Lab Exercise: Examining Water Quality: Most Probable Number & Colilert Test Kit Lab
|
|
- James George
- 6 years ago
- Views:
Transcription
1 Lab Exercise: Examining Water Quality: Most Probable Number & Colilert Test Kit Lab OBJECTIVES 1. Understand the use of MPN to determine likely fecal water contamination. 2. Understand the use of MUG, ONPG and β-galactosidase in the Colilert test kit. 3. Understand the use of positive and negative controls in this experiment. 4. Determine the cleanliness of the experimental water sample. BACKGROUND Most Probable Number (MPN) is a procedure used to estimate bacterial populations in the environment (including bacterial contamination of water). This procedure is based on the application of the Theory of Probability with the following given assumptions. 1. The organisms are randomly and evenly distributed throughout the sample; 2. The organisms exist as single entities, not as chains, pairs or clusters and they do not repel one another; 3. The proper medium, temperature and incubation conditions have been selected to allow even a single viable cell in an inoculum to produce detectable ; 4. The population does not contain viable, sub-lethally injured organisms that are incapable of in the culture medium used. Samples are diluted prior to use in order to reduce the number of positive tubes to a manageable number. The number of tubes prepared is generally based on the expected population contained within the sample. Reliable results occur when all tubes at the lower dilution are positive and all tubes at the higher dilution are negative for, as the dilution scheme has accurately bracketed the population, much as the case in dilution plating. Typically, this results in serial dilutions for MPN analysis in sets of 3, 5 or 10 MPN tube series, with more tubes increasing accuracy. MPN values are an indirect measurement of the number of viable organism in a given sample. However, MPN counts are useful for their technical ease and are particularly useful when low concentrations of organisms (<100/ml) are encountered in such materials as milk, food, soil and water where a low concentration of organisms and/or particulate matter of the matrix may interfere with accurate colony counts. Once dilutions have been performed, samples that are positive for microbial are indicated by gas production and/or visible turbidity. In this lab, you will be inoculating a series of 3 MPN tubes with the sample serial diluted (i.e., undiluted; a 10-1 dilution; and a 10-2 dilution). You will be looking for the presence of aerobic or facultatively anaerobic, Gram negative, non-endospore forming rods that ferment lactose and produce gas within 24 hours at 35 C. Organisms having these characteristics are typically considered indicator organisms for fecal contamination of water. However, there are many organisms that fit this description but are not indicative of fecal contamination. For this reason, positive MPN results are a putative result for fecal contamination. Confirmation of this must be done by further biochemical and physiological analysis of the cultured microorganisms. More sophisticated testing can be directly on water samples through the use of test kits which examine the bacteria s ability to metabolize lactose (and indicator substrates) into specific chemical compounds. Remember from earlier laboratory discussions that when bacterial cells are exposed to lactose, the lac operon is induced, in order that the cells may begin the process of metabolizing this sugar. One of the enzymes coded for in the lac operon is β-galactosidase, which converts the disaccharide lactose to the monosaccharides glucose and galactose. Thus, the catabolism of lactose into glucose and galactose in a given water sample is typically indicative of the presence of fecal contamination. Cells will also produce β- galactosidase when exposed to the nutrient-indicator ONPG (ortho-nitrophenyl-β-d-galactopyranoside), which mimics the lactose molecule and is broken down by β-galactosidase into ONP (ortho-nitrophenyl-β-dpyranoside), which is yellow in color ( Figure 1). In this way, the conversion of ONPG to ONP, which is easily detectable with the naked eye (Figure 2), is an indicator reaction for the presence of coliform contamination. (For reference, the ONPG
2 works in the same manner as IPTG in Lab Exercise 15: pbluescript Transformation: Blue/White Colony Selection.) Figure 1: Chemical representation of the catabolism of either lactose (upper diagram) or ONPG (lower diagram) by the Bacterial enzyme β-galactosidase into their respective catabolites. Figure 2: Left to right: Colilert test results for E. coli (under fluorescent light); a sample containing noncoliform Bacteria; and a sample containing coliform Bacteria (shown under visible light). Additionally, the indicator organism Escherichia coli will use β-galactosidase to convert a second indicatornutrient MUG (4-methylumbelliferyl-β-D-glucuronide) into its fluorescent counterpart (Figure 2). Although this test will confirm the presence of the fecal contaminant, E. coli, the ability to convert MUG to a fluorescent compound is lacking in most pathogenic E. coli strains, and so additional tests are usually required to single them out. A commercially available test kit, Colilert, which contains both ONPG and MUG can be used to easily determine the presence of coliforms as well as E. coli. The sensitivity of this test kit is such that it can detect E. coli and coliforms at concentrations of 1 CFU/ml within 24 hours, with as many as 2x10 6 other bacterial cells/ml present. It is important to note that both of these tests are sensitive only to bacterial water contamination. Fecal contamination can also result in the transmission of viral and eukaryotic pathogens. INTRODUCTION During this laboratory activity, we will be looking for typical biological markers of fecal water contamination, including the enteric bacterium, Escherichia coli. The class will work as a team to investigate the cleanliness of an environmental water sample compared to both positive (water inoculated with E. coli) and negative (sterile water) control samples. We will determine the cleanliness of the experimental water sample using both MPN and Colilert data. Water samples will be inoculated into Durham lactose tubes with Colilert reagents and will be incubated for 48 hours at 35ºC. Data will be collected and analyzed in order to determine the likelihood of fecal contamination of the water. Remember that in lactose broth is only a putative diagnosis, as this is not a selective media- and will allow most bacteria to grow. Further analysis can be done using the more
