In vivo Toxicity Assessment of Refined Red Palm-pressed Mesocarp Olein in Sprague-Dawley Rats.

Refined red palm-pressed mesocarp olein (PPMO) is recovered from palm-pressed mesocarp fiber, which is a by-product from palm oil mill. Its utilization in food industry is extremely limited even though it contains various phytonutrients. Thus, this study aimed to evaluate its toxicity effects by using the male Sprague-Dawley rat model. The rats were administered with a single dose of 2 g/kg PPMO in an acute toxicity study while administered with 2, 1, or 0.5 g/kg PPMO daily for 28 days in a sub-chronic toxicity study. The mortality, oral LD50 value, clinical observation, body and organ weight, hematological and biochemical analyses, pathological and histopathological examinations were assessed. The overall outcomes indicated that PPMO is non-toxic up to 2 g/kg and considered safe to be used in food application, especially as functional food ingredient and supplement attributed to its phytonutrients. Besides, this study provides an insight in alternative utilization of the wastes from palm oil mill.

Acute and chronic oral toxicity assessment of longan sugar extracts derived from whole fruit and from fruit pulp in rats.

ETHNOPHARMACOLOGICAL RELEVANCE: Longan (Dimocarpus longan Lour.) is one of the most popular subtropical fruits. Various parts of longan, including seeds, pericarp and pulp, have long been used in traditional medicine in China, Thailand and other Asian countries. The pulp has high sugar, vitamin and mineral content as well as bioactive components. The seeds and pericarp have also been reported to contain beneficial polyphenolic compounds. Longan sugar extract from pulp (LGSP) is prepared as a conventional sugar product. Longan sugar extract from whole longan fruit (LGSW) is also offered as a health food and as a medicinal product. AIM OF THE STUDY: The objective of this study was to identify and compare potential health hazards of both LGSW and LGSP by testing for acute and chronic oral toxicity in rats. MATERIALS AND METHODS: In acute toxicity testing, an oral dose (20 g/kg) of either LGSW or LGSP was administered to groups of rats. Mortality and clinical signs of toxicity were observed for 24 h, and then daily for a total of 14 days. In the chronic toxicity test, either LGSW (1, 2.5 and 5 g/kg/day) or LGSP (5 g/kg/day) was administered orally for a period of 180 days. After that treatment period, the rats in the satellite groups which received the highest doses of either LGSW or LGSP were observed for an additional 28 days. The rats then underwent clinical observation, body and organ weight measurement, hematological and biochemical analyses, and histopathological examination. RESULTS: In the acute toxicity study, the oral administration of LGSP or LGSW in either pellet or syrup formulations did not cause mortality or any pathological abnormalities. In the chronic toxicity study, neither LGSW nor LGSP resulted in death or in any changes in behavior of the rats. All hematological and serum biochemical values of both the LGSW- and LGSP-treated groups were within the normal ranges. No histopathological abnormalities of any internal organs were observed. CONCLUSION: The safety of longan sugar extract made from whole fruit (pulp, seeds and pericarb) is comparable to that of longan sugar extract made from pulp alone.

A toxicogenomic approach for the risk assessment of the food contaminant acetamide.

Acetamide (CAS 60-35-5) is detected in common foods. Chronic rodent bioassays led to its classification as a group 2B possible human carcinogen due to the induction of liver tumors in rats. We used a toxicogenomics approach in Wistar rats gavaged daily for 7 or 28 days at doses of 300 to 1500 mg/kg/day (mkd) to determine a point of departure (POD) and investigate its mode of action (MoA). Ki67 labeling was increased at doses ≥750 mkd up to 3.3-fold representing the most sensitive apical endpoint. Differential gene expression analysis by RNA-Seq identified 1110 and 1814 differentially expressed genes in male and female rats, respectively, following 28 days of treatment. Down-regulated genes were associated with lipid metabolism while up-regulated genes included cell signaling, immune response, and cell cycle functions. Benchmark dose (BMD) modeling of the Ki67 labeling index determined the BMD10 lower confidence limit (BMDL10) as 190 mkd. Transcriptional BMD modeling revealed excellent concordance between transcriptional POD and apical endpoints. Collectively, these results indicate that acetamide is most likely acting through a mitogenic MoA, though specific key initiating molecular events could not be elucidated. A POD value of 190 mkd determined for cell proliferation is suggested for risk assessment purposes.

