Evaluation of in vitro toxicity information for zebrafish as a promising alternative for chemical hazard and risk assessment.

In vitro assays are widely proposed as a test alternative to traditional in vivo standard acute and chronic toxicity tests. However, whether toxicity information derived from in vitro assays instead of in vivo tests could provide sufficient protection (e.g., 95 % of protection) for chemical risks remain evaluated. To investigate the feasibility of zebrafish (Danio rerio) cell-based in vitro test method as a test alternative, we comprehensively compared sensitivity differences among endpoints, among test methods (in vitro, FET and in vivo), and between zebrafish and rat (Rattus norvegicus), respectively using chemical toxicity distribution (CTD) approach. For each test method involved, sublethal endpoints were more sensitive than lethal endpoints for both zebrafish and rat, respectively. Biochemistry (zebrafish in vitro), development (zebrafish in vivo and FET), physiology (rat in vitro) and development (rat in vivo) were the most sensitive endpoints for each test method. Nonetheless, zebrafish FET test was the least sensitive one compared to its in vivo and in vitro tests for either lethal or sublethal responses. Comparatively, rat in vitro tests considering cell viability and physiology endpoints were more sensitive than rat in vivo test. Zebrafish was found to be more sensitive than rat regardless of in vivo or in vitro tests for each pairwise endpoint of concern. Those findings indicate that zebrafish in vitro test is a feasible test alternative to zebrafish in vivo and FET test and traditional mammalian test. It is suggesting that zebrafish in vitro test can be optimized by choosing more sensitive endpoints, such as biochemistry to provide sufficient protection for zebrafish in vivo test and to establish applications of zebrafish in vitro test in future risk assessment. Our findings are vital for evaluating and further application of in vitro toxicity toxicity information as an alternative for chemical hazard and risk assessment.

Deriving freshwater guideline values for neonicotinoid insecticides: Implications for water quality guidelines and ecological risk assessment.

The increased use of neonicotinoid insecticides in aquatic environments poses a significant threat to non-target freshwater species. However, the existing water quality guidelines (WQGs) for neonicotinoids mainly focus on imidacloprid, and only a few authoritative institutions have established WQGs for other neonicotinoids. There is a critical need to develop WQGs and conduct ecological risk assessment (ERA) of different neonicotinoids in global freshwater environments. In this study, we derived interim acute and chronic guideline values and acute-to-chronic ratios (ACRs) for six neonicotinoids based on publicly available acute and chronic toxicity data. The exposure concentrations of neonicotinoids were obtained from published literature worldwide, and ERA was conducted for neonicotinoids in global freshwater ecosystems using a tiered approach. The derived chronic guideline values (95% confidence interval (CI), ng/L) were 0.63 (0.02-5.47) for thiacloprid (the lowest) and 16.4 for dinotefuran (the highest). The identified ACRs (95% CI) ranged from 90.9 (47.0-180) to 957 (102-3350), which can be used to extrapolate scarce chronic data from the acute data. Neonicotinoid concentrations in global freshwater were predicted from 10.6 (6.88-23.4) (thiacloprid) to 339 (211-786) ng/L (thiamethoxam). The estimated risk quotients ranged from 3.23 (dinotefuran) to 21.73 (thiacloprid), and the probability of exceeding WQGs ranged from 27.1% (dinotefuran) to 77.1% (thiacloprid). The ERA results indicated that the six neonicotinoids posed negligible acute risks but high chronic risks to global freshwater ecosystems, especially acetamiprid (65.8%) and thiacloprid (28.1%). The key findings of this study provide critical scientific information regarding the ecological risks of long-term neonicotinoid exposure and key insights for policy development and water quality control.

Life stage and endpoint sensitivity differences of fathead minnow (Pimephales promelas) to chemicals with various modes of action.

