Inoculation techniques in microbiology pdf

Fundamentals of microbiology, history, kochs postulates. Aseptic techniques are practices that prevent the contamination of growth media. It is one speciic type of cognitive behavioral therapy cbt. A small amount of sample is placed on the side of the agar plate either with a swab, or as a drop from an inoculating loop. Bios 318 microbiology methods manual rice university. Introduction if the human eye could resolve images as well as the light microscope, we would see bacteria and fungi virtually everywhere. In microbiology, the term isolation refers to the separation of a strain from a natural, mixed population of living microbes, as present in the environment, for example in water or soil flora, or from living beings with skin flora, oral flora or gut flora, in order to identify the microbes of interest. To process clinical specimens satisfactorily for bacteriological culture, consideration must be given to. After inoculation, swirl the bottle several times to mix and transport to a microbiology laboratory immediately see section iii. Inoculation is the purposeful introduction of bacteria into a sterile growth medium. Historically, the laboratory techniques of isolation first developed in the field of.

To demonstrate skill in isolation of organisms from a mixed culture using selective and differential media. Historical records suggest that by the 17th century, and perhaps earlier, inoculation also known as variolation was being used to prevent the viral disease smallpox in various parts of the world. The inoculation method could impact the outcome of. Disinfection describes a process that eliminates many or all pathogenic microorganisms, except bacterial spores, on inanimate objects. Growth of methanogens on agar medium, the bottle plate 10 2.

Experimental inoculation in rats and mice by the giant. The microbiology society, in association with the microbiology in schools advisory committee misac. Colonies on an agar plate one very important method in microbiology is to isolate a single type of bacteria from a. Pure culture techniques university of pennsylvania. Absolutely no food, drinks, chewing gum, or smoking is allowed in the laboratory. Live animal inoculation requires an experienced personnel to perform various inoculation techniques. As infection preventionists, you are going to encounter these organisms during your career. The most critical and most expensive piece of equipment in the microbiology laboratory is the microscope. Partially lift the lid of the plate culture and open it just enough to insert the inoculation loop. To distinguish the range of activity of an antibiotic. Insert the inoculating loop into the stock culture tube.

Effect of inoculation techniques and relative humidity on the. Sep 26, 2009 all students working in the microbiology laboratory are responsible for maintaining equipment and materials in proper working condition. Microbiology department carolina biological supply company 2006 carolina biological supply company printed in usa. Common course outline of material included in the course. Introduction if the human eye could resolve images as well as the light microscope, we. Touch the loop to an area of the agar with no growth in order to cool down the loop. To utilize specific monitoring techniques to evaluate the susceptibility of a microbe to different antibiotics. Risk assess and practise simulation of aseptic techniques. Animal inoculation may be used as diagnostic procedure for identifying and isolating a virus from a clinical specimen. To enumerate obligate anaerobes, we could modify the spread plate method by using anaerobic jar after inoculation. Method 1, frozen stock to a plate, a colony picked and added to lb for overnight culture in a test tube, and then inoculation for the experiment. List of equipment apparatus used in microbiology laboratory.

Motility is apparent by the presence of diffuse growth away from the line of inoculation. The laboratory exercises in microbiology, 5e by pollack, et al. Therefore, an article on basic techniques involved in microbiology can help all in many ways. Microbiology module sterilisation and disinfection microbiology 44 notes 1.

A material is sterile when it has no living organisms present. Inoculating needles are used when stabbing into a medium during specific inoculation. Wash your hands with disinfectant soap when you arrive at the lab and again before you leave. Inoculation method could impact the outcome of microbiological. Preparation of 200 ml of anoxic medium by boiling 5 2. To start, aseptic technique is used to introduce a very small sample of cells the inoculum into a receptacle containing nutrient or culture medium. It should grow vigorously after inoculation in seed stage vessels.

Laboratoryresearch skills for science resume list of skills to assist with resumes, cover letters and interviews animal care handle and restrain weigh animals breed various species clean cages administer injections prepare special diets anesthetize maintain and evaluate animal records prepare for sterile surgical procedures. Please read over and follow the instructions listed below. Issued by the standards unit, microbiology services, phe. Added certain substances to the medium can make it useful for selecting different types of organisms and differentiating. Experience has shown that these two groups of microorganisms. Often, having too many bacteria on the loop can cause problems with obtaining a proper dilution, seeing individual cells in a gram stain, and it uses up your culture faster. The use of microtiter plates and automated inoculation and reading systems. Bacteria and fungi are grown on or in microbiological media of different types. Inoculation is done by placing a pinch of leaf meal into whorl of each plant, when plant attains the height of 3045 cm. Use aseptic techniques in the safe inoculation of various forms of media. Molds were isolated from commercial cakes and were identified as aspergillus sydowii, aspergillus ochraceus, penicillium funiculosum, and eurotium herbariorum. In the liver and the omentum, the viable virus was recovered in the first seven days in rats, and viral dna i. A sterile loop is then used to spread the bacteria out in one direction from the initial site of inoculation. The different staining procedures for viewing cells.