3 specific tests where ONPG and MUG are broken down selectively by enteric bacteria generally and E. coli specifically. In order to analyze the data, you will be looking for visible changes in the water samples for: visual color changes, as ONPG is broken down to ONP by fecal/enteric bacterial contaminants; turbidity and gas production, as lactose is consumed and gas is produced by microorganisms; and fluorescence, as MUG is broken down by E. coli. These results can be compared to the expected results in Table 2 below to calculate the most probable number (MPN) of organisms per milliliter of water. Additionally, Table 3 can be used to determine the specific presence of fecal contamination and E. coli as a measure of color change and fluorescence due to the addition of Colilert reagents. Class assignments are as follows: table 1 will be analyzing a positive control, table 2 will be analyzing the experimental water sample and table 3 will be analyzing a negative control. PROTOCOL Team Supplies 1 Colilert pack 100ml water sample (sterile water for 2 teams, environmental water samples for 4 teams) in Milk Dilution Bottles Escherichia coli 3 lactose Durham tubes 1. Experimental teams: add the contents of 1 Colilert pack to the 100ml Milk Dilution Bottle water sample. Label this water experimental. Cap the sample and shake vigorously. 2. Positive and negative control teams: add Colilert pack to 100ml Milk Dilution Bottle sterile water. Label this water control. Cap the sample and shake vigorously. The control water can be shared for the positive and negative control inoculations. Please plan accordingly to share reagents with your neighbors. 3. Collect and label your Lactose Durham tubes. Positive control team: Preparation of Positive Control Samples (Figure 3) 1. Add 10ml control water to 3 Lactose broth tubes. Label these tubes positive control. 2. Add 1 loopful of E. coli to each of the positive control tubes. 3. Incubate the sample at 37 C for 48 hours. 4. Incubate the Milk Dilution bottle at 37 C for 48 hours Negative control team: Preparation of Negative Control Samples (Figure 3) 1. Add 10ml control water to 3 Lactose broth tubes. Label these tubes negative control. 2. Incubate the sample at 37 C for 48 hours. 3. Incubate the Milk Dilution bottle at 37 C for 48 hours
4 Figure 3: Diagram of dilution protocol for preparation of Control Samples. Experimental sample teams: Preparation of Experimental Samples (Figure 4) Undiluted Samples 1. Aseptically pipette 10ml of the experimental water into 3 Lactose Broth tubes Samples 1. Add 1ml experimental water to each of the tubes Samples 1. Add 100 l (0.1ml) experimental water to each of the tubes. 2. Incubate all 9 of the sample at 37 C for 48 hours. 3. Incubate the Milk Dilution bottle at 37 C for 48 hours
5 Figure 4: Diagram of dilution protocol for preparation of Experimental Samples. DATA AND OBSERVATIONS 1. Collect florescence data (MUG) by reading the absorbance of the samples at 260nm using the Spec Collect visual color change data (ONP) by reading the absorbance of the samples at 570nm using the Spec Collect turbidity data by reading the absorbance of the samples at 600nm using the Spec Input your data into the appropriate table below, and then get class data for the other 5 samples (total of 6: 4 experimental, 1 positive, 1 negative) Experimental sample 1: Sample site Table 1: Data collection for each of the sample test tubes for the experiment. = average value. Experimental sample 2: Sample site Experimental sample 3: Sample site
6 Experimental sample 4: Sample site Positive control Negative control 5. Count the number of tubes for the experimental water sample that have turbidity and/or gas production. Each turbid tube is considered a positive result. Record these data in a table similar to that shown in Table 2. Experimental sample 1: Table 2: Data collection for each of the sample test tubes for the experiment for turbidity. Mark each tube with with a + and each tube with no with a are the undiluted samples. Experimental sample 2: Experimental sample 3: Experimental sample 4:
7 Positive control: Negative control: 6. Calculate the average absorbance for both visual and ultraviolet wavelengths of light for each of the three sample dilutions (i.e. undiluted; 10-1 ; 10-2 ). Input these data into Table 2 Table 1 above. 7. Using Table 4 below, determine the MPN of the samples. 8. Analyze your colorimetric data according to Table 3. appearance less yellow than the positive control more yellow the positive control more yellow & florescent the positive control result negative for total coliforms and E. coli positive for total coliforms positive for E. coli Table 3: Colorimetric changes of water samples due to the addition of the Colilert reagents ONPG and MUG and their subsequent catabolism by bacteria. DISCUSSION 1. Determine the MPN for the water sample using the data collected in Table 2 and analyzing it according to the MPN table (Table 4). Experimental sample 1: Experimental sample 2: Experimental sample 3: Experimental sample 4: Positive control: Negative control: 2. Was the water sample you collected contaminated? 3. What do both yellow color change and fluorescence indicate about a water sample?
8 positive tubes MPN/ml lower 95% CI a < upper 95% CI positive tubes MPN/ml lower 95% CI a upper 95% CI a CI is an abbreviation for Confidence Interval, the range of values within which you have a statistically accurate chance of predicting the actual population number. Table 4: MPN Table. Data are collected for tubes displaying for each dilution series. As an example, 3 positive undiluted tubes, 2 positive 10-1 tubes and 0 positive 10-2 tubes would yield an MPN of 93 with lower & upper confidence limits of 18 and 420 respectively.
