Determining Microbes in Food By Amar Ch Das
Determining Microbes in Food By Amar Ch. Das Ghosh
Culture and Microscopic Methods A microbiological culture, or microbial culture, is a method of multiplying microbial organisms by letting them reproduce in predetermined culture medium under controlled laboratory conditions. Microbial cultures are foundational and basic diagnostic methods used as a research tool in molecular biology.
Standard Plate Count The standard plate count method consists of diluting a sample with sterile saline or phosphate buffer diluent until the bacteria are dilute enough to count accurately. That is, the final plates in the series should have between 30 and 300 colonies.
Introduction The standard plate count (SPC) is suitable for estimating bacterial populations in most types of dairy products, and it is a reference method specified in the Grade A Pasteurized Milk Ordinance to be used to examine raw and pasteurized milk. This procedure is also recommended for application in detecting sources of contamination by testing linesamples taken at successive stages in the processing.
Principle The test employs a serial dilution technique for easy quantification of the micro-organisms. The appropriate dilutions of the milk sample are mixed with a sterile nutrient medium that can support the growth of the micro-organisms, when incubated at a suitable temperature. Each bacterial colony that develops on the plate is presumed to have grown from one bacterium or clump of bacteria in the inoculums. The total number of colonies counted on the plates multiplied by the dilution factor to represent the number of viable micro-organisms present in the sample tested.
Most Probable Number (MPN) is a method used to estimate the concentration of viable microorganisms in a sample by means of replicate liquid broth growth in ten-fold dilutions.
What is MPN in 100 m. L? A: CFU stands for 'Colony Forming Units' and refers to the number of viable bacterial cells in a sample per unit of volume. For example: 50 CFU/100 m. L means 50 Colony Forming Units per 100 m. L of sample. . For example: 50 MPN/100 m. L means that the Most Probable Number of viable cells in 100 m. L of sample is 50.
How do you find that most probable number? Calculation Using Table 4 MPN Index, it is possible to estimate the number of organisms from any combination of positive and negative test results. The MPN Table (Table 4) is based on 3 dilutions of decreasing decimal increments. . Calculation Equation= MPN Index per 100 m. L value x the Lowest dilution factor. How is MPN calcuted? MPN value is calculated as follows: . . . The MPN value is expressed as 2. 2 per 100 ml. These numbers are given by Maccardy based on the laws of statistics.
Direct Microscopic Count (DMC) is a quantitative test and is helpful in assessing the actual number of bacteria present in milk. DMC is used as a platform test to assess the microbiological quality of milk received at the Raw Milk Receiving Dock.
What are immunological Techniques Immunological techniques are the wide varieties of methods and focused experimental protocols developed by immunologists for inducing, measuring, and characterizing immune responses. . They allow the immunologists to alter the immune system through cellular, molecular and genetic manipulation.
What are the microbiological techniques Microbiology techniques are methods used for the study of microbes, including bacteria and microscopic fungi and protists. They include methods to survey, culture, stain, identify, engineer and manipulate microbes.
ELISA Principle. Enzyme-linked Immunosorbent Assays (ELISAs) combine the specificity of antibodies with the sensitivity of simple enzyme assays, by using antibodies or antigens coupled to an easily-assayed enzyme. ELISAs can provide a useful measurement of antigen or antibody concentration.
What are the applications of immunology Immunology is the study of the immune system and is a very important branch of the medical and biological sciences. The immune system protects us from infection through various lines of defence. If the immune system is not functioning as it should, it can result in disease, such as autoimmunity, allergy and cancer.
Sterility testing for pharmaceuetical products Sterility Testing is defined as a testing which confirms that products are free from the presence of viable microorganisms. Sterility testing is very important for medical devices, pharmaceuticals, preparations, tissue materials and other materials that claim to be sterile or free from viable microorganisms.
What is pharmaceutical sterility? Sterility can be defined as the freedom from the presence of viable microorganisms. . In pharmaceutical practice, a container is defined as sterile when the probability is less than one out of one million that it is contaminated with replicating microorganisms.
What are the 3 types of sterilization There are several types of sterilization equipment. Steam sterilizers (autoclaves), dry heat sterilizers, heated chemical vapor sterilizers, and gas sterilizers. How is sterility testing performed? Sterility testing is conducted on bulk or final products by direct inoculation or membrane filtration methods and can be performed in an isolator or cleanroom environment. A bacteriostasis/fungistasis test is also performed to assess hether or not the test article inhibits the growth of microorganisms.
Definition of Nucleic acid probe Nucleic-acid probe. n. A nucleic-acid fragment that is complementary to another nucleicacid sequence and thus, when labeled in some manner, as with a radioisotope, can be used to identify complementary segments present in the nucleic-acid sequences of various microorganisms.
