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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