Phytoplankton Diversity in Tumalaong River

A Project Proposal Presented to the

Biology Department, Xavier University
Cagayan de Oro City

In Partial Fulfillment
of the Requirements for the Course
Limnology Laboratory (Bio 23L)


Submitted By:
Arjay Lorenz L. Layawan

Submitted To:
Astrid Lara Sinco, PHD

January, 2014


Project Title: Phytoplankton Diversity in Tumalaong River
Proponent: Arjay Lorenz L. Layawan
Project Duration: 10 months (June 2013-March 2014)
Project Budget: Php 65,233.00

















TABLE OF CONTENTS
Chapter Page
Title Page A i
Title Page B ii
TABLE OF CONTENTS iii
LIST OF TABLES v
LIST OF FIGURES v
I. Introduction 1
II. Objectives 2
III. Scope and Limitations of the Study 2
IV. Significance of the Study 2
V. Literature Review 2
VI. Work Plan 5
A. Study Area 5
B. Duration and Frequency of Investigation 6
C. Methods 7
1. Collection 7
2. Preparation and Mounting of Phytoplanktons 8
3. Enumeration and Identification of Phytoplanktons 9


D. Data Analysis 9
VII. Financial Requirements 10
VIII. Working References 13
IX. Biodata 16

















LIST OF TABLES
Table Page
1 Gantt Chart of the schedule of activities from June 2014 to 7
March 2015
2 Shannon-Weiner Diversity Index 10
3 Simpson Diversity Index 10
4 Financial Requirements Table 10

LIST OF FIGURES
Figure Page
1 Map of Baungon showing Tumalaong River with the sampling 6
sites







1

I. Introduction
The falling of rain, melting of snow, or even moisture condensing may
accumulate enough freshwater to form a stream system (Moss, 2010). In this
system, a river can be born. Rivers are defined as a relatively large volume of
freshwater moving within a channel, including subsurface water moving in the
same direction and the associated floodplain and riparian vegetation (Naiman
and Bill, 1998). Systems of rivers are providing transport for water, chemicals,
and sediments downstream (Callow, 1992; Likens, 2010). Some ecosystems
are formed within rivers and they play a part in nutrient cycling (Moss, 2010).
Rivers are also important in ecological terms. Although 0.8 percent of the
surface area of the planet is occupied by the aquatic ecosystems, 12 percent
of all animal species live in fresh waters (Brierly and Fryirs, 2008).
Animal species that live in the freshwaters need food or energy for
them to survive. Phytoplanktons play an important part of the food web and
provide new particulate matter for other organisms to consume and continue
live (Moustaka-Gouni, 2006). Phytoplanktons are the primary producers and
constitute the first level trophic status for the aquatic food chain (Shanthala,
2009). Diversity indices of these organisms would give results on how many
species are found and how relative are their abundances to one another. The
diversity indices of all the species can explain if water is polluted or not based
on the results of its diversity (Shanthala, 2009).
The Tumalaong River of Bukidnon, Northern Mindanao, Philippines is a
tributary river to the main Cagayan de Oro River. This study will primarily list
the species of phytoplankton found in the said river together with its diversity
indices.
2

II. Objectives
The general objective of the study is:
To determine the diversity of phytoplankton in Tumalaong River
The specific objectives of the study are to:
1. Identify the phytoplanktons in the area
2. Determine the diversity indices of the phytoplankton in the area

III. Scope and Limitations of the Study
The study will only include three sites in the river situated in Imbatug,
Lingating, and Bayanga. Study will not be temporal as sampling will only be
done once.

IV. Significance of the Study
This study will provide information on diversity of phytoplankton
species in Tumalaong River. This study will also provide baseline information
for future studies involving planktons in Tumalaong River.