Pulmonary toxicity in rats following inhalation exposure to poorly soluble particles: The issue of impaired clearance and the relevance for human health hazard and risk assessment.

Intensive discussions are ongoing about the interpretation of pulmonary effects observed in rats exposed to poorly soluble particles. Alveolar clearance differs between rats and humans and becomes impaired in rats at higher exposure concentrations. Some have doubted the human relevance of toxic effects observed in rats under impaired clearance conditions and have suggested that experimental exposures should stay below concentrations inducing impaired clearance. However, for regulatory purposes, insight in potential health effects at relatively high concentrations is needed to fully understand the hazard. Many aspects of impaired particle clearance remain unclear, hampering human health hazard and risk assessment. For an adequate evaluation of the impact of impaired clearance on pulmonary toxicity, a clear definition of alveolar clearance is needed that enables to quantitatively relate the level of impairment to the induction of adverse pulmonary health effects. Also, information is needed on the mechanism of action and the appropriate dose metric for the pulmonary effects observed. In absence of these data, human hazard and risk assessment can only be performed in a pragmatic way. Unless available data clearly point out otherwise, rat pulmonary toxicity including lung inflammation and tumour formation, needs to be considered relevant for human hazard and risk assessment.

Use of scanning and image recognition technology to semi-automate larval development assessment in toxicity tests with a tropical copepod.

There is a high demand for the development of reliable chronic toxicity tests using tropical marine species for subsequent use in tropical risk assessment. However, many chronic test endpoints can be laborious and time-consuming to assess, particularly if the endpoints require measurements of individuals (e.g. growth, size) or advanced taxonomic expertise (e.g. differentiating between larval development stages). In this study, we used scanning and image recognition (SIR) technology to develop and validate a chronic toxicity test with larvae of the tropical euryhaline copepod, Acartia sinjiensis. Optimisation steps are described, and included egg age, and effect of algal food type and salinity on toxicity. Comparisons were made between traditional endpoints measured using microscopy and those measured using SIR. Traditional endpoints of larval development ratio (LDR) and survival achieved using microscope examination and SIR were almost identical (R2 = 0.96-0.97). Additional endpoints made possible by SIR included larval development index (LDI; based on the number of animals at different stages of development), and a range of size measurements (e.g. surface area, perimeter and length) for individual animals and for total populations (i.e. a proxy for biomass). The SIR-derived endpoints were based on measurements that had concentration-dependant responses to tested toxicants (copper, nickel, ammonia), and were a sub-set of the full range of metrics provided by the software. Toxicity values based on SIR-measurements were similar to or more sensitive than the traditional LDR endpoint. SIR technology provides a major opportunity to improve and modernise larval development tests for a range for species, but comes at a cost of increased data size and complexity. Therefore, as a research tool, SIR has significant advantages over traditional microscope methods, but for routine toxicity testing, SIR incorporation into invertebrate toxicity testing will benefit from further improvements to the associated software and data management systems.

Assessment of whole-sediment chronic toxicity using sub-lethal endpoints with Monocorophium insidiosum.

A whole-sediment test with the infaunal amphipod Monocorophium insidiosum has been developed to assess the long-term effects exerted by sediment contamination on survival, growth rates and attainment of sexual maturity. Juvenile amphipods were exposed for 28 days to a control sediment (native sediment) and three sediment samples collected in sites of the Venice Lagoon, characterized by contamination levels ranging from low to moderate, and absence of acute toxicity toward amphipods. Growth rate was estimated as daily length (µm d-1) and weight increments (µg d-1). The long-term exposure to the test sediments affected significantly both growth rate and attainment of sexual maturity of the females of M. insidiosum. In contrast, survival was high and uniform among all the samples, despite the contamination gradient. The results suggest growth to be the more reliable and statistically relevant endpoint. Attainment of sexual maturity, although allowed the identification of detrimental effects, was affected by a higher among-replicates variance as compared with growth rates, and thus less reliable than growth for the identification of impairments. The significant impairments observed both on growth and attainment of maturity evidenced the need to address the monitoring, also in the Lagoon of Venice, towards the assessment of the long-term effects on benthic species.

