Sengupta, M. and Dalwani, R. (Editors). 2008 Proceedings of Taal2007: The 12th World Lake Conference: 484-489

Biodiversity of Protozoa in Freshwater Case Study in the Chao-Praya River, Thailand Yuwarad Polameesanaporn Department of Environmental Science, Rajabhat-Suan-Dusit University, 295 Rachaseema Rd., Dusit, Bangkok 10300 Email: muayyai@hotmail.com, www.dusit.ac.th

ABSTRACT Chao-Praya River is a major river flowing through many provinces in the central region including Bangkok and Nonthaburi. This river is very important for Thai people as a water resource for Agricultural, Residential and Household used. The researcher selects Nonthaburi as a case study because the area usage of this province has changed from agricultural usage in the past to residential and commercial usage these days. Some area has also been arranged for industrial zone.

The water samples have been collected from nine areas classified by land usage and season, follow the Nonthaburi Land Usage Map in 2005. There are two seasons, which are drought (from December 2005 to February 2006) and rainy season (from June 2006 to August 2006). We studied the relationship between the diversity of protozoa and physical/chemical properties of water. The results showed that the diversity of protozoa in agricultural area is higher than other areas. We also found greater number of protozoa in rainy season than in drought. The diversity of protozoa in Class Euglena and Class Phacus could be used as bio-indicators for Mesosaprobic Zone water resources follows Saprobic System. From the result we can conclude that this area is medium deteriorate water resource. Keywords: Biodiversity/ Bio-indicator/ BOD/ Chao-Praya River/ Do/ Protozoa/ Saprobic System

INTRODUCTION The Chao-Praya River is the major river located in the central region of Thailand. This river, which has 20,125 squared kilometers, has more than 22 billion cubic meters water flowing through 11 provinces covering some part of Nonthaburi province. This paper discusses about the biodiversity of protozoa in this river located in Nonthaburi. This province is located close to Bangkok, thus there is a lot of water utilization such as transportation purposes, industrial usage, household usage and agricultural usage. All these usages affect to the quality of water in the river.

Current status of water in this river depends on the utilization. There is a lot of waste in form of organic, inorganic and pollutant from mentioned activities. Therefore, the quality of water has direct impact to the diversity of protozoa in the water. The researcher chose protozoa as an indicator of the water quality because of their genome, which is in Eukaryote cell, is similar to the genome of higher level animal than the genome of Prokaryote cell. Protozoa has high growing rate and has thin call wall. This makes them response to the changes of environment quickly and need specified environment for survival. Farmer (1980) found that Synura Uvella and Stentor Polymorphus are protozoa in high level oxygen. Bopit and Nuntaporn (1989) studied class of protozoa which could be used as indicator of water quality in Bung Magasun, an area in Bangkok. They confirmed the existing of protozoa class Metopus in

Polysaprobic zone, or so called low-oxygen water resource. As researchers know that Protozoa has fast response to the changes of environment, therefore the use of protozoa as bio-indicator is very popular in this field. They can benefit from the cheap investment, the accuracy of result and save time.

There are many researchers in Thailand which had done water quality survey using Protozoa diversity as indicator. Poowadon (2003) studied the diversity of protozoa in Nam-Pong-river, Khonkhan province during July 2001 to July 2002 (12 months). He found 13 classes of protozoa such as Class Euglenoidea and Class Lobosea. Intira (1999) studied the diversity of protozoa and quality of water in Klong-Mae-Kar, Chiang-Mai province during December 1997 to November 1998. She found 4 classes of protozoa such as class Ciliata, class Mastigophora and class Sarcodina. Chidchai (2000) studied the quality of water and found protozoa in Chula-Longkorn-university. He found more than 60 classes. He classified the water quality based on Saprobic system. He divided the water quality into 4 zones which are Polysaprobic zone (BOD 10 – 50 milligrams/liter), α- mesosaprobic zone (BOD 5-10 milligrams/liter), β- mesosaprobic zone (BOD 2.5 – 5 milligrams/liter) and Oligosaprobic zone (BOD 1-2.5 milligrams/liter). The result showed the BOD is around 7.6 – 35.1 milligrams/liter. Thus, he can conclude that this water resource is in α-mesosaprobic Zone and Polysaprobic Zone. They also found Euglena and Phacus as imminent class.

The result corresponded to previous research such as Margulis, Corliss, Melkonian, Chapman, 1990, Curds 1992 (refer to Chidchai (2000)) found that Euglena and Phacus are bio-indicators of α-mesosaprobic Zone and Polysaprobic Zone METHODOLOGY The Chao-Praya River is the major river of Thailand, which flows pass Nonthaburi province, has various usage and many kinds of waste water flowing into

the river and impact to the quality of water. Therefore, the sample collection was done follow the land usage map (2005) from the civil department of Nonthaburi as shown in figure 1. Sample Collection Areas There are 9 stations to collect the sampled water from Rama VII bridge to Wat-Poramaiyikawas-Worawihan and Klong Bangkok-Noi to Klong Aomnon Nonthaburi as shown in the table 1.

Station 5Wat Yai Sawang Arrom

Station 4 Wat Chueng LaneStation 3 Phra-Nang Klaw Bridge

Station 6 Wat Polamaiyikawasworawiharn

Station 1 Rama VII BridgeStation 7 Wat Chalor

Station 8 Bang Muang

Station 2 Rama V Bridge

Station 9 Wat Molee

Figure 1 Water sample collecting area of Chao-praya river in Nonthaburi province

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Table 1: Water sample collecting areas and their location Sampled Area Location (Lat/Lon)

13.814360 1. Rama VII Bridge

100.514210 13.8316903

2. Rama V Bridge 100.4945988 13.8699395

3. Phra-Nang-Klao Bridge 100.4761356 13.8992270

4. Wat Chueng Lane 100.4880912 13.9121661

5. Wat Yai Sawang Arrom 100.4665662 13.9142558 6.Wat

Poramaiyikawasworawiharn

100.4886777

13.8189489 7. Wat Chalor

100.5013255 13.8080530

8. Bang Muang 100.4765500 13.8497379

9. Wat Molee 100.4239779

Water Quality Study Process The sampled water is collected at described stations by collecting water at the center of Chao-praya river at the depth around center of the river. The sample is collected during drought season (December 2005 – February 2006) and rainy season (June 2006 – August 2006). They have been collected once a month and analyzed for parameters such as transparency and collect the sampled water in 1 liter bottle stored in controlled temperature container. This container is controlled at 4 degree celsius. After that sampled water is analyzed in laboratory to find the Biochemical Oxygen Demand (BOD) by Alkali-Iodide-Azide, Total Phosphorus and Ascorbic acid

methods. They use UV-Spectrophotometer/UV 1601 machine, Total Nitrogen by Kjeldahl method and Dissolved oxygen in water (DO) in the analysis. Protozoa Studied Method Water was horizontally sampled collected on water surface. We used 40 microns plankton net and keep the water sample using 4% concentration formaldehyde and tighten the container. Protozoa was examined using 10 x – 40 x microscope and record the shape, structure and movement organ. The organelles were also recorded for classification using method in Patterson (1996), Farmer (1980), Nantaporn Jarupan (2004) and Bopit and Nantaporn Jarupan (2006) RESULTS The results of water colleted at Chao-Phaya River at Wat Poramaiyikawasworawihan to Rama VII bridge and Bangkok-Noi to Klong Oam-nont in drought season showed that the transparency index was 36-70 centimeters, dissolved oxygen 1.80 – 6.0 milligrams/liter, BOD 2.0-4.6 milligrams/liter, Total phosphorus 0.109 – 0.267 milligrams/liter and total nitrogen 1.007-1.095 milligrams/liter In rainy season, the transparency index is 32-53 centimeters, dissolved oxygen is 2.1-6.5 milligrams/liter, BOD is 2.3 – 6.3 milligrams/liter, Total phosphorus is 0.1.2 – 0.260 milligrams/liter and total nitrogen is 1.119-2.394 milligrams/liter

We found both Autotroph and Hetrotroph protozoa in the river. We found 4 classes, 32 species which are class Phytomastigophora (23 species), class Lobosea (8 species), class Sprortricsea (1 species) and class Oligohymenophorea (1 species) In addition, 7 classes, 50 species protozoa were found in the rainy season which are class Phytomastigophorea (36 species), class Lobosea (7 species), class Spirotricsea (2 species), class Oligophymenophorea (1 species), class Heliozoea (2 species), class Prostomatea (1 species) and class Litostomata (1 species).

0

5

10

15

20

25

30

35

40

1 2 3 4 5 6 7 8 9

Station

Spec

ies Drought Season

Rainy Season

Figure 2: Biodiversity of Protozoa

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Table 2: Protozoa species found and water quality studies

DO BOD Transparency Index

Total phosphorus

Total nitrogen

(mg/l) (mg/l) (centimeter) (mg/l) (mg/l)2.2-3.9 2.2-2.3 65-67 0.122-0.132 1.034-1.039(3.05) (2.25) (66) (0.13) (1.04)2.1-2.4 3.2-3.6 34-36 0.236-0.252 1.132-1.134(2.25) (3.4) (35) (0.24) (1.13)

2.7-5.15 2.0-2.1 50-55 0.156-0.159 1.007-1.036(3.92) (2.05) (52.5) (0.16) (0.52)2.4-2.7 2.8-3.2 39-42 0.184-0.234 1.123-1.127(2.55) (3) (40.5) (0.21) (1.13)

3.7-5.05 2.1-2.4 39-42 0.110-0.121 1.029-1.035(4.38) (2.25) (40.5) (0.12) (1.03)2.2-2.7 2.8-3.1 39-42 0.102-0.201 1.119-1.123(2.45) (2.95) (40.5) (0.15) (1.12)2.8-5.4 2.3-2.5 62-70 0.114-0.115 1.034-1.037(4.1) (2.4) (66) (0.11) (1.04)

5.6-6.4 2.9-3.2 50 0.193-0.245 1.129-1.142(6) (3.05) (50) (0.22) (1.14)

3.2-5.2 2.1-2.3 57-65 0.109-0.120 1.019-1.025(4.2) (2.2) (61) (0.11) (1.02)

4.8-6.1 2.3-2.9 33-53 0.152-0.244 1.132-1.210(5.45) (2.6) (43) (0.2) (1.17)3.8-6.0 2.3-2.5 54-56 0.182-0.202 1.035-1.042(4.9) (2.4) (55) (0.19) (1.04)

5.2-6.5 2.7-3 32-38 0.159-0.258 1.173-1.310(5.85) (2.85) (35) (0.21) (1.24)

2.55-3.35 4.2-4.3 55-56 0.245-0.267 1.047-1.050(2.95) (4.25) (55.5) (0.26) (1.05)2.6-3.2 5.4-6.1 34-36 0.250-0.260 2.225-2.231(2.9) (5.75) (35) (0.26) (2.23)

1.80-3.7 4.2-4.6 36-37 0.257-0.260 1.037-1.095(2.75) (4.4) (3.65) (0.26) (1.07)2.2-3.8 5.1-6.3 35-39 0.235-0.260 2.263-2.281

(3) (5.7) (37) (0.25) (2.27)2.55-3.95 4.2-4.5 56-58 0.195-0.210 1.015-1.036

(3.25) (4.35) (57) (0.2) (1.03)2.8-3.2 5.6-6 37-40 0.235-0.254 2.368-2.394

(3) (5.8) (38.5) (0.24) (2.38)

Station

Wat Molee

Wat Yai Sawang Arrom

Wat Poramaiyikawas

Wat Chalor

Bang Muang

Rama VII Bridge

Rama V Bridge

Phra Nang Klao Bridge

Wat Cheung Lane

Drought season 22

Rainy season 28

Drought season 15

Rainy season 20

Drought season 17

Rainy season 24

Drought season 32

Rainy season 34

Drought season 31

Rainy season 33

Drought season 31

Rainy season 38

Drought season 25

Rainy season 32

Drought season 26

Rainy season 30

Protozoa Species

Drought season 21

Rainy season 26

Season

Numbers in the bracket are estimated values. CONCLUSIONS AND DISCUSSIONS Seasonal Effect The diversity of protozoa found in the rainy season is greater than found in the drought season as shown in figure 2. This is because of the amount of phosphorus and nitrogen in water in the rainy season is greater than in drought season as shown in table 2. These two substances are the major substances for Protozoa (self-food-generated: ic organisms). In addition, light plays an important role in food generation of Phytoflagellate, which need light in food generation process. The food generation process of Autotroph is affected by the transparency of water because of water in the drought season is transparent than in the rainy season as shown in figure 3. Each protozoa species also have different amount of light needed in food generation process. For example Euglena and Volvox like minded sun light. It will flow on water surface when sun shine in small amount and it will go in deep water when the sun light is strong. This is the nature of protozoa.

There also has much micro-organism which is food for protozoa which cannot generate food (Hetorotroph). The micro-organism in water can breed well in the environment by decompose the

organic substance in the water. Thus, it could be food for protozoa and indirectly help protozoa to breed. This makes BOD value, as shown in figure 4, in the rainy season greater than in drought season. The more organic decomposition, the more bacteria could be found in the water and also reflected by the low DO number. However, the bio-diversity does not relate to this. Land Usage Effects

From the results, as shown in figure 2 and table 2, we can conclude that the diversity of protozoa depends on the land usage and the quality of water. In the station 4 (Wat Chueng Lane), station 5 (Wat Yai Sawang Arrom) and station 6 (Wat Poramaiyikawasworawiharn), which are in agricultural area, the quality of water is better than other areas. The DO number is around 5 milligrams/liter, BOD is 2.58 milligrams/liter. Therefore, the diversity of protozoa is greater than other areas. In the station 2 (Rama V bridge) and station 3 (Phra Nang Klao bridge), which are medium populous-residential areas, have DO number around 3.3 milligrams/liter, BOD is 2.8 milligrams/liter.

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0

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50

60

70

1 2 3 4 5 6 7 8 9

Station

Tran

spar

ency

Inde

x

Drought Season

Rainy Season

Figure 3: Transparency Index

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1

2

3

4

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1 2 3 4 5 6 7 8 9

Station

BO

D/D

O (

mg

/l) DO (Drought Season)

DO (Rainy Season)

BOD (Drought Season)

BOD (Rainy Season)

Figure 4. BOD and DO Value

The diversity of protozoa is less than the first group area. The last group which has less diversity of protozoa includes station 1 (Rama VII bridge), Station 7 (Wat Chalor), Station 8 (Bang Muang) and Station 9 (Wat Molee). These areas are commercial area and crowded residential area. The DO number is approximately 2.8 milligrams/liter and BOD is 4.5 milligrams/liter. This shows that the usage of water has direct effect to the diversity of protozoa. The researcher analyzes the results follows the Saprobic system, which use BOD value in the classification. The results show that the quality of Chao-Praya River in Nonthaburi area can be classified as Mesosaprobic zone, which has BOD around 2.5-10 milligrams/liter. We can found Euglena class and Phacus class in every station which could be use as bio-indicator for this kind of water.

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