V. Literature Review
A community is comprised of different populations interacting with one
another (Leo et al., 2010). Communities play a huge role in the ecosystem
through interactions and cycling of nutrients and energy (Moss, 2010).
Amongst communities, there are groups that comprise them which are called
3

populations. These populations are then composed of a number of individuals
whose membership of the group is determined by their relationship to the rest
of the said group (Hardwood, 2009). Populations are very important since the
interactions of the individuals within the group help each other grow positively
or negatively (Sandholm, 2010). These groups of particular species may have
different geographical distribution and are important for a variety of
applications for ecology (Phillips, 2008). Specific gradients such as latitude
and altitude are addressed as causes of diversity patterns (Korhonen, 2011).
These species then have genetic differences from one other which in time
would eventually lead to speciation and more diversity amongst all species
(Elith, 2009).
Diversity is a compound quantity which is composed of species
richness and evenness components (Jost, 2010). Through diversity, the
interactions and benefits per species would either be maximized or not
(Phillips, 2008). Values of diversity can be derived from the Shannon-Weiner
Index or the Simpson Diversity Index (Shanthala, 2009).
Diversity of planktons is studied since these organisms are on the
bottom of the food chain and all other aquatic organisms depend on their
availability. Planktons can be classified into three groups; the phytoplanktons,
zooplanktons, and the microbial loop. The phytoplanktons are then subdivided
into picophytoplanktons, nanophytoplanktons, and the filamentous diatoms.
Herbivorous zooplanktons are composed of protozoa, rotifers, and calanoid
copepods. The microbial loop is then represented by the heterotrophic
bacteria and two groups of detritivores (Beninica, 2008).
4

Within the three groups, the diversity phytoplanktons are particularly
focused on this study. Phytoplanktons are defined as free-floating unicells and
colonies that grow through photosynthesis in aquatic environents (Vaulot,
2008). One photosynthetic pigment present to all phytoplanktons is
chlorophyll a which is commonly used proxy for total phytoplankton biomass
(Gregor, 2004). Eukaryotic and prokaryotic species are both present in
phytoplanktons, they colonize the upper part of the water column, down to
where penetration of light into the water is at its maximum value (Vaulot,
2008). Accordingly, the structure and abundance of phytoplankton populations
are mainly controlled by the availability of inorganic nutrients such as nitrogen,
phosphorus, silica, and iron (Vaulot, 2008). Phytoplanktons can be found in
either marine or freshwater habitats or areas where both of them meet (Harris,
2013).
Phytoplanktons are also seen in freshwater habitats and are abundant
depending on seasonal changes in mean temperature, radiations, hydrology,
and nutrient availability (Offem, 2011). Three major groups of these
phytoplanktons are the diatoms, dinoflagellates, and the coccolithophorids
(Cermeo, 2010). Diatoms are unicellular, eukaryotic organism that are either
pennate or centric and play a role as bioindicators for biotic and abiotic events
(Gordon, 2009). Dinoflagellates are single celled microorganisms that are
free-living or symbiotic with other animals. Half of the dinoflagellates are
heterotrophic and the other halves are photosynthetic species and belong to
the phytoplankton group (Steidinger, 2011). The last of the three groups are
the coccolithophorids. Coccolithophorids mostly belong to the marine area
and produce minute calcite plates called coccoliths (Obaje, 2013).
5

Phytoplanktons can be found in freshwater environments and one
study conducted in Spain studied trade-offs in phytoplankton species richness
arising from drought. Results of the study showed that annual average
phytoplankton species richness increased from the wet to dry years and taxon
richness diminished again when the drought in that area was severe (Rojo,
2012). Phyoplankton diversity also is a good indicator of environmental
stresses (Paerl, 2010). Another study in a freshwater environment in Belgium
tackled on the factor of water turbidity in relation with phytoplanktons. It shows
that the less light penetrating the water due to its turbidity caused less
phytoplankton development in the area and prevented them to fully use the
available nutrients (Muyalert, 2005).
The study of phytoplankton diversity in Tumalaong River would then
add to the less knowledge known on that said area. Results would give
baseline information for future studies involving taxonomy, diversity, as use of
bioindicators and many other more available for studying.