What is the function of the probe? The purpose of the probe routine is to detect devices residing on the bus and to create device nodes corresponding to these devices. What is a probe in molecular biology? probe. (Science: molecular biology) general term for a piece of DNA or RNA corresponding to a gene or sequence of interest, that has been labelled either radioactively or with some other detectable molecule, such as biotin, digoxygenin or fluorescein.
What does a probe do? The Dissolved Oxygen Probe is great for biology, chemistry, ecology, or integrated science courses. It can be used to perform a wide variety of experiments to determine changes in dissolved oxygen levels, one of the primary indicators of the quality of an aquatic environment.
What is DNA probe test? DNA probe tests are now available to the clinical microbiology laboratory for the rapid diagnosis of infectious disease. A unique method for DNA probe tests has been developed that allows the test to be performed in a single test tube in four basic steps within two hours.
What is probe in Biotechnology? Hybridization probe is a fragment of DNA or RNA of variable length (usually 1001000 bases long) which is radioactively labeled. It can then be used in DNA or RNA samples to detect the presence of nucleotide sequences (the DNA target) that are complementary to the sequence in the probe.
What is an RNA probes? Definition. RNA probes are stretches of singlestranded RNA used to detect the presence of complementary nucleic acid sequences (target sequences) by hybridization. RNA probes are usually labelled, for example with radioisotopes, epitopes, biotin or fluorophores to enable their detection.
What is the function of a probes in a PCR Probes are fluorescently labelled DNA oligonucleotides. They are designed to bind downstream of one of the primers during the PCR reaction and to give a fluorescent signal during the reaction. The 5' end of the probe is labelled with a fluorescent reporter molecule.
Can a person be identified by a DNA probes? Nucleic acid probes are able to detect the presence of particular sequences in a sample down to the level of a few hundred molecules. . Nucleic acid 'fingerprints' provide an unequivocal identification of the origin of cells which may be applied in criminal law, civil law, and in the follow up to bone marrow transplants.
PCR based detection PCR-based tests have allowed detection of small numbers of disease organisms (both live or dead), in convenient samples. . Viral DNA can be detected by PCR. The primers used must be specific to the targeted sequences in the DNA of a virus, and PCR can be used for diagnostic analyses or DNA sequencing of the viral genome.
What is PCR testing? Polymerase chain reaction (PCR) tests are used to detect HIV's genetic material, called RNA. These tests can be used to screen the donated blood supply and to detect very early infections before antibodies have been developed. This test may be performed just days or weeks after exposure to HIV.
How is PCR used to identify bacteria? The principle of the method is simple; when a pure PCR product of the 16 S gene is obtained, sequenced, and aligned against bacterial DNA data base, then the bacterium can be identified. . A selected PCR band from each of 40 isolates was sequenced and the bacterium identified to species or genus level using BLAST.
What diseases can PCR detect? Detecting infectious agents PCR is extensively used in analysing clinical specimens for the presence of infectious agents, including HIV, hepatitis, human papillomavirus (the causative agent of genital warts and cervical cancer), Epstein. Barr virus (glandular fever), malaria and anthrax.
What is PCR used for? Sometimes called "molecular photocopying, " the polymerase chain reaction (PCR) is a fast and inexpensive technique used to "amplify" - copy - small segments of DNA. What is the function of PCR? PCR is typically used to amplify a specific gene, or portion of gene, so that we can study the function of that gene or gene region. Primers are used to flank the region you want to amplify. Each primer will amplify the gene sequence on both strands, creating a doublestranded gene product.
Is PCR test a blood test? A polymerase chain reaction (PCR) test may also be performed to determine whether the virus itself is present in the patient's blood. A sample of the patient's blood is taken and sent to a laboratory. If the virus' genetic makeup (DNA) is present, a positive diagnosis is made.
How accurate is PCR test? PCR viral load is the most sensitive test for detecting HIV infection in seroconversion, though greater sensitivity may be achieved when used in conjunction with the proviral DNA test. . This test is very reliable for detecting HIV in someone recently exposed to virus and will be highly accurate within 48 to 72 hours.
What is biosensors? Biosensors are devices used to detect the presence or concentration of a biological analyte, such as a biomolecule, a biological structure or a microorganism. Biosensors consist of three parts: a component that recognizes the analyte and produces a signal, a signal transducer, and a reader device.
What is the use of biosensors? Biosensors are devices comprising a biological element and a physiochemical detector that are used to detect analytes. These instruments have a wide range of applications ranging from clinical through to environmental and agricultural. The devices are also used in the food industry.
Thank You for your attention
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