INTRODUCTION water-soluble Figure 1.
INTRODUCTION Natural waters contain bacteria. The aerobic gram negative bacillus of the genera Psedomonas, Alcalignes, and Flavobacterium are common in natural waters. Many of these bacteria are able to
More informationIDEXX is an ISO certified facility. Gil Dichter World Wide Technical Support Manager
USEPA ATP Process for Approval of Colilert-18 and Quanti-Tray for the Detection of Fecal Coliforms in Waste Water; What is a Number: Comparison of MPN and CFU Gil Dichter World Wide Technical Support Manager
More informationExercise 13 DETERMINATION OF MICROBIAL NUMBERS
Exercise 13 DETERMINATION OF MICROBIAL NUMBERS Introduction When biologists discuss the growth of microorganisms (microbial growth), they are actually referring to population size rather than to the size
More informationAdapted from Biology 15 Laboratory Manual Supplement: Wrightsman, Ininns and Cannon-Moloznic, Saddleback College, CA 92692
Biology 4B Laboratory Bacteriological Examination of Water Adapted from Biology 15 Laboratory Manual Supplement: Wrightsman, Ininns and Cannon-Moloznic, Saddleback College, CA 92692 Objectives Carry out
More informationA Discovery Laboratory Investigating Bacterial Gene Regulation
Chapter 8 A Discovery Laboratory Investigating Bacterial Gene Regulation Robert Moss Wofford College 429 N. Church Street Spartanburg, SC 29307 mosssre@wofford.edu Bob Moss is an Associate Professor of
More informationSELECTED QUESTIONS F ROM OLD MICRO 102 QUIZZES PART I EXPERIMENTS 1 THROUGH 7
SELECTED QUESTIONS F ROM OLD MICRO 102 QUIZZES PART I EXPERIMENTS 1 THROUGH 7 Question numbers refer to the applicable experiment. Questions with blanks are multiple true-false questions unless otherwise
More informationENV H 433 LABORATORY EXERCISE 1
ENV H 433 LABORATORY EXERCISE 1 Multiple tube fermentation and presence/absence methods to detect total coliforms, fecal coliforms and Escherichia coli I. LABORATORY GOAL To determine concentration of
More informationINTRODUCTION Contaminated serial dilution countable plates
INTRODUCTION In recent days, the foods that we consume are usually pre-processed in a facility removed from our home, cities, countries, and even continents. It is now more than ever important to be aware
More informationModified Colitag Test Method for the Simultaneous Detection of E. coli and other Total Coliforms in Water (ATP D ) August 28, 2009
Modified Colitag Test Method for the Simultaneous Detection of E. coli and other Total Coliforms in Water (ATP D05-0035) August 28, 2009 1 Rev 08/28/2009 Modified Colitag Test Detection of E. coli and
More informationCARBOHYDRATE FERMENTATION TEST
Microbiology Laboratory (BIOL 3702L) Page 1 of 6 Principle and Purpose CARBOHYDRATE FERMENTATION TEST Microorganisms need to generate energy in order to grow, divide, and survive. In any given environment,
More informationCOMPARATIVE STUDY ON THE MICROBIOLOGICAL QUALITY OF WATER COOLER DISPENSERS AND TAP WATER
COMPARATIVE STUDY ON THE MICROBIOLOGICAL QUALITY OF WATER COOLER DISPENSERS AND TAP WATER research paper ELENA TRAISTARU 1 GemAnalysis Ltd, Nicosia, Cyprus Abstract: The microbiological quality of water
More informationMICROBIOLOGICAL DETECTION OF E. COLI WITH UNPARALLELED SENSITIVITY. RUG A novel beta-glucuronidase substrate
MICROBIOLOGICAL DETECTION OF E. COLI WITH UNPARALLELED SENSITIVITY RUG A novel beta-glucuronidase substrate E. coli detection Escherichia coli (E. coli) is a Gram negative bacterium that inhabits the intestines
More information3 8 COLIFORM BACTERIA AS INDICATOR ORGANISMS Laboratory tests for disease-producing bacteria, viruses, and protozoa are difficult to perform
3 8 COLIFORM BACTERIA AS INDICATOR ORGANISMS Laboratory tests for disease-producing bacteria, viruses, and protozoa are difficult to perform Most utilities have neither qualified personnel nor laboratories
More informationDAIRY WATERS. (Coliform Group and Escherichia coli) [E. coli verification required only on source water] IMS #24
DAIRY WATERS (Coliform Group and Escherichia coli) [E. coli verification required only on source water] IMS #24 [Unless otherwise stated all tolerances are ±5%] 1. Laboratory Requirements a. Cultural Procedures
More informationSerial dilution and colony count (Viable count) Pour plate. Spread plate Membrane filtration. Turbidity. Microscopic cell count
Aljawharah Alabbad 2016 Serial dilution and colony count (Viable count) Pour plate Spread plate Membrane filtration Turbidity Microscopic cell count Many studies require the quantitative determination
More informationLab Activity #14 - Bacteriological Examination Of Water and Milk (Adapted from Lab manual by Dr. Diehl)
Lab Activity #14 - Bacteriological Examination Of Water and Milk (Adapted from Lab manual by Dr. Diehl) Some of the diseases that humans can contract from drinking polluted water include typhoid, dysentery,
More informationLab Exercise 13: Growth Curve
Lab Exercise 13: Growth Curve OBJECTIVES 1. Know the different phases of a standard growth curve. 2. Understand and perform direct measurement of bacterial growth through serial dilutions and standard
More informationKARL F. ECKNER* KM Lab AB, Helsingborg, Sweden. Received 6 January 1998/Accepted 7 May 1998
APPLIED AND ENVIRONMENTAL MICROBIOLOGY, Aug. 1998, p. 3079 3083 Vol. 64, No. 8 0099-2240/98/$04.00 0 Copyright 1998, American Society for Microbiology. All Rights Reserved. Comparison of Membrane Filtration
More information01/08/2018. Counting Microorganisms. Counting microorganisms. Turbidity Measurements. Relative abundance. Direct counts.