VI. Work Plan
A. Study Area
The study area is located in Baungon, Bukidnon, in the province of
Misamis Oriental. There are three sampling sites and is 1-5km away from the
provincial road. On Figure 1, the sites are shown within the province of
Baungon and specified on which barangay it is located. The sampling sites
6

are on: (1) Bayanga (8 21' 31N; 124 36' 5E), (2) Lingating (8 21' 24N; 124
37' 32E) (3) Imbatug (8 19' 1N; 124 40' 58E).

Figure 1. Map of Baungon showing Tumalaong River with the sampling
sites
B. Duration and Frequency of Investigation
Collection of samples will be done on the month of August while
storage, identification, and enumeration of phytoplanktons will be in the
months of August to January (Table 1).
7

Table 1. Gantt Chart of the schedule of activities from June 2014 to March
2015
Activities J J A S O N D J F M
Research Topic Brainstorming & Submission
Literature Review Submission
Reconnaissance
Submission of Project Proposal
Project Proposal Defence
Preparation of Materials
Collection of Samples
Storage for Identification and Enumeration
Enumeration and Identification of
Phytoplankton

Data Analysis
Writing and Submission of Progress Report
Fabrication and Submission of First Draft
Fabrication and Submission of Second Draft
Poster Making
Oral Defence
Submission of Final Paper

C. Methods

1. Collection
8

A 200-meter sampling reach will be first established in each of the 3
sampling sites measured by a calibrated rope. Each reach would then be
divided into 4 equal replicate sites having a length of 50 meters. Samples will
then be taken from the center of the river using a Hose-pipe Depth Sampler
having a length of 4 meters and a diameter of 20 millimeters. The hose will be
weighted on one end with a string attached to the collar. A cork will then be
placed on the opposite end of the weight right after the hose is dipped in the
water at a depth of 1 meter. After the end is sealed, the string will then be
pulled pulling the collar and the other end of the hose along with the water
sample. The collected water will be dispensed in a 1000 mL beaker and
shaken thoroughly. A 1000 mL of the collected sample will be placed in a 1.5
liter dark brown glass bottle. Those sampling bottles will then be placed in ice
and within 8 hours, 3 mL of Lugols solution will be added along with 25 mL of
20% buffered formalin. The sample bottle will be mixed thoroughly and proper
labelling will be done. This process will be repeated for all other replicates and
sites.
2. Preparation and Mounting of Phytoplanktons
Using a chamber called the Lund cell, the phytoplanktons will then be
observed. Two thin pieces of shim brass will be glued to the longer edges of
the slide. A rectangular coverslip will be laid on top to form the chamber. The
volume of the cell chamber would then be calibrated by weighing it before and
after filling it with deionized water. This will be repeated ten times and the
mean volume will be solved. The area of the chamber will then be measured.
A subsample of the original sample will then be placed in the chamber and
after short sedimentation, placed under an upright microscope.
9

3. Enumeration and Identification of Phytoplanktons
Before the identification and enumeration of the phytoplanktons, the
eyepiece graticule should be calibrated with a stage micrometer per objective.
Identification and enumeration of the phytoplankton would be performed and
the visible desired organisms will be counted per random field of view.
Pictures will then be taken and notes will be written as well.
D. Data Analysis
1. Number of algae present

2. Shannon-Weiner Index (H)

Where: p
i
= number of individuals/total number of individuals
ln = natural log or log base e
Shannon-Weiner Index value interpretation (Shanthala, 2009) can be
found on table 2.
3. Simpson Index (D)

Where: p
i
= number of individuals/total number of individuals
Simpson Index value interpretation (Shanthala, 2009) can be found on
table 3.
10

Table 2. Shannon-Weiner Diversity Index
Diversity Level Shannon-Weiner Index
High 3.0-4.5
Moderate 2.0-3.0
Less 1.0-2.0
Very Less 0.0-1.0

Table 3. Simpson Diversity Index
0 (0 to 0.5) index value Lowest possible diversity (when
species are same)
1 (0.5 to 1) index value Highest possible equal number of
different species

