INTRODUCTION

Geologists tend to attribute observable mineral changes to abiotic processes while biologists try
to explain everything in terms of biotic mechanisms (Barton et al. 2001), the actual mechanisms
are often a combination of abiotic and biotic. Earlier it is belived that formation of dripstones is
an abiotic mechanism but recent studies have proved that microbes play important role in the
formaton of dripstones.

Geomicrobiology recognizes that microorganisms are important active and passive promoters
of redox reactions that can influence geological formation (Ehrlich1996). Geomicrobiology is a
combination of geology and microbiology, which includes the study of interaction of
microorganisms with their environment, such as in sedimentary rocks. Geomicrobiology
examines the role that microbes have played in the past and are currently playing in a number
of fundamental geological processes. The temporal and spatial scales of geomicrobiological
processes vary greatly from minutes to eons and microniche to global, yet the contributions
made by microbes to most processes are unseen and unknown due to very small size of typical
microbial cell.

Scientists have determined that despite abundant life on the surface, most of the of the
biomass of life on earth is below the ground and primarily microbial (Whietman et al.
1998).Microbes form metabolic products and selectively reduce or oxidize mineral constituents
of rocks and soils. All sedimentary and igneous rock is susceptible to microbial-mediated
dissolution and diagenesis (Ehrlich 1998).

In the present report, the Speleothems (Stalactites, Stalagmites, Flowstones etc) from the
Shastradhara caves have been studied in detail with special reference to Sedimentological,
Geomicrobiological, Paleoclimatic and Paleomonsoonal aspects.

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1. GEOLOGY AND STRATIGRAPHY

The caves are located at Sahastradhara in Dehradun valley which are formed in the carbonates of
the Krol belt (Tewari, 2007, 2012 and the references therein). The caves at Sahastradhara, are
situated in the Dehradun Valley, a crescent shaped intermountain valley formed within the
Siwalik Formations in Garhwal Himalaya (Figure1a, b; Plate1,2,7). They are situated on the Krol
carbonates between the Rivers Ganga in the east and Yamuna in the west. Both the rivers have
their origin from two different Himalayan glaciers, namely Gangotri for Ganga and Yamunotri
for Yamuna. Doon Valley falls under the temperate area due to the elevation of 600-800 meters
above mean sea level. (Fig. 1c, after Tewari, 2012).

The Doon Valley is separated by the Lesser Himalayan Neoproterozoic Krol Belt carbonate
sediments in the north (Tewari, 2012) by Main Boundary Thrust (M.B.T.).The Siwalik Foreland
Basin sediments are found in the south of the Doon Valley Thrust over the Indo-Gangetic Plains
along Himalayan Frontal Thrust (H.F.T.). Shastradhara lies between 300 23’ 07.6’’ N and 780 07’
44.9’’ E with altitude of 830.5 meter above the sea level.

The Krol belt carbonates are well developed in and around the Shasatradhara cave system.

The Upper Krol carbonates are the host for the modern caves. The age of the Krol belt has been
very controversial and now has been established as Neoproterozoic (Ediacaran ; Tewari 2012
and the references therein ; Fig. 1c).

The caves are rather small in size (10 m long, 2 m wide), and are well known for their pH
neutral springs that are believed to cure various kinds of skin diseases. The caves are located at
Sahastradhara in Dehradun valley which are formed in the carbonates of the Krol belt (Tewari,
2007, 2012 and the references therein). The revised lithostratigraphy of the Lesser Himalaya
showing the facies variations, depositional environment, paleoclimate and age of the formations
is shown in the figure 1, c.

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3
Plate 1
Plate 2
2. SHASTRADHARA CAVE SYSTEM

Shastradhara is situated around 15 kms east of Dehra Doon Bus stand in the state of Uttarakhand.
As its name depicts “Thousand fold springs” in English. It is a lovely waterfall, with a depth of λ
meters. Its name derives from the fact that the water literally drips down from limestone
stalactites numbering thousands formed in a sort of a grotto or cave making it a famous tourist
destination.

The presence of a sulphur spring near the falls has medicinal and healing properties for any skin
cure diseases or ailments. The sulphur in the water has its therapeutic value strangely enough the
many sulphur springs and falls ebb into a channel which is the source of the Baldi River which
flows along the villages and continues towards Lacchiwala (river song) and finally falls into the
holy river of the Ganges.

Besides medicinal merits, the spring is also known for its beautiful views. The beauty of this
spring is further enhanced by the surrounding caves and Baldi River. The water of this spring has
lime, which contributes towards the formation of projecting ridge that eventually takes the shape
of a cave. Due to its medicinal properties and beautiful views that can be enjoyed at this place,
Sahastradhara is popular among a large number of tourists.
Seven caves of Shastradhara are arranged in a typical way and making an arch. This type of
structure making it distinct from other cave systems. It is a complex type of cave system with a
variety of caves. Following seven caves have been recognized in the Sahastradhara.

 CAVE I: This cave shows the developing stages of the stalactite formation in which the
water is dripping slowly and the microbial process is also taking place simultaneously.
 CAVE II (DRONACHARYA CAVE): An age old temple and Drona cave-named after the
Guru Dronacharya of Mahabharata period- is present here which acts as an abode of prayer
and worship for devotees across India. That is why sampling from this cave is not permitted.
 CAVE III and IV: It shows best developed speleothems such as stalactites, stalagmites,
pillars and flowstones. This cave also shows maximum percolating water from the ceiling
and the stalctities and stalagmites are showing carbonate precipitation. Dripping water
samples have been collected from this cave for the isotopic studies.
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 CAVE V: The cave is dry, there is no activity of the percolating water at present and
therefore the spelothems are not being formed.
 CAVE VI: This cave is approximately 2 m. long and shows good development of
stalagmites and thin and long stalactites. Drip water sample was collected.
 CAVE VII: This cave shows very good development of the speleothems. Sampling has been
done from this cave for various studies.

The speleothems found in various caves in Sahastradhara, Dehradun, (Pt.1, 2; Figs. a,b,c,d)
(Tewari, 2009, 2011). Speleothems found in three caves in Sahastradhara (Baskar R., 2005),
however we have collected speleothem from four caves of Shastradhara and have been studied
for petrography mineralogy, geochemistry and geomicrobiology to better understand the role of
microorganisms in mineral formation. Calcite is the dominant mineral in the stalactites.

3. SPELEOTHEMS

A speleothem is commonly known as a cave formation, is a secondary mineral deposit formed

in caves. Speleothems are typically formed in limestone or dolostone solutional caves.The term
Speleothem originate from the Greek words spelaion means cave and thema means deposit
(Robertson, 2004).

3.1 ORIGIN AND COMPOSITION

Water seeping through cracks in a cave's surrounding bedrock may dissolve certain compounds,
usually calcite and aragonite (both calcium carbonate), or gypsum (calcium sulphate). The rate
depends on the amount of carbon dioxide held in solution, on temperature, and on other factors.
When the solution reaches an air-filled cave, a discharge of carbon dioxide may alter the water's
ability to hold these minerals in solution, causing its solutes to precipitate. Over time, this may
span tens of thousands of years, the accumulation of these precipitates.

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3.2 TYPES AND CATEGORIES

Speleothems take various forms, depending on whether the water drips, seeps, condenses, flows,
or ponds. Many speleothems are named for their resemblance to man-made or natural objects.
Types of speleothems include

Dripstone is calcium carbonate in the form of stalactites or stalagmites

 Stalactites are pointed pendants hanging from the cave ceiling, from which they
grow

 Soda straws are very thin but long stalactites having an elongated
cylindrical shape rather than the usual more conical shape of stalactites
 Helictites are stalactites that have a central canal with twig-like or spiral
projections that appear to defy gravity
 Include forms known as ribbon helictites, saws, rods, butterflies, "hands",
curly-fries, and "clumps of worms"
 Chandeliers are complex clusters of ceiling decorations

 Stalagmites are the "ground-up" counterparts of stalactites, often blunt mounds

 Broomstick stalagmites are very tall and spindly

 Totem pole stalagmites are also tall and shaped like their namesakes
 Fried egg stalagmites are small, typically wider than they are tall

 Columns result when stalactites and stalagmites meet or when stalactites reach the
floor of the cave.

 Flowstone is sheet like and found on cave floors and walls

 Draperies or curtains are thin, wavy sheets of calcite hanging downward
 Bacon is a drapery with variously colored bands within the sheet.
 Rimstone dams, or gours, occur at stream ripples and form barriers that may contain
water.
 Stone waterfall formations simulate frozen cascades.
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  Cave crystals

 Dogtooth spar are large calcite crystals often found near seasonal pools
 Frostwork is needle-like growths of calcite or aragonite
 Moonmilk is white and cheese-like
 Anthodites are flower-like clusters of aragonite crystals

 Speleogens (technically distinct from speleothems) are formations within caves that
are created by the removal of bedrock, rather than as secondary deposits. These include:

 Pillars
 Scallops
 Boneyard
 Boxwork

Others

 Cave popcorn, or cave coral, are small, knobby clusters of calcite

 Cave pearls are the result of water dripping from high above, causing small "seed"
crystals to turn over so often that they form into near-perfect spheres of calcium
carbonate
 Snottites are colonies of predominantly sulfur oxidizing bacteria and have the
consistency of "snot", or mucus[1]
 Calcite rafts are thin accumulations of calcite that appear on the surface of cave
pools

Speleothems may also occur in lava tubes. Although sometimes similar in appearance to
speleothems in caves formed by dissolution, these are formed by the cooling of residual lava
within the lava tube. Speleothems formed from salt, sulfur and other minerals are also known.
Speleothems made of pure calcium carbonate are a translucent white color, but often
speleothems are colored by minerals such as iron, copper or manganese, or may be brown
because of mud and silt particulate inclusions.

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3.3 GEOCHEMISTRY AND MECHANISM

Cave minerals are secondary minerals formed inside a cave resulting from one or several of the
following processes: reprecipitation of the bedrock, supersaturation of solutions, dehydration,
biogenic processes (or human activity), hydrothermal processes, hypergenic processes
(weathering), reaction of karst solutions with minerals of non-karstic bedrock, or by eruptive
processes (fumarolic or magmatic) due to crystallization of volcanic gases or their reactions with
minerals or solutions.

Most speleothems (dripstones, including stalagmites and stalactites) are formed by

hydrocarbonate reprecipitation of carbonate bedrock. Groundwater saturated with carbonate
dioxide dissolves calcium carbonate (CaCO3 from the bedrock and reprecipitates it inside the
cave when carbonate dioxide volatilizes. Most caves are developed in limestone or marble, so
CaCO3 forms most speleothems. Ninety-five percent of all speleothems are formed by calcite,
2% by aragonite (the second polymorphic form of CaCO3) and less than 2% are formed by the
rest of about 250 cave minerals, most frequently gypsum (as some caves are developed in
gypsum bedrock).

3.4 SIGNIFICANCE OF SPELEOTHEMS

Cave minerals and speleothems are so unique in appearance that they are considered natural
heritage objects, and laws in most countries prohibit their collection, mining and selling.

Stalagmites (speleothems growing on the cave floor) are powerful paleoclimatic archives, as they
contain a multitude of environment-dependent proxies (e.g. δ18O, δ13C, trace elements, humic
acids). They have been established as one of the best terrestrial archives of past climate because
they can be accurately and precisely dated by absolute U-Th methods.

Speleothems (stalagmites, stalactites, flowstones etc.) grow in caves as the result of precipitation
of calcium carbonate (either as calcite or aragonite) from drip waters that enter the cave's
atmosphere. The ratios of the stable isotopes oxygen and carbon of the carbonate (CaCO3) are
analyzed as they depend on environmental factors, such as rainfall amount, temperature,
vegetation cover and type, etc. As speleothems grow they incorporate trace elements into their

10
structure, and the concentration and ratios of those might be related to environmental conditions
(atmosphere, soil, host rock, cave air) at the time of deposition. Where banding in stalagmites
can be demonstrated to be annual by dating methods, stalagmites have the potential to record
long-term, high-resolution climatic data. Based on previous work we selected one speleothem
(YOK I) for high resolution dating and isotope work. YOK I is 56cm long and was harvested
from Yok Balum cave in 2006. This speleothem was selected because it had the highest
probability of providing a continuous record during the period of interest and preliminary oxygen
isotopic work suggested that it contained a clear climate signal.

One-hundred and two cave minerals are known to form coatings and crusts, 57 form stalactites
and stalagmites, 23 form moonmilk, 15 form anthodites, 14 form helictites, 12 form Angel hair,
7 form coralloides and pearls, and 6 form cave balloons.

4. LABORATORY STUDIES

Samples collected from the field have been studied for the Sedimentological (petrography
microfacies and diagenetic changes), Mineralogical and geochemical (X-RAY Diffraction ,
Scanning Electro Microscopy ( SEM ) , SEM – EDX ( SEM – Energy Dispersive X-Ray
Microanalyser ), XRF (X- Ray Flourescence) and GeomicrobiologicaL (interaction of microbes
with carbonate sediments) investigations. The details of the each laboratory studies have been
described below:

4.1 SEDIMENTOLOGICAL STUDIES

The petrography reveals replacement of the original micrite by Mg rich sparry dolomite
(radiaxial fibrous microstructure: Pt. 3; Fig. a). Alternating light (carbonate) and dark laminae
(organic) are well preserved, the lighter interlaminal space shows diagenetic changes and the
thickness of the darker lamellae decreases upwards ( Pt. 3 ; Fig. b). The wavy discontinuous
organic laminae in sparry calcite indicate micro hiatus during the formation (Pt. 3; Fig. c). The
sparry dolorhombs showing pinkish colour and twinkling suggest dolomitization process (Pt. 3;
Fig. d)

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Plate 3
Plate 4
Plate 5
Different types of microbiallamintes have been observed in the caves 4 and 6 (Pt. 4; Figs. a, b,
d). The microbiallamites are crenulated , wavy and discontinuous (Pt. 4; Fig b, d).The wavy
laminated bands are thicker and the interlaminar space shows sparry calcite ( Pt 4; Fig. d), The
thinner layers also show partial recrystallisation.( Pt. 2; Fig b), The sharp contrast with
microbial lamination and the sparry calcite is observed in some thin sections ( Pt. 4 ; Fig. a ).
Some thin sections do not show any microbiallaminites, however, linearly distribution sparry
calcite is also observed (Pt 4; Fig. c).

The petrographic thin sections of cave 6 shows a network of the large dolorhombs (maximum
size more than 200 µ; Pt. 5; Fig. a). The recrystallisation seems intense which resulted in such
larger grains. The biomicrite microfacies are preserved in the thin sections of the cave 7. The
black patches represent the organic matter whereas the pinkish small patches indicate
recrystallization from micrite to sparite (Pt. 5; Fig. b). Wavy microbial laminated microfacies is
well preserved in the cave 3 (Pt. 5; Figs. c, d). Dark (organic) laminae alternate and pinkish
white (carbonate) laminae and the interlaminar space between them shows highly recrystallised
sparite. The crest of the wavy structures shows micritic grains whereas the trough contains
sparitic grains (Pt. 5; Figs c, d).

4.2 MINERALOGICAL AND GEOCHEMICAL STUDIES

Mineralogical studies (XRD and SEM-EDAX) of the stalactite and stalagmite samples revealed
that calcite is the dominant mineral. An abundant microbial community (9 × 105 cells, g sed-1)
was detected by direct microscopic observation after DAPI staining. Application of fluorescence
in situ hybridization techniques (FISH), based on the presence of rRNA, demonstrates the
presence of a large number of active microbial cells (around 55 % of the total cell number). The
microbial community is dominated by Eubacteria, mainly sulphate-reducing bacteria
(representing 10% of the total microbial community), but Archaea are also present. A significant
fraction of these cells are active, indicating the high probability of their participation in
biomineralization processes involved in the stalactite formation. This conclusion is at variance
with the established classical model for stalactite formation based entirely on inorganic processes
associated with carbonate solubility’s dream destination, where one avail a tranquil holiday

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Plate 6
4.3 X-RAY POWDER DIFFRACTION (XRD)

X-ray powder diffraction (XRD) is a rapid analytical technique primarily used for phase
identification of a crystalline material and can provide information on unit cell dimensions. The
analyzed material is finely grounded, homogenized, and then average bulk composition is
determined. X-ray powder diffraction is most widely used for the identification of unknown
crystalline materials (e.g. minerals, inorganic compounds). Determination of unknown solids is
critical to studies in geology, environmental science, material science, engineering and biology.
The sample preparation for XRD analysis involves following steps-

 Take a small quantity (few tenths of a gram) of the material, as pure as possible.
 Grind the sample to a fine powder, typically in a fluid to minimize inducing extra
strain (surface energy) that can offset peak positions, and to randomize orientation.
Powder less than ~10 m (or 200-mesh) in size is preferred.
 There are two methods of filling in the sample holder i.e. upper filling and side filling.
In upper filling fill the powder sample from the upper side of the sample holder.
Upper filling is done for the rocks which do not contain any linear structure such as
foliation.
 The rocks containing linear structure such as foliation due to Mica minerals align
themselves in a particular direction so that we do not get random orientation of the
atoms and the random orientation of the atoms is needed for the X-ray Diffraction.
 Place into a sample holder using glass slide to smooth the surface of the sample. Care
must be taken to create a flat upper surface and to achieve a random distribution of
lattice orientations unless creating an oriented smear.

4.3.1 DATA COLLECTION AND DATA ANALYSIS

X-pert Data Collector software is used for data collection and to analyse the data we use the
software X-pert High Score Plus.

The intensity of diffracted X-rays is continuously recorded as the sample and detector rotate
through their respective angles. A peak in intensity occurs when the mineral contains lattice
planes with d -spacing appropriate to diffract X-rays at that value of θ. Although each peak

17
consists of two separate reflections (Kα1 and Kα2), at small values of 2θ the peak locations
overlap with Kα2 appearing as a hump on the side of Kα1. Greater separation occurs at higher
values of θ. Typically these combined peaks are treated as one. The 2 position of the diffraction
peak is typically measured as the center of the peak at 80% peak height.

4.3.2 RESULTS

By the above procedure XRD analysis has been done, 2θ is 300 which suggest dominance of
magnesium bearing calcite in the samples.

4.4 GEOMICROBIOLOGICAL AND SEM- EDX STUDIES OF MICROBES

Generally caves host microbially mediated mineral precipitation; however, mineral precipitation
commonly considered abiogenic inspite of the fact microbes are present in the thin sections of
the speleothems. In the present study, the possible cellular structures and diatoms of
microbiological origin have been found in the petrographic thin sections and further confirmed in
the Scanning Electron Microscopy (SEM , Pt 6; Figs. a, b). The microbial fossils observed are
semi circular filamentous structure of cyanobacteria (Pt 6; Fig. a) and a well preserved
spheriodal cell (Pt 6; Fig. b). Combines SEM and Energy Dispersive X- ray Microanalysis
(EDX) of these microbes has been done and shown in Plate 6; figs b and d respectively. The
predominance of Ca is recorded in the microbes in EDX analysis, however Si, K, S, Mg and
Cl is also present in minor amount in some of the microbes such as diatoms etc. Earlier
researchers have shown experimentally in the laboratory that the microbial precipitation in the
stalactites of the Sahastradhara cave is possible under standard conditions (Baskar et al. in
Tewari and Seckbach, 2011, Tewari 2011).

5. BIODIVERSITY IN AND AROUND SHASTRADHARA CAVES

Sahastradhara represents higher biodiversity of schrubs and trees around the cave and the
adjoining forests (Pt. 7; Figs. a,b,c). Soni et al. (2011) have studied the biodiversity in
Mussoorie-Dehradun area and have interpreted the biodiversity index in these areas.
Sahastradhara is located about 18km from Dehradun town on the bank of river Baldi.