Counting Microorganisms 1 Counting microorganisms 2 Relative abundance Turbidity measurements Direct counts Absolute counts Viable counts Absolute number of growing bacteria Most probale number (MPN) Probable
More informationSample for use a Guide Revised IDEXX COLILERT -18 TEST METHOD FOR THE SIMULTANEOUS DETECTION OF TOTAL COLIFORMS AND E.
IDEXX COLILERT -18 TEST METHOD FOR THE SIMULTANEOUS DETECTION OF TOTAL COLIFORMS AND E. COLI IN WATER 1 IDEXX COLILERT -18 TEST METHOD FOR THE SIMULTANEOUS DETECTION OF TOTAL COLIFORMS AND E. COLI IN WATER
More informationSome Industrially Important Microbes and Their Products
2 Some Industrially Important Microbes and Their Products 2.1. ENZYME PRODUCING MICROBES Type of enzyme Substrate Microorganism Amylase Starch Saccharomyces diastaticus Protease Proteins Bacillus sp. Lipase
More informationApplied Environmental Microbiology. Copyright McGraw-Hill Global Education Holdings, LLC. Permission required for reproduction or display.
43 Applied Environmental Microbiology Copyright McGraw-Hill Global Education Holdings, LLC. Permission required for reproduction or display. Water Purification and Sanitary Analysis Microbial containment
More informationMicrobiology Methods for Drinking Water Laboratories. Erica Fox
Microbiology Methods for Drinking Water Laboratories Erica Fox Origins of Drinking Water Bacteriological Testing 1854 Cholera epidemic in London Dr. John Snow determined source of Cholera to be public
More informationINS 099 Data Analysis Summer 2007
INS 099 Data Analysis Summer 2007 Day 1: Solutions, Dilutions, and Exponential Growth Due Tues. July 17, 2007 Microbial population counts Exponential Growth If N 0 is the initial population and the growth
More informationAseptic Techniques. A. Objectives. B. Before coming to lab
Aseptic Techniques A. Objectives Become familiar with 1. The ubiquity of microorganisms (see Note 1) 2. Aseptic techniques (see Note 2) 3. Standard methods for growing/observing microorganisms (see Note
More informationSCHEDULE. Friday: Pet Investigations: Plate counts - how to know how many clones of your pet you have (pg. 9-10)
SCHEDULE Wednesday: Pet Investigations: Phenol Red Broth with Durham tubes (pg. 3-4) Oxidation/Fermentation Agar (pg. 5-6) Anaerobic Growth (pg. 7) Growth in Liquid Culture (pg. 8-9) Friday: Pet Investigations:
More informationLab Date Experiment Reports, Midterms, Reminders
Lab Date Experiment Reports, Midterms, Reminders Lab 1 Lab 2 Jan 5-6 Jan 7-8 o Registration, introductory remarks, safety lecture, etc. Sterile technique. o Expt 1: Microbes in the environment o Expt 2:
More informationKit Information 4 Introduction. 4 Kit Contents, Storage, and Testing Conditions. 4 Principle. 5 Applicability. 5 Precautions: 5
Contents 3. Kit Information 4 Introduction. 4 Kit Contents, Storage, and Testing Conditions. 4 Principle. 5 Applicability. 5 Precautions: 5 Sample Preparation 5 Dairy (Liquid Cream). 5 Solid Dairy...........................
More informationBacterial Counts - Quantitative Analysis of Microbes
Bacterial Counts - Quantitative Analysis of Microbes Introduction: It is often important to know not only what types of bacteria are in a sample but also how many of them are present. Food manufacturers
More informationGROWTH AND SURVIVAL OF PATHOGENIC E. COLI DURING CURDLING OF MILK
Int. J. LifeSc. Bt & Pharm. Res. 2014 Aryya Mitra and Sanjib Ghoshal, 2014 Research Paper ISSN 2250-3137 www.ijlbpr.com Vol. 3, No. 1, January 2014 2014 IJLBPR. All Rights Reserved GROWTH AND SURVIVAL
More informationCOUNT METHOD 5.0 OBJECTIVES 5.1 INTRODUCTION 5.2 PRINCIPLE. Structure
Food Microbiology EXPERIMENT 5 STANDARD PLATE COUNT METHOD Structure 5.0 Objectives 5.1 Introduction 5.2 Principle 5.3 Materials Required 5.4 Procedure 5.4.1 E-coli Culture 5.4.2 Food Samples 5.5 Observations
More informationBiology Lab Activity 4-5 DNA Transformation
Biology Lab Activity 4-5 DNA Transformation Scientists can insert genes into bacteria. The genes inserted in the Indo-Blu process (this lab) are on a circular piece of DNA called a plasmid. (The plasmid
More informationLaboratory Procedure October 1999 HEALTH PROTECTION BRANCH OTTAWA ANALYSIS OF SPROUTS FOR COLIFORMS, ESCHERICHIA COLI, AND KLEBSIELLA PNEUMONIAE..
Government of Canada Gouvernement du Canada Laboratory Procedure MFLP-64 October 1999 HEALTH PROTECTION BRANCH OTTAWA ANALYSIS OF SPROUTS FOR COLIFORMS, ESCHERICHIA COLI, AND KLEBSIELLA PNEUMONIAE.. Don
More informationMost Probable Number (MPN) & Biological Oxygen Demand (BOD)
Most Probable Number (MPN) & Biological Oxygen Demand (BOD) Part : Presumptive Coliform Test (MPN) Introduction This lab exercise will employ a commonly used multi-tube fermentation technique. The results
More informationThe Biotechnology Education Company. Water Quality Testing I: Chromogenic Analysis of Water Contaminants. See Page 3 for storage instructions.
The Biotechnology Education Company REVISED & UPDATED EDVO-Kit 951 Water Quality Testing I: Chromogenic Analysis of Water Contaminants See Page 3 for storage instructions. EXPERIMENT OBJECTIVE: The objective
More informationBacterial Abundance. Objective Measure bacterial numbers and mass per unit volume. Note, we are not concerned with identification here.
Bacterial Abundance Objective Measure bacterial numbers and mass per unit volume. Note, we are not concerned with identification here. Why do we want to know abundance? Allows determination of biomass
More informationBiology 322 Fall 2010 Transfer of genetic information in the bacterium Escherichia coli: Part I
Biology 322 Fall 2010 Transfer of genetic information in the bacterium Escherichia coli: Part I REQUIRED Reading Assignments: Superbugs on the Hoof http://fire.biol.wwu.edu/trent/trent/superbugs.pdf Triple
More informationSample for Use as a Guide Revised IDEXX ENTEROLERT TEST METHOD FOR THE DETECTION OF ENTEROCOCCI IN WATER
IDEXX ENTEROLERT TEST METHOD FOR THE DETECTION OF ENTEROCOCCI IN WATER 1 IDEXX ENTEROLERT TEST METHOD FOR THE DETECTION OF ENTEROCOCCI IN WATER 1. Scope and Application 1.1. This method is intended for
More informationSoleris system evaluation of testing applications for UHT/aseptic packs
Summary system evaluation of testing applications for UHT/aseptic packs September 2012 Neogen Corporation (Lansing, Mich.) conducted internal studies to validate optimal test procedures for the ultrahigh
More informationColiform bacteria are quantitated by the fractional gram pour plate technique (Note 1). Test tubes containing gas collector tubes (Durham Tubes)
Microbiological Methods IV-A- 1 (STANDARD PLATE COUNT METHOD) PRINCIPLE SCOPE Coliform bacteria are quantitated by the fractional gram pour plate technique (Note 1). The method is applicable to starches,
More informationLarge Volume Serial Dilutions:
Serial Dilutions All three bacterial plate count methods described in lab require you to serially dilute your samples until you have 30-300 colony forming units (CFU) on the plate. Plates with more than
More informationEXPERIMENT. Biochemical Testing for Microbial Identification Methyl Red, Voges- Proskauer, and Catalase Testing
EXPERIMENT Biochemical Testing for Microbial Identification Methyl Red, Voges- Proskauer, and Catalase Testing Hands-On Labs, Inc. Version 42-0246-00-02 Review the safety materials and wear goggles when
More informationENUMERATION OF COLIFORMS AND ESCHERICHIA COLI BY IDEXX (COLILERT 18) QUANTI-TRAY TM
NATIONAL STANDARD METHOD ENUMERATION OF COLIFORMS AND ESCHERICHIA COLI BY IDEXX (COLILERT 18) QUANTI-TRAY TM W 18 Issued by Standards Unit, Evaluations and Standards Laboratory Specialist and Reference
More informationM. Dalbey/Bio 105M Isolation of E. coli - Isolation of E. coli from an Environmental Sample
Isolation of E. coli from an Environmental Sample We want to expand our horizons a bit beyond the domesticated lab strains of E. coli. In this exercise you will isolate "wild" E. coli strains from an environmental
More informationKit Information 4 Introduction. 4 Kit Contents, Storage, and Testing Conditions. 4 Principle 4 Applicability. 5 Precautions Sample Preparation 6
Contents 3. Kit Information 4 Introduction. 4 Kit Contents, Storage, and Testing Conditions. 4 Principle 4 Applicability. 5 Precautions.......................... 5 Sample Preparation 6 Test Procedure 7
More informationI January 23-January 26 INTRODUCTION; SAFETY; ASEPTIC TECHNIQUE; USE OF MICROSCOPES; OBSERVATION OF PREPARED SLIDES; ENVIRONMENTAL SAMPLE; EPIDEMIC
MICROBIOLOGY LABORATORY (BIOL 310L) SCHEDULE Spring 2017 Lecture Professor: Dr. Susan Morrison Lab Instructors: Ms. Tracy Hirsch Dr. Susan Morrison Required: (1) Leboffe & Pierce, Microbiology Laboratory
More informationDiagnostic Microbiology
Diagnostic Microbiology Identification of Microbes Lecture: 1 Out lines What is expected out of this course??? At the end of this course, you will be able to apply Conventional/ Molecular diagnostic methods
More informationENVIRONMENTAL PARAMETERS OF GROWTH
ENVIRONMENTAL PARAMETERS OF GROWTH The growth and survival of microorganisms are affected by the chemical and physical conditions of the external environment. Environmental factors which have significant
More informationBacterial Abundance. Objective Measure bacterial numbers and mass per unit volume. Note, we are not concerned with identification here.
Bacterial Abundance Objective Measure bacterial numbers and mass per unit volume. Note, we are not concerned with identification here. Why do we want to know abundance? Allows determination of biomass
More informationENVIRONMENTAL PARAMETERS OF GROWTH
ENVIRONMENTAL PARAMETERS OF GROWTH The growth and survival of microorganisms are affected by the chemical and physical conditions of the external environment. Environmental factors which have significant
More informationMicrobiology sheet (6)
Microbiology sheet (6) Made by marah marahleh corrected by : abd. Salman DATE :9/10/2016 Microbial growth / control of microbial growth 1 The method of counting bacteria is divided into: 1) direct 2) indirect
More informationDiscussion Items. Microbial Indicators of Water Quality
Discussion Items! Announcements! Group Project topics! Discuss previous lab (Microbes in food) Results Lab report Isolates streak isolate by Thursday.! Microbial analysis of water (Tuesday) MPN and MF!
More informationResult:COMPLETE Report Date: December 28 th, 2015
Send to: Clean Water Environmental, LLC 1939 Talamore Court Southeast, Grand Rapids, MI 49546 Dr. Dale Williams Result:COMPLETE Report Date: December 28 th, 2015 Customer Name: Clean Water Environmental,
More informationLab 6. API System 320 MBIO PRACTICAL MIC AMAL-NORA-ALJAWHARA 1
Lab 6. API System 320 MBIO PRACTICAL 2018 320 MIC AMAL-NORA-ALJAWHARA 1 (API 20E) Analytical Profile Index System for Identificationof Enterobacteriaceae 2018 320 MIC AMAL-NORA-ALJAWHARA 2 Identification
More informationENVR 421 Laboratory #1: Basic Bacteriology Techniques
ENVR 421 Laboratory #1: Basic Bacteriology Techniques Introduction The purpose of this laboratory exercise is to familiarize you with two fundamental bacteriology techniques: the streak plate and the spread
More informationBacterial Transformation: Unlocking the Mysteries of Genetic Material
PR009 G-Biosciences 1-800-628-7730 1-314-991-6034 technical@gbiosciences.com A Geno Technology, Inc. (USA) brand name Bacterial Transformation: Unlocking the Mysteries of Genetic Material Teacher s Guidebook
More informationSOP-C-124 Determination of Total Coliform, Escherichia coli, and Enterococci by IDEXX Colilert and Enterolert
i. Identification of the method a. (2012) b. SM 9223 and 9223B (approved 2004) ii. Applicable matrix or matrices a. Nonpotable water and Drinking water iii. Limits of detection and quantitation a. From
More informationChapter 6: Microbial Growth
Chapter 6: Microbial Growth 1. Requirements for Growth 2. Culturing Microorganisms 3. Patterns of Microbial Growth 1. Requirements for Growth Factors that affect Microbial Growth Microbial growth depends
More informationNATIONAL STANDARD OF THE PEOPLE S REPUBLIC OF CHINA. National Food Safety Standard
GB NATIONAL STANDARD OF THE PEOPLE S REPUBLIC OF CHINA GB 4789.38-2012 National Food Safety Standard Microbiological Examination of Food Hygiene - Enumeration of Escherichia Coli 食品安全国家标准食品微生物学检验大肠埃希氏菌计数
More information7.02 Microbial Genetics in Lab Quiz. Fall, September 27, 2001 ANSWER KEY
7.02 Microbial Genetics in Lab Quiz Fall, 2001 September 27, 2001 ANSWER KEY This quiz contains 4 questions worth a total of 48 points. Be sure to write your name, Bench letter and Undergraduate TA s (UTA)
More informationIdentifying Enterobacter aerogenes from a Mixed Culture of Unknown Gram Positive and Gram Negative Bacteria Kevin Le November 13, 2013
Identifying Enterobacter aerogenes from a Mixed Culture of Unknown Gram Positive and Gram Negative Bacteria Kevin Le November 13, 2013 PURPOSE The focus of this study is to be able to identify an unknown
More informationIntroduction. Michael J. Miller, Ph.D. RMM»
RMM» Case Study of a New Growth-based Rapid Microbiological Method (RMM) that Detects the Presence of Specific Organisms and Provides an Estimation of Viable Cell Count Michael J. Miller, Ph.D. President
More information2009 Summary Report for Municipalities
2009 Summary Report for Municipalities (As per Schedule 22 of O. Reg. 170/03) The Onaping Potable Water System - #220003519 Certificate of Approval 1414-0696HD3 0014-6MQQFQ Prepared by: Jean-Marc Joliat
More informationPathogenic Bacteria. culture media. Components of the Typical Culture Medium: Culture Media Importance:
Level4 Lab2: Pathogenic Bacteria culture media Microorganisms, like all other living organisms, require basic nutrients for sustaining their life. All microorganisms have the same basic requirements but
More informationBACTERIAL GENETICS: Labs I & II
BACTERIAL GENETICS: Labs I & II The Bacterial Genetics Labs will extend over two laboratory periods. During the first lab, you will set up two different experiments using the bacterium Escherichia coli.
More informationOutline. This week s plans Introduction to Kirby Bauer and Antibiotics Introduction to Biochemical analysis of strains
Outline This week s plans Introduction to Kirby Bauer and Antibiotics Introduction to Biochemical analysis of strains 1 This week 2 Monday Finish Exercise 10, part C. Run MspI restriction digest on a gel
More informationM I C R O B I O L O G Y
ninth edition TORTORA FUNKE CASE M I C R O B I O L O G Y a n i n t r o d u c t i o n 6 Microbial Growth PowerPoint Lecture Slide Presentation prepared by Christine L. Case Microbial Growth Microbial growth
More information2/25/2013. Psychrotrophs Grow between 0 C and C Cause food spoilage Food Preservation Temperatures
3 4 5 6 7 8 9 0 Chapter 6 Microbial Growth Microbial Growth Increase in number of cells, not cell size Populations Colonies The Requirements for Growth Physical requirements Temperature ph Osmotic pressure
More informationigem 2018 InterLab Study Protocol Before You Begin
igem 2018 InterLab Study Protocol Before You Begin Read through this entire protocol sheet carefully before you start your experiment and prepare any materials you may need. In order to improve reproducibility,
More informationDetermination of MIC & MBC
1 Determination of MIC & MBC Minimum inhibitory concentrations (MICs) are defined as the lowest concentration of an antimicrobial that will inhibit the visible growth of a microorganism after overnight
More informationDetermination of MIC & MBC
1 Determination of MIC & MBC Minimum inhibitory concentrations (MICs) are defined as the lowest concentration of an antimicrobial that will inhibit the visible growth of a microorganism after overnight
More informationMicrobiology Chapter 2 Laboratory Equipment and Procedures 2:1 The Light Microscope MICROSCOPE: any tool with a lens to magnify and observe tiny
Microbiology Chapter 2 Laboratory Equipment and Procedures 2:1 The Light Microscope MICROSCOPE: any tool with a lens to magnify and observe tiny details of specimens Micro tiny, small Scope to see SIMPLE
More informationMICROBIAL PLATE COUNT USING IMAGE PROCESSING
MICROBIAL PLATE COUNT USING IMAGE PROCESSING By PAWANKUMAR FAKATKAR Department of Electronics & Telecommunication Engineering Sardar Patel Institute of Technology Munshi Nagar, Andheri(W), Mumbai-400058
More informationIsolation and Characterization of Escherichia coli
Chapter-4 Isolation and Characterization of Escherichia coli 4.1 Sample source and collection of samples: Escherichia coli is known to be a colon bacteria which shows ubiquitous presence in many ecological
More informationConfirming the Phenotypes of E. coli Strains
Confirming the Phenotypes of E. coli Strains INTRODUCTION Before undertaking any experiments, we need to confirm that the phenotypes of the E. coli strains we intend to use in the planned experiments correspond
More informationDetection of Escherichia coli in Drinking Water Sources of Filter Units and Supply Water
Bangladesh Pharmaceutical Journal 19(2): 206-210, 2016 Detection of Escherichia coli in Drinking Water Sources of Filter Units and Supply Water Arman Chowdhory 1, Nafisa Kabir 2, Md. Mazharul Islam Chowdhury
More informationMicroorganisms In Our Environment
PR015 G-Biosciences 1-800-628-7730 1-314-991-6034 technical@gbiosciences.com A Geno Technology, Inc. (USA) brand name Microorganisms In Our Environment Teacher s Guidebook (Cat. # BE 106) think proteins!
More informationMarine Bacteria Cause False-Positive Results in the Colilert-18 Rapid Identification Test for Escherichia coli in Florida Waters
APPLIED AND ENVIRONMENTAL MICROBIOLOGY, Feb. 2002, p. 539 544 Vol. 68, No. 2 0099-2240/02/$04.00 0 DOI: 10.1128/AEM.68.2.539 544.2002 Copyright 2002, American Society for Microbiology. All Rights Reserved.
More informationBiology 2180 Laboratory #7. Bacterial Growth and Transformation
Biology 2180 Laboratory #7 Name Bacterial Growth and Transformation Introduction: Most aspects of molecular biology require the use of basic microbiological methods. This section is included for those
More informationLab Exercise #4 Microbial Control Lab Exercise #4 Control of Microorganisms: Physical, Chemical and Chemotherapeutic
Lab Exercise #4 Control of Microorganisms: Physical, Chemical and Chemotherapeutic I. OBJECTIVES: Investigate the effectiveness various agents of control. Assess the effectiveness of heat in killing vegetative
More informationMarkerGene TM Fluorescent Bacterial Detection and Quantification Kit
Product Information Sheet MarkerGene TM Fluorescent Bacterial Detection and Quantification Kit Product M1460 Marker Gene Technologies, Inc. University of Oregon Riverfront Research Park 1850 Millrace Drive
More informationLactose Assay Kit. Catalog Number KA assays Version: 04. Intended for research use only.
Lactose Assay Kit Catalog Number KA1672 100 assays Version: 04 Intended for research use only www.abnova.com Table of Contents Introduction... 3 Intended Use... 3 Principle of the Assay... 3 General Information...
More informationActivity 5.1.5: Student Resource Sheet
Activity 5.1.5: Student Resource Sheet Biochemical tests are the most definitive way to identify bacterial species. Each biochemical test helps determine a property or characteristic specific to a certain
More informationCell Growth and DNA Extraction- Technion igem HS
Growing Cells and DNA Extraction Goals 1. Become familiar with the process of growing bacteria 2. Get to know the DNA extraction process 3. Perform miniprep in the lab Keywords 1. Growth stages 6. Techniques
More informationLactose Assay Kit. Catalog Number KA assays Version: 05. Intended for research use only.
Lactose Assay Kit Catalog Number KA1672 100 assays Version: 05 Intended for research use only www.abnova.com Table of Contents Introduction... 3 Background... 3 Principle of the Assay... 3 General Information...
More informationLABORATORY #2 -- BIOL 111 BACTERIAL CULTIVATION & NORMAL FLORA
LABORATORY #2 -- BIOL 111 BACTERIAL CULTIVATION & NORMAL FLORA OBJECTIVES After completing this exercise you should be able to: 1. Identify various types of media 2. Isolate bacteria using aseptic technique.
More information--> Buy True-PDF --> Auto-delivered in 0~10 minutes. GB Translated English of Chinese Standard: GB4789.
Translated English of Chinese Standard: GB4789.36-2016 www.chinesestandard.net Sales@ChineseStandard.net NATIONAL STANDARD OF THE GB PEOPLE S REPUBLIC OF CHINA National Food Safety Standard - Microbiological
More informationGB Translated English of Chinese Standard: GB NATIONAL STANDARD OF THE
Translated English of Chinese Standard: GB4789.36-2016 www.chinesestandard.net Sales@ChineseStandard.net NATIONAL STANDARD OF THE GB PEOPLE S REPUBLIC OF CHINA GB 4789.36-2016 National Food Safety Standard
More informationLab 5/5a Transformation of E. coli with a Recombinant Plasmid
Lab 5/5a Transformation of E. coli with a Recombinant Plasmid Lab 2 Pre Lab Readiness Familiarity and Proper use of micropipettes Remember the 1 st and 2 nd stops Aseptic Technique Antibiotic Resistance
More informationMICROBIAL GROWTH. Dr. Hala Al-Daghistani
MICROBIAL GROWTH Dr. Hala Al-Daghistani Microbial Growth Microbial growth: Increase in cell number, not cell size! Physical Requirements for Growth: Temperature Minimum growth temperature Optimum growth
More informationChapter 3 SCREENING AND SELECTION OF STRAIN FOR ALKALINE PROTEASE PRODUCTION BY SUBMERGED FERMENTATION
Chapter 3 SCREENING AND SELECTION OF STRAIN FOR ALKALINE PROTEASE PRODUCTION BY SUBMERGED FERMENTATION - 42 - 3.1 MATERIAL AND METHODS 3.1.1 Isolation of bacterial strains for alkaline protease production
More informationBioBuilding: Synthetic Biology for Teachers: itune device
Lab 2: itune Device Teacher Considerations This lab examines the role of parts, such as promoters and ribosome binding sites, in predicting the output of a genetic device. The students measure b- galactosidase
More informationHeterotrophic Bacteria
, m-hpc, 8242 DOC316.53.01225 Pour Plate Method Method 8242 m-hpc Scope and Application: For water and wastewater. Test preparation Introduction Before starting the test: The Pour Plate Method, also known
More informationControl of Metabolic Processes
Control of Metabolic Processes Harriet Wilson, Lecture Notes Bio. Sci. 4 - Microbiology Sierra College As described earlier, the metabolic processes occurring within living organisms (glycolysis, respiration,
More informationBlue Flag Beaches and Recreational Water Testing for and Enterococci E.coli using Enterolert & E. coli
Blue Flag Beaches and Recreational Water Testing for and Enterococci E.coli using Enterolert & E. coli Gil Dichter World Wide Technical Support Manager, Water www.idexx.com/water 1 OBJECTIVES 2 Blue Flag
More informationWHY DO THEY PUT MINT IN TOOTHPASTE? WOULD GARLIC BE BETTER?
Activity 4.22 Student Sheet WHY DO THEY PUT MINT IN TOOTHPASTE? WOULD GARLIC BE BETTER? Purpose To investigate the antibacterial properties of plants. To develop practical skills. YOU NEED Agar plate seeded
More informationCOMPARITIVE STUDY OF THE EFFECT OF SINGLE WALLED CARBON NANOTUBES ON Escherichia coli IN CULTURES AND BIOFILMS. A write-up on the proposed Study
COMPARITIVE STUDY OF THE EFFECT OF SINGLE WALLED CARBON NANOTUBES ON Escherichia coli IN CULTURES AND BIOFILMS A write-up on the proposed Study Prepared by: Indumathy Jayamani For: Ene 806 Laboratory Feasibility
More informationMicrobial Nutrition and Growth
PowerPoint Lecture Presentations prepared by Mindy Miller-Kittrell, North Carolina State University C H A P T E R 6 Microbial Nutrition and Growth CHNO Growth Requirements Nutrients: Chemical and Energy
More informationHiPer Transformation Teaching Kit
HiPer Transformation Teaching Kit Product Code: HTBM017 Number of experiments that can be performed: 10 Duration of Experiment: 4 days Day 1- Preparation of media and revival of E. coli Host Day 2- Inoculation
More informationCh 6. Microbial Growth
Ch 6 Microbial Growth Student Learning Outcomes: Classify microbes into five groups on the basis of preferred temperature range. Explain the importance of osmotic pressure to microbial growth. Provide
More information