International Journal for Research in Applied Science & Engineering Technology (IJRASET) ISSN: 2321-9653; IC Value: 45.98; SJ Impact Factor:6.887 Volume 5 Issue IX, September 2017- Available at www.ijraset.com 50 ©IJRASET (UGC Approved Journal): All Rights are Reserved Novel Techniques to Study Ancient Micro Amber from Tropical Beach Sand Reveal a Treasurehouse of Exceptionally Well Preserved Fossilized Microfungi Sujata Dabolkar 1 , Nandkumar Kamat 2 1, 2, Mycological Laboratory, Department of Botany, Goa University, Taleigao, Goa, 403206, India. Abstract: Simple, novel techniques developed for separation and simultaneous direct morphometric study of Amber Micro Fragments (AMF) from tropical beach sand are reported yielding rich information on unidentified fossilized microfungi. Sieves of different mesh sizes were used to separate AMF from tropical beach sand. Fractions below 150 μm which proved rich in AMF were used for manual retrieval using stereomicroscope. A handprinted slide microarray having 4 X 12 squares used for microscopic examination of multiple AMF mounts revealed AMF having either rough or smooth surfaces and with or without microinclusions. The microinclusions could be morphologically attributed to fungi. The potential for systematic and comprehensive studies to retrieve and examine AMF at high frequency from tropical beach sand in the world and especially those which are threatened due to sea level rise due to climate change was demonstrated. The potential of retrievable AMF from tropical beach sand in microbiological, metagenomic studies and as biological proxies to reconstruct bygone biospheres has been highlighted. Keywords: Amber micro fragments (AMF), tropical beach sand, sea level rise, sieving, fossilized fungi, microarray I. INTRODUCTION Amber (Succinate=C6H6O4) is the fossilized resin produced from the trunks and the roots of certain trees mainly belonging to family Pinaceae, found in Russia, France, Germany, Lebanon, Spain, Dominican Republic, Austria, USA, Myanmar, Japan and Mexico [1] in various environments such as in lignite mines, silts, sediments (Table 1). The age of amber varies from 4 to 300 Million years. Size of amber ranges from sand grains to several centimeters [2]. Recent advances in spectrometry allow the development of physical characterization of amber to confirm that amber originates from various kinds of plants such as conifers and Fabaceae [3]. Various methods have been used (Table 2) to process the amber specimens. Differences between “true/natural/biogenic” and false/synthetic/artificial amber can be detected by some classical tests such as production of sweet, pine smell when burnt and insolubility in acetone and salt water test indicating flotation displayed by true, authentic amber [4]. Other tests include fluorescence test, refractive index test, IR spectroscopy, polarized light test [5]. Since gaps in knowledge about detection of amber in tropical beach sand samples were found in literature [6,7], the present study was aimed at development of novel techniques for relatively rapid high frequency isolation and microscopic visualization of amber using samples of tropical beach sand collected from coast in Goa, India. The locations primarily included local beaches facing threats due to sea level rise as it was felt that a degree of urgency is required to retrieve these locally and globally useful bioresources, before coastal erosion/submergence.