Prospective environmental risk assessment of mixtures in wastewater treatment plant effluents - Theoretical considerations and experimental verification.

The aquatic environment is continually exposed to a complex mixture of chemicals, whereby effluents of wastewater treatment plants (WWTPs) are one key source. The aim of the present study was to investigate whether environmental risk assessments (ERAs) addressing individual substances are sufficiently protective for such coincidental mixtures. Based on a literature review of chemicals reported to occur in municipal WWTP effluents and mode-of-action considerations, four different types of mixtures were composed containing human pharmaceuticals, pesticides, and chemicals regulated under REACH. The experimentally determined chronic aquatic toxicity of these mixtures towards primary producers and the invertebrate Daphnia magna could be adequately predicted by the concept of concentration addition, with up to 5-fold overestimation and less than 3-fold underestimation of mixture toxicity. Effluents of a municipal WWTP had no impact on the predictability of mixture toxicity and showed no adverse effects on the test organisms. Predictive ERAs for the individual mixture components based on here derived predicted no effect concentrations (PNECs) and median measured concentrations in WWTP effluents (MCeff) indicated no unacceptable risk for any of the individual chemicals, while MCeff/PNEC summation indicated a possible risk for multi-component mixtures. However, a refined mixture assessment based on the sum of toxic units at species level indicated no unacceptable risks, and allowed for a safety margin of more than factor 10, not taking into account any dilution of WWTP effluents by surface waters. Individual substances, namely climbazole, fenofibric acid and fluoxetine, were dominating the risks of the investigated mixtures, while added risk due to the mixture was found to be low with the risk quotient being increased by less than factor 2. Yet, uncertainty remains regarding chronic mixture toxicity in fish, which was not included in the present study. The number and identity of substances composing environmental mixtures such as WWTP effluents is typically unknown. Therefore, a mixture assessment factor is discussed as an option for a prospective ERA of mixtures of unknown composition.

Current pesticide dietary risk assessment in light of comparable animal study NOAELs after chronic and short-termed exposure durations.

Dietary risk assessment (DRA) of pesticides includes the estimation of chronic and acute exposures from crop residues, but assesses acute exposures only for pesticides with an acute reference dose (ARfD). Acute estimation uses high percentiles of food consumption surveys which are considerably higher than per capita lifetime averaged food consumption values which are used for chronic estimations. Assessing acute risks only for pesticides with an ARfD tacitly assumes that chronic risk assessment covers also intermittent occurring exposures which could significantly exceed chronic estimates. The present investigation conducted on 2200 rat studies from 436 pesticides provides evidence demonstrating that pesticides with and without ARfD have no-observed-adverse-effect levels (NOAELs) which remain statistically unchanged in developmental, subacute, subchronic, reproductive and chronic toxicity studies covering exposure durations between 2 and 104 weeks. DRA of pesticides without ARfD needs reconsideration in light of equally high toxic dose levels after short- and long-term exposures, suggesting that intermittent exposures could be toxic, if they repeatedly exceed the acceptable chronic daily intake (ADI; conceptually the human counterpart of chronic animal NOAEL). As such risks are currently not assessed for pesticides without ARfD, the current DRA concept, which automatically presumes the use of low chronic exposure estimates entirely covers the risks of not acutely toxic pesticides, needs reconsideration. Furthermore, risks to intermittent occurring high exposures are probably also insufficiently assessed for pesticides where the ARfD is significantly higher than the ADI. As an example, the maximum residue limit for bifenazate in peaches is discussed.

Role of chronic toxicology studies in revealing new toxicities.

Chronic (>3 months) preclinical toxicology studies are conducted to support the safe conduct of clinical trials exceeding 3 months in duration. We have conducted a review of 32 chronic toxicology studies in non-rodents (22 studies in dogs and 10 in non-human primates) and 27 chronic toxicology studies in rats dosed with Merck compounds to determine the frequency at which additional target organ toxicities are observed in chronic toxicology studies as compared to subchronic studies of 3 months in duration. Our review shows that majority of the findings are observed in the subchronic studies since additional target organs were not observed in 24 chronic non rodent studies and in 21 chronic rodent studies. However, 6 studies in non rodents and 6 studies in rodents yielded new findings that were not seen in studies of 3-month or shorter duration. For 3 compounds the new safety findings did contribute to termination of clinical development plans. Although the incidence of compound termination associated with chronic toxicology study observations is low (∼10%), the observations made in these studies can be important for evaluating human safety risk.

