Microbio Lab Ex 1 PDF

04/10/2023 Biological Safety Cabinet...

04/10/2023 Biological Safety Cabinet Microbiological safety cabinet It is an enclosed, ventilated laboratory workspace for Introduction to safety working with materials contaminated pathogens requiring a defined biosafety level. Microbiology To serve as the primary means to protect the laboratory worker and the surrounding environment from pathogens. Centro Escolar University - Makati Biological Refrigerator Deep Freezer It used to preserve cultures of bacteria, yeast, and other Use: used to preserve plasma, related blood microbes at temperatures between 2-8 °C. components, vaccines, microorganism, and testing of For most living things, as the temperature decreases, materials and electronic components at desired low metabolic rates also decrease. temperature. It differs from a refrigerator as it does not have a freezer Description: it is a micro controller based temperature controller with a digital temperature display of set value and process value. Use for storage at lower than 2°C Incubator An apparatus with a chamber used to provide controlled environmental conditions especially for the cultivation of microorganisms. The temperature is set at 35-37°C Types of Incubator It has a glass cover to provide protection against microbes and prevent diffusion of foul odor. Example E. Coli, this organism grows well in this condition. Saccharomyces cervisiae 30°C 1  04/10/2023 Types of incubator Types of incubator Carbon dioxide incubator Non Carbon Dioxide incubator – Used in the cultivation of mammalian cells. - Is a incubator or containing room air, thus, it contains An incubator to which a cylinder of carbon dioxide is attached, CO2 is periodically introduced into the about 20-21% oxygen. incubator to maintain CO2 conc. of about 5-10%. The relative humidity is typically >95% and a slightly acidic pH. Anaerobic incubator - It is used to isolate capnophiles (organisms that grow - Containing an atmosphere devoid of oxygen. best in atmosphere containing increased CO2). It is important to keep in mind that CO2 incubator contains oxygen (about 15%-20%) in addition to CO2. Thus CO2 incubator is not anaerobic incubator. Hot air oven / Oven sterilizer Hot air oven / Oven sterilizer It is an electrical devices use in sterilization. Mechanism of action is via oxidation. It follows the principle of “dry-heat sterilization” This is used for sterilizing metals, glasswares, cotton They were originally developed by Pasteur. The plugs, and cotton swabs, all must be wrapped using a standard settings for a hot air oven are: clean manila paper. – 1.5 to 2 hours at 160-165 °C – 1 hour at 170-180 °C – 6 to 12 minutes at 190 °C Autoclave Autoclave An apparatus whose mechanism follows the principle Invented by Charles Chamberland of “moist heat sterilization” The name comes from Greek auto –ultimately Mechanism of action is by coagulation of proteins. meaning self, and Latin clavis meaning key – a self- Autoclaving at a pressure of 15psi at temp of 121 °C locking device. for 15-20mins, kills vegetative microorganisms, bacterial endospores, and viruses, as long as they are Coagulation or denaturation of proteins by cleavage not protected by pus, feces, vomitus, blood, or other of H bonds resulting in the loss of the 3D structure of proteinaceous materials. the proteins. Operates by Steam under pressure. Used for sterilizing culture media and for killing grown up or cultured microorganisms. 2  04/10/2023 Loop incinerator Weighing balance Is a device used in microbiology laboratories to Use: Measuring instrument for determining the sterilize instruments particularly the loop. weight or mass of an object. Also known as bactoincinerator. Description: Balance or scales can be calibrated to Any combustible hazardous or non-hazardous read in units. material may be incinerated by the practice of the present invention. Microincinerator can reach a temp of 800 deg., sufficient enough to incinerate any organic material on the inoculating loop. Uses: Sterilizing the loop and heat fixing the bacterial smear on the microscope slide. Compound microscope Rotary shaker Also known as biological or high power microscope. Is ideal for developing of cultures, chemicals, They are used for viewing transparent specimens that solvents, assays etc. are not typically visible to the naked eye. Microscopic It’s of an important use for the production of basic means invisible to the eye unless aided by a chemicals, pathological work and various other microscope. application for shaking solutions. An instrument consisting of a combination of lenses Rotating apparatus with a platform on which liquid such that the image formed by the lens or set of media or cultures can be continuously shaken. lenses nearest the object (called the objective) is magnified by another lens called the ocular or eyepiece Colony Counter Transfer pipets/Pasteur pipets An instrument used to count colonies of bacteria or Are used to transfer small quantities other microorganisms growing on an agar plate. Such counters are used to estimate the density of of liquids. microorganisms within a liquid culture. They are usually glass tubes tapered It usually consists of an illuminated to a narrow point, and fitted with a rubber bulb at the top. Named after Louis Pasteur. Made up of glass or plastic (disposable). 3  04/10/2023 Inoculating loops and needles Is a simple tool used mainly by Usually made from either nichrome or platinum microbiologists to retrieve an wire. inoculum from a culture of a. k. a. “smear loop”, “inoculation wand”, microorganisms. The loop is used “microstreaker”. in the cultivation of microbes on Loop diameter is 5mm. Nichrome is cheaper. plates by transferring inoculum for streaking. Petri Dish Petri dishes are a shallow glass or plastic After the agar cools and solidifies, the dish is ready to cylindrical lidded dish, often used to make agar plates for microbiology studies. These receive a microbe-laden sample in a process known are covered dishes used to culture as inoculation or “plating.” microorganisms. The dish is partially filled with warm liquid containing agar and a mixture of specific ingredients that may include nutrients, blood, salts, carbohydrates, dyes, indicators, amino acids and antibiotics. Screw-capped test tubes Durham tubes/Fermentation tubes Test tubes with screw caps are suitable for Used in microbiology to detect production of gas by transporting and growing of cultures. microorganisms. It is used for handling, sealing after autoclaving, They are simply smaller test tubes inserted upside and culturing all kinds of microorganisms. down in another test tube. This small tube is initially filled with the solution in which the microogarnisms is Common piece of laboratory glassware consisting to be grown. If gas is produced after inoculation and of a finger-like length of glass or clear plastic incubation, a visible gas bubble will be trapped inside tubing, open at the top, usually with a rounded U- the small tube. shaped bottom. To detect production of gas by microorganisms. The With enamel marking spot for labelling using initial air gap produced when the tube is inserted pencil. upside down is lost during sterilization. 