Author/Authors :
Zare Pourshob ، Somayeh Department of Parasitology - Faculty of Veterinary Medicine - University of Tehran , Meshgi ، Behnam Department of Parasitology - Faculty of Veterinary Medicine - University of Tehran , Shayan ، Parviz Department of Parasitology - Faculty of Veterinary Medicine - University of Tehran , Hassanpour ، Gholam Reza Center for Research of Endemic Parasites of Iran - Tehran University of Medical Sciences
Abstract :
BACKGROUND: A large part of our country s waste is organic matter, so researchers are studying fertilizer production from organic waste in various ways, including compost and vermicompost. On the other hand, glycolipoprotein extract of Eisenia fetida (G-90) has been defined to show numerous biological activities, e.g., anticoagulation, fibrinolysis, and anti-oxidative, etc. OBJECTIVES: The purpose of the present study was to determine phylogenetic relationship of the Iranian isolate of Eisenia fetida (E. fetida) with the other available taxa and to determine the G-90 protein complex for evaluation of its biological activities. METHODS: A piece of 1×1cm clitellum was separated and used for DNA extraction after homogenate preparation (by two different methods). The second internal transcribed spacer of the nuclear ribosomal DNA (rDNA-ITS2) and cytochrome C oxidase subunit 1 of the mitochondrial DNA (mtDNA-COX1) from adult E. fetida were amplified using polymerase chain reaction (PCR). Subsequently, PCR products were sequenced and their phylogenetic relationships were determined. Further-more, The G-90 protein complex was extracted from adult worms and electrophoretic pattern of proteins was obtained by sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE). RESULTS: The electrophoretic pattern of glycolipoprotein G-90, a protein complex, showed 10 bands with molecular weights of 14-130 kDa. ITS2 and COX1 sequences were 516 bp and 277 bp long, respectively. The amplified DNA sequences from both ribosomal and mitochondrial sequences had 88%-99% and 99% similarity to relevant sequences in GenBank, respectively. CONCLUSIONS: mtDNA-COX1 showed no considerable sequence variations as compared to other isolates in Gen-Bank, while rDNA-ITS2 exhibited more variations, in comparison with other isolates, indicating more variations among some of these isolates. Our results revealed that rDNA-ITS2 of our E. fetida was in a separate subclade, showing the greatest similarity with EF534709.1 isolate (99%) provided in GenBank, followed by JX531618.1 (94%), and KU708469.1 (88%). Our COX1 sequence demonstrated the high similarity of 99% when compared with five isolates from GenBank (MH475674.1, MH475673.1, MH475672.1, MH475670.1, and MH475666.1). The G-90 glycolipoprotein showed different proteins that can be assessed for their potential biological activities in medicinal properties.
Keywords :
Earthworm , Eisenia fetida , G , 90 , Molecular characteristics
