Mammalian Chromosomal Aberration Test Research Articles

Toxicological Safety Assessment of Heat-Treated Lactiplantibacillus plantarum OLL2712 as a Food Ingredient.

Lactiplantibacillus plantarum is a Gram-positive, bacilli-shaped bacterium commonly found in fermented foods and the human gastrointestinal tract. Heat-treatment of L. plantarum OLL2712 prior to consumption has been found to have positive health effects. However, a full toxicological evaluation of heat-treated L. plantarum OLL2712 cells (heat-treated OLL2712) has not been conducted. In this regard, heat-treated OLL2712 was evaluated for use as an ingredient in food and dietary supplements using a panel of toxicological tests including genotoxic and subchronic studies. Heat-treated OLL2712 did not show any genotoxicity based on an invitro mammalian chromosomal aberration test and invivo mouse micronucleus assay up to 8.0 × 109 cells/mL and 6.4 × 1012 cells/kg, respectively. In the acute oral toxicity study, no toxic effects were observed at 5 × 1011 cells/kg body weight. In the 13-week subchronic toxicity study, rats were administered (via gavage) heat-treated OLL2712 at concentrations of 0.5 × 1011, 1.5 × 1011, and 5 × 1011 cells/kg body weight. Administration did not significantly impact mortality, body weight, food consumption, ophthalmology, hematology, clinical chemistry, coagulation, urinalysis, and macroscopic or microscopic findings. The no-observed-adverse-effect level of heat-treated OLL2712 in both sexes was 5 × 1011 cells/kg body weight/day, the highest dose tested.

Assessment of genotoxicity and acute oral toxicity of a standardized Ocimum tenuiflorum extract (HolixerTM)

Ocimum tenuiflorum, commonly referred to as holy basil or Tulsi, has a long history of use in traditional medicine systems, particularly Ayurveda, due to its various health benefits, including anti-inflammatory, antioxidant, and adaptogenic properties. In contemporary contexts, this plant is progressively incorporated into dietary supplements and nutraceuticals. Given its widespread use and potential health beneficial properties, it is imperative to scientifically evaluate the safety of Ocimum tenuiflorum. This study presents comprehensive safety assessments of a standardized extract of Ocimum tenuiflorum. We conducted a series of genotoxicity studies, including the bacterial reverse mutation test (BRMT), in-vitro mammalian chromosomal aberration (CA) test, and in-vivo mammalian erythrocyte micronucleus (MN) test in Swiss Albino mice. Additionally, an acute oral toxicity study was performed using Sprague Dawley rats, adhering to OECD guidelines in a GLP-compliant laboratory. The results showed no mutagenic effect with O. tenuiflorum extract up to a dose of 5000 µg/plate in BRMT. The results of CA test revealed the non clastogenic activity of O. tenuiflorum extract up to a dose of 500 µg/mL with and without metabolic activation (S9). Ocimum tenuiflorum extract was found to be non-clastogenic at the highest tested dose of 2000 mg/kg bodyweight in invivo MN test. In acute oral toxicity study, O. tenuiflorum extract was found to be safe up to 5 g/kg bodyweight in Wistar rats. Collectively, these findings suggest that Ocimum tenuiflorum extract is non-genotoxic and safe for oral consumption up to 5000 mg/kg body weight in Sprague Dawley rats.

Safety evaluation of alpha-glycerylphosphorylcholine as a novel food

Safety evaluation of alpha-glycerylphosphorylcholine as a novel food

Lack of genotoxicity and subchronic toxicity in safety assessment studies of Akkermansia muciniphila formulation

Lack of genotoxicity and subchronic toxicity in safety assessment studies of Akkermansia muciniphila formulation

Safety evaluation of aqueous extract from Valeriana officinalis L. roots: Genotoxicity, acute, subchronic and teratology toxicity

Safety evaluation of aqueous extract from Valeriana officinalis L. roots: Genotoxicity, acute, subchronic and teratology toxicity

Safety assessment of enzymatically converted chicken bile as a novel food material: Genotoxicity, teratogenicity, and acute and subchronic toxicity studies.

