Aquat Ecol (2022) 56:573–584

https://doi.org/10.1007/s10452-021-09921-x (0123456789().,-volV)
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89().,-volV)

Changes in species composition of cyanobacterial

and microalgal communities along a temperature gradient
in Tapovan Hot Spring, Garhwal Himalaya, Uttarakhand,
India
Sana F. Ikram . Vaishnavi Uniyal . Dhananjay Kumar

Received: 28 May 2021 / Accepted: 29 October 2021 / Published online: 25 November 2021
Ó The Author(s), under exclusive licence to Springer Nature B.V. 2021

Abstract To investigate how the water temperature
influences diversity and distribution pattern of
cyanobacteria and microalgae in a thermal spring,
algal samples were collected from Tapovan Hot
Spring of Garhwal Himalaya, Uttarakhand, India,
along the gradient of 52–30 °C. A total of 71 algal
species belonging to different algal groups like
Cyanophyta (34), Bacillariophyta (31) and Chloro-
phyta (6) were identified from the collected samples.
Among cyanophytes, five species were heterocystous,
while others belonged either to non-heterocystous
filamentous (20) or to unicellular forms (9). The
species richness decreases as temperature increases
towards the source point of the spring (i.e. from subsite
S5 to S1). However, Shannon’s measured diversity
varied little across the subsites except witnessing a
sudden drop at S2. Whittaker’s components of diver-
sity, such as beta and gamma diversity, successfully
reflected the major changes in the species composition
of cyanobacterial and microalgal communities resid-
ing in different thermal subsites. Some green algae and
diatoms were found thriving well at elevated
Handling editor: Ted Harris.
S. F. Ikram
V. Uniyal
D. Kumar (&)
Laboratory of Algal Biotechnology, Department of
Botany and Microbiology, School of Life Sciences,
H.N.B. Garhwal University, Srinagar,
Garhwal 246 174, India
e-mail: drdkumar83@gmail.com
temperatures ([ 38 °C). These species need to be
studied further for their under-operating cellular
mechanisms and encompassing robust biomolecules
that might be proven useful in diverse biotechnolog-
ical processes.
Keywords Algae
Beta diversity
Cyanobacteria

Gamma diversity
Shannon diversity
Thermal spring
Introduction
Cyanobacterial and microalgal communities inhabit-
ing thermal spring ecosystems are in limelight these
days as they can be a potential source of ther-
mostable biomolecules and bioactive compounds
(van Den Burg 2003; Dobretsov et al. 2011; Malavasi
et al. 2020; Singh and Kumar, 2021). Since these
communities perform oxygenic photosynthesis effi-
ciently at elevated temperatures, deciphering their
diversity would be of immense importance in the
context of global warming and the gradually increas-
ing concentration of CO2 in the environment. These
communities might be the source of genes suitable for
raising transgenic crops having resistance to various
abiotic stresses including thermal stress (van Den Burg
2003; Dobretsov et al. 2011; Malavasi et al. 2020;
Ikram et al. 2021). However, exploring these organ-
isms for any such applications must require an