Nonclinical and pharmacokinetic assessments to evaluate the potential of tedizolid and linezolid to affect mitochondrial function.

Prolonged treatment with the oxazolidinone linezolid is associated with myelosuppression, lactic acidosis, and neuropathies, toxicities likely caused by impairment of mitochondrial protein synthesis (MPS). To evaluate the potential of the novel oxazolidinone tedizolid to cause similar side effects, nonclinical and pharmacokinetic assessments were conducted. In isolated rat heart mitochondria, tedizolid inhibited MPS more potently than did linezolid (average [± standard error of the mean] 50% inhibitory concentration [IC50] for MPS of 0.31 ± 0.02 µM versus 6.4 ± 1.2 µM). However, a rigorous 9-month rat study comparing placebo and high-dose tedizolid (resulting in steady-state area under the plasma concentration-time curve values about 8-fold greater than those with the standard therapeutic dose in humans) showed no evidence of neuropathy. Additional studies explored why prolonged, high-dose tedizolid did not cause these mitochondriopathic side effects despite potent MPS inhibition by tedizolid. Murine macrophage (J774) cell fractionation studies found no evidence of a stable association of tedizolid with eukaryotic mitochondria. Monte Carlo simulations based on population pharmacokinetic models showed that over the course of a dosing interval using standard therapeutic doses, free plasma concentrations fell below the respective MPS IC50 in 84% of tedizolid-treated patients (for a median duration of 7.94 h) and 38% of linezolid-treated patients (for a median duration of 0 h). Therapeutic doses of tedizolid, but not linezolid, may therefore allow for mitochondrial recovery during antibacterial therapy. The overall results suggest that tedizolid has less potential to cause myelosuppression and neuropathy than that of linezolid during prolonged treatment courses. This, however, remains a hypothesis that must be confirmed in clinical studies.

Risk assessment's insensitive toxicity testing may cause it to fail.

BACKGROUND: Risk assessment of chemicals and other agents must be accurate to protect health. We analyse the determinants of a sensitive chronic toxicity study, risk assessment's most important test. Manufacturers originally generate data on the properties of a molecule, and if government approval is needed to market it, laws globally require toxicity data to be generated using Test Guidelines (TG), i.e. test methods of the Organisation for Economic Cooperation and Development (OECD), or their equivalent. TGs have advantages, but they test close-to-poisonous doses for chronic exposures and have other insensitivities, such as not testing disease latency. This and the fact that academic investigators will not be constrained by such artificial methods, created a de facto total ban of academia's diverse and sensitive toxicity tests from most risk assessment. OBJECTIVE: To start and sustain a dialogue between regulatory agencies and academic scientists (secondarily, industry and NGOs) whose goals would be to (1) agree on the determinants of accurate toxicity tests and (2) implement them (via the OECD). DISCUSSION: We analyse the quality of the data produced by these incompatible paradigms: regulatory and academic toxicology; analyse the criteria used to designate data quality in risk assessment; and discuss accurate chronic toxicity test methods. CONCLUSION: There are abundant modern experimental methods (and rigorous epidemiology), and an existing systematic review system, to at long last allow academia's toxicity studies to be used in most risk assessments.

A docking-based receptor library of antibiotics and its novel application in predicting chronic mixture toxicity for environmental risk assessment.

