However a significant decrease in food consumption was observed during the 4th week in rats of the group treated with EASPA at the dose of 14 mg/kg body weight () compared to the control group. According to the results, a gradual increase in the mean body weight was observed in rats in treated and control groups during the 28-day study period. The change in hematological parameters was within the normal range as showed elsewhere [39, 42]. This study financially supported by UMRG Grant (RG422/12HTM), University of Malaya, Malaysia and FRGS Grant (FP030-2015A) from the Ministry of Education Malaysia. This Guideline is intended primarily for use with rodents (rat preferably). Sub-acute toxicity study of ARELE was conducted on Wistar albino rats according to the protocols described in OECD guidelines 407 . Before conducting the experiment, the animals were randomly selected and grouped into three group (n=4) and then kept in their cage for 5 days prior to dosing to allow acclimatization to the laboratory conditions. The organs were cleaned of fat and blotted with clean tissue paper, and then weighed on balance. Therefore, sub-chronic toxicity should be proceeded based on the oral doses of DC resin methanol extract in sub-acute oral toxicity test. Cocos nucifera L. variety aurantiaca is commonly known as king coconut in English and “Thembili” in Sinhalese [3]. Similarly, normal structure and histology of the spleen also observed in all the rats of acute and sub-acute oral toxicity tests. The DC resin methanol extract was administered to the female rats under overnight fasting by using oral gavage in a volume of 10 ml/kg body weight. I. U. Kariyawasam of Department of Botany and voucher specimen (Assess. Konelab 20XT auto analyzer analyzed biochemical parameters. ; El-Hadiyah, T.M. Regul Toxicol Pharmacol. and A.B. Adopted: 3 October 2008: OECD Publishing; 2008. The resin of DC collected from Socotra Island (Yemen) in May 2013. Necropsy and gross examination on internal organs (liver, heart, kidney, uterus, and spleen) were carried out. Nine rats (females) were used in an acute oral toxicity test, whereas 60 rats (30 males and 30 females) used in sub-acute oral toxicity test (the repeated dose 28 day). There was no significant difference in biochemical parameters in rats of all test groups compared to the control group in both acute and subacute toxicity studies. exposed 5 days per week but exposure for 7 days per week is also allowed. 28 (OECD,1987) The body weights of animals recorded shortly before the administration of the tested substance and at the end of each week. In the satellite group, no significant difference showed between the two groups (Table 11). https://doi.org/10.1186/s12906-018-2110-3, DOI: https://doi.org/10.1186/s12906-018-2110-3. Female Wistar albino rats (Species—Rattus norvegicus, Origin—CLEA Japan) approximately 12 weeks old, weighing 112 g–144 g were purchased from the Animal Center, Medical Research Institute (MRI), Sri Lanka for toxicity studies. At the end of each experiment (at 15th day for acute oral toxicity and at 29th day for sub-acute oral toxicity tests except the satellite groups which were at 43rd day), the rats generally anesthetized by intraperitoneal injection of 80 mg/kg of ketamine 100 mg/ml + 7 mg/kg of xylazine 100 mg/ml (Troy Laboratories PTY. Cookies policy. Effect of EASPA on relative organ weights in the subacute toxicity study. A. Salawu, B. ; Muruganandam, A.V. Organs excised were saved in 10% formalin for further histological analysis [, Data of the present study was expressed as means ± SEM (five replicates). Liver: black arrow – portal vein; white arrow – portal triad. Prior to the start of the experiment, body weight of animals was recorded individually for calculating proper treatment dosage. In the acute toxicity study, all haematological parameters in rats treated with EASPA at the dose level of 2000 mg/kg body weight were similar to those of the control group rats. The current study performed to evaluate the safety of the DC resin methanol extract after a single or 28 consecutive daily oral administrations. BMC Complement Altern Med 18, 50 (2018). The statements, opinions and data contained in the journals are solely Subacute toxicity study was designed according to the Organisation for Economic Co-operation and Development (OECD). Under microscopic examination, the liver of DC treated animals showed with normal cellular architecture and binucleation and was without any distortions similar to the control groups. Finally, they were fixed in 10% buffered formalin solution for histopathological examination similar to the procedure described under Section 2.6.2. They were acclimatised to laboratory conditions for 7 days before the experiments and housed in groups of three for acute oral toxicity and in groups of five for sub-acute oral toxicity. This result was consistent with the previous literature that showed the Genus, Physical and behavioural examination revealed no adverse effects on mice given 400 mg/kg/day and 2000 mg/kg/day when compared to the control group (, After 28 days of extract administration at 400 mg/kg and 2000 mg/kg body weight (, The results of the biochemical parameters analyzed such as urea, creatinine, and aminotransferases (ALAT and ASAT) are shown in, Although the liver excised from all treated animals after 28 days of dosing was subjected to histological examination for analyzing the following injuries, bile ducts, hepatic vein, the artery in the portal area, and hepatic fat had no modification detected when compared to the control mice (. An experienced pathologist who was unaware of the experimental groups to which each section belonged conducted the analysis. ; Vaidya, R.A.; Vaidya, A.B. PubMed Central This revised Test Guideline 412 (TG 412) is designed to fully characterize test chemical toxicity by the inhalation route for a subacute duration (28 days), and to provide robust data for quantitative inhalation risk assessments. A histopathological study carried to confirm biochemical findings and to identify any structural changes. The haematological parameters of rats of all treated groups were similar to those of control group rats, with the exception of mean corpuscular haemoglobin (MCH) and mean corpuscular haemoglobin concentration (MCHC) of test group rats treated with EASPA at the dose level of 14 mg/kg body weight) (). A raw data of hematological parameters, is available in the Additional file 1. The 13C NMR spectrum was similar to what is reported previously and showed signals characteristic for epicatechin units indicating that EASPA is composed mainly of epicatechin units [5] and was used for toxicity studies. This research was funded by DSR King Saud University under RG No RG-1440-009. 2013;1(2):123–9. Kidney: black arrow – cortex; white arrow – medulla. 1. Preliminary phytochemical screening of the IC revealed that it contains a high amount of proanthocyanidins. Exp Toxicol Pathol. A post hoc Tukey test showed significant differences between the DC treated (male and female) and control groups as shown in Table 8 as p < 0.05. Furthermore, effects of EASPA on oestrogen and progesterone levels in female rats have also been reported. ; Weerasooriya, T.R. There was no significant difference in ROW between test group and control group rats. Therefore, it can be suggested that the oral administration of EASPA to rats daily for 28 days at the above-mentioned dose levels did not produce any treatment related body weight change during the study period. The procedures used to perform this study are in agreement with the international guidelines used for the use of laboratory animals. C. P. Ekanayake, M. G. Thammitiyagodage, S. Padumadasa, B. Seneviratne, C. Padumadasa, A.M. Abeysekera, "Acute and Subacute Toxicity Studies of the Ethyl Acetate Soluble Proanthocyanidins of the Immature Inflorescence of Cocos nucifera L. in Female Wistar Rats", BioMed Research International, vol.
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