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574
adequate understanding of their diversity, distribution
patterns and ecological adaptations along the temper-
ature gradient.
Waring et al. (1965) categorized thermal springs
into different categories like hottest, hot and warm
types. The hot springs may further be grouped into
high-, moderate- and low-temperature categories
depending on their respective water temperature at
the source sites (Dimri 2013; Poddar and Das 2017).
The high-temperature hot springs are well known to
sustain the growth of cyanobacteria as and when their
source water cools down with the downstream flow
and attains a temperature of moderate range (i.e.
75–55 °C) (Ward et al. 2012). Moreover, other
photosynthetic forms, such as green microalgae and
diatoms, can also contribute a considerable share in
constituting the photosynthetic communities, particu-
larly in low-temperature hot springs and in similar
thermal clines of high- and moderate-temperature hot
springs (Franks and Stolz 2009; Ghozzi et al. 2013;
Jonker et al. 2013). Although cyanobacterial and
bacterial diversity of high- and moderate-temperature
hot springs and clines has often been studied by
previous workers (Lau et al. 2009; Ward et al. 2012),
similar reports for low-temperature hot springs are
limited. The studies conducted so far remained
confined to documenting only cyanobacterial strains
along the temperature gradient. The quantitative
estimates of biodiversity and relative abundance of
different cyanobacterial taxa along the temperature
gradient have been sporadically investigated (Som-
pong et al. 2005; Ghozzi et al. 2013; Jonker et al.
2013). Likewise, the species richness and distribution
patterns of green microalgae and diatoms in these
habitats have been overlooked by researchers except
for a few reports (Franks and Stolz 2009; Ghozzi et al.
2013; Jonker et al. 2013). Thus, it is imperative to
determine the microalgal and cyanobacterial diversity
of low-temperature hot springs.
Uttarakhand Himalaya has several low-temperature
hot springs (Sharma 2012; Dimri 2013). In the recent
past, some of them have been studied for their
bacterial diversity (Arya et al. 2014; Pandey et al.
2015), but their microalgal diversity hitherto remained
unexplored. Some earlier workers (Prasad and Srivas-
tava 1965; Vasishta 1968; Bhardwaj and Tiwari 2010;
Bhardwaj et al. 2011) studied the distribution of
cyanobacterial taxa in a few of these thermal springs
but failed to offer an adequate quantitative estimate of
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Aquat Ecol (2022) 56:573–584
algal diversity along the temperature gradient. Such
kinds of information, however, help to devise appro-
priate strategies to culture these organisms in the
laboratory and explore them for large-scale biotech-
nological applications (Ikram et al. 2021; Singh and
Kumar 2021). Therefore, the main aim of this study
was to investigate the distribution and structural
composition of cyanobacterial and microalgal com-
munities inhabiting the different thermal clines of
Tapovan Hot Spring (hereafter referred as to THS) of
Garhwal Himalaya, Uttarakhand, India.
Materials and methods
Study site
The study site, THS, is located approximately 16 km
from Joshimath on the Joshimath-Malari Road in the
vicinity of Shiva-Parvati temple in Tapovan
(30.491663 N, 79.637189 E), Garhwal Himalaya,
Uttarakhand, India. The approachable stretch of the
stream was divided into five subsites, namely S1
(52–49°C), S2 (\ 49–45°C), S3 (\ 45–41°C), S4
(\ 41–38°C) and S5 (\ 38–30°C), considering the
downstream water temperature gradient. Figure 1
provides the map of the study area that clearly shows
the upstream and the downstream positioning of
different subsites. Sample collection was done during
April 2018.
Physicochemical parameters
Physical parameters of thermal spring water, such as
temperature, pH, EC and TDS, were recorded using
mercury bulb thermometer, pocket pH tester
(HANNA Instruments), electrical conductivity meter
(Multi-Parameter TestrTM 35 Series, Eutech Instru-
ments) and TDS metre (HM Digital TDS-3), respec-
tively. Calibration of the pocket pH tester was
performed before each use using standard buffer
solutions of pH 4.01, 7.01 and 10.01. The water
samples were collected in clean autoclaved bottles,
and analyses of various cations and anions were
performed in the Central Laboratory of ICAR-Indian
Institute of Soil and Water Conservation, Dehradun,
for measuring the concentration of iron (atomic
absorption spectrophotometry), nitrate (cadmium
reduction method), chloride (Mohr’s method),
Aquat Ecol (2022) 56:573–584
Fig. 1 Location map of Tapovan Hot Spring (THS), Garhwal Himalaya, Uttarakhand, India
calcium and magnesium (versenate EDTA method),
sulphate (indirect EDTA titration), phosphate (ascor-
bic acid method) and sodium and potassium (flame
photometry) following standard protocols.
Identification, relative abundance
and categorization
The algal samples were scraped from stones, pebbles
and adhering soils using forceps and spatula by
randomly laying 10 9 10 cm2 quadrats (Ikram et al.
2021). The morphological features, such as cell shape,
presence or absence of heterocyst, constrictions at
cross-walls, length and width of cells, heterocyst,
presence or absence of sheath around the filaments,
were considered for the identification of genera and
species from the collected samples. A compound light
microscope (CX21i-LED; Olympus) attached with a
digital camera (Magnüs, MIPS-5 MP, MagCam DC
Series, Noida, India) was used for this purpose. The
standard literature (Desikachary 1959; Phillipose
1967; Prescott 1978; Rippka et al. 1979; Cox and
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575
Cox 1996), Algaebase (http://www.algaebase.org) and
ANSP algal image database (http://diatom.acnatsci.
org/AlgaeImage/) were consulted for the identification
of algal genera and species. The relative abundance of
chronicled taxa was calculated in terms of cell count,
which was done using Spencer’s Bright-Line haemo-
cytometer. Unicellular alga and diatoms were counted
as a unit, whereas each cell of the filament was con-
sidered a single unit in the case of filamentous algae
and chain-forming diatoms. The essential rectification
to count the total number of cells of the filaments for
their curve placing on haemocytometer grids was
made following corrections suggested by Olson
(1950) and Lawton et al. (1999). An area of 5 9 5 lm2
was counted as a single unit for colonial and aggregate
forming species. For small cyanobacterial forms, with
no clear transverse walls, a 5 lm length was taken as
one unit employing the methodology described else-
where by other workers (DeNicola et al. 2006; Passy
and Larson 2011; Yadav et al. 2018). Based on the
relative abundance of the recorded taxa, along with the
different subsites, algal/cyanobacterial species were
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categorized as absent, rare, frequent, abundant and
dominant forms following Singh et al. (2008). The
Berger–Parker dominance index (Berger and Parker
1970) and the Shannon diversity index were calculated
as described elsewhere (Ikram et al. 2021).
Estimation of Whittaker’s components of diversity
(b and c diversity)
Whittaker’s components of diversity, such as beta
diversity (b) and gamma diversity (c), were estimated
considering subsequent pairs of thermal subsites
according to the methodology described by Titmus
(1981). Here, the alpha (a) diversity of each subsite
was calculated using the Shannon function. Subse-
quently, the gamma diversity (c) of successive pairs
was also computed using the same function, but this
time the compiled data of two successive subsites were
used. The beta (b) diversity, which reveals the degree
to which the species composition changed between the
thermal subsites, was calculated simply by taking the
difference between c and a diversity values.
Results and discussion
The variations in the physicochemical parameters of
THS water are given in Table 1. Water temperature of
the approachable portion of the hot spring, from which
algal samples were collected, varied from 52 to 30 °C.
However, excluding upstream site S1, all the subsites
(i.e. S2 to S5) also comprised some intermittent
patches of low-temperature water due to the intermix-
ing of a cold water stream running side-by-side of the
main hot spring. The pH of the spring water increased
from 6.4 to 7.4 along the thermal gradient of 52–30 °C.
The luxuriant growth of cyanobacterial mats was seen
in the spring, which is obvious as these photosynthetic
systems are well known for inhabiting such aquatic
habitats (Ward et al. 2012). Similarly, Bhardwaj et al.
(2011) and Ikram et al. (2021) reported the growth of
cyanobacterial forms in the hot spring of Badrinath,
Uttarakhand, India. Bhattacharya et al. (2016) and Roy
et al. (2017) documented cyanobacterial growth in hot
water springs located in some other states of India like
West Bengal and Jharkhand. Likewise, hot springs of
Thailand (Sompong et al. 2005), Tunisia (Ghozzi et al.
2013), South Africa (Jonker et al. 2013), Sri Lanka
(Rajapaksha et al. 2014) and Greece (Zarikas et al.
2014) have also been studied and reported for their
dwelling cyanobacterial diversity.
The EC and TDS of THS ranged from 185 to 201 lS
cm-1 and 267 to 279 mg L-1, respectively. These
observations matched well with the ranges mentioned
elsewhere for the Himalayan hot springs (Vasishta
1968; Bhardwaj et al. 2011). The concentration of
sulphate in THS water varied slightly, from
25.00 to 26.78 mg L-1, across the thermal gradient,

