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      Sub-Acute Toxicity Study of Tiger Milk Mushroom Lignosus tigris Chon S. Tan Cultivar E Sclerotium in Sprague Dawley Rats

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          Abstract

          Lignosus also known as “Tiger Milk Mushroom,” is classified in the family Polyporaceae and mainly consumed for its medicinal properties in Southeast Asia and China. The sclerotium is known as the part with medicinal value and often used by the natives to treat a variety of ailments. Lignosus tigris Chon S. Tan, one of the species of the Malaysia Tiger Milk mushroom, has recently been successfully cultivated in laboratory. Earlier studies have demonstrated the L. tigris cultivar E sclerotia exhibited beneficial biomedicinal properties. This study evaluated the potential toxicity of L. tigris E sclerotia in a 28-day sub-acute oral administration in Sprague Dawley (SD) rats. L. tigris E sclerotial powder was administered orally at three different doses of 250, 500, and 1000 mg/kg to the SD rats once daily, consecutively for 28-days. Body weight of the rats was recorded and general behavior, adverse effects, and mortality were observed daily throughout the experimental period. At the end of the experiment, blood hematology and biochemistry, relative organ weights, and histopathological analysis were performed. Results showed that there were no mortality nor signs of toxicity throughout the 28-day sub-acute toxicity study. Oral administration of the L. tigris E sclerotial powder at daily dose up to 1000 mg/kg had no significant effects in body weight, relative organ weight, blood hematological and biochemistry, gross pathology, and histopathology of the organs. L. tigris E sclerotial powder did not cause any treatment-related adverse effect in the rats at different treatment dosages up to 1000 mg/kg. As the lethal dose for the rats is above 1000 mg/kg, the no-observed-adverse-effect level (NOAEL) dose is more than 1000 mg/kg.

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          Acute and chronic toxicological studies of Ajuga iva in experimental animals.

          Ajuga iva (L.) Schreber (AI), is widely used in the Moroccan pharmacopoeia as a panacea (cure-all), and specifically for gastrointestinal disorders and diabetes, and as an anthelmintic. No toxicological investigations have been carried out on this plant. We have previously observed that single oral doses (2-14 g/kg) of a lyophilised aqueous extract of AI (AI-extract) in mice or daily oral administration of 10 mg/kg of AI-extract in rats for 2 weeks did not result in any adverse effects. We have now evaluated AI-extract for its behavioural and pharmaco-toxicological effects after acute and chronic administration by the oral and intraperitoneal routes in rats and mice. No toxicity was observed in mice after single oral doses of as high as 14 g/kg of the AI-extract. However, single intraperitoneal injections of the AI-extract (1500-5500 mg/kg BW) produced a dose-dependent increase in adverse effects in the general behaviour and the mortality rate; the LD50 of acute intraperitoneal dose was 3.6 g/kg. In chronic toxicological studies in rats, the AI-extract (administered orally at daily doses of 100, 300 and 600 mg/kg for 3 months), did not cause any changes in haematological and biochemical parameters, with the exception of a transient rise in platelet counts and a short-term decrease in serum glucose levels. Histopathological examination of the brain, liver and the kidneys at the end of the study (3 months) showed normal architecture suggesting no morphological disturbances.
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            Clinical chemistry and haematology historical data in control Sprague-Dawley rats from pre-clinical toxicity studies.

            The purpose of this paper is to provide historical data pertaining to clinical chemistry and haematology parameters, obtained from control Sprague-Dawley rats, used in pre-clinical toxicity studies. Mean, standard deviation, minimum and maximum values for haematological and coagulative profiles, haemato-biochemistry and urine analysis data, and the differences per sex and study duration, 4 versus 13 weeks, are presented. The studies were conducted in agreement with the GLP (Good Laboratory Practice) regulations. Statistically significant differences, at the confidence level of 99%, for the red blood cell (RBC) parameters, the white blood cell (WBC) series parameters, plasmatic albumin/globulin (A/G), alanine amino-transferase (ALT), alkaline phosphatase (ALP), creatinine, globulin, glucose, sodium, total protein, tryglycerides, urea and urine volume were observed in males, when 4-week study values were compared with those obtained from 13-week studies. Female rats showed statistically significant variations, at the confidence level of 99% for RBC number and mean corpuscular haemoglobin (MCH), mean red blood cell volume (MCV), WBCs count and lymphocytes percentage, A/G, albumin, ALT, AST, ALP, creatinine, globulin, and sodium, when 4-week study values were compared to 13-week studies. Similar differences were observed comparing the female with male haematological and biochemical data for the two different times of the sample collection. These data could be useful as a reference for evaluation of background pathology in Sprague-Dawley rats, when used in studies performed to evaluate the toxicological profile of a new chemical entity (NCE) in agreement with requirements from international regulatory agencies.
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              Preliminary toxicity and phytochemical studies of the stem bark aqueous extract of Musanga cecropioides in rats.

