Integration of biological responses from a suite of bioassays for the Venice Lagoon (Italy) through sediment toxicity index - Part A: Development and comparison of two methodological approaches

Marine and coastal quality assessment, based on test batteries involving a wide array of endpoints, organisms and test matrices, needs for setting up toxicity indices that integrate multiple toxicological measures for decision-making processes and that classify the continuous toxicity response into discrete categories according to the European Water Framework Directive.Two toxicity indices were developed for the lagoon environment such as the Venice Lagoon. Stepwise procedure included: the construction of a database that identified test-matrix pairs (indicators); the selection of a minimum number of ecotoxicological indicators, called toxicological core metrics (CMs-tox) on the basis of specific criteria; the development of toxicity scores for each CM-tox; the integration of the CMs-tox into two indices, the Toxicity Effect Index (TEI), based on the transformation of Toxic Unit (TU) data that were integrated as logarithmic sum, and the Weighted Average Toxicity Index (WATI), starting from toxicity classes integrated as weighted mean. Results from the indices are compared; advantages and drawbacks of both approaches are discussed.     

Uptake and depuration of the anti-depressant fluoxetine by the Japanese medaka (Oryzias latipes)

The selective serotonin reuptake inhibitor (SSRI) class of anti-depressants is among the most widely prescribed groups of pharmaceuticals. Consequently, aquatic ecosystems impacted by municipal wastewater discharges are predicted to receive substantial annual loadings of these compounds. Although SSRIs have been detected in fish tissues, little is known of their uptake and depuration in freshwater fish species. In this study, Japanese medaka (Oryzias latipes) were exposed to fluoxetine at a nominal concentration of 0.64 microg L(-1) for 7d and subsequently allowed to depurate in clean water over a 21d period. Fluoxetine uptake by medaka was observed within the first 5h of exposure and the biologically active metabolite, norfluoxetine, was also detected in medaka tissues during this timeframe. A maximum fluoxetine concentration was measured in medaka by the third day of the uptake phase, yielding an uptake rate constant (k(1)) of 5.9+/-0.5 (d(-1)). During the depuration phase of the experiment, a half life of 9.4+/-1.1d was determined for fluoxetine. Using these data, bioconcentration factor (BCF) values of 74 and 80 were estimated for fluoxetine and a pseudo-BCF (the ratio of the concentration of norfluoxetine in medaka and the aqueous fluoxetine concentration) of 117 was calculated for norfluoxetine. These results indicate longer persistence and greater potential for the bioaccumulation of fluoxetine and norfluoxetine in fish tissues than would be predicted from prior half life estimates derived using mammalian species.     

Effects of formulations of the fungicide, pentachloronitrobenzene on early life stage development of the Japanese medaka (Oryzias latipes)

Quintozene is a fungicide containing the active ingredient, pentachloronitrobenzene (PCNB) that is used to control "snow mold" on golf courses in temperate regions of North America. In this study, quintozene and a formulation of quintozene widely used on golf courses, FFII were tested for toxicity to early life stages of the Japanese medaka, Oryzias latipes. For medaka exposed in static non-renewal assays to quintozene for 17d from the fertilized egg stage to yolk resorption at the fry stage, the LC(50) for mortality was a nominal concentration of 707 microgl(-1) and the effective concentration for 50% hatch (i.e. EC(50)) was a nominal concentration of 71 microgl(-1). Eggs and fry showed developmental abnormalities, including ocular malformations and retarded development of the brain, notochord, organs and body segmentation, which were interpreted as teratogenic responses to exposure to PCNB. For medaka exposed to quintozene, the LOECs for abnormalities of the eye and all other developmental abnormalities were 750 and 100 microgl(-1), respectively. In medaka exposed to the FFII formulation, similar patterns of mortalities, reduced hatching success and developmental abnormalities were observed, but at higher test concentrations that were consistent with the proportion of quintozene in the formulation. For medaka exposed to the formulation, the LOECs for abnormalities of the eye and all other developmental abnormalities were 10,000 and 1,000microgl(-1), respectively. Overall, these data indicate that studies should be conducted to assess the risk of exposure of early life stages of fish to quintozene in watersheds impacted by golf courses.     