4  04/10/2023 Graduated flask Beaker USE: used to measure the volume of a USE: simple container for stirring, liquid. mixing and heating liquids in laboratory. DESCRIPTION: is usually narrow and high and has a plastic or glass bottom. DESCRIPTION: are generally cylindrical in shape, with a flat bottom. L-rod Anaerobic jar It is used to spread the sample over An instrument used in the production of an anaerobic microorganisms. Example the surface of an agar plate. Clostridium species (anaerobic gram (+) spore-forming bacteria) Alternative to bending glass rods or It has a thermometer and pressure pipets for spreading samples evenly gauge to monitor the temperature and an agar surfaces and filters in petri initial pressure inside the jar. dishes. Permits oxygen to combine with hydrogen that comes from a gas generator producing CO2 and H2. Gas-pak jar Membrane filter/Syringe filter It is used to culture bacteria which die or fail Membrane filters or “membranes” are microporous plastic films with specific pore size to grow in presence of oxygen (anaerobes). It ratings. Also known as screen, sieve, or is a much simpler technique than the microporous filters. Mclntosh and Filde’s anaerobic jar where Used to trap bacteria from air and water one needs to pump gases in and out. samples, can be transferred directly to agar plates, and the quantity of bacteria in the sample Use-and-throw sachets containing a dry can be determined. powder or pellets. These filters are only 0.1 mm thick. The pores of membrane filters include, for example, 0.22µm The indicator changes color if there is a and 0.45µm sizes, which are intended for presence of oxygen. bacteria. 5  04/10/2023 UV-VIS Spectrophotometer Alcohol lamp Indirect method of determining Alcohol lamps are usually found in bacterial population. Turbidity biology labs where they do inoculations indicates bacterial growth. of bacteria cultures. It heats the wire Measure the growth of cultures of loop on a stick that they use to spread bacteria by comparing the light the bacteria on the growth medium, intensity between the reference which will prevent contamination to and test sample. the cultures. Microtiter plates ELISA reader Microplate or microwell plate. Enzyme-linked immunosorbent Is a flat plate with multiple “wells” assay (ELISA) the basis of most used as small test tubes. It is used in modern medical diagnostic testing culturing small volume of in humans and animals. microorganisms or cells. ELISA is a test that uses antibodies This is used when performing ELISA. and color change to identify a substance. Universal Precaution Importance of Good Personal Hygiene Refers to the practice of avoiding contact with Personal hygiene in the laboratory is directed mainly patient’s bodily fluids, by means of the wearing of toward the prevention of occupational acquired non-porous articles such as medical gloves, goggles, disease or physical injury. and face shields. It can raise the quality of laboratory work by reducing Protocols used to maintain an aseptic field and to possibilities for contamination of experimental prevent cross-contamination and cross-infection material. between health care providers and patients. 6  04/10/2023 Some of Biologically-active Specimen with Significance to Spills and Breakage Pharmacy Protamine – polypeptide from sperma of fish species 1. Wear gloves and face masks, as Botulinum toxin – protein from Clostridium botulinum protective barriers. Heparin – anticoagulant Enoxaparine – anticoagulant to treat deep vein thrombosis or 2. For small spills and breakages, pulmonary embolism remove excess materials using Pancreatin towel/rag. Place towel/rag in Lactobacillus Acidophillus Lyophilisatus disposable plastic for sterilization. Streptomyces – Tetracycline Bacillus subtilis – Bactracin Disinfect the area with germicide Fungi or molds – β-LACTAMS or household bleach. Spills and Breakage Chemical spill on a person 3. For large spills, cover with Over the body towel/rag and then pour liquid Within seconds, quickly remove all contaminated clothing germicide or household over while person is under safety shower. Flood the affected the covered spill. Allow body area with cold water for at least 15 minutes. If pain disinfection for at least 10 continues or resumes, flood with more water. Wash off minutes, then clean the area as chemicals with a mild detergent solution. Do not apply any stated previously. materials such as neutralizing agents to the area. Obtain medical assistance immediately. Chemical spill on a person Safety gears and apparatus In the eyes Complete laboratory attire You will need to assist the person who has chemical Fire extinguisher spattered in the eyes. Immediately drench the eyes at the nearest emergency eyewash station. Force the eye Safety shower or eyes open to get water into them. The speed of your First aid kit response to this emergency is extremely important. Eye wash station Notify the laboratory instructor of the accident immediately. 7  04/10/2023 Definition of terms Isolation – to separate (a pure stain) from mixed bacterial and pure culture. Incubation – the development of infectious diseases from the entrance of the pathogen to the appearance of clinical symptoms Inoculation – is the placement of something that will grow or reproduce and is most commonly used in respect to introduction of serum, vaccine or antigenic substance into the body of a human or animal. Inoculum – the material used in an inoculation, also called “inoculant”. Coagulation – the process by which blood forms clots (thrombogenesis). 8