Bio Micro

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

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.

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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

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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).

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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

(Cermeño, 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).

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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

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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).

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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

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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 Lugol’s 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.

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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

Number of planktonml−1=Number of organisms countedNumber of replicates

2. Shannon-Weiner Index (H)

H=−∑pi ¿

Where: pi = 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)

D=∑i=1

s

p i2

Where: pi = number of individuals/total number of individuals

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Simpson Index value interpretation (Shanthala, 2009) can be found on

table 3.

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

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B. Operating Expenses

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. Lugol’s Reagent 500.00/50 mL 1000.00

1 Liter 37% Formalin 1000.00/Liter 1000.00

1 pc. Qualpex ½ 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

Quantity and Unit Particulars Price/Unit Amount

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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

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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). 1st 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.

Cermeño, 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. Maršálek. 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: 765–768. doi: 10.1111/j.1365-294X.2008.04066.x

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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. 1st 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. 4th 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. 1st 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.

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Phillips, S. J. and M. Dudík .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. Álvarez-Cobelas, J. Benavent-Corai, M. M. Barón-Rodríguez, 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 μm) in marine ecosystems. FEMS microbiology reviews, 32(5), 795-820.

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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: [email protected]

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