Pdf advanced techniques in diagnostic microbiology. Q 5 inoculation of culture media for bacteriology apsi. The methods are well referenced and represent a good minimum standard for clinical and public health microbiology. Insert the loop into the sterile broth and inoculate it by gently moving the loop back and forth in order to disperse the cells. The culture medium volume and the inoculation method. Pdf the main dilemma when performing microbiology challenge testing, is the selection and validation of a method of inoculation. The manual contains a variety of interactive activities and experiments that teach students the. Proper inoculation technique is critical in microbiology.

If unable to collect 3 ml of urine, collect in sterile specimen container 37777 or yellow top tube 104184 and transport urine specimens to the microbiology laboratory or refrigerate within 30 minutes. Contrary to most expectations, these cultures are not homogeneous. Uk standards for microbiology investigations issued by the standards unit, public. Key concepts n many soil conditions make it necessary to inoculate legume crops to obtain maximum yields. Introduction although techniques for growing methanogens vary in detail among laboratories, these procedures follow the legacy of hungate who. Culture tube flaming prior to inserting a cooled transfer instrument into a culture tube, you need to remove the cap and flame the lip of the tube. To demonstrate good aseptic technique in culture transfer or inoculation and in. Inoculation techniques pour, streak, and lawn plates. By teaching you coping skills, sit can help you ind new ways to deal with ptsd symptoms. Pick up a small quantity of bacteria, yeast, or fungal spores.

Grow each of the bacterial and fungal test strains separately as described in table 1. You can download the microorganisms cheat sheet pdf by clicking on the download. Many of the techniques and strategies that you learn in this laboratory will be useful if you conduct any type of biological laboratory investigation in the future. Samples can then be taken from the resulting colonies and a microbiological culture can be grown on a new plate so that the organism can be identified, studied, or tested. Pdf inoculation techniques in microbiology challenge testing. Live inoculation was first used on human volunteers for the study of yellow fever virus.

Laboratory safety adapted from the american society of microbiology website. This smi should be used in conjunction with other smis. Choose an isolated colony and scrape off a very small amount of culture with the loop. Find, read and cite all the research you need on researchgate. Historically, the trend toward automation in clinical pathology laboratories has largely bypassed the clinical microbiology laboratory. Mice provide a reliable model for studying viral replication. Inoculation application is explained in detail, including seed coating by various methods, soil inoculation, and the handling and storage of inoculant. Do not completely open the lid and expose the surface to the air. Inoculation of agar plates by pour, streak and lawn methods. Pdf lab report of microbiology akbar haqi academia. Used for the study of immune responses, epidemiology and oncogenesis. Effect of inoculation techniques and relative humidity on. The labs are introduced in a clear and concise manner, while maintaining a studentfriendly tone. This manual, which explains the basic techniques necessary to carry out microbiology experiments safely and effectively, is intended as a guide for teachers and technicians.

Spiral system and pour plating techniques correlated very well for all species, as did the spiral system and spread plating techniques r greater than 0. The modern streak plate method has progressed from the efforts of robert koch and other microbiologists to obtain microbiological cultures of bacteria in order to study them. Microbiology is a popular option for practical work in schools. Gives unique insight into viral pathogenesis and host virus relation. Samples can then be taken from the resulting colonies and a microbiological culture can be grown on a new plate so that the organism can be identified, studied, or tested the modern streak plate method has progressed from the efforts of robert koch and other. In microbiology lab, it is used for maintaining ph of the medium and diluents. Different inoculation methods used in bacteriology. Microbiology has some exciting perhaps even scary years ahead of it. The anaerobic jar will create an anaerobic environment for the obligate anaerobes. Pick up a tube of sterile nutrient broth with your free hand, carefully remove the cap cotton plug with your little finger of the hand holding the loop, and flame the mouth of the tube. Should produce a single valuable product, and no toxic by products. The risk of contamination should be minimal under the optimum performance conditions. Tubes must be filled to 3 ml to prevent inhibition of bacterial growth. Full text get a printable copy pdf file of the complete article 469k, or click on a page image below to browse page by page.

Quantification of medically important candida species and. By the late 18th century, englishman edward jenner was inoculating patients with cowpox to prevent smallpox, a technique he coined vaccination. Stress inoculation training what type oftreatment is this. This quality guidance describes the methods of inoculating culture media and subculturing of organisms using aseptic techniques. Chapter 3 methods of culturing microorganisms five basic techniques. Agar media with insoluble phosphates are used for semiquantitative screening of phosphatesolubilizing bacteria based on the solubilization halo formed around. When inoculating, one should barely be able to see any bacteria on the loop.

These skills can also help you manage other stressful situations or events in your life. In microbiology, the term isolation refers to the separation of a strain from a natural, mixed population of living microbes, as present in the environment, for example in water or soil flora, or from living beings with skin flora, oral flora or gut flora, in order to identify the microbe of interest. Stress inoculation training, or sit, is a psychotherapy for ptsd. Automation in clinical microbiology journal of clinical. A second inoculation may be made five to ten days later. Refer to our experiment result, the pour plate method shows that there are 10900 viable cellsml in the dilution of 10 2, while 93000 viable cells. Various cultures and media are used in studying microorganisms. Laboratory exercises in microbiology, 5th edition wiley. For the past 150 years, bacteria have been investigated primarily in liquid batch cultures. Register to obtain the microbiology concept inventory. General biochemical tests microbiology resource center.

Follow oral and written instructions and manage time in the lab efficiently. List and discuss the basic principles of microbial growth, metabolism, host interaction, and pathogenesis. The medium that is used to culture the microorganism depends on the microorganism that one is trying grow. The parts of a microscope and how they are used for magnifying cells and their parts. Standard resistant and susceptible checks need to be included in all evaluations so that the level of infection can be compared with previous inoculations. According to apic text of infections control and epidemiology. Animals of choice for cultivating viruses include monkeys, rabbits, guinea pigs, rats, hamsters, and mice. This will help prevent contamination with transient and airborne microbes. Bd labo a newsletter providing the latest microbiology news and ideas to the clinical laboratory.

Microbiology module methods of isolation of bacteria microbiology 116 notes streak plates 1. Microbiologists use aseptic technique for a variety of procedures such as transferring cultures, inoculating media, isolation of pure cultures, and for performing microbiological tests. Historically, the laboratory techniques of isolation first developed in the field of bacteriology and parasitology, before those in virology during the 20th century. Incubate at a temperature and duration appropriate for the organism being tested. Inoculation techniques in microbiology pdf squarespace. Laboratory exercises in microbiology cuny academic works. Demonstrate mastery of and apply aseptic techniques in the laboratory setting. Routes of viral inoculation in embryonated eggs theory. Read through booklet on safe handling of microorganisms.

The method of inoculating the solid culture media is as shown in the figure. Microbiology skills are clearly much in demand and the work of microbiologists is central to developments in. Four inoculation techniques were compared for initiation of growth on cake surfaces. Unicellular vegetative microorganisms bacteria and yeasts. Seedlot culture maintenance techniques seedlot systems are used so that the viable microorganisms used for inoculation are not more than 5 passages removed from the original master seedlot. Please note, you must be an educator in higher ed or maybe high school to qualify to recieve the mci. Microbiology and its basic techniques are related to some other sciences like plant physiology, genetics, evolution, molecular biology. In microbiology, streaking is a technique used to isolate a pure strain from a single species of microorganism, often bacteria.

This method of inoculation will be ineffective if dry weather prevails following application of the leaf meal. Inoculating loops are the most common method of transferring bacteria. Apply correct terminology regarding microbiological techniques, instruments, microbial growth, biochemical testing, media types and forms and epidemiology when making observations. Inoculate with growth from an 1824 hour culture by stab inoculation with a needle.

To achieve this goal microbiologists use culture media and aseptic transfer techniques. Microbes are managed and characterized by using the five is inoculation, incubation, isolation, inspection and identification. Objectives to demonstrate good aseptic technique in culture transfer or inoculation and in handling sterile materials. Inoculation meaning in microbiology is that transfer from culture for their growth. Hot water below boiling point moist heat sterilization involves the use of steam in the range of 1214c. Aseptic technique and the transfer of microorganisms. Basic techniques for the cultivation of methanogenic archaea in anoxic media, where the or potential is maintained below 330 mv under a pressurized atmosphere of 20% carbon dioxide, are described. Laboratory animals are widely used for routine cultivation of virus. To inoculate a slant culture in a fishtail inoculation technique w. Microbiology is used as a generic term to include the two gmcrecognised specialties of medical microbiology which includes bacteriology, mycology and parasitology and medical virology. This involves the progressive dilution of an inoculum of bacteria or yeast. Basic practical microbiology why microbiology matters.

Steam under pressure is used to generate high temperature needed for sterilization. Cake surfaces were inoculated with mold spores and incubated under three. Frontiers microbial inoculants for improving crop quality. Do not put anything in your mouth such as pencils, pens, labels, or fingers.