Toxicological Evaluation of the Repeated Dose Administration of the Ethanolic Extract of Azolla microphylla in Wistar Albino Rats

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  • Department of Biochemistry, Karpagam Academy of Higher Education, Coimbatore – 641021, Tamil Nadu ,IN
  • Department of Biochemistry, Karpagam Academy of Higher Education, Coimbatore – 641021 ,IN
  • Department of Biochemistry, Karpagam Academy of Higher Education, Coimbatore – 641021, Tamil Nadu ,IN
  • Department of Biochemistry, Karpagam Academy of Higher Education, Coimbatore – 641021, Tamil Nadu ,IN



Azolla microphylla, Hematology, Lethal Dose, Toxicology
Toxicological evaluation


Azolla microphylla is an easily cultivable aquatic plant with the commendable nutritious property. Recent reports on Azolla species emphasize the therapeutic potential of the plant extracts. Moreover, the same genus of plant also had displayed antioxidant potential owing to its free radical scavenging tendency. Although these attributes were identified, a study investigating the toxicological property of different dosages of ethanolic extract of A. microphylla (EAM) is not yet reported. Thus the present study aims for the in vivo toxicological evaluation of the EAM in Wistar strain of rats. Daily doses of 0, 250, 500, 1000 and 2000 mg/kg body weight of EAM were administered orally to group-I, group-II, group-III, group-IV & group-V rats, respectively for 14 days. Biochemical and histopathological studies were established through standard methods. The acute toxicity results suggest the non-toxic nature of the extract supported with the absence of mortality and toxic symptoms until 72 h of observation. The results of sub-acute toxicity study in the extract-treated rats (group-II to group-IV) indicate non-significant changes to the biochemical (total protein, AST, ALT, LDH, albumin, globulin, urea, creatinine, cholesterol, & triglycerides), hematological (Hemoglobin, RBCs, WBCs, platelets, monocytes, lymphocytes, & neutrophils), and histopathological observations when compared to the control group of rats. However, group-V rats were treated with 2000 mg/kg b.w. exhibited statistically significant variations to most of the biochemical and hematological parameters although no mortality/physical toxic signs were reported till the end of the experimental period. Thus, the sub-acute toxicity results suggest that the extracts were non-toxic and safe to rats between 250-2000 mg/kg b.w. concentration under 14 days observational period. Moreover, as there was no mortality upto 2000 mg/kg b.w., 50% lethal dose (LD50) could not be determined, and hence it is considered to be greater than 2000 mg/kg/day.


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How to Cite

Kunnathupara Bhaskaran, S., Kannappan, P., Muneeswari, P., & Madathil, R. (2021). Toxicological Evaluation of the Repeated Dose Administration of the Ethanolic Extract of <i>Azolla microphylla</i> in Wistar Albino Rats. Toxicology International, 28(1), 39–48.



Original Research
Received 2020-09-30
Accepted 2020-10-06
Published 2021-03-24



Vikrant A, Vivek KG. Chemistry and pharmacology of plant cardioprotectives: A review. Int. J. Pharm. Sci. and Res. 2011; 2(11):56–1167.

Gurib-Fakim A. Medicinal plants: Traditions of yesterday and drugs of tomorrow. Mol. Aspects Med. 2006; 27: 1–93. PMid:16105678 DOI:

Chen F, Jiang Y. Algae and their Biotechnological Potential. Springer. 2001; 316. DOI:

Sreenath KB, Sowmya S, Gopalakrishnan VK, Poornima K. Quantitative phytochemical analysis, In vitro antioxidant potential and GC–MS studies in ethanolic extract of Azolla microphylla. Asian J. Pharm. Clin. Res. 2016; 9:318–23.

Abraham G. Evaluation of antimicrobial activity of methanolic extracts of Azolla microphylla. Vegetos- Int. J. Plant Res. 2013; 26: 200–204. DOI:

Indira D, Rao KS, Suresh J, Naidu KV, Ravi A. Azolla pinnata as feed supplement in buffalo calves on growth performance. Indian J. Anim. Nutr. 2009; 26: 345–8.

Pereira AL, Bessa LJ, Leí£o PN, Vasconcelos V, da Costa PM. Bioactivity of Azolla aqueous and organic extracts against bacteria and fungi, Symbiosis. 2015; 65:17–21. DOI:

Kumar A, Kumari J, Kumar H, Nath A, Singh JK, Ali M, Kumar, R. Hepatoprotective and antioxidant effect of Azolla filiculoides on Profenofos induced hepatotoxicity in Swiss Albino mice. Caribb. J. Sci. Technol. 2014; 2:372–7. DOI:

Sreenath KB, Anandan R, Sowmya S, Gopalakrishnan VK, Poornima K. Effect of aqueous extract of Azolla filiculoides in gastric mucosa of ulcerated rats. Int. J. Pharm. Pharm. Sci. 2015; 7: 355–8.

Sreenath KB, Poornima K. Protective Effect of Azolla microphylla on biochemical, histopathological and molecular changes induced by Isoproterenol in rats, Biomed. Pharmacother. 2017; 89:473–81. PMid:28249249 DOI:

Organization for Economic Cooperation and Development (OECD), 2002. Guidelines for the testing of chemicals/Section 4: Health Effects Test No. 423, Acute oral toxicity-acute toxic class method, Paris.