VII. Financial Requirements
Table 4. Financial Requirements Table
A. Renumeration of Personnel
Quantity Personnel Duration Rate Amount
1 Adviser 15 times 2,475.00/ consultation 37,125.00
2 Field
Assistants
2 times 300.00/day 1,200.00
SUBTOTAL 38,325.00
B. Operating Expenses
11

1. Operating Materials
Quantity and Unit Particulars Price/Unit Amount
35 pcs. 1.5 Liters Amber
Glass Bottles
500.00/pc 17,500.00
2 pcs. 1000 mL Beakers 1250.00/pc 2,500.00
60 meters Nylon Rope 20.00/meter 1200.00
2 pcs. Lugols Reagent 500.00/50 mL 1000.00
1 Liter 37% Formalin 1000.00/Liter 1000.00
1 pc. Qualpex inch 6
meter roll
1800.00/pc 1800.00
1 pc. Qualpex insert 60.00/pc 60.00
10 pcs. Glass Slides 30.00/pc 300.00
10 pcs. Cover Slip 20.00/pc 200.00
1 pc. Wash Bottle 50.00/pc 50.00
TOTAL 25,610.00
2. Office supplies
Quantity and Unit Particulars Price/Unit Amount
1 pc. Notebook 12.00/pc 12.00
1 roll Masking tape 80.00/roll 80.00
2 pcs. Markers 40.00/pc 80.00
2 pcs. Ballpens 8.00/pc 16.00
2 pcs. Pencils 5.00/pc 10.00
TOTAL 198.00
3. Travel Expenses
12

Quantity and Unit Particulars Price/Unit Amount
1 unit Car Service(rent) 500.00/day 500.00
10 Liters Gas 60.00/Liter 600.00
TOTAL 1,100.00
SUBTOTAL 26,908.00
GRAND TOTAL 65,233.00














13

VIII. Working References
Brierly, G. J. and K. A. Fryirs. 2008. River Futures: An Integrative Scientific
Approach to River Repair (The Science and Practice of Ecological
Restoration Series). 1
st
ed. Island Press. 2-3 pp.

Callow, P. P. 1992. Rivers Handbook: The Science and Management of River
Environments. John Wiley and Sons. Volume 1. 2-3 pp.

Cermeo, P., C. de Vargas, C. F. Abrantes and P. G. Falkowski. 2010.
Phytoplankton biogeography and community stability in the
ocean. PloS one,5(4), e10037.

Elith, J., and J. R. Leathwick. 2009. Species distribution models: ecological
explanation and prediction across space and time. Annual Review of
Ecology, Evolution, and Systematics, 40, 677-697.

Gertraud, H. and R. Croome. 1999. A Phytoplankton Methods Manual for
Austrailian Freshwaters. LWRRDC Occasional Paper 22/99. ISSN
1320-0992

Gordon, R., D. Losic, M. A. Tiffany, S. S. Nagy and F. A. Sterrenburg. 2009.
The glass menagerie: diatoms for novel applications in
nanotechnology. Trends in Biotechnology, 27(2), 116-127.

Gregor, J., and B. Marlek. 2004. Freshwater phytoplankton quantification by
chlorophyll a : a comparative study of in vitro, in vivo and in situ
methods. Water Research, 38(3), 517-522.

Harris, J. M., & P. Vinobaba. 2013. Assessment the Present Status of
Batticaloa Lagoon, Sri Lanka by means of Water Quality, Fish
Diversity Indices and Pollution Indicating Planktons. Journal of
Biodiversity & Endangered Species.

Jost, L. 2010. The relation between evenness and diversity. Diversity, 2(2),
207-232.Hardwood, T. D. 2009. The circular definition of populations
and its implications for biological sampling. Molecular Ecology,
18: 765768. doi: 10.1111/j.1365- 294X.2008.04066.x

14

Korhonen J.J., J. Wang and J. Soininen. 2011. Productivity-Diversity
Relationships in Lake Plankton Communities. PLoS ONE 6(8): e22041.
doi:10.1371/

Likens, G. E. 2010. River Ecosystem Ecology: A Global Perspective. 1
st
ed.
Academic Press. xi-xii pp.

