Repeated Dose Oral Toxicity Study Research Articles

Bacillus subtilis ZB423: 90-Day repeat dose oral (gavage) toxicity study in Wistar rats.

The novel genetically modified probiotic Bacillus subtilis ZB423 was assessed in a 90-day repeated-dose oral toxicity study adhering to Good Laboratory Practice (GLP) and Organization for Economic Cooperation and Development (OECD) guidelines. Spray-dried spores at a concentration of 1.1E12 CFU/g were administered at doses of 130, 260, and 519 mg/kg body weight/day correlating to 1.43 × 1011, 2.86 × 1011, and 5.71 × 1011 CFU/kg/day, respectively, by oral gavage to Wistar rats for a period of 90 consecutive days. Results showed no toxicologically relevant findings for B. subtilis ZB423 from measured parameters. The no observed adverse effect level (NOAEL) of B. subtilis ZB423 is 519 mg/kg body weight/day corresponding to 5.71 × 1011 CFU/kg/day for lyophilized B. subtilis ZB423 spores under the test conditions employed.

Safety of a feed additive consisting of vitamin B2/riboflavin produced with Eremothecium ashbyi CCTCCM 2019833 for all animal species (Hubei Guangji Pharmaceutical Co., Ltd).

Following a request from the European Commission, the Panel on Additives and Products or Substances used in Animal Feed (FEEDAP) was asked to deliver a scientific opinion on the safety of a feed additive consisting of vitamin B2/riboflavin produced with Eremothecium ashbyi CCTCCM 2019833 intended for use as a nutritional additive (functional group: vitamins, pro-vitamins and chemically well-defined substances having similar effects) for all animal species. The characterisation, safety and efficacy of the additive have been already assessed previously; however, the FEEDAP Panel could not conclude on the safety of the additive for the target species, consumers and the users due to lack of reliable toxicological data. In the present assessment, the applicant submitted new genotoxicity and repeated dose oral toxicity studies. After the assessment of the data newly submitted, the FEEDAP Panel concluded that the use of the feed additive in animal nutrition under the conditions of use proposed is of no safety concern for the target species and the consumer. The additive is not a skin/eye irritant nor a skin sensitiser, but it is considered a respiratory sensitiser.

Re-evaluation of the reduced heart weights in male rats in a 28-day oral repeated-dose toxicity study of tetramethylammonium hydroxide

We recently conducted a detailed hazard assessment of tetramethylammonium hydroxide (TMAH), a priority chemical substance under the Japan Chemical Substances Control Law. During this assessment, there was debate regarding the reduced heart weight observed in the treated male groups in the 28-day rat oral repeated-dose toxicity study. This finding was not observed in females in this study and in both sexes of oral toxicity studies for tetramethylammonium chloride (TMAC) or tetramethylammonium hydrogen phthalate (TMAHP). Unpublished individual data from the oral TMAH developmental and reproductive toxicity (DART) screening study were also obtained; no effect on heart weight was observed. In addition, background data on rat heart weight from six 28-day oral toxicity studies conducted in the same facility, year, strain, age, and breeder as the TMAH study were obtained from the Japan Existing Chemical Substances Database (JECDB). These investigations suggest that the statistically significant lower heart weight in the treated males in the 28-day toxicity study is likely caused by an incidental skewing of individuals with heavier heart weights toward control male groups and is not due to TMAH treatment. Thus, it is worthwhile to include as much relevant data as possible to confirm or refute unexpected findings in toxicity studies.

PSIII-27 Blood biochemistry, histology, and metabolomic profiling of contaminated with mycotoxin: A repeated dose oral toxicity study in rats

