Naoki Hashizume

Prediction of the bioconcentration factor in common carp (Cyprinus carpio L.) using data from the dietary exposure bioaccumulation fish test

An approach to predicting the bioconcentration factor (BCFpre ) from the predicted uptake rate constant (k1 pre ) and the depuration rate constant measured in the dietary exposure bioaccumulation fish test (k2 dietary ) [BCFpre  = k1 pre /k2 dietary ] is proposed in test guideline 305 of the Organization for Economic Cooperation and Development Guidelines for Testing of Chemicals. Data were collected on the BCFs of 197 test chemicals from Japans Chemical Substances Control Law database. To demonstrate how the BCFpre compares with experimentally derived BCF under optimum conditions, 48 of 197 test chemicals, including a number of studies that could be considered problematic, were excluded from the analysis. The k1 pre was calculated by using 22 published prediction methods: the correlations between experimental uptake rate constants (k1 aqueous ) and k1 pre for all prediction methods were very low and were statistically nonsignificant (p > 0.05). Three prediction methods were also selected that gave relatively good values for the geometric mean of k1 pre /k1 aqueous and calculated values of BCFpre for 12 test chemicals. Linear relationships (p < 0.05) are presented between logarithm of experimental and predicted BCF. The correlation coefficients of growth-corrected experimental and predicted BCF tended to be higher than values that were not growth corrected. For some test chemicals, use of predicted BCF led to a bioaccumulation classification different from that of existing regulatory criteria.

Categorization of nano-structured titanium dioxide according to physicochemical characteristics and pulmonary toxicity

A potentially useful means of predicting the pulmonary risk posed by new forms of nano-structured titanium dioxide (nano-TiO2) is to use the associations between the physicochemical properties and pulmonary toxicity of characterized forms of TiO2. In the present study, we conducted intratracheal administration studies in rats to clarify the associations between the physicochemical characteristics of seven characterized forms of TiO2 and their acute or subacute pulmonary inflammatory toxicity. Examination of the associations between the physicochemical characteristics of the TiO2 and the pulmonary inflammatory responses they induced revealed (1) that differences in the crystallinity or shape of the TiO2 particles were not associated with the acute pulmonary inflammatory response; (2) that particle size was associated with the acute pulmonary inflammatory response; and (3) that TiO2 particles coated with Al(OH)3 induced a greater pulmonary inflammatory response than did non-coated particles. We separated the seven TiO2 into two groups: a group containing the six TiO2 with no surface coating and a group containing the one TiO2 with a surface coating. Intratracheal administration to rats of TiO2 from the first group (i.e., non-coated TiO2) induced only acute pulmonary inflammatory responses, and within this group, the acute pulmonary inflammatory response was equivalent when the particle size was the same, regardless of crystallinity or shape. In contrast, intratracheal administration to rats of the TiO2 from the second group (i.e., the coated TiO2) induced a more severe, subacute pulmonary inflammatory response compared with that produced by the non-coated TiO2. Since alteration of the pulmonary inflammatory response by surface treatment may depend on the coating material used, the pulmonary toxicities of coated TiO2 need to be further evaluated. Overall, the present results demonstrate that physicochemical properties may be useful for predicting the pulmonary risk posed by new nano-TiO2 materials.

Effects of pentobarbital, isoflurane, or medetomidine-midazolam-butorphanol anesthesia on bronchoalveolar lavage fluid and blood chemistry in rats.

Bronchoalveolar lavage fluid (BALF) is commonly examined for pulmonary toxicity in animal studies. Two common means of anesthesia before euthanasia and bronchoalveolar lavage in rats are intraperitoneal injection of pentobarbital and inhalation of isoflurane. Medetomidine-midazolam-butorphanol is an alternative anesthesia to pentobarbital for animal welfare; however, the effect of this combination on BALF and blood chemistry is unknown. Here, we compared the effects of anesthesia by intraperitoneal injection of pentobarbital or one of two combinations of medetomidine-midazolam-butorphanol (dose, 0.375-2.0-2.5 or 0.15-2.0-2.5 mg/kg) or by inhalation of isoflurane on BALF and blood chemistry in rats with or without pulmonary inflammation. In BALF, we determined total protein, albumin, lactate dehydrogenase, total cell count and neutrophil count. In serum, we conducted a general chemistry screen. After anesthesia with pentobarbital or isoflurane, there were no significant differences between any of the BALF or blood chemistry parameters with or without inflammation. After anesthesia with either of the combinations of medetomidine-midazolam-butorphanol, lactate dehydrogenase, total cell count, neutrophil count, and almost all of the blood chemistry parameters were comparable with those observed after pentobarbital or isoflurane; however, BALF albumin and serum glucose were significantly increased in rats without inflammation. After the combination of low-dose medetomidine in rats with inflammation, BALF parameters were comparable with those observed after pentobarbital or isoflurane. Our results show that, of the anesthetics examined, inhalation of isoflurane is the most appropriate means of anesthesia when examining BALF or serum for toxicity studies in rats.

Resampling the bioconcentration factors data from Japan's chemical substances control law database to simulate and evaluate the bioconcentration factors derived from minimized aqueous exposure tests

Existing standard bioconcentration tests (e.g., the Organization for Economic Cooperation and Development [OECD] test guideline 305) require large numbers of test animals and resources. The minimized aqueous exposure test is a new approach based on the standard bioconcentration test but allows estimation of bioconcentration factor (BCF) by minimized sampling of the test fish. The authors collected BCF data (298 curves from 155 chemicals, using common carp as test species) from Japans Chemical Substances Control Law database and resampled the data to simulate the calculation of BCF that would be obtained if studies had been designed to obtain kinetic BCF derived from minimized aqueous exposure tests (BCF(km)). The correlation was high (r(2)  = 0.967) between BCF derived from standard bioconcentration tests (BCF(full)) and BCF(km). The average value of the BCF(full) to BCF(km) ratio (BCF(full):BCF(km)) was 1.04 and ranged from 0.54 to 1.93, the 5th and 95th percentiles being 0.74 and 1.45, respectively. The results based on the 5th and 95th percentiles of the BCF(full):BCF(km) ratio suggest that BCF(full) 2,000 corresponds to BCF(km) 1,400 to 2,700, whereas BCF(full) 5,000 corresponds to BCF(km) 3,400 to 6,800. The authors also emphasize that the standard bioconcentration test should be performed when the resulting BCF(km) is in the region of regulatory concern.