As organisms are typically exposed to chemical mixtures over long periods of time, chronic mixture toxicity is the best way to perform an environmental risk assessment (ERA). However, it is difficult to obtain the chronic mixture toxicity data due to the high expense and the complexity of the data acquisition method. Therefore, an approach was proposed in this study to predict chronic mixture toxicity. The acute (15 min exposure) and chronic (24 h exposure) toxicity of eight antibiotics and trimethoprim to Vibrio fischeri were determined in both single and binary mixtures. The results indicated that the risk quotients (RQs) of antibiotics should be based on the chronic mixture toxicity. To predict the chronic mixture toxicity, a docking-based receptor library of antibiotics and the receptor-library-based quantitative structure-activity relationship (QSAR) model were developed. Application of the developed QSAR model to the ERA of antibiotic mixtures demonstrated that there was a close affinity between RQs based on the observed chronic toxicity and the corresponding RQs based on the predicted data. The average coefficients of variations were 46.26 and 34.93 % and the determination coefficients (R (2)) were 0.999 and 0.998 for the low concentration group and the high concentration group, respectively. This result convinced us that the receptor library would be a promising tool for predicting the chronic mixture toxicity of antibiotics and that it can be further applied in ERA.

The toxicity of molybdate to freshwater and marine organisms. II. Effects assessment of molybdate in the aquatic environment under REACH.

The REACH Molybdenum Consortium initiated an extensive research program in order to generate robust PNECs, based on the SSD approach, for both the freshwater and marine environments. This activity was part of the REACH dossier preparation and to form the basis for scientific dialogues with other national and international regulatory authorities. Chronic ecotoxicity data sets for the freshwater and marine environments served as starting point for the derivation of PNECs for both compartments, in accordance with the recommended derivation procedures established by the European Chemicals Agency (ECHA). The HC(5,50%)s that were derived from the generated Species Sensitivity Distributions were 38.2 mg Mo/L and 5.75 mg Mo/L for the freshwater and marine water compartment, respectively. Uncertainty analysis on both data sets and available data on bioaccumulation at high exposure levels justified an assessment factor of 3 on both HC(5,50%) leading to a PNEC(freshwater) of 12.7 mg Mo/L and a PNEC(marine) of 1.92 mg Mo/L. As there are currently insufficient ecotoxicological data available for the derivation of PNECs in the sediment compartment, the equilibrium partitioning method was applied; typical K(D)-values for both the freshwater and marine compartments were identified and combined with the respective PNEC, leading to a PNEC(sediment) of 22,600 mg/kg dry weight and 1980 mg/kg dry weight for freshwater and marine sediments, respectively. The chronic data sets were also used for the derivation of final chronic values using the procedures that are outlined by the US Environmental Protection Agency for deriving such water benchmarks. Comparing PNECs with FCVs showed that both methodologies result in comparable protective concentration levels for molybdenum in the environment.

Safety assessment of a solid lipid curcumin particle preparation: acute and subchronic toxicity studies.

Curcumin, a polyphenol, is obtained from turmeric, the ground rhizomes of Curcuma longa L. Extensive research over the past half century has revealed several health benefits of curcumin. The objective of the present study was to investigate potential adverse effects, if any, of a novel solid lipid curcumin particle (SLCP) preparation in rats following acute and subchronic administration. The oral LD50 of the preparation in rats as well as in mice was found to be greater than 2000 mg/kg body weight (bw). In the subchronic toxicity study, Wistar rats (10/sex/group) were administered via oral gavage 0 (control), 180, 360, and 720 mg/kg bw/day of SLCP preparation for 90 days. Administration of the curcumin preparation did not result in any toxicologically significant treatment-related changes in clinical (including behavioral) observations, ophthalmic examinations, body weights, body weight gains, feed consumption, and organ weights. No adverse effects of the curcumin preparation were noted on the hematology, serum chemistry parameters, and urinalysis. Terminal necropsy did not reveal any treatment-related gross or histopathology findings. Based on the results of this study, the No Observed-Adverse-Effect Level (NOAEL) for this standardized novel curcumin preparation was determined as 720 mg/kg bw/day, the highest dose tested.

Toxicological assessment of dihydroergotamine after chronic inhalation in dogs.