Fish Embryo Acute Toxicity (FET) test was proposed as an alternative to the traditional test methods using larval or adult fish. However, whether fathead minnow (Pimephales promelas) embryo is appropriate for FET remains uncertain. In the present study, ecological threshold of toxicological concern (ecoTTC) values and uncertainty factors (UFs) for each Verhaar et al. category in P. promelas were identified by employing probabilistic ecological risk assessment (PERA) approach with chemical toxicity distributions (CTDs). The sensitivity among different life stages and toxicity among different mode of actions (MOAs) classes were comprehensively compared by CTD comparisons. The results showed that embryo exhibited the less or similar sensitivity compared to larva or adult for Verhaar et al. MOA classes (1-4) while adults were more sensitive, followed by embryo than larval for non-classified chemicals. Considering growth effect as endpoint to class 1, class 3, and non-classified chemicals on P. promelas embryo and larva was more sensitive than mortality. Non-classified chemicals especially inorganic compounds were most toxic to P. promelas embryo for the four concerned Verharr et al. MOA-specific chemical classes. This study also derived uncertainty factors (UFs) as 26.5 (9.8, 109) for embryo-to-larva, 6.26 (3.94, 11.0) for embryo-to-adult, 15.6 (10.1, 36.1) for mortality-to-growth, and 3.03 (1.86, 7.08) for mortality-to-reproduction, which can be applied for extrapolations of life stage-to-life stage and effect-to-effect to reduce the underestimating and overestimating risk by the use of default UF such as 10, 100 or 1000. Our findings are vital for feasibility of FET test of P. promelas for ecotoxicity testing and ecological risk assessment for chemicals with different MOAs.

Species and Life-Stage Sensitivity of Chinese Rare Minnow (Gobiocypris rarus) to Chemical Exposure: A Critical Review.

Chemical production and consumption in Asia are increasing at an unprecedented rate, calling for regulations on chemical management. Under the New Chemical Substance Notification in China, information on ecotoxicological effects of chemicals is mandatory for the simplified registration of chemicals with the requirement that one ecotoxicological test is conducted locally. It is now mandatory to use the native fish species Chinese rare minnow (Gobiocypris rarus). However, its chemical sensitivity compared to that of fathead minnow (Pimephales promelas) or zebrafish (Danio rerio) is still unclear. We performed a holistic literature review on toxicity data with G. rarus from 1997 to 2020. Species sensitivity among G. rarus, P. promelas, and D. rerio and life-stage sensitivity of G. rarus were systematically investigated for various chemicals using both chemical ratio distribution and probabilistic chemical toxicity distribution approaches. Comparatively, the Chinese native fish species G. rarus was more sensitive than D. rerio, particularly to metals. Juvenile and adult G. rarus were more sensitive than its larvae and embryos. The observed lower sensitivity of G. rarus embryo was likely due to the thick embryonic chorion, discrepant methods of collecting embryos, and the paucity of toxicity data, implying the necessity to standardize G. rarus embryo tests and validate the sensitivity with various types of chemicals. This unique review allows us to conclude that G. rarus studies could be used in worldwide registrations and that further investigations are needed to use G. rarus embryos as alternatives to the fish test. Environ Toxicol Chem 2021;40:2680-2692. © 2021 SETAC.

Comparative influences of dermal and inhalational routes of exposure on hazards of cleaning product ingredients among mammalian model organisms.

Health risks resulting from dermal or inhalational exposures are frequently assessed based on rodent oral toxicity information due to a lack of species- or route-specific toxicity data. Default uncertainty factors (UFs; e.g., 10-fold) are also applied during risk assessments to account for variability such as inter-species, intra-species, exposure duration, dose-response, and route-to-route extrapolations. However, whether rodent oral data and a default UF approach can provide adequate protection for other mammalian species under dermal or inhalational exposure scenarios remains understudied, particularly for cleaning product ingredients. Therefore, we collated and examined publicly available median lethal dose (LD50), no-observed-adverse-effect level (NOAEL) and lowest-observed-adverse-effect level (LOAEL) values from different types of standard mammalian toxicity studies for rats (dermal and inhalational), mice, rabbits, guinea pigs, and dogs (oral, dermal and inhalational) using the Cleaning Product Ingredient Safety Initiative (CPISI) database. Probabilistic hazard assessments using chemical toxicity distributions (CTDs) were subsequently conducted, and threshold concentrations (TCs) and 95% confidence intervals (95% CIs) were derived to identify thresholds of toxicological concern (TTCs). Relative sensitivities among or between mammalian species, exposure routes, and chemical classes were also compared based on calculated TC5s and 95% CIs to support future toxicology studies and hazard and risk assessments. We then identified uncertainty factors (UFs) using both CTD comparisons and individual UF probability distributional approaches. Based on available rodent inhalational data, chemical category-specific UFs were derived for ethers. Additionally, we also determined whether default UFs of 10 or 100 would be protective for various distributions of cleaning product ingredients. Our novel observations among these routes of exposure and common mammalian model organisms appear particularly useful for read across and screening-level health hazard and risk assessments when limited data exists for specific chemicals.