              These studies were designed to determine the preliminary oral toxicity profile of the crude aqueous stem bark extract of Musanga cecropioides (MCW) in adult Sprague-Dawley rats and its active chemical constituents by way of phytochemistry. The acute oral toxicity study was conducted using limit dose test of Up and Down Procedure according to the OECD/OCDE Test Guidelines on Acute Oral Toxicity (AOT425statPgm, version: 1.0) at a limit dose of 3,000 mg/kg body weight/oral route. Repeat dose oral toxicity studies were conducted by daily oral dosing of 750 mg/kg body weight of MCW dissolved in 1 ml of 0.9% saline and 1 ml of 0.9% saline to rats in the test and control groups, respectively, for 28 days. On day 29, blood samples for bioassays were collected by cardiac puncture under diethyl ether anesthesia. The phytochemical analysis was conducted using standard procedures. The LD(50) estimate of the extract was calculated to be greater than 3,000 mg/kg body weight/oral route. The extract caused a significant (P<0.05) decrease in weight gain, differential eosinophil count and increase in serum creatinine but did not affect the organ weights, other serum electrolytes (Na(+), K(+), HCO(3)(-)), liver enzymes and other hematological indices in test rats. Its phytochemical analysis showed it contains saponins, flavonoids, alkaloids, tannins, phlobatannins, glycosides, reducing sugars and anthraquinones. These results show that the aqueous extract of Musanga cecropioides is relatively safe toxicologically when administered orally. Thus, its use in folkloric medicine as an oral antihypertensive is relatively safe when used over the tested period.
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                Author and article information

                Contributors
                Journal
                Front Pharmacol
                Front Pharmacol
                Front. Pharmacol.
                Frontiers in Pharmacology
                Frontiers Media S.A.
                1663-9812
                09 August 2016
                2016
                : 7
                : 246
                Affiliations
                [1] 1Medicinal Mushroom Research Group, Department of Molecular Medicine, Faculty of Medicine, University of Malaya Kuala Lumpur, Malaysia
                [2] 2Department of Pathology, Faculty of Medicine, University of Malaya Kuala Lumpur, Malaysia
                Author notes

                Edited by: Adolfo Andrade-Cetto, National Autonomous University of Mexico, Mexico

                Reviewed by: Benedict Green, United States Department of Agriculture-Agricultural Research Service, USA; Manoj Gajanan Kulkarni, University of KwaZulu-Natal, South Africa

                *Correspondence: Shin-Yee Fung fungshinyee@ 123456gmail.com

                This article was submitted to Ethnopharmacology, a section of the journal Frontiers in Pharmacology

                Article
                10.3389/fphar.2016.00246
                4977358
                da3fc840-565a-4b85-9b7f-94ff47379747
                Copyright © 2016 Kong, Tan, Fung and Pailoor.

                This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

                History
                : 09 June 2016
                : 25 July 2016
                Page count
                Figures: 3, Tables: 4, Equations: 0, References: 25, Pages: 10, Words: 6584
                Funding
                Funded by: Kementerian Sains, Teknologi dan Inovasi 10.13039/501100003200
                Award ID: FP029-2014A
                Funded by: Universiti Malaya 10.13039/501100004386
                Award ID: PG144-2014B
                Categories
                Pharmacology
                Original Research

                Pharmacology & Pharmaceutical medicine
                lignosus tigris,tiger milk mushroom,sclerotium,sub-acute toxicity,sprague-dawley rats

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