Calawen, Raquel P.

Alalag, Denzel L.

Ayan, Wendy B.

Effect of Effluent from Selected Creeks of Pinsao Pilot Project on Daphnia magna

RESEARCH PLAN

A. Problem being addressed

Wastewater pollution has been a major problem throughout the world. Water

pollution has a negative effect on the delicate balance of the ecosystem. It destroys

organisms that provide foods for other animals. Another effect of water pollution is water

shortage due to pollute water supplies. Harmful substances in water can cause human

illnesses and disease like cholera, typhoid fever or damage to kidney and liver.

Wastewater coming from the different residents will be carried and disposed to thee

bodies of water which include related activities like washing clothes, bathing, (source of

irrigation and like). People also depend on water bodies that may cause complicated

problems since natural water bodies are very important to human. B. Goals

A bioassay will be conducted to determine the toxic effects of effluent to the

mortality of Daphnia magna. This study is important because it provides the complete

response of the test organism to toxic compounds present in wastewater. The effect of the

toxic compound to test organisms could represent the living organisms (especially fauna
and flora) in the bodies of water because of their physiological similarities. Specifically,

the project will focus on the following objectives.

1. Determine if EC50 will be established by any treatment from the two sources of

wastewater.

2. Determine if there are significant difference in the EC50 or mortality means between

and among the treatments (after 48 hours) by the sources of wastewater.

3. Determine which treatment will have the highest and lowest mortality for Daphnia

magna (after 48 hours).

4. Determine which source will have higher and lower mortality for Daphnia magna

(after 48 hours).

C. Methods

Experimental Design

Arranged the experiment in a completely randomized design (CRD) with 2

replications per treatment. The treatments are randomly assigned to the experimental

subject without restriction.

Bioassay Experiment:

Acute toxicity test will be the procedure to be used for the experiment. Acute

toxicity is a short term lethal or other effect, usually defines as occurring within 48 hours

for Daphnia. The standardized protocol for conducting Daphnia magna Toxicity

Bioassays by Beisinger, Williams and Van der Schalie by the EPA (1987) will be
adopted with slight modifications. Wash twenty-four (300 ml) clear glasses that contain

80-ml solution with tap water and then distilled water. Properly label the bottles. Filter

the effluents from Source 1 and 2 separately using filter paper and test solutions should

be prepared by making appropriate dilutions. The treatments were:

T0 - ( as positive control with pure distilled water)

T1 (100 %) – (80 ml wastewater + 0ml distilled water)

T2 (50%) – (40 wastewater + 40ml distilled water)

T3 (25%) – (20 wastewater + 60ml distilled water)

Fill the test bottles with appropriate test solutions. Daphnia (24-hour-old) should

be transferred with a medicine dropper (with an inside diameter of 1.5 times the size of

the daphnids) into the bottles. The randomly add the daphnids into each test bottle until it

contain 4 daphnids. The procedure should be accomplished in less than 1 hour.

During the experiment proper, the daphnids should not be fed as according to the

protocol. The bottles were covered to avoid any organisms to enter the bottle on the time

of the experiment and to minimize the evaporation of the test solutions. More

importantly, the contamination of volatile compounds and air particulates should be

avoided because these are harmful to the test organisms and might cause experimental

errors. This will not deprive the daphnids of oxygen because the test solution didn’t touch

the rim of the bottle leaving a space for the oxygen needed by the test organisms.

Then observations, pH and temperature should be recorded after 5 minutes, 10

minutes, 20 minutes, 1 hour, 2 hours, 3 hours, 24 hours, 48 hours.

 Data Collection Parameters

1. Mortality – number of dead daphnids. Daphnia is identified as dead if it’s immobile.

Immobilization means that no visible movement of appendages when gently prodded.

2. EC50 - a statistically estimated toxicant concentration at which 50 percent of exposed

organisms would be killed a specific time of the observation (48 hours).

3. pH - power of Hydrogen. pH should be measured using a pH paper.

4. Temperature – the hotness or coldness of the solution. Measure the temperature of the

solution using a laboratory thermometer.

Statistical Analysis

Raw data from Source 1 and 2 in the form of mortality means were subjected to Analysis

of Variance (ANOVA) to determine any significant difference between and among

treatments.

D. Bibliography

Akpor O. B. Wastewater Effluent Discharge: Effects and Treatment Processes.

Institute for Economic Research on Innovation, Tshwane University of

Technology, 159 Skinner Street,Pretoria, South Africa, 2011 3rd International

Conference on Chemical, Biological and Environmental Engineering IPCBEE

vol.20 (2011) © (2011) IACSIT Press, Singapore

Biesinger, Kenneth, Williams, Llewellyn, and van der Schalie, March 1987, Procedure
 for conducting Daphnia magna Toxicity Bioassays , Environmental Monitoring

Systems Laboratory: Las Vegas Nevada, Office of Research and Development,

U.S. Environmental Protection Agency

Elenbaas, M. 2013. “Daphnia magna”(On-line), Animal Diversity Web. Accessed

August 8, 2013 at http://animaldiversity.ummz.umich.edu/accounts/....

Marinella Farre and Barcelo Damia. “Toxicity testing of wastewater and

sewage sludge by biosensors, bioassays and chemical analysis .”

<http://www.science direct.com/science/article /pii/S0165993603005041>

Movahedian, Bina, and Asghari. Toxicity Evaluation of Wastewater Treatment Plant

Effluents Using Daphnia magna.

2005.http://journals.tums.ac.ir/upload_files/pdf/_/2015.pdf