Abstract Mycotoxins causes diseases and physiological responses in humans and livestock. This study was performed to estimate the effects of binary mixtures mycotoxin {e.g., deoxynivalenol (DON) and fumonisin (FB)] on blood biochemistry, histology, and metabolic profiles in rats fed with different toxin concentrations. Experimental animals were administered by oral gavage with 0.9% saline, DON (1, 5, and 10 mg/kg BW), FB (1, 5, and 10 mg/kg) at 8 wk of age. Histological changes, metabolomic profiling, and blood biochemical analysis were performed Masson’s trichrome stain kit, liquid chromatography mass spectrometry, and VetTest chemistry analyzer, respectively. The final BW of the rats contaminated with mixtures mycotoxin significantly decreased compared with the control group. Biochemistry results and fibrosis were also significantly differentially regulated according to mycotoxin concentrations. Additionally, we observed collagen fibers in the liver and kidney showed blue staining after toxin contamination. For metabolomic profiling, data were analyzed using MetaboAnalyst 4.0. Results were analyzed by multivariate analysis to elucidate the potential compounds. The toxin-treated groups observed discriminating metabolites in the liver and kidney tissue but no differences in ileum. The profiling revealed potential metabolites such as glycine, serine and threonine metabolism. Phosphatidylethanolamine and tryptophan were identified with high sensitivity as candidate biomarkers via variable importance in the projection plot in piglets fed with mycotoxin (FDR < 0.05; P < 0.05). The differently treated groups showed discriminating metabolites in different concentrations. In conclusion, these findings suggest that mycotoxin exposure disturbs histological changes and metabolites. Dietary toxins in rats may better understand the physiological effects consistent with metabolism, which are commonly reported in animals.

Fragile Guts Make Fragile Brains: Intestinal Epithelial Nrf2 Deficiency Exacerbates Neurotoxicity Induced by Polystyrene Nanoplastics.

Oral ingestion is the primary route for human exposure to nanoplastics, making the gastrointestinal tract one of the first and most impacted organs. Given the presence of the gut-brain axis, a crucial concern arises regarding the potential impact of intestinal damage on the neurotoxic effects of nanoplastics (NPs). The intricate mechanisms underlying NP-induced neurotoxicity through the microbiome-gut-brain axis necessitate further investigation. To address this, we used mice specifically engineered with nuclear factor erythroid-derived 2-related factor 2 (Nrf2) deficiency in their intestines, a strain whose intestines are particularly susceptible to polystyrene NPs (PS-NPs). We conducted a 28-day repeated-dose oral toxicity study with 2.5 and 250 mg/kg of 50 nm PS-NPs in these mice. Our study delineated how PS-NP exposure caused gut microbiota dysbiosis, characterized by Mycoplasma and Coriobacteriaceae proliferation, resulting in increased levels of interleukin 17C (IL-17C) production in the intestines. The surplus IL-17C permeated the brain via the bloodstream, triggering inflammation and brain damage. Our investigation elucidated a direct correlation between intestinal health and neurological outcomes in the context of PS-NP exposure. Susceptible mice with fragile guts exhibited heightened neurotoxicity induced by PS-NPs. This phenomenon was attributed to the elevated abundance of microbiota associated with IL-17C production in the intestines of these mice, such as Mesorhizobium and Lwoffii, provoked by PS-NPs. Neurotoxicity was alleviated by in vivo treatment with anti-IL-17C-neutralizing antibodies or antibiotics. These findings advanced our comprehension of the regulatory mechanisms governing the gut-brain axis in PS-NP-induced neurotoxicity and underscored the critical importance of maintaining intestinal health to mitigate the neurotoxic effects of PS-NPs.

90-day oral toxicity study in rats of a protein-rich powder derived from Xanthobacter sp. SoF1.

Xanthobacter sp. SoF1 (SoF1) is an autotrophic hydrogen-oxidizing bacteria that produces protein-rich biomass and has potential to be an alternative protein source that is more environmentally sustainable than animal and plant derived proteins. A protein-rich powder derived from SoF1 was the test material in a 90-day repeated-dose oral toxicity study to explore major toxic effects, demonstrate target organs, and provide an estimate of a no-observed-adverse-effect level (NOAEL). Daily doses of 0 (vehicle only), 375, 750, and 1500 mg/kg bw/day of the test material were administered by gavage to 10 Han:WIST rats/sex/group. An additional group was administered 1290 mg/kg bw/day whey protein concentrate as positive control. No treatment-related adverse effects were observed, and no target organs were determined after 90/91 days of consecutive administration of the test item. A NOAEL of 1500 mg/kg bw/day was determined.

An evaluation of the genotoxicity and 90-day repeated dose oral toxicity in rats of Porphyridium purpureum.

