In Vivo Acute and Sub Chronic Toxicity of Water and Methanol Extracts of Meretrix meretrix Linnaeus on Sprague Dawley Rats

Abstract

Lamis clam (Meretrix meretrix Linnaeus) is one of the sea shells that have been utilized by the community. The shells are traditionally believed by peoples of Cirebon can increase stamina, lower blood pressure and can treat jaundice. Results of several studies demonstrated that these shells have various activities such as antioxidant, antitumor, hypolipidemic, antineoplastic, modulatory immune activity as well as antihyperglycemic and antihyperlipidemia. A wide range of research on the benefits of lamis clams has been done, but the pharmacological effects of the clam extracts have been less explored. Thus, the need to test the toxicity of the clam is very important. Toxicity tests studied in this research covered acute and sub chronic toxicities performed in vivo in rats. Parameters observed in this study were physical observation, presence or absence of rat deaths due to M. meretrix extract as well as the presence or absence of symptoms of liver and kidney damages. It is also useful in the development of research knowledge about the pharmacological effects of M. meretrix and also expected to be able to provide added value for M. meretrix utilization. The purpose of the study was to characterize the mussel (yield, proximate and heavy metals analyzes as well as qualitative tests of bioactive components contained in M. meretrix), to determine the acute and sub chronic toxicities of water and methanol extracts of M. meretrix in vivo on Sprague Dawley rats. The research was carried out in 3 stages. Phase 1 was the characterization of M. meretrix included: yield measurement, proximate analysis (AOAC 2005), analysis of heavy metals Pb, Cd, Hg and Cu (APHA 1989) and analysis of bioactive components (Harborne 1984). Phase 2 was acute toxicity test (OECD Test Guideline 403:2009). The parameters observed at this stage included: growth and feed intake of rats, observations of physical parameters, histopathology of rats liver and kidney as well as observation of the color changes in the organs of rats. Phase 3 was the final stage, namely the sub chronic toxicity test (OECD Test Guideline 413:2009). Parameters observed in this study included: growth and feed intake of rats, physical parameters (weight of kidney and liver) and blood serum chemistry parameters (urea, bilirubin, creatinine, albumin and cholesterol). Meretrix meretrix consisted of 11.09 % meat and 69.85 % shell. Based on the results of the proximate analysis yielded moisture content of 79.99 %, 1.50 % of ash, 0.22 % of fat, 9.42 % of protein and 8.81 % of carbohydrate. Analysis of heavy metals showed lamis clams meat containing Pb 0.0013 ppm, Cd 0.0088 ppm, Hg 0.0045 and Cu 0.0008 ppm. M. meretrix also contained various components of bioactive compounds. Three types of bioactive compounds were detected in the water and methanol extracts of M. meretrix which were alkaloids, steroids and saponins. Acute toxicity test of water and methanol extracts of M. meretrix was done by giving both extracts to rats with doses of 2, 4, 6 and 15 g/kg of body weight. Based on growth and physical observations on the rats for 14 days, the water and methanol extracts of M. meretrix had no acute toxicity effect (studied up to 15 g/kg body weight). Based on histopathological observations on the liver and kidneys, some cells faced necrosis and degeneration, but in general no significant effect was observed in comparison with control group. This result showed that the M. meretrix was not toxic and safe to be consumed. Sub chronic toxicity test of water and methanol extracts of M. meretrix was done by giving both extracts with doses of 0.1 and 1 g/kg bw for 90 days recurring basis. Test results showed that the water and methanol extracts of M. meretrix were not toxic to rats at the doses administered. There was no indication of metabolic disorders in rats after administration of water and methanol extracts at doses of 0.1 and 1 g/kg body weight. These treatments did not affect the growth of rats. No significant effect on liver and kidney weight of rats (p>0.05) was observed. Blood serum test on chemistry parameters of rats showed that water and methanol extracts of M. meretrix had no significant effect on levels of urea, cholesterol, and creatinine (p>0.05). Albumin and bilirubin levels were significantly different with controls (p<0.05), although the five parameters of the blood serum were still in the normal levels of blood serum chemistry for rats.