Table 1 Gradients of different physicochemical parameters measured for THS across the sampling subsites
Parameter Range Tendency
(from source to downstream)
(S1 ? S5)

WT ( °C) 52 ± 0.8 ? 30 ± 0.64 Decreasing

pH 6.4 ± 0.2? 7.4 ± 0.1 Increasing
EC (lS cm-1) 185 ± 0.85 ? 201 ± 0.92 Increasing
TDS (mg L-1) 267 ± 1.44 ? 279 ± 1.47 Increasing
Na? (mg L-1) 9.68 ± 0.38 ? 11.2 ± 0.2 Increasing
? -1
K (mg L ) 6.42 ± 0.19 ? 8.6 ± 0.35 Increasing
Ca2? (mg L-1) 16 ± 0.5 ? 19.7 ± 0.35 Increasing
Mg2? (mg L-1) 10.56 ± 0.26? 11.8 ± 0.6 Increasing
NO3- (mg L-1) 0.6 ± 0.01? 0.51 ± 0.05 Decreasing
PO43- (mg L-1) 1.48 ± 0.07 ? 1.98 ± 0.07 Increasing
Cl- (mg L-1) 8.52 ± 0.04 ? 9.43 ± 0.04 Increasing
SO42- (mg L-1) 25 ± 0.46? 26.78 ± 0.07 Increasing