Enzymatically converted chicken bile (CB), prepared by converting taurine deoxycholic acid (TCDCA) to taurine ursodeoxycholic acid (TUDCA) in CB, possesses various functional activities. But their nutrient composition and safety assessment have not been fully investigated yet. CB was mainly composed of proteins and steroids. CB did not show genotoxic effects based on Ames test, mammalian erythrocyte micronucleus test, and in vitro mammalian chromosomal aberration test. There were no growth abnormalities or deaths in the acute toxicity test for mice, indicating that CB is nontoxic with an LD50>10g/kg·body weight (BW). Subchronic toxicity test and genotoxicity test were performed based on intake of 0.5g CB per person daily at expanded doses of 33.3, 100, and 300 times (278, 833, and 2500mg/kg·BW). The result indicated that CB at 833mg/kg·BW showed no toxicity on BW, body weight gain, food intake, hematological, serum biochemistry, absolute/relative organ weights, urinalysis, and pathological features of rats in the subchronic toxicity test, while CB at 833mg/kg·BW induced maternal toxicity with no fetus teratogenicity or embryotoxicity in the teratogenicity test. In conclusion, CB did not show toxic effects and a long-term daily intake of CB at 0.5g per person is considered safe, but pregnant women should avoid it. These findings could provide a reference for the safe use of CB in functional food.

Evaluating the toxicity of the roots of Asarum heterotropoides var. mandshuricum extracted using the decoction method: Genotoxicity, single-dose toxicity, and 13-week repeated-dose toxicity studies

Evaluating the toxicity of the roots of Asarum heterotropoides var. mandshuricum extracted using the decoction method: Genotoxicity, single-dose toxicity, and 13-week repeated-dose toxicity studies

Toxicology study profile of Nicotinamide mononucleotide after acute and 90-day sub chronic dosing in Wistar rats and mutagenicity tests

Toxicology study profile of Nicotinamide mononucleotide after acute and 90-day sub chronic dosing in Wistar rats and mutagenicity tests

Genotoxic evaluation of kovir capsules using mammalian bone marrow chromosomal aberration test

The genotoxic potential of KOVIR capsules was investigated in vivo using the mammalian bone marrow chromosomal aberration test. Four groups of mice, seven in each group, were used in the study: Group I served as vehicle control; Group II received the positive control (cyclophosphamide 50 mg/kg; single dose; intraperitoneal route); and Group III to IV were given orally the KOVIR capsules, suspended in distilled water, at a single test dose of 2.16 and 4.32 capsules/kg, respectively. The obtained data disclosed the safety of KOVIR capsules, which did not show substantial genotoxic effects at either dose when compared with the vehicle group.

The Potential for Genotoxicity, Mutagenicity and Endocrine Disruption in Triclosan and Triclocarban Assessed through a Combination of In Vitro Methods.

Triclosan and Triclocarban, preservatives widely used in cosmetics and other consumer products, underwent evaluation using a battery of new-approach methodologies in vitro (NAMs). Specifically, the Microplate Ames Test (MPF™ Test, Xenometrix, Allschwil, Switzerland) was employed to assess mutagenicity, the Comet assay in vitro on the HaCat cell line and the Mammalian Chromosome Aberration Test were utilized to evaluate genotoxicity, and the XenoScreen® YES/YAS assay was applied to investigate endocrine disruption. The chemicals did not exhibit any positive responses for mutagenicity. However, the mammalian chromosome aberration test identified both chemicals as being positive for genotoxicity at 10 µg/mL. In the Comet assay, the percentage of DNA in the tail significantly increased in a concentration-dependent manner (at 5 and 10 µg/mL for Triclosan, at 2.5, 5, and 10 µg/mL for Triclocarban). The positive response depended on the increasing concentration and the duration of exposure. Triclosan, but not Triclocarban in any of the endocrine assays performed, indicated a potential for endocrine activity in the anti-estrogenic and anti-androgenic assays. The positive in vitro results detected were obtained for concentrations relevant to final products. The alarming findings obtained with the use of new-approach methodologies (NAMs) justify the current precautionary regulatory approach, limiting the use of these preservatives.

