In pour plate culture technique, successive dilutions of the inoculum serially diluting the original specimen of old broth culture is added to. Fill water dilution bottles to 80 ml or desired volume with butterfields phosphate diluent note 1. When comparing the accuracy of these two techniques, pour plate has a higher accuracy than the spread plate. Withinoculants prepared from presterilized peat, viable numbers ofrhizobia may be determined by the pour, spread, and drop plate methods. In this method, fixed amount of inoculum generally 1 ml from a brothsample is placed in the center of sterile petri dish using a sterile pipette. The technique makes it easier to quantify bacteria in a solution. A comparison of pour plate and spread plate methods article pdf available in journal american water works association 751. Difference between pour plate and spread plate pediaa. Membrane filter method for the simultaneous detection of total coliforms and escherichia coli in drinking water epa600r000 with one exception, the addition of mi broth. Isolate single bacterial colonies by the streakplating method. Pour plate method for isolating bacteria membrane solutions. The results of the third experiment june 26 listed in table 1 are graphically illustrated in fig. Epa test procedures require samples of 100 ml or more to be analyzed for the presence of bacteria.
The speciality of the pour plate method is that a known volume of the sample is first mixed with agar and then poured into the plate. The modern pour plate culture method was initially developed in the laboratory of the famous bacteriologist and the father of bacteriology, dr. In pour plate technique, successive dilutions of the inoculum serially diluting the original specimen of old broth culture is added to the sterile petri plates containing the melted and. Learn vocabulary, terms, and more with flashcards, games, and other study tools. In pour plate method, in this method of plating or culturing the microbial sample first diluted and pour in empty plate and after that growth media is poured in it and which is then skake firmly.
The key difference between pour plate and spread plate is that a known volume of the sample is spread on the surface of the agar medium in spread plate, while, in pour plate, a known volume of the sample is mixed with agar and then poured into a plate. Because of this, a variety of methods have been developed. Pour plate method is usually the method of choice for counting the number of colonyforming bacteria present in a liquid specimen. Aim for about 2 tablespoons of nuts and seeds or 12 cup of tofu. Spread plate technique is the method of isolation and enumeration of microorganisms in a mixed culture and distributing it evenly. Counting bacterial colonies on agar plates is a simple and effective method for determining the number of viable bacteria in a sample.
This is the most commonly used method for enumeration of bacteria in a wide variety of samples including milk, food, meat, soil etc. Form fdancims 2400a standard plate count and coliform plate count rev. The precision of the pour plates wasthusnotrepeatedwith streak plates. Inoculum refers to microorganisms, bacteria or fungi that grow in or on the nutrient agar. Introduction microorganism is an organism that is microscopic or submicroscopic, which is too small to be seen under naked eyes. Spread plate technique is a method employed to plate a liquid sample for the purpose of isolating or counting the bacteria present in that sample. What are the advantages and disadvantages of pour plate.
The difference between pourplate method and spreadplate method are as follows. It has been also found that pour plate method procedures yield lower bacteria count that spread plate methods 151. Pour plate technique procedure, advantages, limitations. The microorganisms are trapped beneath the surface of medium when it solidifies. Whether its customized environmental isolate controls, process controls, certified reference materials, proficiency samples, or something thats never been done before were up to the challenge. We thrive on creating innovative biological reference materials that make microbiologists lives easier. Pour plate methods yield a count of only the living cells in the sample and thus are a viable count.
For the pour plate and agar droplet colworth droplette techniques, the agar was melted and cooled to 46 0c in a water bath before use. The pour plate method is not widely used but may be more convenient in some instances and some experienced workers prefer the pour plate method for isolating bacteria. The plate method nutrition for weight control protein fill. Mar 22, 2018 pour plate method and spread plate method are used for quantification or enumeration of bacterial sample. Hence, surface as well as subsurface colonies are developed and it is very difficult to isolate and count the subsurface colonies. The difference between pour plate method and spread plate method are as follows. Pour plate technique for the isolation of microorganism. A comparison was made of agar plates spread with glass rods and poured agar plates for the enumeration of bacteria in the waters of fishers island. The spreadplate technique is analogous to the pourplate procedure in its utility as a means to perform viable plate counts. Basic practical microbiology why microbiology matters. A pour plate method, meanwhile, involves the dilution of a mixed population of bacteria into tubes of nutrient agarmedium, which is then poured into a petri dish for incubation.
Pour plate 12 using a spreader spread plate 14 working with moulds 15 incubation 16 in conclusion. The most common method for determining the total viable count is the pourplate method. The pour plate method of counting bacteria is more precise than the streak plate method, but, on the average, it will give a lower count. Full text get a printable copy pdf file of the complete article 209k, or click on a page image below to browse page by page. Colony forming unit calculations using spread plate method.
R5 311 kaiser foundation health plan of the midatlantic states, inc. Microbiology of the food chain horizontal method for the enumeration of microorganisms part 1. What is the difference between the pourplate method and. The pour plate technique is limited to a sample volume of 2 ml and the spread plate technique has a limit of 0. What is the difference between the pourplate method and the. B melted agar equilibrated to a temperature of approximately 48 c is then poured into the petri dish with the sample. Metode pour plate lempeng tuang adalah suatu teknik di dalam menumbuhkan mikroorganisme di dalam media agar dengan cara mencampurkan media agar yang masih cair dengan stok kultur bakteri agar sehingga selsel tersebut tersebar merata dan diam baik di permukaan agar atau di dalam agar. Whenlarge numbers ofsamples are plated, as happens during routine quality controlchecks,the methodchosenshouldeconomizeontheplatingmediaandpetri dishes.