Interest in microalgae products for use in food is increasing, as demands for sustainable and cost-effective food choices grow due to the escalating global population and increase in climate-related struggles with agriculture. Toxicological assessments of some species of microalgae have been conducted, but there were little data available for the oral consumption of the red microalgae Porphyridium purpureum and no data on genotoxicity. This article articulates a genotoxicity assessment and a 90-day repeated dose oral toxicity study in rats performed according to OECD guidelines. Under the experimental conditions applied, the test item did not induce gene mutations by base pair changes or frameshifts in the genome of the strains used in the bacterial reverse mutation test. Similarly, the test item did not induce structural chromosomal aberrations in V79 hamster lung cells. The test item also did not cause chromosomal damage in bone marrow of mice in the mammalian micronucleus test. The no observed adverse effect level (NOAEL) of the 90-day repeated dose oral toxicity study in rats was determined to be the highest dose tested, 3000 mg/kg bw/day. These data add to the body of evidence regarding the safety of P.purpureum for human consumption.

An alkaloid enriched fraction from Murraya koenigii (L.) Spreng. Leaves ameliorate HFD-induced obesity and metabolic complexities in C57BL/6J mice

Ethnopharmacological relevanceMurraya koenigii commonly known as curry leaf, is traditionally used in India to manage various ailments including diabetes mellitus. Curry leaves are well documented in Indian Ayurvedic system of medicine for beneficial effects in skin eruptions, dysentery, emesis, poisonous bites and bruises. The anti-hyperglycemic and anti-hyperlipidemic effects of curry leaf extracts have been demonstrated through several in vitro and in vivo experiments previously. Aim of the studyTo prepare an alkaloid enriched fraction (AEF) from M. koenigii and its evaluation on i) in vitro adipogenesis process and ii) in vivo high fat diet-induced obesity in C57BL/6J mice. Materials and methodsMKME and AEF were prepared from M. koenigii leaves. The four carbazole alkaloids (bioactive markers) isolated from AEF were quantitatively determined in the leaves by RP-HPLC method. MKME and AEF were studied for anti-obesogenic activity in adipocytes in vitro and in HFD-induced C57BL/6J obese mice in vivo. At the termination of the in vivo study, lipid profile, hepatic and renal injury and glucose levels were analyzed in the blood samples. Animal tissues were examined histopathologically to determine any signs of damage. Repeated dose oral toxicity study for 28 days on Sprague-Dawley rats was also performed to determine the safety profile of AEF. ResultsBoth MKME and AEF displayed anti-obesogenic activity at 25 μg/ml concentration in vitro and showed 54.06 ± 3.86% and 37.46 ± 3.17% lipid accumulation, respectively compared to control. Further, supplementation of AEF and MKME in HFD-fed C57BL/6J mice helped in controlling weight gain, improved dyslipidemia and glucose intolerance significantly. AEF showed better anti-obesity activity than MKME both in vitro and in vivo study. Repeated administration of AEF up to 1 g/kg dose for 28 days showed no pathological tissue damage. Both MKME and AEF were standardized using a simple and validated RP-HPLC method. ConclusionPresent study was aimed at preparation of a novel alkaloid-enriched fraction from methanolic extract of M. koenigii leaf and its evaluation for anti-diabesity effect. Our results demonstrated AEF to be a promising plant-based therapy for ameliorating obesity and related metabolic complications in HFD-fed C57BL/6J mice.

TOXICOLOGICAL EVALUATION OF OLEANOLIC ACID (PENTACYCLIC TRITERPENOID) EXTRACTED FROM LANTANA CAMARA ROOTS FOLLOWING ORAL EXPOSURE IN WISTAR RATS

Objective: The aim of the present study was to perform an acute toxicity study to obtain information on the possible adverse effects from a single oral administration of Oleanolic acid in Wistar rats as the onset of toxicity, and to determine the range of exposure (to the LD50 cut-off criteria). Following a sub-chronic 90-days Repeated toxicity study by oral route to determine any potential indication of its dose response relationship and determine no observed effect level (NOEL)/ no-observed adverse effect level (NOAEL)/ low observed effect level (LOEL)/ low observed adverse effect level (LOAEL). Methods: A dose level of 2000mg/kg body weight was employed as step one for single acute study. Based on the survival pattern of the previously dosed animals after 48 hours, same dose 2000 mg/kg body weight) was repeated for Step 2 as a confirmatory test. For the 90-day toxicity study, the highest dose was determined as 1000mg/kg Body weight, and the middle and lower doses were 500 and 250mg/kg Body weight respectively. The rat group was held for a 14-day recovery period after the last dose administration, to observe for any persistence or reversal of toxic effects. Results: Results of the acute toxicity study showed no mortality on the dose level of 2000mg/kg body weight with no significant clinical and body weight changes. During the 90-day Repeated dose oral sub-chronic toxicity study, no rats died. There were no significant clinical changes related to the test item in terms of functional evaluation, body weight, food and water consumption, ophthalmological tests, urine analysis, necropsy findings, or organ weight, Hematology, and biochemistry at the highest dose level of 1000 mg/kg bwt. Conclusion: It is concluded that LD50 Cut-off Value of “Oleanolic acid” in from acute oral toxicity study in Wistar Rats is 2000 mg/kg b.wt. In addition, 90 days study have showed no significant changes with respect to any hematological or blood chemical analyses in 1000, 500 and 250 mg/kg bwt groups. Based on histopathological findings, clinical signs and other parameters, it may be concluded that upon repeated once oral administration for consecutive 90 days, the Oleanolic acid (Pentacyclic Triterpenoid) extracted from Lantana camara roots at the dose level of 1000 mg/kg body weight have caused no adverse effect in both sexes of Wistar Rats.

