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PLANKTON DIVERSITY IN TEMPORARY WATER HABITAT IN UPEH
GULING, TAMAN NEGARA JOHOR, ENDAU ROMPIN, MALAYSIA
USMAN ALHASSAN GABI
A thesis submitted in
fulfillment of the requirement for the award of the degree of
Master of Science
Faculty of Science, Technology and Human Development
Universiti Tun Hussein Onn Malaysia
AUGUST 2015
v
To my beloved father and mother
vi
ACKNOWLEDGEMENT
In the name of Allah the Most Gracious the Most Merciful, All praise to Allah, we
seek His aid and His assistance, and ask His forgiveness and we seek refuge in Allah
from the evil of our souls and from the evil of our actions. Whomever Allah guides, no
one will be able to mislead him. Whomever He leads astray, nobody can guide him. I
thank Allah for the strengths and His blessing bestowed on me in completing this
thesis. May His peace and blesses be upon our noble Prophet, Muhammad (S.A.W),
his households, companions and those that followed his footsteps since and up till the
last day.
I wish to thank my supervisor, Dr. Hazel Monica Matias Peralta, for her
unquantifiable patience and for sharing her expertise, for all the useful and critical
suggestion required. May Allah continued to increase her in knowledge and bless her
family.
My thanks and appreciations also go to Universiti Tun Hussein Onn Malaysia
(UTHM) for giving me scholarship in order to undertake this project effectively. I
will not forget to mention, Ibrahim Badamasi Babangida University (IBBU) for giving
me the opportunity, support and courage in attending my master’s degree in UTHM.
My appreciation goes to my beloved family (Abdullahi Muhammad,
Muhammad Al-amin, Hauwa Sulaiman, Amina Lami Alhassan and Zainabs) for their
love, patience, encouragement and understanding and to my brothers and sisters,
friends and well-wishers for their prayers to see my dreams is actualised. May Allah
reward them abundantly, Amin.
vii
ABSTRACT
The investigation of plankton diversity and their distribution in relation to the
environmental conditions of a rare temporary water habitat (rock pool) in Upeh
Guling, Endau Rompin, Johor, Malaysia, was undertaken. Samples were collected on a
monthly basis for a period of one year. However, due to flash flooding during rainy
season, sample collection was only possible for a period of six months (between
March and September 2013). Plankton samples were collected from four stations
representing two different conditions of the sampling area: (1) river fed rock pools
which receive water primarily from the river (2) rain fed rock pools, which receive
water primarily from rainfall. Physico-chemical parameters: temperature, pH, and
dissolved oxygen were measured in situ. A total of 122 species of phytoplankton were
recorded with 116 species at rain fed rock pools and 84 species at river fed rock pools.
The most diverse phytoplankton genus was Staurastrum with 10 species. On the other
hand, a total of 49 species of zooplankton were recorded with 48 species at rain fed
rock pools and 40 species at river fed rock pools. The most diverse zooplankton genus
was Lecane with 11 species in total. For both the phytoplankton and zooplankton, the
rain fed rock pools recorded higher (P<0.05) variation in total species diversity and
density, compared with river fed rock pools. High species diversity index in both rain
fed and river fed rock pools, ranging from H'=3.14-H'=3.56 as a result of long period
of inundation which suggested healthy condition of the ecosystem. Similarly, the
environmental parameters (24.50 – 29.20 °C, 4.2 – 6.9 mg/L and 5.3 – 8.2 unit) of
both rock pools were within the favourable range and suitable for organisms
conservation. The canonical correspondence analysis ordination revealed that the
water depth, which was controlled by unexpected rain fall, was the major factor in
both rock pools that contributed to plankton diversities. The months of June, July and
August recorded highest species similarity in composition and distribution due to the
pools stability in these months. These all favours the proliferation of diverse species of
phytoplankton and zooplankton.
viii
ABSTRAK
Kajian mengenai kepelbagaian plankton dan taburannya yang berhubungkait dengan
keadaan persekitaran habitat air sementara (kolam batu) di Upeh Guling, Endau
Rompin, Malaysia telah dijalankan. Pengambilan sampel telah dilakukan setiap bulan
bagi tempoh satu tahun. Walau bagaimanapun, disebabkan oleh banjir kilat semasa
musim hujan, pengumpulan sampel hanya boleh dilakukan untuk tempoh enam bulan
(antara bulan Mac dan September 2013). Sampel plankton telah dikutip daripada
empat stesen yang mewakili dua keadaan kawasan persampelan yang berbeza: (1)
kolam batu sumber sungai yang mendapat air terutamanya dari sungai (2) kolam batu
sumber hujan, yang mendapat air terutamanya daripada hujan. Bacaan parameter
fiziko-kimia: suhu, pH, oksigen terlarut telah diambil secara in situ. Sejumlah 122
spesies fitoplankton telah direkodkan, dengan 116 spesies di kolam batu sumber hujan
dan 84 spesies di kolam batu sumber sungai. Fitoplankton yang paling pelbagai adalah
dari genus Staurastrum iaitu sebanyak 10 spesies. Sebaliknya, sejumlah 49 spesies
zooplankton telah direkodkan dengan 48 spesies di kolam batu sumber hujan dan 40
spesies di kolam batu sumber sungai. Zooplankton yang paling pelbagai adalah dari
genus Lecane iaitu sebanyak 11 spesies. Bagi kedua-dua fitoplankton dan
zooplankton, kolam batu sumber hujan merekodken (P <0.05) perubahan yang lebih
tinggi dalam jumlah kepelbagaian spesies dan ketumpatan, berbanding dengan sungai
kolam batu. Indeks kepelbagaian species yang tinggi dalam kedua-dua hujan dan
sungai makan kolam batu, bermula dari H' = 3.14-H' = 3.56 akibat tempoh
masapanjang banjir yang menyunbang kepada keadaan baik ekosistem. Begitu juga,
parameter alam sekitar (24,50-29,20 °C, 4,2-6,9 mg/L dan 5,3-8,2 unit) daripada
kedua-dua kolam batu berada dalam julat yang baik dan sesuai untuk pemuliharaan
organisma. Analisis kanun penyelarasan bersuraf ternadap kedalaman air, yang
dikawal oleh taburan hujan yang tidak dijangka, merupakan faktor utama dalam
kedua-dua kolam batu yang menyumbang kepada kepelbagaian plankton. Bulan Jun,
Julai dan Ogos mencatatkan persamaan species yang tinggi dalam komposisi dan
taburan kerana kestabilan kolam dalam bulan tersebut. Hal ini menggalakkan
pertumpuhan pelbagai spesies fitoplankton dan zooplankton.