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Aquat Ecol (2022) 56:573–584
which is comparable with the concentration range
reported in Saldhar Hot Spring (viz. 24.60–31.3 mg
L-1, unpublished data), located only 3 km away from
this site. Contrarily, sulphate concentration reported in
THS was considerably higher as compared to Badri-
nath Hot Spring (4.5–5.1 mg L-1), which is also
situated in the same geothermal field but secluded by
certain mountains (Ikram et al. 2021). The elevated
level of sulphate in thermal springs is supposed to be
due to the chemical reaction of trapped SO2 with hot
water (Torres-Ceron et al. 2019). Some bacterial
communities inhabiting these systems are known for
reducing sulphate into sulphide, which might have
adverse impacts on oxygenic photosynthesis of dif-
ferent algal forms, including cyanobacteria (Ward
et al. 2012). The detailed scheme of reduction of
sulphate into sulphide by bacterial communities
residing in hot spring environments is well elucidated
elsewhere (Muyzer and Stams 2008; Kushkevych
et al. 2021). Moreover, several cyanobacterial species,
particularly oscillatoriales, are recognized for having
extraordinary tolerance to elevated levels of sulphide
(Ward et al. 2012; Ghozzi et al. 2013). The dominant
presence of various oscillatoriales taxa was also
noticed in THS. These organisms can be investigated
for the mining of genes and proteins responsible for
offering such capabilities. A considerable amount of
Na?, K?, Ca2?, Mg2? and PO43- was present in THS.
The availability of these ions in the hot spring water is
due to the leaching of minerals in the hot water, which
is facilitated by the hydrolysis of adjoining under-
ground rocks (Torres-Ceron et al. 2019). A substantial
concentration of these ions has also been reported in
different hot springs of the world and the Indian
subcontinent (Ghozzi et al. 2013; Jonker et al. 2013;
Rajapaksha et al. 2014; Zarikas et al. 2014; Torres-
Ceron et al. 2019).
Table 2 shows a complete list of algal taxa
chronicled from THS along the downstream temper-
ature gradient of 52 °C to 30 °C. Based on the relative
abundance of the recorded taxa from the different
subsites, algal species have been categorized as
absent, rare, frequent, abundant and dominant forms.
In total, 71 algal taxa belonging to three algal
divisions, namely Cyanophyta (34), Bacillariophyta
(31) and Chlorophyta (6), were identified. Although
the maximum number of algal species was identified
for Cyanophyta (34), the considerable number of taxa
under frequent and abundant categories also belonged
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577
to Bacillariophyta. It reflects that though cyanobacte-
ria are prevailing forms, diatoms also comprise a
significant share in constituting the photosynthetic
communities of this hot spring. This particular obser-
vation is unique in the sense that most of the earlier
studies conducted so far to explore the diversity of
photosynthetic forms in hot springs maintained
cyanophytes as the dominant incumbents (Sompong
et al. 2005; Debnath et al. 2009; Jonker et al. 2013).
Perhaps, moderate temperature and intermittent mix-
ing of cold water of parallel running stream in THS are
the responsible factors for this colonization. In this
context, it seems pertinent to mention that although the
diversity and distribution of diatoms in hot springs
have sporadically been studied by the workers, they
are not unusual life forms in these environments.
Ikram et al. (2021) documented several species of
diatoms from the famous hot spring of Badrinath,
Uttarakhand, India. Other workers, such as Balashova
(2011) and Nikulina et al. (2019), reported diatom taxa
from the hot springs of Kyrgyzstan Mountain and the
Kamchatka Peninsula, Russia, respectively. Likewise,
Pathan (2014) studied the algal flora of thermal springs
of the Konkan region of Maharashtra, India, and
documented several species of bacillariophyceae from
there.
Figure 2 represents the species richness of different
algal divisions along the thermal gradient of THS. The
graph depicts that the species richness of all the three
algal classes decreased considerably when we moved
upstream beyond 41 °C. It is obviously due to the
detrimental effects of elevated temperature on the
growth and colonizing ability of microalgal and
cyanobacterial species. A decline in cyanophycean
species richness with increasing temperature has
previously been reported by various workers (Som-
pong et al. 2005; Debnath et al. 2009; Roy et al. 2014).
However, the adverse effect of temperature, as
revealed in the present study, was more prominent in
the case of green algae as compared to cyanophytes.
The green algae were eliminated from the subsites
where water temperature reached C 45 °C. It supports
the generalized view that green algae are temperature-
sensitive photosynthetic forms. Alternatively, some
cyanobacteria and diatoms were found flourishing in
the elevated-temperature zones. The ecological adap-
tations of cyanophytes to grow at high temperatures,
even up to 72 °C, are reported elsewhere (Ward et al.
2012). However, the ability of diatoms to grow at
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578 Aquat Ecol (2022) 56:573–584