Safety profile of Naga Bhasma prepared by two classical methods

Abstract Background: Naga Bhasma (NB) (incinerated lead) has been indicated as a remedy for the treatment of a wide range of diseases including Prameha (diabetes), Kasa (cough), Raktapradara (menorrhagia), Shukradosha (defects in semen), etc., However, improperly prepared NB can cause harmful effects on the human body. In modern science also, lead consumption has been accepted to produce toxicity. Aim: The present study was undertaken to elucidate and compare the toxicity profile of two samples of NB prepared by two different methods to ascertain the safety aspects. Materials and methods: In the first method, NB was prepared by using Parada (mercury), Gandhaka (sulfur), and Nimbu Swarasa (juice of Citrus medica var acida of Watt). In the second method, NB was prepared by using Ashwattha-Twaka-Churna (Ficus religiosa L.), Manahshila (realgar) (Ariloha), and Nimbu Swarasa. In both methods, 30 calcination cycles were given. Charles foster albino rats were used for the pharmacological study. Acute oral toxicity study was carried out as per organization for economic cooperation and development (OECD) 425 and chronic oral toxicity study was carried out as per OECD 408 (90-day oral repeated dose toxicity study). Acute oral toxicity study was conducted at five dose levels, namely 100, 250, 500, 1000, and 2000 mg/kg body weight. Chronic toxicity was carried out at a dose level of 62.5 mg/kg body weight. Animals were sacrificed on the 91st day by cervical dislocation. Blood was collected from the jugular vein and sent for hematological and biochemical investigations were done. Organs were used for histopathological studies. The data were analyzed by unpaired t-test. Results: No mortality was observed in the acute toxicity study. Chronic toxicity study shows that both the test drugs do not produce significant toxicity, but both were not completely free from adverse effects at the dose level studied. Conclusion: The study shows that both the test drugs do not produce significant toxicity, but they are not completely free from adverse effects at the dose level studied.

Safety assessment of mixtures of 1,9-nonanediamine (NMDA) and 2-methyl-1,8-octanediamine (MODA), for use in food contact materials.

The EFSA Panel on Food Contact Materials, Enzymes and Processing Aids (CEP) assessed the safety of mixtures of 1,9-nonanediamine (NMDA) and 2-methyl-1,8-octanediamine (MODA) when used to produce polyamide food contact materials for contact with all food types for long-term storage at room temperature and below, including heating up to 121°C for up to 2 h. The polyamide material is also intended to be used for repeated use up to 121°C with short contact (up to 30 min). The polymer typically contains ■■■■■ of a low molecular weight fraction (LMWF, < 1000 Da). The specific migration was measured with polyamide samples in a set of migration tests with 3% acetic acid and 10% ethanol. NMDA and MODA were not detected at ■■■■■, respectively. The specific migration of the LMWF consisting of NMDA/MODA-related species was up to ■■■■■. The overall migration in olive oil was below the detection limit (3 mg/dm2). The most abundant migrating LMWF oligomers were identified. Toxicological studies were performed with NMDA, MODA and with polyamide formulations enriched in the LMWF. The results of genotoxicity assays did not raise a concern. From a repeated-dose oral 90-day toxicity study in rats, the Panel identified a no observed adverse effect level (NOAEL) of 1000 mg/kg body weight per day for the migrating LMWF. The CEP Panel concluded that NMDA/MODA mixtures do not raise a safety concern for the consumer when used as comonomer with terephthalic acid to manufacture polyamide articles intended for contact with all food types, except for infant formula and human milk, if the migration of NMDA and MODA does not exceed 0.05 mg/kg food (as a sum of the two substances) and if the migration of the LMWF consisting of NMDA/MODA-related species does not exceed 5 mg/kg food.