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TABLE OF CONTENTS
TITLE Page
DECLARATION ii
DEDICATION v
ACKNOWLEDGEMENT vi
ABSTRACT vii
ABSTRAK viii
TABLE OF CONTENTS ix
LIST OF TABLES xi
LIST OF PLATE xiii
LIST OF FIGURES xiv
LIST OF APPENDIX xv
CHAPTER 1 1
INTRODUCTION 1.1 Background 1
1.2 Statement of the problem 4
1.3 Importance of study 5
1.4 Objectives 6
CHAPTER 2 7
LITERATURE REVIEW 7
2.1 Temporary Water habitats 7
2.2 Characteristic of freshwater rock pools 8
2.3 The Importance of Temporary Water Habitats 9
2.4 Inhabitants of Temporary Water Habitats 9
2.5 Plankton diversity of temporary water habitats 11
2.6 Species diversity in rock pool 13
CHAPTER 3 15
MATERIALS AND METHOD 15
3.1 Study Site 15
3.2 Field sampling 15
3.3 Laboratory analysis 18
x
3.3.1 Sample processing 18
3.3.2 Identification and enumeration 18
3.4 Data analyses 20
CHAPTER 4 21
RESULTS 21
4.1 Phytoplankton Composition and
abundance
21
4.2 Zooplankton composition and
abundance
30
4.3 Diversity indices 37
4.4 Environmental variation 38
4.5 Environmental relationship 39
4.6 Similarity of plankton distribution 46
CHAPTER 5 52
DISCUSSION 52
5.1 Phytoplankton composition and abundance 52
5.2 Zooplankton composition and abundance 58
5.3 Diversity indices 62
5.4 Environmental variation 64
5.5 Environmental relationship 64
5.6 Similarity of plankton distribution 67
CONCLUSION 68
RECOMMENDATION 69
REFERENCES 70
APPENDIX 87
xi
LIST OF TABLES
Table page
2.1 Number of planktonic species inhabitants of temporary rock
pools recorded from previous studies
14
4.1 Average phytoplankton density in rain fed rock pools and river
fed rock pools
22
4.2 Phytoplankton species distribution in rain fed and river fed rock
pools (+ denotes presence; - denotes absence)
23
4.3 The phytoplankton monthly species abundance significant
variation between rain fed rock pools and river fed rock pools.
Superscripts with the asterix “*” are significant while those
without the asterix are insignificant
30
4.4 Average zooplankton density in rain fed rock pools and river fed
rock pools
30
4.5 Zooplankton species distribution in rain fed and river fed rock
pools (+ denotes presence; - denotes absence)
32
4.6 The zooplankton monthly species abundance significant variation
between rain fed rock pools and river fed rock pools.
Superscripts with the asterix “*” are significant while those
without the asterix are insignificant
37
4.7 Diversity indices (Average S.E) of phytoplankton and
zooplankton in the rock pools. Data in row with asterix “*”
denotes significance at p <0.05
38
4.8 Pearson Correlation of physico-chemical parameter in rain fed
and river fed rock pools
39
4.9 The monthly summary of physico-chemical parameters 39
4.10 Relationships between phytoplankton abundance and
physicochemical parameters in rain fed rock pool
40
4.11 Relationships between phytoplankton abundance and
physicochemical parameters in river fed rock pools
41
4.12 Relationships between zooplankton abundance and physico-
chemical parameters in rain fed
43
xii
4.13 Relationships between zooplankton abundance and physico-
chemical parameters in river fed rock pools
44
4.14 The monthly species abundance of highest similarities revealed
not significant different from monthly species abundance of less
similarities with P<0.05.
51
5.1 Comparison of the number of phytoplankton species recorded in
the present study with other studies
53
5.2 Comparison of the number of zooplankton species recorded in
the present study with other studies
56
xiii
LIST OF PLATE
Plate Page
3. 1 Sampling stations (1-2 –rain-fed rock pools; 3-4 – river fed rock
pools) at Upeh Guling, TNJER
17
xiv
LIST OF FIGURES
Figure Page
3.1 A map showing the location of the study site in TNJER 16
4.1 Variation in composition (%) of phytoplankton groups from the
rain fed and river fed rock pools
28
4.2 Phytoplankton group density (cells/mL, in logarithmic scale)
variation according to sampling month between two rock pools
(rain fed rock pools and river fed rock pools)
29
4.3 Variation in percent abundance (%) of zooplankton groups
from the rain fed and river fed rock pools
35
4.4 Zooplankton density (individuals/L, in logarithmic scale)
variation according to sampling month between two rock pools
(rain fed rock pools and river fed rock pools)
36
4.5 Canonical correspondence analysis illustrating the relationship
between phytoplankton composition and physico-chemical
parameters in the river fed and rain fed rock pools, Upeh
Guling, Endau Rompin
42
4.5 Canonical correspondence analysis illustrating the relationship
between zooplankton composition and physico-chemical
parameters in the river fed and rain fed rock pools of Upeh
Guling, Endau Rompin
45
4.6.1 The Cluster analysis of phytoplankton species abundance from
river fed rock pools (RVF)
47
4.6.2 The Cluster analysis of phytoplankton species abundance from
rain fed rock pools (RNF)
48
4.7.1 The Cluster analysis of zooplankton species abundance from
river fed rock pools (RVF)
49
4.7.2 The Cluster analysis of zooplankton species abundance from
rain fed rock pools (RNF) 54
xv
LIST OF APPENDIX
Appendix Page
1 Taxonomic classification of phytoplankton from rare
temporary water habitats in Upeh Guling, TNJER, Malaysia
86
2 Taxonomic classification of zooplankton from rare temporary
water habitats in Upeh Guling, TNJER, Malaysia
93
3 Phytoplankton of Upeh Guling Rock pools 97
4 Zooplankton of Upeh Guling Rock pools 110
CHAPTER 1
INTRODUCTION
1.1 Background
Temporary water habitat is a type of freshwater that experience a recurrence of dry
phase of different length which may be periodical. Temporary water habitats, which
include intermittent streams and ponds, seasonal pools, puddles and water-retaining
structures of rock formation (rock pools), drift wood and hollow forest logs, can be
found throughout the world (Williams, 2005).