Acute toxicity and genotoxicity studies on new melatonergic antidepressant GW117

Acute toxicity and genotoxicity studies on new melatonergic antidepressant GW117

Toxicology and digestibility of Chlamydomonas debaryana green algal biomass.

There is an economic interest, both for food security and for the non-meat-eating population, in the development of novel, sustainable sources of high-quality protein. The green algae Chlamydomonas reinhardtii has already been developed for this purpose, and the closely related species, Chlamydomonas debaryana, is a complementary source that also presents some additional advantages, such as reduced production cost. To determine whether C. debaryana may have a similar safety profile to that of C. reinhardtii, a wild type strain was obtained, designated TS04 after confirmation of its identity, and subjected to a battery of preclinical studies. Genetic toxicity was evaluated using a bacterial reverse mutation test, an in vitro mammalian chromosomal aberration test, and an in vivo mammalian micronucleus test in a mouse model. No genotoxic potential (e.g., mutagenicity and clastogenicity) was observed in these tests under the employed conditions up to maximum recommended concentrations or doses. To assess general toxicity, a 90-day repeated-dose oral toxicity study was conducted in rats. No mortality or adverse effects were observed, and no target organs were identified up to the maximum feasible dose, due to solubility, of 4,000 mg/kg bw/day. The no-observed-adverse-effect level was determined as the highest dose tested. A digestibility study in simulated gastric fluid was conducted and determined that TS04 has low allergenic potential, exhibiting rapid digestion of proteins. Due to the negative results of our evaluation, it is reasonable to proceed with further development and additional investigations to contribute towards a safety assessment of the proposed use in food for human consumption.

Toxicological evaluation of protein powder derived from Cupriavidus necator.

Microorganisms have the potential to produce nutrient-rich products that can be consumed as food or feed. The protein-rich powder derived from heat treatment of the whole-cell biomass of polyhydroxybutyrate-deficient Cupriavidus necator, a metabolically versatile organism that uses elements found in the air, is an example of such a product. To assess the safety of the protein powder for use as a nutritional ingredient in human food, in accordance with internationally accepted standards, its genotoxic potential and repeated-dose oral toxicity were investigated. A bacterial reverse mutation test, an in vitro mammalian chromosomal aberration test, and an in vivo mammalian micronucleus test were performed. No evidence of mutagenicity or genotoxicity was found. Additionally, a 90-day repeated-dose oral toxicity study in rats was completed, in which a total of 100 male and female Wistar rats were exposed by gavage to daily doses of 1000, 2000, or 3000 mg/kg bw/day of the test material. Following 90 days of continuous exposure, no mortality or treatment-related adverse effects were observed and no target organs were identified. Therefore, a no observed adverse effect level was determined at 3000 mg/kg bw/day, the highest dose tested.

Genotoxicity study of 2-methoxyethanol and benzalkonium chloride through Comet assay using 3D cultured HepG2 cells.

Though the key data in identifying carcinogenicity is experience in human, long-term carcinogenicity tests using experimental animals are more realistic. Because carcinogenicity tests require much time and cost, performing the test is minimized through pre-screening. Recently, as bioethics has been strengthened, it is required to minimize animal testing in screening tests as well as carcinogenicity tests. The replacement of the micronucleus assay in experimental animal is the beginning, and the ultimate goal is to replace the carcinogenicity test using experimental animals. The micronucleus assay and the comet assay in 3D culture system of human-derived cells is considered as the most applicable practical measures at this stage. This study was conducted to provide more diverse information in the evaluation of carcinogenicity by establishing the comet test method in a three-dimensional cell culture system. In this study, HepG2 cells were cultured for 4 days in hang-in drop method, and then cultured for 7 days on a low adhesion plate to prepare spheroids. The methods were confirmed by d-mannitol (negative control), ethylmethane sulfonate (positive control), and cyclophosphamide (positive control for metabolite). 2-methoxyethanol and benzalkonium chloride were selected as test substances. Though 2-methoxyethanol is positive in in vivo comet assay and in vitro mammalian chromosome aberration test, it is considered negative in the comprehensive genotoxicity evaluation based on negative in bacterial reverse mutation assay, in vitro mammalian cell gene mutation test and mammalian chromosome aberration test. Benzalkonium chloride has been questioned on carcinogenicity because it is a disinfectant ingredient that has become a social issue in Korea. As a result of the Comet assay for 2-methoxyethanol and benzalkonium chloride in the cultured HepG2 cell line, 2-methoxyethanol was evaluated as positive in the metabolic activation system, but benzalkonium chloride was evaluated as negative in both the presence and absence of the metabolic activation system. Therefore, in order to clarify the carcinogenic potential of 2-methoxyethanol, it is judged that additional studies based on mechanistic studies are needed.