Table 2 Cyanobacteria and microalgae documented from different subsites of THS

S. No. Taxa Subsite
S1 S2 S3 S4 S5
(52–49°C) (\49–45°C) (\45–41°C) (\41–38°C) (\38–30°C)

Cyanobacteria (Blue-green algae)

1. Anabaena circinalis * *
2. Anabaena unispora * *** ***
3. Anabaena variabilis *
4. Aphanocapsa thermalis *
5. Aphanothece minutissima *** *** *** *
6. Arthrospira jenneri * *
7. Chroococcus minor *
8. Chroococcus minutus ** **
9. Chroococcus turgidus **
10. Gloeocapsa livida *
11. Gloeocapsa sp. ** **
12. Leptolyngbya sp. * *
13. Lyngbya arboricola *
14. Nostoc muscorum * *** ***
15. Nostoc linckia **
16. Oscillatoria granulata *
17. Oscillatoria obscura **
18. Oscillatoria princeps * *
19. Oscillatoria pseudogeminata * * ***
20. Oscillatoria sancta *
21. Oscillatoria simplicissima *
22. Oscillatoria amphibia * **
23. Oscillatoria sp. **
24. Oscillatoria subbrevis **** * ** *
25. Oscillatoria subsalsa * *
26. Oscillatoria tenuis * ** * *
27. Phormidium ambiguum * **
28. Phormidium cebennense * * *
29. Phormidium formosum * * * **
30. Planktolyngbya circumcreta *
31. Pseudanabaena limnetica * *
32. Pseudanabaena mucicola *** *
33. Synechococcus bigranulatus * * ***
34. Synechocystis sallensis * ** * **
Bacillariophyta (Diatoms)
35. Achnanthes exigua * ** **
36. Achnanthidium minutissimum *** ** ** * **
37. Aulacoseira granulata **
38. Cocconeis lineata * *
39. Cocconeis placentula ** **
40. Craticula sp. * *
41. Cymbella lanceolata **

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Aquat Ecol (2022) 56:573–584 579

Table 2 continued
S. No. Taxa Subsite
S1 S2 S3 S4 S5
(52–49°C) (\49–45°C) (\45–41°C) (\41–38°C) (\38–30°C)

42. Diadesmis confervacea * *

43. Diatoma ehrenbergii ** ** **
44. Diploneis sp. * ** * *
45. Fragilaria capucina *
46. Fragilaria mesolepta **
47. Fragilaria virescens * ***
48. Gomphonema sphaeophorum **
49. Meloneis akytos * ** **
50. Melosira varians ** ***
51. Navicula lanceolata **
52. Navicula oblonga ** ** **
53. Navicula radiosa *
54. Navicula tripunctata **
55. Navicula veneta ** * **
56. Nitzschia amphibia * **
57. Nitzschia clausii ** ** **
58. Nitzschia obtusa **
59. Nitzschia palea *** ** *
60. Nitzschia sigma *
61. Nitzschia sp. **
62. Pinnularia acrosphaeria **
63. Pinnularia parvulissima *
64. Pinnularia viridis * **
65. Synedra ulna **
Chlorophyta (Green algae)
66. Chlorella sp. **
67. Cladophora vagabunda *
68. Closterium moniliforme **
69. Elakatothrix sp. **
70. Scenedesmus dimorphus **
71. Zygnema sp. *
Asterisks represent different categories of relative abundance [*Rare (less than 1% of the total cell count), **Frequent (1-10%),
***Abundant (10-30%), ****Dominant ([30%)], while empty cells reflect absence of the taxa.

elevated temperatures is an interesting observation of
this study and thus demands more focused investiga-
tions. Diatoms, such as Achnanthes exigua, Achnan-
thidium minutissimum, Cocconeis lineata, Craticula
sp., Diploneis sp., Meloneis akytos, Navicula oblonga,
Nitzschia amphibia, N. clausii, N. palea and
Pinnularia viridis, which were chronicled from high-
temperature subsite (i.e. S1, 52–49 °C) need to be
explored further for their under-operating cellular
mechanisms and robustness of encompassing biomo-
lecules. An abrupt decrease in species richness of
diatoms and cyanophytes was noticed for the subsite

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580
Fig. 2 Species richness of algal groups documented from THS
along the thermal gradient of 52–30 °C and across the subsites
S1 to S5
S2 (\ 49–45 °C). Perhaps, it is due to the severe
shading of this subsite that promotes the overgrowth of
Oscillatoria subbrevis. Some other factors, such as the
leaching of chemical substances from adjacent rocks
and sands, might also be responsible for helping
Oscillatoria subbrevis to outcompete the other
cyanobacterial and microalgal species at this specific
site.
The green algal taxa reported in this study were
Chlorella sp., Cladophora vagabunda, Closterium
moniliforme, Elakatothrix sp., Zygnema sp. and
Scenedesmus dimorphus. All these above-listed forms
were chronicled from the thermal range of 30 to 45 °C.
Similar green algal forms have also been reported
earlier from the hot springs of Bulgaria (Stoyneva
2003), South Africa (Jonker et al. 2013) and Odisha,
India (Bhakta et al. 2016). Insofar as the distribution of
cyanobacterial taxa is concerned, the species richness
(SR) of filamentous forms (SR = 25) in the spring was
considerably higher compared to the unicellular forms
(SR = 09). However, among 25 filamentous species,
only five were heterocystous. The dominance of non-
heterocystous filamentous cyanobacteria in hot
springs has also been reported by several other
workers (Pentecost 2003; Ionescu et al. 2007; Ward
et al. 2012; Ghozzi et al. 2013; Bhattacharya et al.
2016). The important filamentous forms inhabiting
elevated-temperature zones of THS were Anabaena
circinalis, Anabaena unispora, Arthrospira jenneri,
Nostoc muscorum, Oscillatoria pseudogeminata, Os-
cillatoria tenuis and Phormidium formosum, while the
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Aquat Ecol (2022) 56:573–584
non-filamentous forms included Gloeocapsa sp. and
Synechococcus bigranulatus. In the diverse literature,
the highest temperature limit for genus Synechococcus
has been reported to vary from 58 to 72 °C (Ward et al.
2012). In this study, this genus was found to inhabit
from 41 to 52 °C, which is consistent with the findings
reported from Yellowstone National Park (Ward et al.
2012). Nevertheless, it might be attributed to the
different morphotypes of species and the chemical
properties of the spring water.
Figure 3 shows order-wise representation and dis-
tribution of algal species along the temperature
gradient. The size of bubbles increases with an
increase in the number of representative species of a
particular order at a particular thermal subsite. For
different subsites, it varied from 0 to 12. The smallest
visible bubbles (dots) indicate a representation by a
single species, while the largest bubble noticed in the
case of Oscillatoriales for the subsite S4 reflects the
highest number of dwelling species (i.e. 12). More-
over, this graph reveals that in lower-temperature
zones, the species representation of cyanophycean
orders, like Oscillatoriales and Nostocales, is consid-
erably higher than that of the other algal orders. Here,
diatoms and green algal orders were represented only
by a limited number of species. In elevated-temper-
ature clines, the representation of green algal orders
was absent. However, diatoms and cyanobacterial
orders were presented by comparable numbers.
Figure 4 shows the variations in the Shannon
diversity index and the Berger–Parker dominance
index of microalgal and cyanobacterial communities
along the temperature gradient in THS. The changes in
the overall species richness across the thermal gradient
have been mentioned in the circles placed over the
bars. Results reveal that the species richness of the
inhabiting communities declined significantly as the
temperature exceeded 41 °C. However, the Shannon
measured diversity varied little except witnessing an
abrupt decline at the subsite S2 (\ 49–45 °C). On the
other hand, the Berger–Parker index displayed its
highest value for this subsite (S2). It is perhaps related
to the fact that the Berger–Parker dominance index
provides extraordinary weightage to the dominance of
the species, and at this particular subsite, Oscillatoria
subbrevis dominated the community (relative abun-
dance [ 30%; Table 1). All-time low species richness
and the Shannon diversity of subsite S2 (\ 49–45 °C)
Aquat Ecol (2022) 56:573–584 581

Fig. 3 Bubble plot showing species representation of algal species (dot, single species; the largest bubble, noticed in the
orders in different thermal clines of THS across the subsites S1 case of Oscillatoriales, 12 species)
to S5. The size of bubbles reflects the number of representative

Fig. 4 Temperature-dependent changes in the Shannon measured diversity and the Berger–Parker dominance index of photosynthetic
communities inhabiting THS. The numbers given in the circles placed over the bars show species richness

are also attributed to the prominent growth of Oscil-
latoria subbrevis, as explained earlier.
To check the variations in diversity at successive
thermal subsites, values of beta (b) diversity were
plotted against the total diversity (c) of subsequent
pairs of thermal subsites (Fig. 5). The b diversity
decreased significantly from S5-S4 to S4-S3 depicting
the magnitude of multiplicity between the stations.
Conversely, not much difference was seen in c
diversity. It suggests that the total diversity of
inhabiting communities changed little from S5-S4 to
S4-S3. Both b and c diversities dropped precipitously
from S4-S3 to S3-S2. It pinpoints that the species
composition of the S2 subsite is entirely different from
the other three subsites (viz., S5, S4 and S3). This is
exactly what is revealed by all-through data shown in

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582
Fig. 5 Changes in Whittaker’s components of diversity (b and c
diversity) in THS. The values were calculated considering the
subsequent pairs of different thermal subsites, such as S5-S4, S4-
S3, S3-S2 and S2-S1, following Titmus (1981)
Figs. 2, 3 and 4. Further, a steep increase in b diversity
was observed from S2-S3 to S2-S1, which conveys a
considerably diverse species composition of commu-
nities colonizing S2 and S1 subsites. Thus, Fig. 5
enables easy detection of changing patterns of species
composition of the inhabiting communities along the
thermal gradient in THS.
Conclusions
The most abundant photosynthetic forms in the study
hot spring were cyanobacteria. As compared to the
heterocystous and unicellular forms, non-heterocys-
tous ones were particularly abundant in THS. Gener-
ally, the species richness declines as temperature
increases towards the source point of the hot spring.
However, the Shannon measured diversity changed
little except for an unexceptional drop at subsite S2,
which was attributed mainly to the dominant growth of
Oscillatoria subbrevis at this subsite. Besides
cyanobacteria, diatoms also showed a comparable
presence in this hot spring across the thermal gradient
of 52 to 30 °C. Recording green algal species and
diatoms from subsites comprising elevated tempera-
tures is an important observation. These algal forms
can be tested for large-scale outdoor production of
algal biomass feedstock for various biotechnological
applications. Nonetheless, estimations of beta and
gamma diversity for different pairs of thermal sub-
sites, and their subsequent correlative plotting, were
123
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Aquat Ecol (2022) 56:573–584
found suitable in identifying the thermal gradient-
dependent changes in the species composition of the
photosynthetic communities residing in THS.
Acknowledgments The authors are grateful to the Head,
Department of Botany and Microbiology, H.N.B. Garhwal
University, for providing the necessary facilities. SFI thanks the
University Grants Commission, New Delhi, for providing
financial support in the form of University Research
Fellowship. We are thankful to Mrs. Sarita Gupta, Senior
Technician, ICAR-IISWC, Dehradun, for the analysis of water
samples. We thank Dr. Narendra Kumar, Department of
Geography, H.N.B. Garhwal University, for making the study
area map.
Authors’ contributions SFI planned and executed
experiments, prepared a draft of the manuscript and finalized
it for submission. VU helped in conducting experiments and
preparation and finalization of the manuscript. DK was involved
in conceiving the idea, planning the experiments, presenting and
discussing results, and submission of the manuscript. All authors
read and approved the final manuscript.
Funding The study is funded by the University Grants
Commission, New Delhi, India, HNB Garhwal University,
Srinagar, Garhwal, India.
Data availability The data sets analysed during the current
study are available from the first author on reasonable request.
Declarations
Conflict of interests The authors declare that they have no
competing interests.
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