Muhammad K Md., Mustafa S, Sengupta P Md., Sarker MR, Das A, Das SK. Evaluation of the acute and subacute toxicity of the ethanolic extract of Pericampylus glaucus (Lam.) Merr. in BALB/c mice. J. Acute Dis. 2015; 4: 309–15. DOI:

Culling CFA. Handbook of histopathological and histochemical techniques: including museum techniques, third ed., Butterworths, London; 1974. P. 73–159. 8.50012-3 DOI:

Konan NA, Bacchi EM, N. Lincopan N, Varela SD, Varanda EA. Acute, subacute toxicity and genotoxic effect of a hydroethanolic extract of the cashew (Anacardium occidentale L.). J. Ethnopharmacol. 2007; 110:30–8. PMid:17088034 DOI:

Hilaly JEI, Israili ZH, Lyoussi B Acute and chronic toxicological studies of Ajuga iva in experimental animals, J. Ethnopharmacol. 2004; 91:43–50. PMid:15036466 DOI:

Banaee M, Mirvagefei AR, Rafei GR, Majazi AB. Effect of sub-lethal Diazinon concentrations on blood plasma biochemistry. Int. J. Environ. Res. 2008; 2:189–98.

Prakash KM, Naidu PV, Muralidhar P. Promising phytochemicals from medicinal plant Ecliptaalba. Int. J. Pharm. Tech. 2011; 3:2868–73.

Brautbar N, Williams J. II Industrial solvents and liver toxicity: risk assessment, risk factors and mechanisms. Int. J. Hyg. Environ. Health. 2002; 205:479–91. PMid:12455270 DOI:

Shivaraj G, Prakash BD, Vinayak VH, Avinash AKM, Sonal NV, Shruthi SK. A review on laboratory liver function tests, Pan. Afr. Med. J. 2009; 3:17.

Ping KY, Darah I, Chen Y, Sreeramanan S, Sasidharan S. Acute and sub chronic toxicity study of Euphorbia hirta L. methanol extract in rats, BioMed Res. Int. 2013. PMid:24386634 PMCid:PMC3872372 DOI:

Zaias J, Mineau M, Cray C, Yoon D, Altman NH. Reference values for serum proteins of common laboratory rodent strains, J. Am. Assoc. Lab. Anim. Sci. 2009; 48:387–90.

Thapa BR, Walia A. Liver Function Tests and their Interpretation, Indian J. Pediatr. 2007; 74:663–71. PMid:17699976 DOI:

Loeb S. Clinical Laboratory Test: Values and Implication, Springhouse Corporation, Pennsylvania, USA; 1991.

Wurochekke AU, Anthony AE, Obidah W. Biochemical effects on the liver and kidney of rats administered aqueous stem bark extract of Xemenia Americana. Afr. J. Biotechnol. 2008; 7: 2777–80.

Liu M, Li M, Liu J, Wang H, Zhong D, Zhou H, Yang B. Elevated urinary urea by high-protein diet could be one of the inducements of bladder disorders. J. Transl. Med. 2016; 14:53. PMid:26879937 PMCid:PMC4755000 DOI:

Yakubu MT, Akanji MA Oladiji AT. Alterations in serum lipid profile of male rats by oral administration of aqueous extract of Fadogia agrestis stem. Res. J. Med. Plant. 2008; 2:66–73. DOI:

Pendota SC, Yakubu MT, Grierson DS, Afolayan J. Effect of administration of aqueous Extract of Hippobromus Pauciflorus leaves in male Wistar rats, Afr. J. Tradit. Complement. Altern. Med. 2010; 7:40–6. PMid:21304611. PMCid:PMC3005378 DOI:

Adebayo JO, Adesokan AA, Olatunji LA, Buoro DO, Soladoye AO. Effect of ethanolic extract of Bougainvillea spectabilis leaves on haematological and serum lipid variables in rats. Biokemistri. 2005; 17:45–50. DOI:

Vikas SW, Mali VR, Arulmozhi S, Bodhankar SL, Kakasahed R, Mahadik R. Cardioprotective effect of ellagic acid on doxorubicin induced cardiotoxicity in wistar rats. J Acute Med. 2015; 5:1–8. DOI:

Sagar K, Vidyasagar GM. Evaluation of acute toxicities of leaf extract of Caesalpinia bonducella (L.) Flem. Int. J. Pharm. Biosci. 2010; 6:1–15.

Singh T, Sinha N, Singh A. Biochemical and histopathological effects on liver due to acute oral toxicity of aqueous leaf extract of Ecliptaalba on female Swiss albino mice. Indian J. Pharmacol. 2013; 45:61–5. PM id:23543876. PMCid:PMC3608297 DOI:

Damek-Poprawa M, Sawicka-Kapusta K. Histopa thological changes in the liver, kidney and testes of bank voles environmentally exposed to heavy metal emissions from the steelworks and zinc smelter in Poland. Environ. Res. 2004; 96:72–8. PMid:15261786 DOI:

Robaszkiewicz A, Balcerczyk A, Bartosz G. Antioxidant and prooxidative effects of quercetin on A549 cells. Cell Bio. Int. 2007; 31:1245–50. PMid:17583542 DOI: