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FACULTY OF CIVIL AND ENVIRONMENTAL ENGINEERING

DEPARTMENT OF WATER RESOURCES AND
ENVIRONMENTAL ENGINEERING

ENVIRONMENTAL ENGINEERING LABORATORY

LABORATORY INSTRUCTION SHEETS

SUBJECT CODE BFC 3121
EXPERIMENT CODE MA2
EXPERIMENT TITTLE BACTERIA COUNT
COURSE CODE

Saya juga mengaku tidak menerima atau memberi sebarang bantuan dalam menyediakan laporan ini dan membuat ikrar ini dengan kepercayaan bahawa apa- apa yang tersebut di dalamnya adalah benar. KEJ. Matrik :____________________________ Tarikh :________________________________ FACULTY OF CIVIL & ENVIRONMENTAL ENGINEERING . SUMBER AIR & ALAM SEKITAR FAKULTI KEJURUTERAAN AWAM & ALAM SEKITAR KUiTTHO Saya dengan ini mengaku bahawa saya telah menyediakan laporan ini dengan daya usaha saya sendiri. ___________________________ Tandatangan Pelajar Nama : _______________________________ No. KOD ETIKA PELAJAR (KEP) JAB.

3. DATE: 20/11/2007 .OF GROUP GROUP MEMBER 1. 5. 4. DEPARTMENT OF WATER RESOURCES AND ENVIRONMENTAL ENGINEERING ENVIRONMENTAL ENGINEERING LABORATORY SHORT REPORT SUBJECT CODE CODE & EXPERIMENT TITLE COURSE CODE EXPERIMENT DATE NAME OF STUDENT NO.: 03 EFFECTIVE DATE: 01/12/2007 EXPERIMENT : BACTERIA COUNT AMMEND. NAME OF LECTURER/ INSTRUCTOR/TUTOR DATE OF SUBMISSION MARKS ATTENDANCE & /10% DISCIPLINE INTRODUCTION /5% RESULTS /15% DATA ANALYSIS /15% DISCUSSION /25% CONCLUSION /5% REFERENCES /5% TOTAL /80% EXAMINER’S COMMENT APPROVAL RECEIVE FACULTY : CIVIL & ENVIRONMENTAL PAGE NO.: 1/5 ENGINEERING DEPT : WATER RESOURCE & EDITION: MA2 ENVIRONMENTAL ENGINEERING LAB : ENVIRONMENTAL ENGINEERING REVISION NO. 2.