Comparison of laboratory‐derived biomagnification factors for hexachlorobenzene in common carp conducted under 9 test conditions

Nine dietary exposure bioaccumulation fish tests with hexachlorobenzene (HCB) were conducted with common carp to explore how differences in test conditions (different test foods and feeding rates) influenced the lipid-corrected, growth-corrected kinetic biomagnification factor (BMFkgL ) value (BMFkgL  = BMFkg  × lipid content of test food/lipid content of test fish). The BMFkgL values for HCB differed by approximately a factor of 5 among the tests. The average, median, 95% confidence interval, and coefficient of variation of the BMFkgL values were 0.925, 0.998, 0.578 to 1.27, and 49%, respectively. The BMFkgL value differed markedly between tests conducted using test food with lipid contents of approximately 5 and 15%. Different feeding rates (2 or 3% of body weight/d) had comparatively little effect on the BMFkgL of HCB. The present study revealed that the lipid content of test fish was correlated with the growth-corrected kinetic BMF (BMFkg ) value of HCB but the lipid content of test food was poorly correlated with BMFkg . This lack of correlation might explain the large variations of the BMFkgL values observed. The value of the BMFkg normalized to a fish with a 5% lipid content (defined as the 5% lipid-normalized BMFkg in the present study) did not differ markedly between tests conducted using test food with different lipid contents (5-15%). It is therefore useful to report the 5% lipid-normalized BMFkg as well as the BMFkgL when dietary exposure tests are conducted. Environ Toxicol Chem 2018;37:1032-1039.

Combined repeated-dose and reproductive/developmental toxicity screening test of benzene, 1,1′-oxybis-, tetrapropylene derivs. in rats

Abstract We have conducted animal toxicity tests of chemicals for a chemical safety program implemented by the Ministry of Economy, Trade and Industry of Japan. Here we conducted a combined repeated-dose and reproductive/developmental toxicity screening test of benzene, 1,1′-oxybis-, tetrapropylene derivs. (BOTD). BOTD was administered to 9-week-old Crl:CD(SD) male and female rats by gavage at 0, 40, 200, or 1000 mg/kg/day. Males were treated for 42 days including mating period. Females were treated for 42–53 days through the premating, mating, pregnancy, and until Day 4 of lactation periods. Increases in prothrombin time and activated partial thromboplastin time values were observed only in males at 200 and 1000 mg/kg/day. Hypertrophy of centrilobular hepatocytes was observed with increased liver weight in both sexes at 200 and 1000 mg/kg/day, but there was no histologic evidence of hepatotoxicity. Diffuse hypertrophy of follicular cells in thyroid glands was observed in females at 200 mg/kg/day and in both sexes at 1000 mg/kg/day, with an increased blood cholesterol level in females at 1000 mg/kg/day. The conception index was decreased for females at 1000 mg/kg/day; and no abnormalities were detected in the reproductive indices of implantation, delivery, or pups’ condition, although a slight increase in the pups’ body weight was noted at birth. Our data indicate a no-observed-adverse-effect level of 40 mg/kg/day for repeated-dose toxicity on the basis of the prolongation of blood coagulating time, and of 200 mg/kg/day for reproductive/developmental toxicity on the basis of the decreased conception index.

Combined repeated-dose and reproductive/developmental toxicity screening test of 1-tert-butoxy-4-chlorobenzene in rats

Abstract We have carried out animal toxicity tests of chemicals for a chemical safety program implemented by the Ministry of Economy, Trade, and Industry of Japan. Here, we tested 1-tert-butoxy-4-chlorobenzene in a combined repeat-dose and developmental and reproductive toxicity test. The test chemical was administered daily by gavage to 9-week-old Crl:CD (SD) rats at doses of 0, 20, 100, and 500  mg/kg/d. Males were treated for 42 d beginning 14 d before mating. Females were treated from 14 d before mating to day 4 of lactation. Decreased spontaneous locomotion, decreased respiratory rate, and incomplete eyelid opening were observed at 500 mg/kg/d (both sexes), but resolved within 30 min of administration, suggesting central nervous system depression. No notable changes were observed in body weight, food consumption, functional battery tests, or blood test. Increased liver weight with centrilobular or diffuse hepatocyte hypertrophy was observed at 100 and 500 mg/kg/d (both sexes). There were no biochemical or histopathological changes related to hepatotoxicity. Increased kidney weight with basophilic tubules, tubule dilatation, and increased hyaline droplets were observed in males dosed at 100 and 500 mg/kg/d. Immunohistochemical staining indicated α2u-globulin nephropathy, a male rat-specific toxicity. Although kidney weight was also increased in females dosed at 500 mg/kg/d, it was not considered to be an adverse effect because there were no histopathological changes. Pup weights on postnatal day 0 were decreased at 500 mg/kg/d and still decreased on postnatal day 4. Our data indicated the no-observed-adverse-effect-level for repeated-dose and reproductive/developmental toxicity for 1-tert-butoxy-4-chlorobenzene was 100 mg/kg/d.