Immunological Screening of Actinomycetes of khumbu Region of Nepal

Abstract

The preserved isolates of actinomycetes from Lobuche area (5000-5300 meter in height)and Lukla area (2660 meter in height) of Khumbu region of Nepal were taken for thestudy. A total of eighty isolates of actinomycetes were revived on starch casein agar(SCA). Out of them, isolate Lobuche 137.2 was selected for further study. Isolate Lobuche137.2 has already been identified as a new species of actinomycetes in the project work ofNepal Academy of Science and Technology (NAST) named “Biodiversity of Khumburegion: Population study of Actinomycetes”in year 2002. The isolates of Lobuche 137.2 were streaked on starch casein agar (SCA). Isolates ofLobuche 137.2 are observed to be creamy in colour on starch casein agar (SCA) plate.Macroscopically, Lobuche 137.2 has creamy aerial mycelium with creamy substrate.Microscopically, Lobuche 137.2 has been identified by coverslip method. On 1000x ofmicroscope, it has long non-septate hyphae with the branching filaments and its spores arepresent at the end of the filament. Shapes of the spores of Lobuche 137.2 are oval and theyare present in a chain form. To detect the immunological sensitivity of the spore 137.2 as Indirect Enzyme LinkedImmunosorbent Assay (IND-ELISA) is considered more appropriate. Therefore, IndirectEnzyme Linked Immunosorbent Assay (IND-ELISA) for spore of Lobuche 137.2 wasperformed. Three control and a test was set on the ELISA plate. Intense colour wasobserved on the well having 1:20 dilution of test. Spore density of 0.6 × 10 spores/200μlwas detected on 1:20 dilution of test sample. Key words:-actinomycetes, hyphae, Indirect Enzyme Linked Immunosorbent Assay.