Its physical and chemical abiotic factors made the habitat special only for
species fit to survive and reproduce. Those abiotic factors are habitats morphology,
sediment characteristics, nutrient concentration; dissolved oxygen (DO)
concentration, hydrogen ion concentration (pH), light availability and temperature
(Williams, 1987). Temporary water species are generally well adapted to dealing
with water loss. Many species exhibit opportunistic and pioneering traits, and also a
range of drought-survival mechanisms, such as dormancy and seed formation
(Williams 2005).
Temporary water habitats support specialised assemblages and moderate
numbers of rare and endemic species (Bratton, 1990; Baskin, 1994; Kalettka, 1996;
King, Simovich & Brusca, 1996) which comprise of bacteria, protoctists,
vertebrates, fungi, and an abundance of higher and lower organisms (Williams,
2005). Many aquatic species such as algae, bacteria, fungi and invertebrate
organisms transform matter and energy into life. Temporary water species are well
represented by micro- and macro forms from major taxonomic groups, namely:
2
Branchiopoda, Ostracoda, Copepoda, Rotifera, Cladocera, Decapoda and Pericardia
(Williams, 2005). Similarly the insects are well represented in this type of water
bodies, such as the group Hemiptera, Coleoptera, Trichoptera, and Diptera,
especially the Chironomidae, Culicidae, and Ceratopogonidae, (Williams, 2005).
Aquatic mites (Hydracarina) are also common in temporary waters along with
several fish species which migrate in and out of temporary streams, rivers and
floodplains (Welcomme, 1979). Similarly, temporary waters serve as habitat to some
rare species of amphibians and also provide important feeding areas for migratory
birds (Poiani & Johnson, 1991).
The organisms of this habitat are living biomass that becomes food for other
organisms like fish. Insects that migrate from this habitat also provide nutrients and
energy to the neighbouring forest and sustain terrestrial animals like birds, spiders
and lizards. In addition, temporary water are components of landscapes (Nakano &
Murakami, 2001; Sabo & Power, 2002; Baxter & Fausch, 2005) and allow their
communities to perform important ecological services, such as, improving water
quality for consumption, food for commercial fishing, offering leisure and recreation
opportunities (Nedeau 2006; Pamela et al., 2013;). According to Tavernini (2008),
water fluctuation in temporary pools affects its hydrological features, hence,
influenced pools’ colonization and species richness. Likewise, Mahoney, Mort &
Taylor, (1990) and Girdner & Larson (1995) noted that hydroperiod is an important
determinant of planktonic communities. In temperate regions, the extent of water
permanence and the length of the dry phase in temporary waters are usually regulated
by snow and rainfall amount, whereas, in tropical areas it is mainly the amount of
rainfall (Hanes, Hecht & Stromberg, 1990; Fischer et al., 2000). Williams (2005)
believed that trees density/canopy cover may be a controlling agent that influences
pond community structure through a possible filtering effect on colonizing species,
mostly flying insects.
The contributions of temporary waters become even greater with the presence
of some biotas like plankton (phytoplankton and zooplankton). Plankton are minute
floras or faunas that float, drift or inactively swim against water current. In
oligotrophic ecosystem, phytoplankton are the major contributor of biomass and
primary productivity (Campbell, 1997). They produce oxygen during photosynthesis,
which was estimated on a global scale to be 50% – 60% of all photosynthesis
3
performed (Campbell, 1997). Moreover, they form the base of the food chain in
aquatic environments and their diversity and density are generally determined by the
existing water quality (Campbell, 1997). Phytoplankton (algae) are used as bio-filters
for removal of nutrient and other pollutants from wastewaters, to examine water
quality, as indicators of environmental changes, in space technology, and laboratory
research system. They are cultivated for the purpose of pharmaceuticals,
nutraceuticals, cosmetics and aquaculture (Christopher, 2008). The zooplankton in
the second level, transform food energy synthesized by the phytoplankton to the
higher tropic level (Abujam et al., 2011). Moreover, zooplankton is the most vital
biotic components affecting mostly the functional aspects of all aquatic ecosystems,
via; food chains, food webs, energy flow/transfer and cycling of matter. Different
environmental factors that detect the features of water play an essential role on the
growth and abundance of zooplankton (Suresh, Thirumala & Ravind, 2011). As such,
water quality influences zooplankton abundance, clustering and biomass.
Freshwater rock pools (a group of all types of depressions that occur on rocky
substrate and periodically hold water) which are found all over the world in all major
biomes depends mainly on precipitation for filling. The hydro periods ranging from
several days to a whole year and interaction between the climate and geology
determined the morphology and hydrology of this habitat (Jocque, Vanschoenwinkel,
& Brendonck, 2010). It experiences daily changes of water temperature, pH and CO2
(Keeley & Zedler, 1998). They have low conductivity and wide range variation of
pH (from 4.0-11.00) and temperature from freezing point to 40°C. The uncertain
nature of the flooding regime requires high endurable species with adaptation for
surviving the dry phase such as such as dormant eggs, resistance larvae or by active
migration and recolonization (Wiggins, Mackay & Smith, (1980); Brendonck & De
Meester, 2003). The quality of rock pools environments is controlled mainly by
timing and quantity of precipitation, and temperature of water. The temperature is a
vital parameter that determines which species might hatch in a pool, provided the
pool last longer for the time required for a species to reach maturity and produce
eggs to maintain continuity in a population. Slight change in average temperature
may not affect a species diversity and abundance directly but could change pool
durability and reduce the chances of certain species reproduction. Considering the
sensitivity nature of the rock pool flora and fauna to environmental situations, and
4
wide array of adapted species to little different condition, rock pools may be a vital
indicator system where a global climatic change effect can be noticed (Graham &
Wirth, 1996). Rock pools ecosystem provide early indications of biological unstable
climate, and for that research of this ecosystem should be encouraged to improved
our knowledge and understanding of these ecosystem and global climatic change
(Graham & Wirth, 1996).