Genotoxicity test of eight natural color additives in the Korean market.

BackgroundVarious natural color additives are preferred by many consumers over synthetic color additives because they are perceived to be safer. However, most do not have sufficient toxicity data for safety assurance. Color ingredients in particular have some structures suspected of being toxic. Eight natural color additives, gardenia red, blue, and yellow; lac color; cochineal extract; beet red; Curcuma longa Linne extract (Curcuma extract); and Monascus red, currently permitted for use in Korea, were selected and subjected to genotoxicity tests. Acceptable daily intake values have not been allocated to these color additives (except for cochineal extract) due to the lack of toxicity data. We used genotoxicity testing—the bacterial reverse mutation test (Ames test), in vitro mammalian chromosomal aberration test, and in vivo alkaline comet test—for minimum safety assurance.ResultsGardenia red and blue, cochineal extract, lac color, and beet red did not induce mutagenicity or chromosomal abnormalities. Gardenia yellow was mutagenic in the Ames test, but was not positive in the in vitro chromosomal aberration test or in vivo alkaline comet assay. Curcuma extract and Monascus red induced cytotoxicity in the Ames test at high concentrations in Salmonella typhimurium TA1537 and TA100, without showing mutagenicity. On cytotoxicity testing, Curcuma extract and Monascus red showed cytotoxicity at concentrations higher than 313 μg/ml in Chinese hamster ovary CHO-K1 cells and showed equivocal results in chromosomal aberration assay of the same cells. Curcuma extract and Monascus red produced significant increases in DNA damage at a dose of 2000 mg/kg b.w./day, and induced dose-dependent increases in % DNA in the tail and tail moment on in vivo comet assay.ConclusionsSix out of eight food colorants did not cause genotoxicity and cytotoxicity. However, Monascus red and Curcuma extract showed definite cytotoxicity and probable genotoxicity.

Safety evaluation of mutagenicity, genotoxicity, and cytotoxicity of Lactobacillus spp. isolates as probiotic candidates.

BackgroundProbiotics promote a healthy balance of gut bacteria and have many beneficial effects on human digestive physiology. Although, few side effects of probiotics have been reported. This study aimed to assess the safety of five probiotic candidate Lactobacillus strains isolated from healthy individuals by examining mutagenicity, genotoxicity, and oral toxic effects.MethodsFive selected candidate probiotic (SCPs) strains were evaluated for genotoxicity (Ames test with Salmonella typhimurium), in vitro mammalian chromosome aberration test and an in vivo mouse micronucleus assay on peripheral blood of mice. To evaluate the oral dose toxicity, BALB/c mice models were treated repeatedly (2000, 1000, and 500 mg/kg body weight /day) for 28‐days.ResultsThe Ames test performed for two S. typhimurium strains TA 98 and TA100 (both in the absence and in the presence of S‐9 metabolic activation system) did not show an increase in reverse mutation because of exposure to the SCPs in any of the doses (5.0, 2.5, 1.25, 0.625, and 0.3125 mg/plate). There was no genotoxicity in the SCPs treatment in the vitro chromosome aberration assay with Chinese hamster ovary cells (CHO‐K1). In addition, none of the tested strains increased the frequency of micronucleated reticulocytes in reticulocytes, the SCPs with the studied doses caused no substantial variation in the experimental groups compared to the negative control group (p > 0.05). SCPs were not acutely toxic when administered to male and female BALB/c mice by single gavage at (2000, 1000, and 500 mg/kg b.w/day) with no mortality or clinical signs, change in body weight or macroscopic abnormalities were observed in this dose range.ConclusionAs a result, SCPs did not induce mutagenic potential in vitro with bacterial reverse mutation, clastogenicity, and in vivo tests in the ranges of concentrations evaluated in our study.