The objective was to determine the respiratory toxicity of MAP0004, orally inhaled dihydroergotamine (DHE), via inhalation for six months. Forty beagle dogs (twenty females, twenty males) were treated by nose-only inhalation for 182 days. Groups 2 through 5 received MAP0004 (mean doses: 0.045, 0.154, 0.44, 0.825 mg/kg); Group 1 received vehicle only. Groups 1 through 4 received single thirty-minute exposures, whereas Group 5 was exposed twice daily for thirty minutes. Toxicity was assessed from clinical observations, objective evaluations, and clinical and anatomical pathology. Systemic effects were scabbing of ear tips in Groups 3, 4, and 5 and excessive salivation and emesis, observed in Group 5. No changes were observed in the lungs in any dose group. Minimal treatment-related microscopic changes were observed in the respiratory nasal epithelium only in Group 5. No plexiform, vascular media, or fibroproliferative changes in any heart valves were observed in any group. Expected systemic pharmacologic effects were observed only at MAP0004 target doses ≥ 0.224 mg/kg (achieved doses > 0.154 mg/kg), which was more than five times the maximum daily intravenous (IV) human clinical dose of DHE, and more than twenty times the systemic equivalent dose of MAP0004. The no-observed adverse effect level (NOAEL) was the achieved inhaled dose of 0.045 mg/kg, or four times the human clinical dose of MAP0004.

Uterotrophic assay, Hershberger assay, and repeated 28-day oral toxicity study of flumorph based on the Organization for Economic Co-operation and Development draft protocols.

To evaluate the endocrine-mediated effects of flumorph, we performed the uterotrophic assay, the Hershberger assay, and the repeated 28-day oral toxicity study based on the OECD draft protocols. In the uterotrophic assay, female ovariectomized SD rats were subcutaneously injected with flumorph at doses of 0, 50, 150, and 500 mg/kg on each of 3 days, and no changes were observed. In the Hershberger assay, castrated male SD rats were administered with flumorph by oral gavage at doses of 0, 50, 150, and 500 mg/kg/day for 10 consecutive days, and no abnormal changes were observed. However, in the repeated 28-day oral toxicity study , flumorph was orally administered at doses 0, 30, 100, and 300 mg/kg/day for at least 28 days, a significantly increase in T(4) value in male rats and TSH value in female rats were detected in the highest dosage group, respectively. Besides, the flumorph administration increase liver weights, produce hepatocellular diffuse fatty degeneration, and effect biochemical parameters related to liver function in male and/or female rats in dosed groups. Therefore, flumorph is concluded to have thyroid disruption effects and is likely a thyroid disrupter, but the further studies are needed for hazard identification.

Biodegradability and toxicity assessment of bleach plant effluents treated anaerobically.

As part of an experimental project on the treatment of bleach plant effluents the results of biodegradability and toxicity assessment of effluents from a bench-scale horizontal anaerobic immobilized bioreactor (HAIB) are discussed in this paper. The biodegradability of the bleach plant effluents from a Kraft pulp mill treated in the HAIB was evaluated using the modified Zahn-Wellens test. The inoculum came from a pulp mill wastewater treatment plant and the dissolved organic carbon (DOC) was used as the indicator of organic matter removal. The acute and chronic toxicity removal during the anaerobic treatment was estimated using Daphnia similis and Ceriodaphnia silvestrii respectively. Moreover, the evaluation of chromosome aberrations (CA), micronucleus frequencies (MN) and mitotic index (IM) in Allium cepa cells were used as genotoxicity indicators. The results indicate that the effluents from the anaerobic reactor are amenable to aerobic polishing. Acute and chronic toxicity were reduced by 90 and 81%, respectively. The largest CA and MN incidence in the meristematic cells of A. cepa were observed after exposure to the raw bleach plant effluent. The HAIB was able to reduce the acute and chronic toxicity as well as chromosome aberrations and the occurrence of micronucleus.

Toxicity assessment of pramipexole in juvenile rhesus monkeys.