About 460 animal species are recorded from rock pools worldwide and 170
of these are passive dispersers, that is, those mainly dispersed by wind and the
overflow of water between the pools. Freshwater rock pools stand as a source of
freshwater in dry countries, but despite that, they remain unexplored in large parts of
the world (Jocque et al., 2010).
Taman Negara Johor Endau Rompin (TNJER) is a forest containing rock
formations (Upeh Guling) with history traced back to 240 million years. It houses
many species of macro and micro flora and fauna that are not found elsewhere.
TNJER is a proposed site for Geopark with second oldest and largest park in the
whole of Malaysia after Langkawi Geopark (Devapriya et al., 2012). The historically
preserved site with temporary freshwater rock pools located in Upeh Guling, has
rarely been touched, particularly the records of plankton species that existed there.
1.2 Statement of the problem
Despite the fact that freshwater covers only 0.8% of the earth’s surface, yet it
sustains more than 100,000 species, which represents 6% of all known species on
earth out of approximately 1.8 million species. However, temporary water habitat of
this study is a rock pool specifically pothole, which are found worldwide but
received very little attention (Dudgeon et al., 2006; Balian et al., 2008).
Temporary water is very rich in diversity of flora and fauna in which some
cannot be found elsewhere. In Malaysia, increase in human population and
development which led to urbanization, industrialization, and growth in agricultural
irrigation have affected the existing temporary water and its community diversity
(Khoo, et al., 2003). Aquatic fauna and flora, especially plankton extinction by any
factors such as predation, competition, urbanization, agriculture practices, etc. may
directly or indirectly affect the communities of hydrosphere and atmosphere such as
5
in-balance in aquatic nutrients and gasses (Duffey, 1968; Lassen et al., 1975;
Pajunen & Pajunen, 1993; Bengtsson, 1989, 1991, 1993; Schneeweiss & Beckmann,
1999; Altermatt, Hottinger & Ebert, 2007).
Plankton is often used as indicators of environmental and aquatic health
because of their remarkable sensitivities to environmental change and short life span
(Hugo Rodier 2011). The phytoplankton (food producers) and zooplankton (food
consumers) contribute to water quality (Martin & Fitzwater, 1988). They are also
believed to increase the atmospheric oxygen and reduced excess carbon dioxide
(Neospark, 2002). However, temporary water and its planktonic communities that are
known with more efficient active compound than marine water communities are left
untouched. These vital roles played by plankton made it necessary for further studies
in the tropical rainforest of TNJER where rare temporary water habitat is naturally
abundant and conserved.
Since the last two decades, only few researches have extensively focused on
the study of flora and fauna of fresh water rock pools (Withers & Edward, 1997;
Porembski & Barthlott, 2000; Withers & Hopper, 2000; Merlijn et al., 2010; Hazel &
Ing, 2012) and among these researches, only one researcher in Malaysia concentrated
on plankton diversity (Hazel & Ing, 2012). An extensive study by Merlijn et al.
(2010) was recently conducted on diversity patterns of rock pools across the world
continents. The authors compared records of species diversity on fresh water rock
pools and concluded that the lowest species diversity was recorded in Asia continent
which may be due to “the paucity of fresh water rock pools studies”.
With all of the above-mentioned significance of plankton, the study of
temporary water habitat (rock pools) in Malaysia and Asia as a whole is left
untouched by researchers. Hence, this study is timely and necessary.
1.4 Importance of study
The temporary water habitat has always been a medium that support a diverse form
of animals (both invertebrates and vertebrates) and macro/microscopic organisms
including plankton. Indeed, these types of habitats, where important unknown
resources are still left unexplored, and worth studying. Thus, there is a great
6
possibility of discovery in terms of new species for biodiversity records, and
fundamental knowledge may be expected.
1.3 Objectives
This study was undertaken with the following objectives:
1. To determine the distribution and abundance of plankton thriving in temporary
rock pools in a tropical rainforest
2. To assess the differences between the plankton diversity of each rock pool types
(river fed and rain fed).
3. To determine the variations in physicochemical parameters of each rock pool
types and relate them with plankton diversity
CHAPTER 2
LITERATURE REVIEW
2.1 Temporary Water habitats
Temporary water is any "habitat that intermittently has standing water and that once
flooded or over spread holds water long enough for some species to complete the
aquatic phase of their life cycle (Williams, 2005). Such temporary water habitat is
highly varied and widely distributed almost everywhere in the world (Junk, 1993).
They may be intermittent streams and ponds, seasonal pools, puddles and water-
retaining structures of rock formation (rock pools), drift wood and hollow forest logs,
and phytotelmater or wetlands (Williams, 2005). They are characterized by the
alternation of flood and dry phase and whose hydro-regime is mostly independent.
Moreover, their water supplies are usually from precipitation, water run-off and the
ground water table (The Ramsar Convention on Wetlands, 2002a). Temporary water
habitats are ecosystems that contain water during periods that can vary from a few
months to several years (Schwartz & Jenkins, 2000) and can vary from a few square
meters to hundreds of hectares (Schwartz & Jenkins, 2000; Williams, 1987).
Temporary aquatic environments are regionally different in their types and method of
formation but common in their physical, chemical and biological properties
(Fontanarrosa et al., 2009; Williams, 1996). The concentrations of dissolved
substances in temporary waters vary more than in most permanent waters. This is
due largely, to three physical processes to which temporary waters are subjected:
drying out, refilling, and freezing (The Ramsar Convention on Wetlands, 2002a).
8
2.2 Characteristic of freshwater rock pools
In most cases, freshwater rock pools are temporary waters. They are characterized by
highly unpredictable timing and length of the inundation period (Brendonck et al.,
1998; 2000a). The hydro-period in rock pools averagely ranged from several days up
to a month (Brendonck et al., 2000a) and several months in the case of semi-
permanent rock pool (Jocque et al., 2010). Rock pools occur everywhere in the
world, and they all possess very similar structures and abiotic environment. They are
geo-morphologically similar because they originated from weathering and erosion
(Campbell, 1997; Domínguez-Villar, 2006) although vary in surface and depth.
