorial functions of fishes.     

Pesticide effects on freshwater zooplankton: an ecological perspective

The effects of pesticides on zooplankton are reviewed and their ecological significance is discussed. Toxicity is shown to vary depending on animal species, genotype, life stage, and size at birth. Natural stresses such as food shortage, oxygen depletion and odors of potential predators can also affect toxicity. Populations in the growth phase are vulnerable to pesticides but have the potential to recover rapidly from the damage. Pesticides may affect the population dynamics by controlling individual survival and reproduction, and by altering the sex ratio. Furthermore, toxic chemicals may control predation risk by changing swimming behavior and body morphology, and this in turn influences the population dynamics. Many zooplankton display morphological and behavioral responses to predators when exposed to their odor-producing chemicals. However, pesticides induce a maladaptive response to predator odor, and this poses an ecological risk. The following patterns are recognized as effects of pesticides at the community and ecosystem levels: (1) induction of dominance by small species; (2) an increase of species richness and diversity; and (3) elongation of the food chain and reduction of energy transfer efficiency from primary producers to top predators.     

Toxicity of methyl-tert-butyl ether to freshwater organisms

Increased input of the fuel oxygenate methyl-tert-butyl ether (MTBE) into aquatic systems has led to concerns about its effect(s) on aquatic life. As part of a study conducted by University of California scientists for the State of California, the Aquatic Toxicology Laboratory, UC Davis, reviewed existing literature on toxicity of MTBE to freshwater organisms, and new information was generated on chronic, developmental toxicity in fish, and potential toxicity of MTBE to California resident species. Depending on time of exposure and endpoint measured, MTBE is toxic to various aquatic organisms at concentrations of 57-> 1000 mg/l (invertebrates), and 388-2600 mg/l (vertebrates). Developmental effects in medaka (Oryzias latipes) were not observed at concentrations up to 480 mg/l, and all fish hatched and performed feeding and swimming in a normal manner. Bacterial assays proved most sensitive with toxicity to Salmonella typhimurium measured at 7.4 mg/l within 48 h. In microalgae, decreased growth was observed at 2400 and 4800 mg/l within 5 days. MTBE does not appear to bioaccumulate in fish and is rapidly excreted or metabolized. Collectively, the available data suggests that at environmental MTBE exposure levels found in surface waters (< 0.1 mg/l) this compound is likely not acutely toxic to aquatic life. However, more information is needed on chronic and sublethal effects before we can eliminate the possibility of risk to aquatic communities at currently detected concentrations.     

Sublethal effects of monocrotophos on locomotor behavior and gill architecture of the mosquito fish, Gambusia affinis

Subacute studies of monocrotophos [Dimethyl (E)-1-methyl-2-(methyl-carbamoyl) vinyl phosphate] on mosquito fish, Gambusia affinis, were carried out in vivo for 24 days to assess the locomotor behavior, structural integrity of gill, and targeted enzyme acetylcholinesterase (AChE, EC: 3.1.1.7) interactions. Monocrotophos (MCP) can be rated as moderately toxic to G. affinis, with a median lethal concentration (LC(50)) of 20.49 +/- 2.45 mgL(-1). The fish exposed to sublethal concentration of LC(10) (7.74 mgL(-1)) were under stress and altered their locomotor behavior, such as distance traveled per unit time (m min(-1)) and swimming speed (cm sec(-1)) with respect to the length of exposure. Inhibition in the activity of brain AChE and deformities in the primary and secondary lamellae of gill may have resulted in failure of exchange of gases. The maximum inhibition of 95% of AChE activity was observed on days 20 and 24.Morphological aberrations in the gills were also studied during exposure to the sublethal concentration of monocrotophos for a period ranging from 8 to 24 days. The extent of damage in gill was dependent on the duration of exposure. The findings revealed that inhibition in brain AChE activity and structural alteration in gill were responsible for altering the locomotor behavior of exposed fish.     

Cloning of a river pufferfish (Takifugu obscurus) metallothionein cDNA and study of its induction profile in cadmium-exposed fish

We report here the full-length cDNA sequence of metallothionein (MT) gene from an anadromous river pufferfish, Takifugu obscurus (order: Tetradotiformes; family: Tetradontidae). Phylogenetic relationship analysis revealed that the identified MT has high sequence similarity with many Perciformes fish species. The tissue distribution and concentration- and time-dependent expression of MT mRNA were studied in fish exposed to cadmium. Liver showed the highest level of MT gene expression followed by other tissues (brain, gill and kidney) in response to cadmium exposure. Muscle showed a weak expression response of MT gene. Time-course study revealed highest early phase (at 6h) expression in the brain and late persistence of induction in the intestine. MT mRNA expression showed a concentration-dependent expression in all the tissues. However, induction in brain and liver occurred at much lower concentrations as compared to other tissues. Our results demonstrate that MT in T. obscurus is induced by cadmium exposure which indicates that it plays a functionally conserved function of metal detoxification.     

Organochlorine contaminant assessment in great blue herons using traditional and nonlethal monitoring techniques

A nonlethal method is discussed for the evaluation of contaminant concentrations in whole eggs. Concentrations of pentachlorobenzene, hexachlorobenzene, DDE, and the PCB congeners, BZ-60, BZ-118, BZ-138, BZ-180, and BZ-170 were quantified in tissue samples from great blue herons (Ardea herodias). All tissues within whole eggs from two colonies were homogenized together and analysed for these chlorinated contaminants. Contents and chorio-allantoic membranes (CAMs) of additional whole eggs were separated and analysed. Contaminant distributions were determined for the CAM and contents of whole eggs from the same colonies. CAM tissues remaining in hatched eggs were also analysed for comparative purposes. Utilizing the distributions derived for contaminants between CAM and egg contents and the chemical concentrations determined in CAMs from hatched eggs, contaminant burdens in whole eggs were calculated. This process produced concentration estimates that described actual, whole egg burdens of chlorinated contaminants within a factor of 2 (1.03-3.7). Contaminant burdens in eggs from the two colonies were also statistically different for DDE and total PCB concentrations.