Toxicological safety evaluation of an aqueous lemon balm (Melissa officinalis) extract

Melissa officinalis (lemon balm) has a long history of safe use as an aromatic herb, flavoring, tea, food supplement, and traditional medicine. An aqueous extract of the leaves of M. officinalis is intended for use as a food ingredient, however the existing safety database does not contain any high quality toxicological studies to support safe consumer exposure. Therefore, a standard tier 1 genotoxicity battery (bacterial reverse mutation and in vitro mammalian cell micronucleus tests) and a 90-day repeated dose oral toxicity study in rats were conducted in accordance with GLP and OECD guidelines. The genotoxicity studies confirmed that aqueous lemon balm extract is not genotoxic at up to the highest concentrations tested (5000 μg/plate or 5000 μg/mL). A non-GLP 14-day dose range finding study was conducted prior to the 90-day study to confirm dietary administration of aqueous lemon balm extract at concentrations of 0, 0.5, 1.6, or 5.0%. The 90-day study was conducted using the established dietary concentrations and no test substance-related adverse effects on clinical, hematological, biochemical, macroscopic, or histopathologic parameters were reported. Thus, the no-observed-adverse-effect-level was determined to be at least 3046.1 and 3720.9 mg/kg body weight/day (the highest doses tested) for male and female rats, respectively.

Risks for animal and human health related to the presence of polychlorinated naphthalenes (PCNs) in feed and food.

EFSA was asked for a scientific opinion on the risks for animal and human health related to the presence of polychlorinated naphthalenes (PCNs) in feed and food. The assessment focused on hexaCNs due to very limited data on other PCN congeners. For hexaCNs in feed, 217 analytical results were used to estimate dietary exposures for food-producing and non-food-producing animals; however, a risk characterisation could not be performed because none of the toxicological studies allowed identification of reference points. The oral repeated dose toxicity studies performed in rats with a hexaCN mixture containing all 10 hexaCNs indicated that the critical target was the haematological system. A BMDL20 of 0.05 mg/kg body weight (bw) per day was identified for a considerable decrease in the platelet count. For hexaCNs in food, 2317 analytical results were used to estimate dietary exposures across dietary surveys and age groups. The highest exposure ranged from 0.91 to 29.8 pg/kg bw per day in general population and from 220 to 559 pg/kg bw per day for breast-fed infants with the highest consumption of breast milk. Applying a margin of exposure (MOE) approach, the estimated MOEs for the high dietary exposures ranged from 1,700,000 to 55,000,000 for the general population and from 90,000 to 230,000 for breast-fed infants with the highest consumption of breast milk. These MOEs are far above the minimum MOE of 2000 that does not raise a health concern. Taking account of the uncertainties affecting the assessment, the Panel concluded with at least 99% certainty that dietary exposure to hexaCNs does not raise a health concern for any of the population groups considered. Due to major limitations in the available data, no assessment was possible for genotoxic effects or for health risks of PCNs other than hexaCNs.

Safety Evaluation of Vatari Guggulu through Acute and 90-day Repeated Dose Toxicity Study in Experimental Animals

Abstract Background: Vatari Guggulu is an Ayurvedic Medicine used in the treatment of Vata disorders. It contains Vatari taila (Castor oil), Shuddha gandhaka (purified Sulphur), Haritaki (Terminalia chebula Retz.), Vibhitaki (Terminalia bellirica [Gaertn.] Roxb.), Amla (Emblica officinalis Gaertn) and guggul (Purified Commiphora mukul Hook). As per the Ayurvedic texts, it is widely used clinically in the treatment of rheumatoid arthritis, hip pain, sciatica, gout, joint pain, and burning sensation. However, there are no systematic toxicity studies were performed to scientifically evaluate the safety of Vatari Guggulu. Objectives: The present study was designed to investigate the safety of Vatari Guggulu in acute and 90-days repeated dose oral toxicity as per Organization for Economic Cooperation and Development guidelines 423 and 408. Materials and Methods: During the acute toxicity, Vatari Guggulu was orally administered at a single limit dose of 2000 mg/kg. The mortality and clinical signs were observed for 14 days. Similarly, the 90-days repeated dose toxicity was studied by administering the Vatari Guggulu at a limit dose of 1000 mg/kg. Results: In the acute toxicity, there were no signs of toxicity, and the LD50 was calculated as &gt;2000 mg/kg. During the 90 days of repeated dose toxicity, the Vatari Guggulu at the dose of 1000 mg/kg did not show any signs of toxicity as compared to the control. The hematological, serum biochemical, and urine analysis profile of the Vatari Guggulu at the dose of 1000 mg/kg did not show any significant difference as compared to the control (P &gt; 0.05). Further, the histopathological data of control and Vatari Guggulu-treated rats were found to be normal. Conclusions: It was concluded that the test drug Vatari Guggulu is found to be safe upto 2000 mg/kg in acute toxicity and 1000 mg/kg in 90-days repeated dose oral toxicity study. The NOAEL of Vatari Guggulu is established as 1000 mg/kg/day.