The pour plate technique can be used to determine the number of microbes ml in a specimen. According to a consensus of views, it is recommended to keep the molten agar in. Pour plate method definition of pour plate method by. However, because the colonies that form using the spreadplate technique are evenly distributed across the surface of the agar medium, cells from individual colonies can be isolated and used in subsequent experimental.
An introduction to microbiology, aseptic technique and safety. Note that two types of colonies arise in and on the pour plate. Jun 04, 2018 the main difference between pour plate and spread plate is that the molten agar is poured on to the inoculum during the preparation of the pour plate whereas inoculum is spread on the surface of the solidified agar during the preparation of the spread plate. Pour plate technique for bacterial enumeration david b. The pour plate technique can be used to determine the number of microbesml in a specimen. Pour plate definition is a plate prepared by mixing the inoculum with the cooled but still fluid medium before pouring the latter into the petri dish. Label the bottom of the plate with the above data plus initials, seat number and date. How to make a four season room from a porch duration. The pourplate technique requires a serial dilution of the mixed culture by means of a loop or pipette. Visit our website for technical documents, illustrated instructions, instructional videos, faqs and more. Pdf lab report of microbiology akbar haqi academia. Simply add the bacteria to melted agar which is not so hot it will kill the desired bacterium. Since mi broth was approved on november 6, 2001, as a minor modification of the mi agar method, it has also been included in this document. It is assumed that each bacterial colony arises from an.
Pour plate technique is a microbial method to enumerate some viable cells present in a sample. May 11, 2012 the spread plate technique is analogous to the pour plate procedure in its utility as a means to perform viable plate counts. Procedure aseptically weigh 20 g of the sample into a sterile 80 ml water blank and homogenize note 2. Contents 10 foilpacked sterile media vessels for 100 ml of media when hydrated 100 sterile simplate plates with lids mpn table. Sep 09, 2014 this general microbiology practical lecture explains the pour plate method of bacterial culture and culture media. The spread plate technique involves using a sterilized spreader with a smooth surface made of metal or glass to apply a small amount of bacteria. Download a pdf of our pour plate best practices and instructions. This method relies on the growth of a bacterial cell in an agar plate to form a visible colony, only living or viable bacterial cells will be counted. This is the primary dilution and represents a sample dilution factor of 5.
The bacterial colony counter left image is used with pour plates right image to help determine cell numbers in a sample. However, because the colonies that form using the spread plate technique are evenly distributed across the surface of the agar medium, cells from individual colonies can be isolated and used in subsequent experimental. A perfect spread plate technique will results visible and isolated colonies of bacteria that are evenly distributed in the plate and are countable. Jun 01, 2018 side by side comparison pour plate vs spread plate in tabular form 6. It has the advantage of not requiring previously prepared plates and is often used to assay bacterial contamination of foodstuffs. The main difference between pour plate and spread plate is that the molten agar is poured on to the inoculum during the preparation of the pour plate whereas inoculum is spread on the surface of the solidified agar during the preparation of the spread plate. Here are the steps we follow in our laboratory to successfully perform the pour plate method. The spread plate technique involves using a sterilized spreader with a smooth surface made of metal or glass to. Spread plate technique principle, procedure and uses. Pourplate method and spreadplate method are used for quantification or enumeration of bacterial sample. The pour plate procedure was neither as accurate nor as precise as the spread plate procedure for enumerating the heterotrophic plate count population. Pour plate method is neither as accurate nor as precise as the spread plate procedure for heterotrophic plate count enumeration 150. What are the advantages and disadvantages of pour plate method.
Method of isolation of pure culture linkedin slideshare. Aug 15, 2019 spread plate technique is the method of isolation and enumeration of microorganisms in a mixed culture and distributing it evenly. Streak plate 11 pour plate 12 using a spreader spread plate 14. However, the numbers of microorganisms in a given sample are required to know in certain aspect such as dairy industries, diseases investigation, and so on. The plates are then incubated under conditions that permit microbial reproduction so that colonies develop that can be seen without the aid of a microscope. Solid media tested for growth promotion is to be set up using pour plate method in order to determine a microbial plate count cfu which must be. Twenty ml of the primary dilution can be aseptically. Hitung jumlah bakteri metode pour plate nana asriyah. This general microbiology practical lecture explains the pour plate method of bacterial culture and culture media. Mar 08, 2018 download a pdf of our pour plate best practices and instructions.
The most common method for determining the total viable count is the pour plate method. Pour plate definition of pour plate by merriamwebster. For healthy meal planning ucsf diabetes teaching center. Salah satu metode yang digunakan adalah metode pour plate hitung cawan. Microbiological methods xia mesophilic aerobic spore. Best practices for pour plate method according to a consensus of views, it is recommended to keep the molten agar in the water bath for no more than 3 to 4 hours. Comparison pour, spread, plate methods enumeration of. Difference between pour plate and spread plate compare. As well as causing a familiar range of diseases in animals and plants and problems in food spoilage and deterioration of other materials, microbes are also our invisible allies.