Weathering and erosion may result to joining of neighbouring pools that may lead to
more complex shape (Twidale & Corbin, 1963). Rock pools are usually in the form
of the pan, or bucket shape with a cylindrical or ellipsoid surface with different
dimensions (Twidale & Corbin, 1963).
Hydro-period is so important in determining the composition, structure and
diversity of the rock pool bio-community (Hulsmans et al., 2008). The climatic
change in rock pool significantly changes the hydro-regime with decreasing stability
on some biotas. This established the hydrologic sensitivity of rock pool habitats to
precipitation patterns and its potential to predict future climatic change. The
regularity and duration of inundation depend on basin physical factor (shape, size
and structure), area, types of vegetation and local climate. The maximum depth of
basin determines the maximum length of inundation period (Vanschoenwinkel et al.,
2009). Rock pool mostly filled with rain water, which results in the highly diluted
environment at the beginning of the inundation. The conductivities of this water are
below 10 µScm-1 and varying depth ranging from 5cm to 30cm. The temperature of
the water close to the air temperature and showed wide diurnal fluctuation in pH and
DO. The freshwater rock pool is distinct between the pool filled by precipitation and
those fed by rivers and ground water. The ground water fed rock pool (e.g. quarry
pond) and potholes in the river floodplains inhabit biotas that are mainly brought in
with the water. These communities of fauna in these habitats are not often adapted to
this temporary pool condition. However, the precipitation dependent rock pools
accommodate communities of species fit to survive dynamic and unpredicted
flooding cycles.
9
2.3 The Importance of Temporary Water Habitats
Despite the generally small size and seasonal or ephemeral nature, temporary water
habitats can be of critical importance for the maintenance of biodiversity and as
sources of water, food and other products for local communities and indigenous
peoples (Biggs et al., 2008). Temporary ponds contributed most to biodiversity,
supporting considerably more species, more unique species and more scarce species
than other water body types (King et al., 1996; Forró et al., 2003). More so,
temporary water serves great importance to mankind, some of which includes
fulfilling biological significance or needs of man; may be used for agricultural
activities; may harbour some species of organisms that can transmit diseases and
covered a significant portion of the global landscape (Blaustein & Schwartz, 2001;
Williams, 2005). They serve as alternating aquatic and terrestrial phases that are
linked functionally by accommodating diverse biotic communities, various
biogeochemical processes and viable ecosystem society.The rock pools are used in
local population as reservoirs for both human and their cattle (Seine, 2000).
Similarly, in some places where freshwater is scares e.g. Western Australia, the
natural water reservoir on rocky outcrop were used as a source of drinking water
(Laing & Hauck, 1997; Bayly, 1999; Twidale, 2000).
2.4 Inhabitants of Temporary Water Habitats
Temporary fresh water support invertebrate communities ranging from the complex,
with many species (e.g., vernal ponds), to those that support only one or two species
(e.g., ephemeral rock pools and water-filled leaf axils). The biota of temporary
aquatic habitats has evolved mechanisms that re-establish the population when the
habitat becomes available again. The typical inhabitant of temporary habitats (e.g.,
crustaceans, molluscs, rotifers, tardigrades, turbellarians, and hydrozoans) produces
resistant eggs or are themselves able to enter a resistance, resting stage (Williams,
1987; Wiggins et al., 1980), and a majority of their life span may depend on these
diapause stages (Fugate, 1998).
The drying nature of the habitat serve as an element of disturbance because it
causes mortality that lead to a decrease in population of aquatic organisms (Corti,
10
Kohler, & Sparks, 1996) This factor contributed to significant mortality to biota that
are not adapted to survive such conditions (Williams, 2006); as such the organism
may also be influenced to develop adaptive strategies (Blaustein & Schwartz, 2001)
such as morphological, physiological, and/or behavioural nature for their survival in
such a habitat (Williams, 2005). Other environmental features that most likely affect
the biota of temporary waters are the period and range of water flow, the degree and
rate of desiccation of the groundwater table including permeability of the bed
substrate, (Williams, 2006). Due to the desiccative nature of the habitat, interaction
by existing organisms in the habitat increases the population density of biotas. Many
species developed trait and drought-survival mechanism, such as dormancy and seed
formation for their adaption to temporary dry up of the water. Drying events also
prevent the colonization of many large predators, such as fish, even if some other
predaceous taxa can tolerate desiccation (i.e., turbellarians) or leave the pool when it
dries out (i.e., amphibians and insects) (Murdoch, Scott, & Ebsworth, 1984; Bohonak
& Whiteman, 1999; Spencer et al., 1999; Hobæk, Manca & Andersen, 2002).
Overall, species richness in temporary waters should be greater than in
permanent fresh waters, with increasing interest in land waters. Schneider (1999)
concluded that the invertebrate communities of short hydro-period and snowmelt
ponds were structured primarily by species adaptations to the threat of drying (i.e., to
abiotic factors). On the other hand, the communities in ponds with longer hydro-
period were structured more by biotic interactions, particularly predatory and
competition. Predators are one of the major forces in the community structure in
permanent water bodies (Kerfoot & Sih, 1987) although temporary water also houses
predation taxa but they support the community structure. Several studies have shown
that fishless temporary water habitat support higher diversity of zooplankton, macro-
invertebrates and water birds than a lake with fish (Havas & Rosseland, 1995).
Unlike other temporary pools that are characterized as an enemy-free habitat
(Fryer, 1985; Kerfoot & Lynch, 1987), predation is an essential component of rock
pools and considered to be an essential community structuring factor. Examples of
such predators are clawed toad, Turbellaria, notonectids, odonates and ceratopogonid
midge larvae (Hamer & Martens 1998; De Roeck, Artois & Brendonck, 2005;
Vanschoenwinkel et al., 2009). Jocque et al., (2010) mention water mite as common
predators in rock pools that colonize pools after the inundation.
11
2.5 Plankton diversity of temporary water habitats
Temporary water habitat (fresh water rock pools) are unique habitats that
accommodate a high diversity of specialized and endemic species and this highly
contributed to regional diversity (Pinder et al.,2000; Jocque, Graham, & Brendonck,
2007). The freshwater community, such as plankton and micro invertebrate are
sensitive to environmental factors; different species vary with the different season
due to the change in the physico-chemical nature of water. Phytoplankton
community shows high diversity with the seasonal fluctuation, which indicates the
diversity in ecological niches. The zooplankton in the second level, transform food
energy synthesized by the phytoplankton to the higher tropic level (Abujam, 2011).
