UGC SPONSORED NATIONAL CONFERENCE ON CURENT & EMERGING TRENDS IN LIFE SCIENCES 18 & 19 MARCH 2014 ORG, BY DEPT.

OF BOTANY AND ZOOLOGY PARVATIBAI CHOWGULE COLLEGE OF ATRTS & SCIENCE MARGAO

Do Unique Stratospheric Life Forms Get A Piggy Back Ride Inside SW Monsoon Clouds To Leave Signatures In Locally Sampled Rainwater ? Kamat Nandkumar, Kolte Rutuja*, Dabolkar Sujata* Dept of Botany, Goa University, Taleigao, Goa, 403206, India Abstract Morris et al, 2008 while stressing the importance of biometeorology as an upcoming field of research highlighted the need of understanding microbiological dimensions of atmospheric processes. Research in USA (Mohler et al., 2007) and France (Brent, 2012), Canada, Germany, Austria (Deguillaume et al, 2008) has implicated or produced evidence of microbial life in stratosphere, storms and rain clouds. South west tropical monsoon is an important event for ecology and economy in India and other recipient monsoon countries. However scant attention has been paid to the microbiological aspects of tropical stratosphere and the possible role of bioaerosols or stratospheric life forms in this mega tropical disturbance. A humble attempt was made to test the hypothesis regarding detection and visualization of life forms in local samples of rainwater collected directly under standard aseptic conditions at the open, secure Goa University campus on basis of positive indications from a previous time series study which led to refinement of sampling technique. The results of physicochemical and microbiological analysis are presented and discussed with some speculations on possible origin of Inorganic, Organic components and the detected morphological diversity of interesting life forms revealed by bright field and phase contrast microscopy. The ecological, biological, microbiological, pathological and public health / epidemiological implications of the findings are presented and the importance of tropical SW monsoon related basic and advanced biometeorological studies is highlighted to solve the riddle of microbial life forms getting a piggy back ride inside the monsoon clouds.

Kamat Nandkumar, Kolte Rutuja*, Dabolkar Dept of Botany, Goa University, Taleigao, Goa, 403206, India
Abstract Morris et al, 2008 while stressing the importance of biometeorology as an upcoming field of research highlighted the need of understanding microbiological dimensions of atmospheric processes. Research in USA (Mohler et al., 2007) and France (Brent, 2012), Canada, Germany, Austria (Deguillaume et al, 2008) has implicated or produced evidence of microbial life in stratosphere, storms and rain clouds. South west tropical monsoon is an important event for ecology and economy in India and other recipient monsoon countries. However scant attention has been paid to the microbiological aspects of tropical stratosphere and the possible role of bioaerosols or stratospheric life forms in this mega tropical disturbance. A humble attempt was made to test the hypothesis regarding detection and visualization of life forms in local samples of rainwater collected directly under standard aseptic conditions at the open, secure Goa University campus on basis of positive indications from a previous time series study which led to refinement of sampling technique. The results of physicochemical and microbiological analysis are presented and discussed with some speculations on possible origin of Inorganic, Organic components and the detected morphological diversity of interesting life forms revealed by bright field and phase contrast microscopy. The ecological, biological, microbiological, pathological and public health / epidemiological implications of the findings are presented and the importance of tropical SW monsoon related basic and advanced biometeorological studies is highlighted to solve the riddle of microbial life forms getting a piggy back ride inside the monsoon clouds. Introduction
Unusual microbial life forms have been reported in clouds. The aim of our work was to challenge the popular romantic paradigm of rainwater falling directly from overhead clouds as pure and undertake a preliminary empirical investigation of its inorganic and biological components during south west monsoon in Goa taking the benefit of open ground on Taleigao plateau where Goa University campus is located. Clouds can be defined as atmospheric air masses in which water is condensed around particles in solid (ice crystals) or liquid form. Biological matter is found in the atmosphere in the form of living or dead organisms. Dimmick et al. (1979), Fuzziet al. (1997) and Sattler et al. (2001) suggested cloud droplets may provide a medium in which airborne cells can divide. Literature says because bacteria require water for their metabolism, they are supposed to be good cloud condensation nuclei(e.g., Mhler et al.,2007, Schnell and Vali, 1972). In a changing climate, one can furthermore suppose that even more bacteria find viable conditions in the atmosphere and, thus, may become more abundant in clouds and may be transported more widely. Microbes and their metabolic activities could affect meteorological processes in the atmosphere both by changing cloud chemistry (Brent 2012). The field of bioprecipitation is expanding rapidly in rest of the world but there is scanty attention paid in India. Astrobiologists are implicating stratospheric microbial life forms. In this work we demonstrate that one doesnt really need rocket science to sample and analyze rainwater directly and detect interesting inorganic and microbial components- the later probably getting a piggy back ride in the clouds whereas the former may be signature of atmospheric dust, volcanic ash, aeolian processes , air transportation exhaust and we conclude that rainwater falling in Goa during SW monsoon can no more be treated as pure but full of inorganic and some potentially dangerous and pathogenic microbial life forms. These may have wide implications in ecology, hydrology, biogeochemistry, plant and human diseases. A state wide effort has to be launched to collect and analyze rainwater samples periodically and build a cohesive spatiotemporal picture of microbial life forms raining in Goa and promote the field of biometeorology and RW microbiology.