The toxicity of titanium dioxide nanopowder to early life stages of the Japanese medaka (Oryzias latipes)

In this study, fertilized Japanese medaka (Oryzias latipes) embryos were exposed from fertilization to 5 d post-hatch using static non-renewal assays to aqueous suspensions of titanium dioxide nanoparticles (nTiO₂) ranging in nominal concentrations between 0 and 14 μg mL⁻¹. The average size of the nTiO₂ in the stock solution before addition to the test treatments was 87 nm (±14 nm). TiO₂ materials accumulated in a concentration dependent manner on the chorionic filaments of developing medaka embryos with evidence of pericardial edema occurring during embryo development. However, no significant (p > 0.05) increases in mortality relative to control treatments were observed for the nTiO₂ exposed embryos. A concentration dependent increase in cumulative percent hatch was observed at 11 d, indicating that exposure to increasing concentrations of nTiO₂ resulted in the premature hatch of medaka embryos. Post-hatch, a significant proportion of sac fry from the nTiO₂ exposure groups exhibited moribund swimming behavior and these individuals also experienced greater mortality at 15 d post-hatch. Combined, these results demonstrate that exposure to nTiO₂ can impact the development of early life stages of fish.     

Effect of certain chemicals on the reproduction of medaka (Oryzias latipes)

In order to understand the effects of estrogenic chemicals on fish reproduction, we exposed male medaka (Oryzias latipes) to a natural estrogen [17 beta-estradiol (17 beta-E2)] and three estrogenic chemicals [bisphenol-A, nonylphenol (NP) and di(2-ethylhexyl)phthalate (DEHP)]. After two weeks' exposure, one male medaka was kept together with two female medaka for spawning, and the number of eggs and hatchings were compared to those of a negative control group. The results indicated that exposure to 17 beta-E2 caused a significant decrease in the number of eggs and hatchings as compared to the negative control group at and above 3 nmol/l. Also, the highest concentrations of bisphenol-A and NP caused a decrease in the number of hatchings, but no decrease in hatchings was observed in DEHP treatments. In the treatment using these chemicals the decrease in egg numbers was not so much as in hatching numbers. When compared to other in vitro studies, concentrations observed to have adverse effects on reproduction in this study are generally lower. In addition, it was suggested that physical alterations, such as an induction of plasma vitellogenin, were caused at much lower concentrations than those at which a decline in reproductivity was actually induced.     

MFI型纳米分子筛对重金属生物有效性的影响

利用小型鱼日本青鳉考察MFI型纳米分子筛对有毒重金属离子(Cu2+、Cd2+)生物有效性的影响,并通过静态平衡实验探讨MFI型纳米分子筛对重金属的吸附行为.暴露3d,MFI型纳米分子筛可降低Cu2、Cd2+的生物有效性；暴露10 d,MFI型纳米分子筛对Cu2+、Cd2+生物有效性的影响减小；清除7d后,MFI型纳米分...     

Developmental and behavioral effects of embryonic exposure to the polybrominated diphenylether mixture DE-71 in the killifish (Fundulus heteroclitus)

Exposures to penta polybrominated diphenylether (PeBDE) cause neurobehavioral toxicity in developing mice and rats. As levels of these ubiquitous contaminants are increasing in the environment, this raises concern that wildlife may also suffer such effects, with consequences for their ability to catch prey and avoid predators. PeBDE levels in wild-caught fish have been steadily escalating over the past fifteen years. To our knowledge, behavioral consequences of piscine embryonic exposure to PeBDE has not yet been studied. The objectives of this investigation were to characterize effects on development in an environmentally relevant fish model, and test for latent behavioral effects following cessation of exposure. Embryos from the estuarine minnow, Fundulus heteroclitus, were exposed from day 0-7 post fertilization to the industrial PeBDE mixture, DE-71 (0.001 to 100 microg l(-1)). Embryos were assayed for hatching success, development, and microsomal enzyme cytochrome P4501A (CYP1A) activity, which was determined by analysis of in ovo ethoxy-resorufin-O-deethylase (EROD) activation in embryos. Larval fish were assayed for predation ability, activity level, and fright response to a simulated predator. Juvenile fish were assayed for learning ability in a three-chambered fish maze. No induction of embryonic EROD activity was observed, nor was a high dose of DE-71 able to inhibit EROD activity induced by beta-naphthoflavone. No deformities were detected, but a subtle developmental asymmetry with respect to tail curvature direction was observed, and a hatching delay of up to 4.5 days was noted. Behavioral test results suggest that embryonic exposure to DE-71 may alter activity level, fright response, predation rates, and learning ability in subsequent life stages.     