Phytoplankton co-occur based on their physiological need and the environmental
constraints (Ariyadej et al., 2004).The environmental constraints and physiological
requirements determine the phytoplankton succession in relation with environmental
parameters, particularly temperature, light, nutrient availability and mortality factors
such as grazing and parasitism (Ariyadej et al., 2004). The later succession is
strongly linked to meteorological and water stratification mixing processes (Wetzel,
2001). The dynamics of plankton are a function of many environmental processes
that affect species diversity (Roelke & Buyukates, 2002).
Patterns in a temperate ecosystem differ considerably from those of tropical
water (Wetzel, 2001). In tropical regions, the water temperature of temporary pools
approaches the upper limit for biological processes. The high temperature in these
habitats influenced the rapid growth of algae that may serve as food for some of the
fauna in the habitat. The consequence is the excessive growth of algae leading to
depletion of dissolved oxygen in the water, especially at night. High temperature
increase level of photosynthesis, which changes the pH level of water and it affect
the transportation of materials across the cell membrane (Williams, 2006). Variations
in plankton community structure depend on the availability of nutrients, temperature,
light intensity and other limnological factors (Vaulot, 2001). It is found that any
slight alteration in environmental status can change diversity until there is no
adaptation or gene flow from non-adaptive sources. A high diversity count suggests a
healthy ecosystem; the reverse of this indicates a degraded environment. Hence,
12
phytoplankton fluctuation and density are used in the determination of water quality
in temporary pools (Palmer, 1977; Shubert, 1984).
Like other types of temporary water habitat, rock pools faunas are
categorized into two, namely: permanent resides (as resistant life stages) and
migrates (when pools are dry) (Wiggins et al., 1980). The high variability in
environmental condition connected with relatively unpredicted flooding regime limit
for specialized species with high tolerance to stress and specific feature for surviving
the dry phase. Most pool biota survives the desiccation through resting stages such as
dormant eggs, resistance larvae or by active migration and recolonization (Wiggins
et al., 1980; Brendonck & De Meester, 2003). Economically, zooplankton is the
major primary consumer or intermedia of energy transfer between phytoplankton and
other aquatic animals including fish (Hammer, 1985; Telesh 1993; Davies, Abowei,
& Otene., 2009). Moreover, zooplankton is the most vital biotic components
affecting mostly the functional aspects of all aquatic ecosystems, via; food chains,
food webs, energy flow/transfer and cycling of matter. Different environmental
factors that detect the features of water play an essential role on the growth and
abundance of zooplankton (Suresh et al., 2011). As such, water quality influences
zooplankton abundance, composition and biomass. Physico-chemical, biological and
microbiological parameters analysis in water quality assessment reveals abiotic and
biotic status of the ecosystem (Rajagopal et al., 2010). The biotic communities of
temporary water tolerate a wide range of environmental variation that influences
their evolutionary processes.
Previous studies revealed that rare species are found on rock pool
communities, which have never been recorded from open water (Dethier, 1980;
Jonsson, 1994). Such rock pool species possess physiological or behavioral features
that can help population persist in the pool (Blackwell & Gilmour, 1991; Jonsson,
1994). The plankton community in rock pool may be used as a model system to
study many ecological concepts such as community assemblage, spatial population
dynamic and local extinction (Pfiester & Terry, 1978). Copepods, ostracods,
dinoflagellates, chlorophytes and various diatoms were the common taxa recorded in
the rock pools (Johnson, 2000).
13
2.6 Species diversity in rock pool
The invertebrate and vertebrate species diversity and density in rock pools varies
considerably among studies. It is a common agreement that diversity increases
stability in communities and ecosystem (McCann, 2000). Dodson (1987) studied in
Utah and reported 23 species of macro-vertebrates and amphibians (Table 2.1). The
study surveyed of 50 pools. Baron, LaFrancois, & Kondratieff (1998) study was
conducted where 59 species of macro-invertebrate and vertebrate were recorded
based on weekly sampling from 20 rock pools. A 66 species of invertebrate were
recorded from 92 pools on two outcrops (Jocque et al., 2007) and 230 species of
aquatic invertebrate were recorded. Sampled were taken from 90 pools equally
divided in nine different outcrops (Pinder et al., 2000). Bayly (1997) research
recorded 88 species of invertebrate in 36 rock pools on 17 granite outcrops (Table
2.1). Rock pools inhabited remarkable high diversity of passive disperser when
compared with other temporary water bodies like phytotelmata (water held in the
plant). This may be attributed to temporary stability and physical properties of the
habitats together with the low exchange rate of individual species between cluster
habitats usually isolated from the different outcrop. The differences in the number of
rocks and pools together with sampling intensity of individual pool may probably
explain part of the variation in recorded diversity.
Indeed, it is an established fact that the rock pools biota is fully dependent on
length and abundance of inundation and for that, the active communities will reflect
the current climatic conditions (Jocque et al., 2010). The rock pool communities
thus may be fit as a proper monitoring system for identifying environmental changes
on both short and long time basis and learning the climatic changes effect on bio
communities. The rock pools habitat is unique for accommodating specialized and
endemic species and, therefore, contribute essentially to regional diversity (Pinder et
al. 2000; Jocque et al., 2007). Manson (1990) stated that the establishment of
conservation strategies may not be straight forward in fresh water habitat, but the
protection of this habitat is essential.
14
Table 2.1: Number of planktonic species inhabitants of temporary rock pools
recorded from previous studies
Rock pools Species number recorded Author
1 23 Dodson (1987)
2 88 Bayly, (1997)
3 59 Baron et al. (1998)
4 230 Pinder et al. ( 2000)
5 66 Jocque et al. (2007)
CHAPTER 3
MATERIALS AND METHOD
3.1 Study Site
Field sampling was carried out in Upeh Guling, Taman Negara Johor Endau Rompin
(TNJER) (Figure 3.1). Four sampling stations were established at geographical
locations and elevations as follows: N 02 30.711” E 103 20.984” at 69m (St. 1); N
02 30.723” E 103 20.995” at 80m (St. 2); N 02 30.722” E 103 20.985” at 80m
(St. 3); N 02 30.740” E 103 20.996” at 75m (St. 4). Stations (rock pool 1 and 2)
received water primarily from rainfall and secondarily fed by the river water flow
during the rainy seasons (designated as rain-fed rock pools), whereas, stations (rock
pool 3 and 4) received water mainly from river water and secondarily from rainfall
(river fed rock pools).