18
Plate 7
The altitude of the Sahstradhara is 800m. The GPS location is between 30. 2λ’ N and 78. 8’ E.
The natural forest of Sahastradhara are characterised by tropical mixed deciduous scrubby
vegetation (Pt.1; Fig.b,, Pt.7; Figs.a,b,c). The main dominating herbaceous species at
Sahastradhara is Erianthus rufipilus and Micromeria biflora. In Sahastradhara site diversity
index is 1.44 (Soni et al. , 2011). The main shrubs of the Sahastrdhara are Agave sisalana,
Lantana camara , Ficus palmate, In adjoining natural forest the species Murraya koenigii and
Lantana camara are predominant. The main species of trees in Sahastradhara (Pt.7; Figs. b and
c) Acacia catchu and Grevillea robusta. It has been observed that species diversity increases
during spring season and declines thereafter. In summer season new species diversify during
autumn and winter species number decline due to adverse climatic conditions. In Shastradhara
the shrubs diversify higher. Among the animals, crabs are seen inside the cave and monkeys and
birds are common around the caves (Pt. 7; Fig. d).

6. DISCUSSION AND CONCLUSIONS

Speleothems potentially contain the most complete record of past climate change and monsoon
in the Peninsular and Himalayan caves from NW to NE (Deshmukh , 1994; Tewari , 2009, 2011,
2012 , Gupta , 2010). The oxygen and carbon isotopic analysis of the speleothems document
intensity of Indian Summer Monsoon rainfall. The annual temperature ranged between 1.80C in
January to 400C in June (Negi and Chauhan, 2002). The meteorological data of last 75 years
shows that the average maximum temperature for Doon Valley is27.650 c average minimum
temperature is 13.790 C (Source: FRI Meteorology).

The U- Th dating along speleothem growth axis of cave deposits or Speleothems are natural
archives of the paleoclimate record and abundantly found in the Himalaya from NW to NE India,
where calcium carbonate is precipitated as flowstones, stalactites and stalagmites. Recently,
much attention has been focused on cave calcites (speleothems) as archives of climatic and
paleoenvironmental changes in terrestrial environments. Factors contributing to make
speleothems valuable paleoenvironmental indicators include: (1) their high preservation potential
shielded from Earth-surface erosional processes; (2) their crystalline structure, frequently
composed of coarsely crystalline calcite (CaCo3) that records geochemical signatures of global
and regional events transferred through atmospheric processes; and (3) the resistance of
speleothems to post-depositional recrystallisation and digenesis.
20
In the present summer project , Sahastradhara caves well developed in the Doon valley (Fig. 1)
have been studied with reference to sedimentological, mineralogical- geochemical and
geomicrobiological parameters of the cave deposits stalactitites, stalagmites, flowstones etc ) and
following conclusions have been drawn.

Microscopic studies of the stalactites and stalagmites show normal, wavy as well as crenulated
laminations of precipitation. The microfacies of the speleothems show micrite, sparite sparry
dolomite, radiaxial fibrous calcite (Pts. 4,5,6 ; Figs. a, b, c, d). Possibly the Mg has triggered the
formation of radiaxial fabric. Various types of light (carbonate) and dark (organic) laminae is
related to the microclimatic decadal scale seasonal variations (Pt. 3; Figs. a, b; Pt. 4; Figs. a, b, c,
d; Pt. 6; Figs b, d). The thickness of the dark and light laminae is variable. The interlaminar
spaces are highly recrystallsed due to intense diagenetic process. More than 200 µ dolorhombs
have been found in the diagenetic change.

The petrographic identification of the Mg calcite is further confirmed by the XRD peaks of
calcite. The SEM- EDX combined study of the filamentous and spheroidal microbes have shown
the presence of mainly Ca in the cells and suggest microbial origin (Pt. 6 ; Figs b ,d).

Study of modern cave drip waters and stalagmites demonstrates that the stalagmites were
deposited in or very near to isotopic equilibrium. The δ18O isotope data of drip water from
Sahastradhara cave in Dehradun (Pt.2 ; Fig. a) during the monsoon season (August and
September, 2007-10) varies from - 4. 58 ‰ to 5.14 ‰ (VPDB; Tewari 200λ, 2011, 2012).
Tewari (2009, 2011) have interpreted that the western part of Indian Summer Monsoon (ISM) in
Oman and Yemen, the oxygen isotope ratios of stalagmite calcite primarily show variations in
the amount of rainfall with more negative δ 18O indicating higher monsoon rainfall. Therefore, it
is concluded that the speleothems are very significant for the paleoenvironmental and
paleocliamtics records in addition to the petromineralogical and geomicrobiological process.

21
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