* Sujata

Results and Discussion

Monsoon was active in first week of July 2013 when rainwater samples were obtained. Rainfall intensity was about 30 mm per hour

SW Monsoon clouds July 2013 at GU campus

Previous work on microbiology of atmospheric processes, clouds, rainwater etc. Authors Work done Cantrell and Heymsfield, 2005; Morris et Reported Biological particles, including al., 2004; Szyrmer and Zawadzki, 1997 certain bacteria, pollen, and decayed organic material Ohler ( Germany), DeMott and Vali Microbiology and atmospheric (USA) and Levin (Israel) 2007 processes: the role of biological particles in cloud physics Duce et al. 1980; Parrington et al. 1983; The intercontinental transportation of Betzer et al. 1988; Uematsu et al. 2002; millions of tons of desert dust annually Jickells et al. 2005 influences ecosystems on a global scale Kellogg and Griffin, 2006; Burrows et al., Most airborne biomass emanates from 2009a; Frohlich-Nowoisky et al., 2009; arid topsoil or marine sea spray Womack et al., 2010

Bioprecipitation research is in infancy in India

Four locations and six samples yielding total 1200 ml rainwater on microscopic analysis produced evidence of inorganic and microbial forms as indicated by photomicrographs in Figures A, B, C (Inorganic) and D, E (Microbial)
Figures A, B, C:- The inorganic dimension of rainwater samples produced some striking results indicating Aeolian quartz from desert storms and signature of unknown volcanic ejecta in the forms of auriferous quartz, metal sulphides and magnificent Gold microparticles which showed a1 characteristic Surface Plasmon Resonance (SPR). Results are consistent with other reports (See ref.) Figure A Microdebris from volcanic ejecta- silica and black sulphide assemblages Figure D:- Microbial forms recovered from rainwater samples showed diverse cell morphologies- Coccoid, Bacilloid bacterial cells; cyanobacteria and numerous fungal spores and hyphae- all these getting piggy back ride in monsoon clouds and landing with rainfall
b1 b2 b3

Several meteorological factors make microbiological studies of rainwater from SW monsoon interesting

a5

b4

a3 b5 b8

Rare view of formation of SW Monsoon clouds in ITCZ- Ocean-Atmosphere interface

Figure B Stunning forms of Quartz possibly or Aeolian origin from desert storms recovered in rainwater consistent with results elsewhere of intercontinental transport of such forms

b6

Figure E Time lapse and BF, DF images indicate motility of unidentified microbial form perhaps unique to its ephemeral existence in clouds. Such forms raining down may have implications for public health
b7

Materials and methods

Leads from previous unpublished work on rainwater in GU campus (2006-12)

Assembly of sterile, dust & contamination free PVC containers (6 sets) Selection of clear vegetation free open spaces permitting direct collection Direct Sampling in SIX sterile sealed containers at 4 different locations & Sample processing under aseptic conditions to obtain slides Microscopy and photomicrography (Bright field, dark field, phase contrast ) followed by Identification of sample components from morphology, optical properties & published literature

Rainfall during collection period July 2013 at GU campus

Figure C This is first report in India of recovery of auriferous Quartz and Gold microparticles of Desert/volcanic/meteoritic/cosmic origin having vast implications in biogeochemistry and astrobiology

Rainwater sampling points

Rainwater sample collection assembly in field

References
W.D. Hamilton and T.M. Lenton (1998). Spora and Gaia: how microbes fly with their clouds. Ethology Ecology & Evolution 10: 1-16, 1998. Mohler et al. (2007) Microbiology and atmospheric processes:the role of biological particles in cloudphysics, Biogeosciences Discuss., 4, 25592591. Ngoc-Phuc Hua et al. (2007) Detailed identification of desert-originated bacteria carried by Asian dust storms to Japan, Aerobiologia, 23:291298DOI 10.1007/s10453-007-9076-9. Deguillaume (2008), Microbiology and atmospheric processes: chemical interactions of primary biological aerosols, Biogeosciences, 5, 10731084. David J. Smith (2013), Microbes in the Upper Atmosphere and Unique Opportunities for Astrobiology Research, Astrobiology Volume 13, Number 10.

Our results indicate that one doesnt need rocket science to sample rainwater anywhere in Goa/India directly and aseptically and subject it to microscopic analysis for rapid detection of inorganic and microbiological components and these preliminary investigations are broadly consistent with exciting and mind blowing findings from other parts of the world which should alert and educate us that there is need for a radical paradigm shift in the way we take the rainwater for granted. India has huge scope to boost Biometeorology, Monsoon cloud and rainwater microbiology and we intend to continue this work with isolation and identification of the microbes and total rainwater DNA sampling for estimating culturable, non culturable microdiversity

Acknowledgements
This contribution was supported by UGC-SAP Phase II Biodiversity, Bioprospecting programme. We thank 17 other student colleagues, M.Sc. Part I, Sem. I, 2013-4, of Dept. of Botany for help in sampling. Ms. Rosy DSouza, DST Inspire fellow for assistance in Sampler assembly and Dr. Goswami of IITM , Dr. Jayant Naralikar , Astrophysicist for inspiration.