3.2 Field sampling
Samples were collected on a monthly basis for one year. However, due to flash
flooding during rainy season, the sampling area was not accessible; hence, sample
collection was only possible for a period of six months (March, April, June, July,
August and September 2013). The physicochemical parameters (temperature, pH,
dissolved oxygen, depth) were measured in situ.
16
Source: (http://go2travelmalaysia.com/maps/images/msia1c.jpg).
Figure 3.1: A map showing the location of the study site in TNJER
17
Phytoplankton samples were collected using Van Dorn water sampler. Triplicate
samples were transferred in 1L plastic sample bottles and fixed immediately with
Lugol’s iodine solution after which, were taken to the laboratory for sample
processing and taxonomic identification and enumeration.
Zooplankton samples were collected by filtering triplicate 10L pooled water
samples using zooplankton net with 67 µm mesh size and preserved with 5%
buffered formalin solution immediately. All samples collected were taken to the
laboratory for taxonomic identification and enumeration.
Plate 3.1: Sampling stations (1-2 –rain-fed rock pools; 3-4 – river fed rock pools) at
Upeh Guling, TNJER
Rock pool 3 Rock pool 4
Rock pool 1 Rock pool 2
18
3.3 Laboratory analysis
3.3.1 Sample processing
Upon arrival in the laboratory, phytoplankton samples were allowed to settle for at
least three days without disturbing. After which, the water was siphoned out until
only 100ml of the sample remained. The remaining samples were then transferred to
a labelled opaque plastic bottle and kept in the dark until further analysis. On the
other hand, zooplankton samples were identified and enumerated without prior
processing as that of the phytoplankton.
3.3.2 Identification and enumeration
Identifications of phytoplankton were done using a compound microscope
(ModelYS2-H Nikon Japan) under 40x magnification or higher depending on the
size of the species and where the species can be identified comfortably and
confidently (McAlice, 1971; Bellinger & Sigee, 2010). A Sedgewick-Rafter counting
chamber was used for species enumeration. Triplicate samples were counted each
time. The species found were identified to the lowest possible taxa using
phytoplankton identification key (Huber-Pestalozzi, 1938; Huber-Pestalozzi, 1941;
Prescott, 1970; Yamagishi & Hirano 1973; Huber-Pestalozzi, 1982; Peerapornpisal,
1996; Yinxin & Minjuan, 2005; Shukla et al., 2008; Stamenković & Cvijan, 2008;
Bellinger & Sigee, 2010; ITIS, 2010; Sayers et al., 2011; Guiry & Guiry, 2014; Pal
& Choudhury, 2014; Parr, et al., 2014). Phytoplankton densities were expressed as
cells/mL (Bellinger & Sigee, 2010).
The cell concentration was expressed as number of cell per milliliter for each
species and was calculated as:
cell/ml = N * 1000mm3/A*D*F
Were N = number of cell counted
A = area of field (mm2)
D = depth of a field (chamber depth)(mm)
F = number of fields counted
19
The enumerations were done by counting individual cell. In addition, those species
of phytoplankton which have countable filaments (e.g. Micrasterias) were counted
by individual cell. However, species which have uncountable filaments (e.g.
Oscillatoria spp) and species that form colony (e.g. Botryococcus sp.) were
estimated following Findlay & Kling (2001). For filamentous (Oscillatoria) and
colonial (Botryococcus) species, cell/ml were determined by L*M/y and Z
respectively;
Where L = length of the species (μm)
y= species average length(μm)
M = species field(μm)
D = diameter of the species colony on stage (μm)
r = radius (μm)
Z = cell number in colony (μm)
Triplicate samples of zooplankton were counted each time using a
zooplankton counting chamber under the compound microscope (Model YS2-H
Nikon Japan) with low-power objective (10 x magnifications). The zooplankton were
identified to the lowest possible taxon using freshwater zooplankton identification
keys (Pennak 1979; Idris 1983; ITIS, 2010; Sayers et al., 2011). Estimation of
zooplankton densities expressed as individuals/L (Bellinger & Sigee 2010), follows
the equation below (Warren & Horvatin, 2003):
Where
NA = Number of individual in 1ml aliquots examined (Three 1ml
aliquots throughout)
Vs = Volume of sub-samples from which aliquots were removed.
V = Volume of water filtered (10liters)
20
3.5 Data Analysis
The plankton densities were used to determine the plankton community structure
using statistical software. The community structures were observed by multivariate
techniques using a PRIMER Software package (version 6.1.9, PRIMER-E Ltd),
(Clarke & Warwive, 2001). Data (species abundance) were log (x+1) transformed to
balance some rare species that are very few in abundance (Clarke & Warwive, 2001).
A similarity matrix was obtained using the Bray-Curtis similarity which gives the
basis for 2-d ordination plot using non-metric multidimensional scaling (NMDS).
Samples were also grouped by sampling stations and on monthly basis, using
t-test to determine the significance difference (P<0.05) between two pools but for
months (6 months) one way ANOVA was used. The community structure such as
species number (s), total number of species (N), the Shannon Wiener diversity were
determined
H' = pi ln pi
H' = index of diversity
In = the natural logarithm
pi = proportion of the ith species to the total count
Margalefs species richness [(d=(S-1)/log (N)] and Pielou’s species evenness
[(J=H'/log e (s)] were calculated for each sample to study the impact in community
structure. (Heip, et al., 1998).
Correlations between species and environmental factors of rain fed and river
fed rock pools were obtained by correspondence component analysis (CCA) using
CANOCO Software Package (version 4.5) (Braak & Šmilauer, 2002). The simple
ordination was constructed and the percentage differences were revealed (Braak &
Šmilauer, 2002).
Regression analysis was run to calculate the Pearson correlation coefficient
between phytoplankton/zooplankton densities and physicochemical factors. The
linear regression model formula (y = a+bX1+bX2+bX3+e) was developed to support
the correlation, Where: y= diversity variation, a = constant b = coefficient variable
and e= error term. The correlation relationship level is = -1≤ r ≤ 1 while 0.05 indicate
the level of p value significant difference (Pallant, 2010).