2. called absorbance or optical density. and this is indicated on a galvanometer.0 OBJECTIVE Students will be able to measure the bacteriological quality of water sample by performing total plate count. A wide series of dilutions (e.0 LEARNING OUTCOMES At the end of the laboratory courses. Be more proficient at dilutions. the number of colonies should give the number of bacteria that can grow under the incubation conditions employed. in arctic ice and glaciers. This method is faster than the standard plate count but it has limitation where sensitivity is restricted to bacterial suspensions of 107 cells or greater. the standard plate count method is an indirect measurement of cell density and reveals information related only to live bacteria. in hot springs. or viable. The spectrophotometric analysis is based on turbidity and indirectly measures all bacteria (cell biomass). there are distinct differences.. 10-4 to 10-10) is normally plated because the exact number of bacteria is usually unknown.: 03 LAB : ENVIRONMENTAL ENGINEERING EFFECTIVE DATE: 01/12/2007 EXPERIMENT : BACTERIA COUNT AMMEND. The standard plate count method consists of diluting a sample with sterile saline or phosphate buffer diluent until the bacteria are diluted enough to be counted accurately. The assumption is that each viable bacterial cell is separate from all others and will develop into a single discrete colony (CFU).1. Our understanding of bacteria and their metabolic processes has been expanded by the discovery of species that can live only deep below the earth's surface and by species that thrive without sunlight or in the high temperature and pressure near hydrothermal vents on the ocean floor. plate count method and spectrophotometric (turbidimetric) analysis. By using a spectrophotometer. the amount of transmitted light decreases as the cell population increases. and even in the stratosphere. DATE: 20/11/2007 . There are more bacteria. dead and alive. PREPARED BY: BALKIS A. the final plates in the series should have between 30 and 300 colonies. The transmitted light is converted to electrical energy. For example. students will be able to : 1.5 billions of bacteria in one gram of fertile soil. Increased turbidity in a culture is another index of bacterial growth and cell numbers (biomass). Greater accuracy is achieved by plating duplicates or triplicates of each dilution. there can be as many as 2.0 THEORY Bacteria are remarkably adaptable to diverse environmental conditions: they are found in the bodies of all living organisms and on all parts of the earth–in land terrains and ocean depths. Many studies require the quantitative determination of bacterial populations.: 2/5 ENGINEERING DEPT : WATER RESOURCE & EDITION: MA2 ENVIRONMENTAL ENGINEERING REVISION NO. and more than 300 colonies on a plate are likely to produce colonies too close to each other to be distinguished as distinct colony-forming units (CFUs).g. Hence. than any other type of organism. indirectly reflects the number of bacteria. Thus. Fewer than 30 colonies are not acceptable for statistical reasons (too few may not be representative of the sample). as separate individuals. The two most widely used methods for determining bacterial numbers are the standard. The reading. 3. Although the two methods are somewhat similar in the results they yield. TALIP SIGNATURE: DATE: 20 NOVEMBER 2007 FACULTY : CIVIL & ENVIRONMENTAL PAGE NO. 2. Be more proficient at performing a standard plate count and determining bacterial counts in a sample.

2 Sample preparation: Prepare the serial dilution of the water sample using the appropriate dilution factor.: 03 LAB : ENVIRONMENTAL ENGINEERING EFFECTIVE DATE: 01/12/2005 EXPERIMENT : BACTERIA COUNT AMMEND. Sterilizer 9.: 4/5 DEPT : WATER RESOURCE & ENVIRONMENTAL ENGINEERING EDITION: MA2 REVISION NO. Bunsen burner 6. Glass rod 5. Microscope 10. FACULTY : CIVIL & ENVIRONMENTAL ENGINEERING PAGE NO.4. Agar=15g. Incubator 7.1 Media preparation: Please prepare the nutrient media using the microbiology standard method. Pipette 3. Petri plate 2. Ethanol 95% @ methanol 8.0 EQUIPMENTS & MATERIALS 1. Distilled water=600 mL 5. Test tube 4. Bacteria medium: Peptone = 5g. DATE: 20/12/2005 .3 Plating procedures: Using the pour plate and spread plate method.0 PROCEDURES 5. 5. Beef Extract=3g. 5.

3. 2. lake water. 4. 2.0 QUESTIONS 1. swimming pool water and rainbarrel water). Show the calculation for each of the plating method and fill in the above table. Explain your findings. Explain the difference of bacteria count for each type of water sample? 4. the results give different readings for both methods. Analyze the results by using appropriate method. 3. Usually. What the meaning of TNTC and the significant amount due to the TNTC? Give the formula for determining bacteria count. Based on this experiment what is the suitable control? How will the control affect 1/100 1/10 your findings? . 8. in some cases. Explain the meaning of a phrase “two times ten to the eight cells per mL” in your own convenient terminology.0 RESULTS & CALCULATIONS / ANALYSIS Plating Average Dilution Total Method Colony bacteria 1/10 1/100 1/1000 1/104 1/105 1/106 /plate / mL Pour 119 190 175 155 85 75 35 Plate Spread 125 200 180 160 90 80 40 Plate 7. However.0 DATA ANALYSIS 1. Design an experiment to compare the bacteria counts in different water samples (tapwater. State the systematic bias error that could occur during this experiment. Explain why the results are indistinguishable. both methods produce the same results. 6. In many experiments there are 2 types of control used which are positive and negative control.