Avoidance by olfaction in a fish, medaka (Oryzias latipes), to aquatic contaminants

Behavioral tests using a fish, medaka (Oryzias latipes), from which the olfactory organs were resected confirmed that fish avoid aquatic contaminants such as surfactants and pesticides mainly by olfaction. Control medaka (non-resected) exhibited significant avoidance to 10, 20 and 30 microg liter(-1) of LAS (sodium linear laurylbenzene sulfonate), 90 and 100 microg liter(-1) of fenitrothion (dimethyl 4-nitro-m-toryl phosphorothionate), and 500 microg liter(-1) of POE-ether (polyoxyethylene lauryl ether). By contrast, medaka with bilateral nose resections (BNRM) exhibited no avoidance to 10-50 microg liter(-1) of LAS, 90 and 100 microg liter(-1) of fenitrothion, and 500 microg liter(-1) of POE-ether. Although medaka with unilateral nose resection (UNRM) avoided some concentrations of these toxicants, the UNRM exhibited no avoidance to 10 and 40 microg liter(-1) of LAS and 90 microg liter(-1) of fenitrothion. That is, the avoidance response of the UNRM was less distinct than that of the controls. This difference may have been caused by the deterioration of detecting ability and the paralyzation or adaptation of olfaction.     

Bioaccumulation of polychlorinated biphenyls in the eel (Anguilla anguilla) at the Camargue Nature Reserve - France

Fish consumption is a potential source of human exposure to pollutants. Here, we study residue levels of PCBs in the eel, Anguilla anguilla, from the Nature Camargue Reserve in southern France. Chromatographic analysis (GC-ECD) found seventy identifiable congeners, among which, 10 are considered as dioxin-like PCBs, such as the non-ortho PCB 81 and the mono-ortho chlorobiphenyls PCB105, 114, 118, 123, 156, 157, 167, 170, 180. Toxic Equivalents (TEQ, WHO 2005 TEF-Toxic Equivalent Factors) varied among sites with a maximum in eels from Mornès (29.6pgg(-1) dry weight). Indicator PCBs (28, 52, 101, 118, 138, 153 and 180) were 22% and 29% of the total PCBs in livers and muscles respectively. Greater homogeneous bioaccumulation in muscle than that in liver suggests an increase risk for humans due to fish consumption.     

Effects of estrone on the early life stages and expression of vitellogenin and estrogen receptor genes of Japanese medaka (Oryzias latipes)

The fertilized eggs of Japanese medaka (Oryzias latipes) were exposed to estrone (E1) at 5–5000 ng L⁻¹ for 15 d, and the hatched fry were exposed continuously to the same concentrations for the additional 15 d. Adverse effects on hatchability, time to hatching, and gross abnormalities occurred at 50 ng L⁻¹ or above. Then the fry were divided into a continual exposure group, and a water recovery group. When the fry were exposed to E1 for another 60 d, there was a decrease in the hepatosomatic index (HSI) of males and the influence disappeared in the water recovery group. The gonadosonatic index (GSI) of females at 500 ng L⁻¹ decreased significantly in another 60 d exposure. While the fry were maintained in dechlorinated tap water for 60 d, a significant decrease in female GSI was observed at 50 ng L⁻¹ or above. An increased GSI was found in males in both continual exposure and water recovery groups at all E1 treatments. Quantitative RT-PCR showed that vitellogenin-I (Vtg-I) gene expressions in the female liver were significantly down-regulated at 50 ng L⁻¹ in the continual exposure group, and at 500 ng L⁻¹ in the water recovery group, while male Vtg-I genes were significantly up-regulated for all E1 treatments. In addition, all E1 treatments caused sex reversal of males. These results suggest that E1 at 5 ng L⁻¹ or above have unrecoverable impacts on the gonadal growth and development of medaka, even if only early life stages were exposed to E1.     