Toxicological assessment of chondroitin sulfate oligosaccharide

Toxicology studies were conducted to evaluate the safety of chondroitin sulfate oligosaccharide (CSO) produced by the enzymatic hydrolysis of shark cartilage by a bacterial lyase. Although the safety of chondroitin sulfate, which is used as a dietary supplement, has been investigated, no safety studies have been conducted on CSO. CSO was evaluated in bacterial reverse mutagenesis, mammalian cell gene mutation using mouse lymphoma cells, in vivo mammalian erythrocyte micronucleus assays, and a ninety-day repeated dose oral toxicity study in rats. The study showed that CSO was not genotoxic. The No Observed Adverse Effect Level (NOAEL) was determined to be 4,000 mg/kg bw/day, the highest dose tested.

Safety and efficacy of a feed additive consisting of phenylcapsaicin (aXiphen) for chickens for fattening (aXichem AB).

Following a request from the European Commission, EFSA was asked to deliver a scientific opinion on the safety and efficacy of phenylcapsaicin (aXiphen®) as a zootechnical additive (functional group: physiological condition stabilisers) in feed for chickens for fattening. The additive under assessment, phenylcapsaicin, is safe for chickens for fattening up to the maximum proposed use level of 15 mg/kg complete feed. A margin of safety could not be established. Phenylcapsaicin is not genotoxic. The reference point for phenylcapsaicin derived from a 90-day repeated dose oral toxicity study in rats is 37.2 mg/kg body weight (bw) per day, the lowest of the model averaged BMDL20 values for alanine aminotransferase increase in plasma. The metabolic similarity in the laboratory animals and the target species was not demonstrated and the identity of the marker residue could not be established. In the absence of such data, the safety for the consumers could not be evaluated. The inhalation exposure of phenylcapsaicin (as liquid) for the user was considered unlikely. The FEEDAP Panel considered the additive irritant to the eyes but not to the skin and it is not a dermal sensitiser. In the absence of appropriate data, the environmental risk assessment for phenylcapsaicin could not be performed. It is unlikely that phenylcapsaicin bioaccumulates in the environment and the risk of secondary poisoning is considered low. The FEEDAP Panel could not conclude on the efficacy of the additive in chickens for fattening at the proposed conditions of use.

Effects of Acute and Repeated Dose Toxicity Profiling of Chelidonic Acid in Rats: in Silico and in Vivo Evidence.

Chelidonic acid is a phytoconstituent found in rhizomes of the perennial plant celandine. The current study aims to evaluate the acute and repeated dose oral toxicity study of chelidonic acid as per the OECD guidelines 425 and 407. The pharmacokinetic and toxicity profile of chelidonic acid was predicted using online servers and tools. A single dose of chelidonic acid (2000 mg/kg) was administered to female Wistar rats in an acute toxicity study, and the animals were monitored for 14 days. We studied the toxicity profile of chelidonic acid at 10, 20, and 40 mg/kg doses in Wistar rats for repeated dose toxicity (28 days). Clinical biochemistry, haematological, and urine parameters were estimated. A gross necropsy and histopathology were performed. A single oral dose of chelidonic acid (2000 mg/kg) showed no signs of toxicity or mortality. The Administration of chelidonic acid showed no significant alterations in haematological, biochemical, and urine parameters. The histopathology showed normal structure and architecture in all the vital organs. A gross necropsy of vital organs showed no signs of toxicity. The chelidonic acid was found to be safe at all selected dose levels in the acute and repeated dose toxicity study in rats.