Evaluation of the genotoxic potential of protoporphyrin IX and the safety of a protoporphyrin IX-rich algal biomass.

Chlamydomonas reinhardtii is a nonpathogenic, nontoxigenic green algae used as a sustainable source of protein in foods. In order to mimic meat-like qualities, a strain rich in protoporphyrin IX (PPIX), an endogenous heme/chlorophyll precursor, was developed using an evolution and selection strategy, and investigations were carried out to evaluate the safety of the novel strain, C. reinhardtii (red), strain TAI114 (TAI114). Digestibility and proteomic evaluations were conducted to determine whether any potentially allergenic or toxic proteins occurred as the result of the mutation process. The genotoxic potential of pure PPIX was evaluated using a bacterial reverse mutation test, an in vitro mammalian chromosomal aberration test, and an in vivo mammalian micronucleus test. Finally, the novel TAI114 biomass was evaluated for general toxicity and identification of target organs in a 90-day repeated-dose oral toxicity study in rats. All proteins were rapidly degraded in pepsin at pH 2.0 suggesting low allergenic potential. The proteomic evaluation indicated that TAI114 biomass contains typical C. reinhardtii proteins. PPIX was unequivocally negative for genotoxic potential and no target organs or adverse effects were observed in rats up to the maximum feasible dose of 4000 mg/kg bw/day TAI114 biomass, which was determined to be the no-observed-adverse-effect-level (NOAEL). These results support the further development and risk characterization of TAI114 biomass as a novel ingredient for use in the meat analogue category of food.

In vitro genotoxicity assessment and 28-day repeated dose oral toxicity study of steady-calcium formula in rats

In vitro genotoxicity assessment and 28-day repeated dose oral toxicity study of steady-calcium formula in rats

Evaluation of Subchronic Toxicity and Genotoxicity of Ethanolic Extract of Aster glehni Leaves and Stems

Aster glehni, a traditional plant on Ulleung Island in the Republic of Korea, has been recognized for its multiple medicinal properties. However, potential toxicity and safety analyses of A. glehni have not been previously investigated. Therefore, this study aimed to evaluate the safety profile of ethanolic extract of A. glehni leaves and stems (EAG) in terms of genotoxicity and subchronic oral animal toxicity under OECD guidelines and GLP conditions. Toxicological assessments were performed at doses of 1,250, 2,500, and 5,000 mg/kg/day in a 13-week oral repeated-dose toxicity study of EAG in male and female SD rats. In addition, an Ames test, an in vitro mammalian chromosomal aberration test, and a micronucleus test were performed. No toxicological changes in clinical signs, body weights, water and food consumption, urinalysis, hematology, clinical biochemistry, gross findings, and histopathological examinations were observed in subchronic oral animal toxicity. In addition, EAG gave negative results when evaluated using in vitro and in vivo genotoxicity tests. In conclusion, the no-observed-adverse-effect level (NOAEL) of EAG was considered to be 5,000 mg/kg/day, and no target organs were identified in both sexes of rats. EAG was also classified as nonmutagenic and nonclastogenic in genotoxicity testing. Collectively, these results show a lack of general toxicity and genotoxicity for EAG that supports clinical work for development as a herbal medicine.

An assessment of mutagenicity, genotoxicity, acute-, subacute and subchronic oral toxicity of paraxanthine (1,7-dimethylxanthine)

An assessment of mutagenicity, genotoxicity, acute-, subacute and subchronic oral toxicity of paraxanthine (1,7-dimethylxanthine)