CHAPTER 4
RESULTS
4.1 Phytoplankton Composition and abundance
This finding showed significant variation between phytoplankton species richness
of rain fed rock pools and river fed rock pools (Table 4.1). A higher (P< 0.05)
phytoplankton density was recorded in rain fed rock pools (1470.02 cells/mL)
compared to river fed rock pools (781.38 cells/mL).. A total of 122 species of
phytoplankton belonging to 54 genera, 36 families, 27 orders, 10 classes and eight
groups were identified (Table 4.2). The green algae group had the highest (P<
0.05) species diversity with 60 species from 25 genera, 12 families, and three
classes. Diatoms followed in dominance after green algae with 39 species from 15
genera, and 14 families and one class. The phytoplankton groups which
contributed the least in terms of number of species were cryptomonads and
golden-brown algae, in which both consisted of two species (Table 4.2). On the
other hand, a total of 116 species, from 53 genera, 35 families, 26 orders, and 8
groups of phytoplankton were present in rain fed rock pools (Table 4.2).
22
Table 4.1: Average phytoplankton density in rain fed rock pools and river fed
rock pools
Rain fed River fed
Phytoplankton Average & S. E
Significant level
1470.02 ± 212.03
cells/mL
0.034
781.38 ± 180.8403
cells/mL
F 4.559
Thirty eight species were found only on rain fed rock pools and some of these
species are Stigonema mirabile, Stigonema sp. 1, Euastrum sp. 1, Actinotaenium
cucurbita, Micrasterias ceratofera, Micrasterias lux, Micrasterias rotate,
Staurodesmus sp. 1, Staurodesmus sp. 2, Triploceras gracile, Netrium sp. 1,
Netrium sp. 2 , Spirogyra sp, Spirogyra weberi, Zygnema himalayense,
Staurastrum alternans, Staurastrum anatinum, Staurastrum avicula, Staurastrum
gracile, Staurastrum manfeldtii, Staurastrum orbiculare, Staurastrum
pseudopelagicum, Staurastrum sp. 2 Dictyosphaerium sp., Pediastrum duplex,
Scenedesmus sp. 1, Scenedesmus sp. 2, Scenedesmus sp. 3, Binuclearia tatrana,
Oedogonium sp, Botryococcus sp,Centritractus sp, Rhizosolenia sp, Epithemia
argus Nitzschia sp. 2, Nitzschia sp. 4, Nitzschia sp. 5., Melosira aequalis, Neidium
affine (Table 4.2). Whereas, a total of 84 species from 46 genera, 33 families, 26
orders and 8 groups of phytoplankton were present in river fed rock pools. Six
species were found present only in the river fed rock pools during the study; these
include Pleurotaenium sp, Nitzschia obtusa, Nitzschia sp. 3, Nitzschia sp. 6,
Surirella sp. 1 and Surirella sp. 2. Both rock pools recorded common
phytoplankton consisting of 78 species (Table 4.2).
23
Table 4.2 Phytoplankton species distribution in rain fed and river fed rock pools
(+ denotes presence; - denotes absence)
No Group Species Rain
fed
River
fed
1 Blue-green algae Geitlerinema sp. 1 + +
2 Geitlerinema sp. 2 + +
3 Oscillatoria sp. 1 + +
4 Oscillatoria sp. 2 + +
5 Phormidium sp. + +
6 Stigonema cf. mirabile + -
7 Stigonema mamillosum + +
8 Stigonema sp. + +
9 Green algae Actinotaenium cucurbita + -
10 Ankistodesmus sp. + +
11 Arthrodesmus michiganensis + +
12 Binuclearia tatrana + -
13 Botryococcus sp. + -
14 Closterium acerossum + +
15 Closterium ehrenbergii + +
16 Closterium kutzingii + +
17 Closterium leibleinii + +
18 Closterium sp + +
19 Closterium tumidum + +
20 Coelastrum pulchrum + +
21 Cosmarium dispersum + +
22 Cosmarium gotlandicum + +
23 Cosmarium granatum + +
24 Cosmarium hammeri + +
25 Cosmarium quinarium + +
26 Cosmarium sp. 1 + +
27 Cosmarium sp. 2 + +
28 Cosmarium sp. 3 + +
29 Dictyosphaerium sp. + -
30 Euastrum sp. 1 + -
31 Euastrum sp. 2 + +
32 Genicularia sp. + +
33 Gonatozygon sp. 1 + +
34 Gonatozygon sp. 2 + +
35 Gonatozygon sp. 3 + +
36 Micrasterias ceratofera + -
24
Table 4.2 continued
No Group Species Rain
fed
River
fed
37 Micrasterias lux + -
38 Micrasterias rotate + -
39 Mougeotia sp. 1 + +
40 Mougeotia sp. 2 + +
41 Mougeotia sp. 3 + +
42 Netrium sp. 1 + -
43 Netrium sp. 2 + -
44 Oedogonium sp + -
45 Pediastrum duplex + -
46 Pleurotaenium ehrenbergii + +
47 Pleurotaenium sp. - +
48 Scenedesmus sp. 1 + -
49 Scenedesmus sp. 2 + -
50 Scenedesmus sp. 3 + -
51 Sphaerozosma granulatum + +
52 Spirogyra sp. + -
53 Spirogyra weberi + -
54 Staurastrum orbiculare + -
55 Staurastrum alternans + -
56 Staurastrum avicula + -
57 Staurastrum gracile + -
58 Staurastrum manfeldtii + -
59 Staurastrum pseudopelagicum + -
60 Staurastrum pseudopelagicum + -
61 Staurastrum smithii + +
62 Staurastrum sp. 1 + +
63 Staurastrum sp. 2 + -
64 Staurodesmus sp. 1 + -
65 Staurodesmus sp. 2 + -
66 Staurodesmus triangularis + +
67 Triploceras gracile + -
68 Zygnema himalayense + -
69 Yellow green algae Centritractus sp. + -
70 Tribonema affine + +
71 Golden-brown algae Dinobryon divergens + +
72 Dinobryon sp. + +
73 Diatoms Achnanthidiun sp. + +
70
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