Effect of 17beta-estradiol on the reproduction of Japanese medaka (Oryzias latipes)

Estrogenic compounds such as 17beta-estradiol (E2) and its analogs are present in the aquatic environment and can adversely affect the reproductive systems of aquatic organisms. Although E2 has been detected at a maximum concentration of 64 ng/l in effluents of sewage treatment works (STWs), few reports address the associated effects on reproduction in fish. Therefore, we exposed adult medaka (Oryzias latipes) to mean measured E2 concentrations of 29.3, 55.7, 116, 227, and 463 ng/l for 21 d and assessed the effects on the egg number and fertility of paired medaka during the exposure period. In addition, we determined the hepatic vitellogenin (Vtg) concentration and histologically assessed the gonads of these fish. The number of egg produced and fertility of the paired medaka exposed to 463 ng/l E2 were significantly less compared with those of the control fish. Males in all treatment groups had developed testis-ova. Males treated with E2 concentrations = 55.7 ng/l contained relating great concentrations of hepatic Vtg. Therefore, although only the greatest E2 concentration tested in our study affected fecundity and fertility, effects of E2 were observed on induction of Vtg and testis-ova in male medaka exposed to environmentally relevant concentrations of E2.     

Identification of potential biomarkers for diazinon exposure to Japanese Medaka (Oryzias latipes) using annealing control primers

A new differential display-polymerase chain reaction (PCR) method based on annealing control primers was used to screen and identify potential biomarkers from differentially expressed genes (DEGs) in medaka exposed to sub-lethal concentration of diazinon (100 ppb). Among the differentially expressed genes identified, the majority were in functional categories of protein biosynthesis, transport and metabolism according to the gene ontology classification. The differential expression of ribosomal protein genes was quantified by real time PCR. The genes encoding ribosomal proteins including L3 and S17 were selected as potential biomarkers for diazinon exposure in medaka fish.     

Toxicity of o,p'-DDE to medaka d-rR strain after a one-time embryonic exposure by in ovo nanoinjection: an early through juvenile life cycle assessment

The toxicity of o,p'-DDE (1,1-dichloro-2-(p-chlorophenyl)-2-(o-chlorophenyl) ethylene) was evaluated in embryos of medaka (Oryzias latipes) following a one time exposure via nanoinjection. Medaka eggs (early gastrula) were injected with 0.5 nl of triolein (vehicle control) or 0.5 nl of 4 graded doses (0.0005-0.5 ng/egg) of o,p'-DDE in triolein. Embryos were allowed to develop, and fry were reared. Embryonic survival was monitored daily during the first 10 d until hatching and thereafter, on a weekly basis until day 59, at which time the fish were monitored for sexual maturity until day 107. In general, o,p'-DDE caused a dose- and time-dependent mortality. No changes in mortality were observed between the last two time points (day 38 and 59, respectively), and hence a 59 day-LD50 of 346 ng o,p'-DDE/egg was derived from the linear dose-response relationship. Prior to late stage death, only isolated cases of cardiovascular lesions and spinal deformities were observed, but were not dose-dependent. The lowest observable adverse effect level (LOAEL), based on upper 95% CI for regression line=0.0018 mg/kg, and the LOAEL based on exposure doses=0.5 mg/kg. Likewise, the no observable adverse effect level (NOAEL) based on linear extrapolation to 100% survival=0.0000388 mg/kg, while the NOAEL based on exposure doses=0.05 mg/kg. The nanoinjection medaka model has potential in the study of hormonally active compounds in the environment.     

Identification of potential biomarkers for diazinon exposure to Japanese Medaka (Oryzias latipes) using annealing control primers

A new differential display-polymerase chain reaction (PCR) method based on annealing control primers was used to screen and identify potential biomarkers from differentially expressed genes (DEGs) in medaka exposed to sub-lethal concentration of diazinon (100 ppb). Among the differentially expressed genes identified, the majority were in functional categories of protein biosynthesis, transport and metabolism according to the gene ontology classification. The differential expression of ribosomal protein genes was quantified by real time PCR. The genes encoding ribosomal proteins including L3 and S17 were selected as potential biomarkers for diazinon exposure in medaka fish.     

