Isolation and Characterization of Psychrophilic Fungi from Batura, Passu and Siachen Glaciers, Pakistan

Abstract

A large portion of Earth consists of cold environment below ~5°C. This cold environment harbor micro and macro-organisms. The Psychrophilic fungi widely exist in Antarctica, Arctic, European alpines, cold deep Sea and the highest mountain range of Asia, HKKH (Hindu Kush, Karakoram and Himalaya). There are several large and small glaciers located in HKKH mountain ranges in the northern area of Pakistan. The samples were collected in the form of ice, glacier melt water and sediments from Batura glacier and Passu glacier (Karakoram) and Siachen glacier (Himalaya). The isolation and total viable count (CFU/ml) was done by spread plate method at 4°C and 15°C. Total 77 fungal strains isolated at 4°C (37 isolates) and 15°C (40 isolates). Fungal isolates were identified morphologically and microscopically and confirmed by 18S rRNA gene sequencing. For the microscopic identification, the slides were staind with lactophenol cotton blue and examined under 40 X magnification while phylogenetic analysis was done for molecular level identification and it has found that mostly, fungal isolates belong to the genus Penicillium, followed by Cladosporium, Mrakia, Geomyces, Leotiomycetes, Trichoderma, Thelebolus, Pseudeurotium, Fontanospora, Beauveria, Cordyceps, Cadophora, Tetracladium, Trametes, Mortierella, Candida, Eupenicillium, Scopulariopsis, Pseudogymnoascus, Antrodia, Sporobolomyces, Phoma, Periconia, Cryptococcus, Dothideomycetes class and Thelebolaceae family. The growths of isolates were checked on the basis of their tolerance to NaCl, pH and temperature. The range for the salt concentration for all isolates was (2 – 26%), pH (1-13) and temperature range for the strains isolated at 4°C was 15, 37, 45 and 50°C while that for strains isolated at 15°C was 4, 37, 45 and 50°C. Few isolates grown at up to 26% NaCl, some showed viability at 45°C, while majority of the isolates were able to grow at both pH 1 and pH 13. All isolates were screened for their antimicrobial activity against clinically isolated bacterial and fungal strains but they showed good antimicrobial activity against gram (+) bacteria. Fungal isolates also screened for extra-cellular enzymes production (including amylase, cellulase, deoxyribonuclease, lipase, phosphatase and protease). Different isolates shown greater abilities to produce extracellular enzymes but they were good producer of cellulases, lipases and phosphatases.