Leo, P., L. Al-Gazali, S. Anand, A. Bittles, J. J. Cassiman, A. Christianson & J.
Schmidtke. 2010. Community genetics. Its definition 2010. Journal of
community genetics, 1(1), 19-22.

Moss, B. 2010. Ecology of Freshwaters: a view for the twenty-first century. 4
th

ed.UK: Blackwell Publishing. 89-91 pp.

Moustaka-Gouni, M., E. Vardaka, E. Michaloudi, K. A. Kormas, E. Tryfon, H.
Mihalatou, and T. Lanaras .2006. Plankton food web structure in a
eutrophic polymictic lake with a history in toxic cyanobacterial
blooms.Limnology and Oceanography, 51(1), 715-727.

Muylaert, K., Tackx, M., & Vyverman, W. (2005). Phytoplankton growth rates
in the freshwater tidal reaches of the Schelde estuary (Belgium)
estimated using a simple light-limited primary production
model. Hydrobiologia, 540(1-3), 127-140.

Naiman, R. J. and R. E. Bill, 1998. River Ecology and Management: Lessons
from the Pacific Coastal Ecoregion. 1
st
ed. Springer-Verlag. 1-2 pp.

Obaje, S. O. and E. A. Okosun. 2013. Taxonomic Notes on Coccolithophorids
from Tomboy Field, Offshore Western Niger Delta,
Nigeria. International Journal of Science and Technology, 2(11).

Offem, B. O., E. O. Ayotunde, G. U. Ikpi, F. B. Ada and S. N. Ochang. 2011.
Plankton-Based Assessment of the Trophic State of Three Tropical
Lakes. Journal of Environmental Protection. doi:10.4236

Paerl, H. W., K. L. Rossignol, S. N. Hall, B. L. Peierls and M. S. Wetz. 2010.
Phytoplankton community indicators of short-and long-term ecological
change in the anthropogenically and climatically impacted Neuse
River Estuary, North Carolina, USA. Estuaries and Coasts, 33(2),
485-497.
15

Phillips, S. J. and M. Dudk .2008. Modeling of species distributions with
Maxent: new extensions and a comprehensive
evaluation. Ecography, 31(2), 161-175.

Rodrigo. 2012. Trade-offs in plankton species richness arising from drought:
insights from long-term data of a National Park wetland (central
Spain).Biodiversity and Conservation, 21(10), 2453-2476.

Rojo, C., M. lvarez-Cobelas, J. Benavent-Corai, M. M. Barn-Rodrguez,
and M. A.

Sandholm, W. H. 2010. Population games and evolutionary dynamics (Vol.
88). Cambridge: MIT press.

Steidinger, K. A., J. H. Landsberg, J. L. Flewelling and B. Kirkpatrick.
2011.Toxic dinoflagellates. Elsevier Science Publishers: New York, NY.

Vaulot, D., W. Eikrem, M. Viprey and H.Moreau. 2008. The diversity of small
eukaryotic phytoplankton ( 3 m) in marine ecosystems. FEMS
microbiology reviews, 32(5), 795-820.







16

IX. Biodata
Name: Arjay Lorenz Legaspi Layawan
Gender: Male
Age: 18
Birthdate: September 18, 1995
Citizenship: Filipino

Educational attainment:
Primary: Shekinah Glory Christian Academy
J.R. Borja, Corrales St., CDO City (S.Y. 2001-2006)
Secondary Shekinah Glory Christian Academy
J.R. Borja, Corrales St., CDO City (S.Y. 2006-2011)
Tertiary Xavier University Ateneo de Cagayan
Corrales Avenue, CDO City
B.S. Biology (S.Y. 2011-Present)
Contact information:
Address: 92-A M.H. Del Pilar Street, CDO City
Mobile Number: 0905-334-4283
Email Address: arjaylayawan@gmail.com