Safety Assessment of AYUSH SC-3 through Acute and 90 Days Repeated Dose Oral Toxicity Study

The present study is focused on establishing the safety of the formulation through acute and 90 days of repeated oral dose toxicity as per the Organization for Economic Cooperation and Development (OECD) guidelines. During the acute toxicity test, the drug was orally administered at a limited test dose of 2000 mg/kg. Clinical signs, feed and body weight were recorded. At the end of 14 days, the animals were euthanized and subjected to a detailed post-mortem examination (necropsy). As per OECD, 408 the 90 days repeated dose oral toxicity study was carried out with three different doses of test drug i.e., 1500, 1000 and 500 mg/kg. Cage side observations, body weight and feed intake were recorded. Upon termination of the study, urine analysis, haematology and clinical biochemical examinations were performed. Finally, the rats were subjected to euthanasia, a gross necropsy was conducted and vital organs were weighed and made prone for histopathological evaluation. Cage-side observation of AYUSH-SC-3 treated animals showed no signs of toxicity, and the mortality or moribund state was observed in both of the studies. No significant change in body weight and feed intake was seen in AYUSH-SC-3 treated animals. Gross morphology and necropsy findings of the animals revealed no treatment-related. Haematological and biochemical parameters of rats treated with AYUSH-SC-3 were found to be non-significant when compared to the control group. Necropsy findings and relative organ weights did not change significantly. Further, the histopathological analysis of major organs showed no major lesions and treatment-related changes. The LD50 of AYUSH SC-3 is greater than 2000 mg/kg and NOAEL is up to 1500 mg/kg.

Therapeutic Effects of Geranium Oil in MPTP-Induced Parkinsonian Mouse Model.

Parkinson's disease (PD) is an incurable neurodegenerative disease characterized by motor and non-motor disabilities resulting from neuronal cell death in the substantia nigra and striatum. Microglial activation and oxidative stress are two of the primary mechanisms driving that neuronal death. Here, we evaluated the effects of geranium oil on 1-methyl-4-phenyl-1,2,3,6-tetra-hydropyridine (MPTP) mouse model for PD, on microglial activation, and oxidative stress. We demonstrate that oral treatment with geranium oil improved motor performance in this model. The therapeutic effects of geranium oil were observed as a significant increase in rotarod latency and distance among the mice treated with geranium oil, as compared to vehicle-treated MPTP mice. Geranium oil also prevented dopaminergic neuron death in the substantia nigra of the treated mice. These therapeutic effects can be partially attributed to the antioxidant and anti-inflammatory properties of geranium oil, which were observed as attenuated accumulation of reactive oxygen species and inhibition of the secretion of proinflammatory cytokines from geranium oil-treated activated microglial cells. A repeated-dose oral toxicity study showed that geranium oil is not toxic to mice. In light of that finding and since geranium oil is defined by the FDA as generally recognized as safe (GRAS), we do not foresee any toxicity problems in the future and suggest that geranium oil may be a safe and effective oral treatment for PD. Since the MPTP model is only one of the preclinical models for PD, further studies are needed to confirm that geranium oil can be used to prevent or treat PD.

Subchronic oral toxicity assessment of a cannabis extract

Cannabidiol (CBD) is present in Cannabis Sativa L. and has been used in medicines and foods to deliver beneficial health effects. Despite this, research on CBD safety utilising modern testing methods is lacking. Therefore three separate safety experiments were performed on a CBD isolate. Sprague-Dawley rats were used to investigate prenatal development, a 14-day toxicity sighting study, and an OECD compliant 90-day subchronic oral toxicity trial, with 35-day off-dose recovery. The prenatal screening study demonstrated reduced body weights and food consumption in the highest dose group, but no substance-related changes in pregnancy rate, maternal or placental gross abnormalities, or premature deliveries. The 14-day study indicated tolerance up to 460 mg/kg bw/d of CBD isolate. Based on these findings, a 90-day repeated dose oral toxicity study was performed at doses of 0, 30, 115, 230, and 460 mg/kg bw/d of CBD, followed by a 35-day off-dose recovery period. In the 90-day study, some non-adverse organ and tissue changes were observed. With the exception of the high dose group, these fully reversed during the recovery period. Based on these findings, sub-chronic consumption of highly purified isolate results in a CBD NOAEL of 460 mg/kg bw/d for males and 230 mg/kg bw/d for females.