Fertilization success and sexual behavior in male medaka, Oryzias latipes, exposed to tributyltin

We evaluated the effects of tributyltin (TBT), polychlorinated biphenyls (PCBs), and a mixture of TBT and PCBs on reproduction, gonadal histology, and sexual behavior in Japanese medaka (Oryzias latipes). We administered TBT (1 microg g(-1) body weight [b.w.] daily), PCBs (1 microg g(-1) b.w. daily), or both to medaka for 3 weeks. We assessed reproductive success during week 3 and the sexual behavior of male medaka after the exposure period. Fertilization success was significantly decreased in groups that received TBT. The profiles of fertility developed a bimodal distribution in the TBT and TBT+PCBs groups, and the numbers of males that performed "following" and "dancing" were significantly decreased in the TBT+PCBs group. The frequency of "dancing" also decreased after treatment with TBT, whereas fish that received PCBs only showed no difference in sexual behavior. Testes that had oocytes showed normal spermatogenesis, even though two of the five males treated with TBT+PCBs had some indication of testis-ova, which did not occur in any other group. Our results indicate that TBT, but not PCBs, affects sexual behavior and reproduction in medaka.     

Reproductive effects and bioconcentration of 4-nonylphenol in medaka fish (Oryzias latipes)

The objective of this study was to investigate the reproductive success (fertility and fecundity) of medaka (Oryzias latipes) exposed to 4-nonylphenol (4-NP) in water and the viability of their F(1) offspring. In addition, we measured the bioconcentration of 4-NP in eggs. After a 21-d exposure to 100microg/l 4-NP, medaka showed reduced egg production and fertility. Hepatic vitellogenin levels were increased significantly in males treated with 10, 50 and 100microg/l of 4-NP. In the F(1) generations, the hatchability and time to hatching of embryos in the 100microg/l treatment group were adversely affected, and 2-7microg 4-NP/g egg was found in spawned eggs (the bioconcentration factor: 30-100). These results indicate that (1) 4-NP adversely affected the reproduction (fecundity and fertility) of adult medaka, (2) 4-NP accumulated in eggs through maternal transfer and (3) these levels of 4-NP were associated with adverse effects in the F(1) offspring.     

Assessing the inhalation cancer risk of particulate matter bound polycyclic aromatic hydrocarbons (PAHs) for the elderly in a retirement community of a mega city in North China

Assessment of the health risks resulting from exposure to ambient polycyclic aromatic hydrocarbons (PAHs) is limited by the lack of environmental exposure data among different subpopulations. To assess the exposure cancer risk of particulate carcinogenic polycyclic aromatic hydrocarbon pollution for the elderly, this study conducted a personal exposure measurement campaign for particulate PAHs in a community of Tianjin, a city in northern China. Personal exposure samples were collected from the elderly in non-heating (August–September, 2009) and heating periods (November–December, 2009), and 12 PAHs individuals were analyzed for risk estimation. Questionnaire and time-activity log were also recorded for each person. The probabilistic risk assessment model was integrated with Toxic Equivalent Factors (TEFs). Considering that the estimation of the applied dose for a given air pollutant is dependent on the inhalation rate, the inhalation rate from both EPA exposure factor book was applied to calculate the carcinogenic risk in this study. Monte Carlo simulation was used as a probabilistic risk assessment model, and risk simulation results indicated that the inhalation-ILCR values for both male and female subjects followed a lognormal distribution with a mean of 4.81?×?10^?6 and 4.57?×?10^?6, respectively. Furthermore, the 95 % probability lung cancer risks were greater than the USEPA acceptable level of 10^?6 for both men and women through the inhalation route, revealing that exposure to PAHs posed an unacceptable potential cancer risk for the elderly in this study. As a result, some measures should be taken to reduce PAHs pollution and the exposure level to decrease the cancer risk for the general population, especially for the elderly.