Pramipexole (PPX) is a dopamine agonist approved for the treatment of the signs and symptoms of idiopathic Parkinson's disease as well as restless leg syndrome. The objective of this study was to investigate the toxicity of PPX when administered orally to juvenile rhesus monkeys once daily for 30 weeks, and to assess the reversibility of toxicity during a 12-week recovery. Rhesus monkeys (N=4 males and 4 females/group; 22-24 months of age) were orally treated daily for 30 weeks with 0.0, 0.1, 0.5 or 2.0 mg/kg PPX, and subjects were assessed daily using the NCTR Operant Test Battery (OTB). Clinical chemistry, hematology, ophthalmology and other standard postmortem toxicological evaluations, including histopathology and neuropathology as well as toxicokinetics were performed. The systemic exposure to PPX was higher than that at therapeutic doses in man and AUC(0-24 h)-data increased proportionally to dose. Blood pressure significantly decreased over time in all groups including control. Near the end of treatment, there were statistically significant decreases in heart rate for the 0.5 and 2.0 mg/kg/day groups compared to control. After 4 weeks of dosing, serum prolactin was significantly decreased in all treatment groups compared to control. This decrease remained at the end of treatment in the 0.5 and 2.0 mg/kg/day groups. In summary, administration of PPX at doses of up to 2.0 mg/kg/day for 30 weeks to juvenile rhesus monkeys produced adverse findings which were attributable to its pharmacological properties, including hypoprolactinemia.

Assessment of the cytotoxicity and genotoxicity of haloacetic acids using microplate-based cytotoxicity test and CHO/HGPRT gene mutation assay.

Haloacetic acids (HAAs) are the second most prevalent class of disinfection byproducts found in drinking water. The implications of HAAs presence in drinking water are a public health concern due to their potential mutagenic and carcinogenic effects. In the present study, we examined the cytotoxic and genotoxic effects of six common HAAs using a microplate-based cytotoxicity test and a hypoxanthine-guanine phosphoribosyltransferase (HGPRT) gene mutation assay in Chinese hamster ovary K1 (CHO-K1) cells. We found that their chronic cytotoxicities (72h exposure) to CHO-K1 cells varied, and we ranked their levels of toxicity in the following descending order: iodoacetic acid (IA)>bromoacetic acid (BA)>dibromoacetic acid (DBA)>chloroacetic acid (CA)>dichloroacetic acid (DCA)>trichloroacetic acid (TCA). The toxicity of IA is 1040-fold of that of TCA. All HAAs except TCA were shown to be mutagenic to CHO-K1 cells in the HGPRT gene mutation assay. The mutagenic potency was compared and ranked as follows: IA>DBA>BA>CA>DCA>TCA. There was a statistically significant correlation between cytotoxicity and mutagenicity of the HAAs in CHO-K1 cells. The microplate-based cytotoxicity assay and HGPRT gene mutation assay were suitable methods to monitor the cytotoxicity and genotoxicity of HAAs, particularly for comparing the toxic intensities quantitatively.

Risk assessment for side-effects of neonicotinoids against bumblebees with and without impairing foraging behavior.

Bombus terrestris bumblebees are important pollinators of wild flowers, and in modern agriculture they are used to guarantee pollination of vegetables and fruits. In the field it is likely that worker bees are exposed to pesticides during foraging. To date, several tests exist to assess lethal and sublethal side-effects of pesticides on bee survival, growth/development and reproduction. Within the context of ecotoxicology and insect physiology, we report the development of a new bioassay to assess the impact of sublethal concentrations on the bumblebee foraging behavior under laboratory conditions. In brief, the experimental setup of this behavior test consists of two artificial nests connected with a tube of about 20 cm and use of queenless micro-colonies of 5 workers. In one nest the worker bees constructed brood, and in the other food (sugar and pollen) was provided. Before exposure, the worker bees were allowed a training to forage for untreated food; afterwards this was replaced by treated food. Using this setup we investigated the effects of sublethal concentrations of the neonicotinoid insecticide imidacloprid, known to negatively affect the foraging behavior of bees. For comparison within the family of neonicotinoid insecticides, we also tested different concentrations of two other neonicotinoids: thiamethoxam and thiacloprid, in the laboratory with the new bioassay. Finally to evaluate the new bioassay, we also tested sublethal concentrations of imidacloprid in the greenhouse with use of queenright colonies of B. terrestris, and here worker bees needed to forage/fly for food that was placed at a distance of 3 m from their hives. In general, the experiments showed that concentrations that may be considered safe for bumblebees can have a negative influence on their foraging behavior. Therefore it is recommended that behavior tests should be included in risk assessment tests for highly toxic pesticides because impairment of the foraging behavior can result in a decreased pollination, lower reproduction and finally in colony mortality due to a lack of food.