Practical 1 Microbiology PDF

INTRODUCTION TO PRACTICAL COURSE IN MICROBIOLOOGY AND IMMUNOLOGY Reasons for conducting practical course in microbiology To teach students basic knowledge of microbiology in general. To teach practical applications of aseptic, technique, disinfection and sterilization. To...

INTRODUCTION TO PRACTICAL COURSE IN MICROBIOLOOGY AND IMMUNOLOGY Reasons for conducting practical course in microbiology To teach students basic knowledge of microbiology in general. To teach practical applications of aseptic, technique, disinfection and sterilization. To teach proper methods of handling materials contaminated with pathogenic organisms. To study the main biological characteristic of more important bacteria causing diseases.  Cont… To emphasize throughout the course the relationship between microorganisms and diseases It should be understood to students and all staff entering or working in microbiology laboratory that many of the microorganisms studied are able to produce diseases, thus the aseptic technique must be practiced all the time. Students should be aware of epidemiology and pathogenesis(spread and causing of diseases), Student be aware of host and parasite relationship, development of resistance to infection(immunity). They should be familiar to communicable diseases, morbidity and mortality due to the diseases and through the course have to realize that prevention is the best way rather than cure.  GENERAL LABORATORY RULES HINT: THINK BIOHAZARDS!!!!!!! Eating, drinking and smoking in the laboratory is not permitted, Mouth pipe ting and placing object in your mouth is prohibited in microbiology laboratory. You must wear personal protective equipments (PPE) such as Laboratory coats and gloves when working in the laboratory. Do not enter in the laboratory with bare feet. Cover your feet with shoes, Open shoes and long heels shoes are not permitted. Report any breakage of equipments to instructors. Keep your work bench free from clusters of objects. Before begging your laboratory exercises, wash off the bench top with disinfectant provided. When exercises are complete, wash off bench top again. Always wash your hands with soap and water before leaving the laboratory. Cultures are not to be removed from the laboratory unless the instructor gives you permission  Cont… Making noise in the laboratory is totally prohibited. Read the instructions carefully before you start any procedure in the microbiology laboratories Flame the inoculating loop or needle immediately before and after use. If viscous materials are present on the loop or needle, dry it at the flame before placing it directly in the flame. Long hair should be tied back; also loose clothes should be tied. Wash hands before leaving the laboratory and when you know or suspect that they have been contaminated with pathogenic microorganisms.  ASEPTIC PROCEDURES IN MICROBIOLOGY LABORATORY These are procedures which should help you not to contaminate yourself, working place and bacterial cultures with pathogenic bacteria when inoculation procedure is taking place. A: Inoculating needles and loops must be sterilized in flame of gas burner before and after use. B:You should keep bottles and tubes stoppers/tops except when in actual use. The plugs and tops must be held in hand without touching the part that might be contaminated. You should also NEVER put them on bench(try to exercise by working with both hands I.e. One hand holding the bottle while the other hand is holding a loop and a bottle top, Flame the lips of the bottles and tubes immediately before and after use  Aseptic technique in microbiology Flame the loop in Bunsen burner/spirit lamp till red hot before and after use, whenever large colon adhere on loop, introduce it slowly into the flame to avoid spraying of materials before the organisms have been killed. Leave loop to cool before picking culture/specimen. Hold the culture plate or tube with left hand and grasp the cap or cotton plug between palm and little finger of right hand, flame mouth of container passing three times through flame. Insert the loop in culture and touch the colon not the wall and tops of the tube. flame the mouth of the tube Immediately and replace cotton plug or cap and lastly sterilize the loop to red hot. NB: Everything found in the microbiology laboratory is potentially infectious. BE CAREFULL!!!  B.IMPORTANT EQUIPMENTS IN MICROBIOLOGY LABORATORY Microbiology is a fascinating field of science. It is not only field of examining microorganisms under microscope but it involves generating them through invitro growing (microbial culture),isolation, biochemical identification, drug sensitivity tests and even Genetic identification. Therefore it needs basic and miscellaneous equipments and reagents to perform the task as tabulated in the next slide.  Cont… AUTOCLAVE Used for sterilization of glassware and media. It is effective at temperature of 121°C, Pressure of 15 Psig for 15 minutes. INCUBATOR Used for bacterial and fungal culture. (Prokaryotic cells and eukaryotic cells). MICROSCOPE To observe tiny organisms that cannot be seen by naked eye. (Magnification and resolution purposes).  Cont…. REFRIGRATOR For preservation of sample, cultures, media and many sensitive materials. FREEZER Similar to refrigerator but preserve materials below freezing point of water HOT AIR OVEN Used for sterilization of materials which cannot be sterilized by moist heat, it is effective at 160-300°c. BIOSAFETY CABINETS To prevent infections especially on very infectious bacteria such as B.anthrax, mycobacterium spp,aspergillus spp etc  Cont… Petri dishes/culture plates/agar plates Supporting containers to hold culture medium. Sterile disposable swabs Specimen collection Eg.Vaginal swab Serological test slides For immunoassay/serological analysis. Universal bottle For sample collection and and media preparation. Vortex For mixing liquids kept in test tubes. Magnetic stirrer For mixing liquids in beakers/conical flasks PH METER To measure PH of media and reagents.  Cont…. Candle jar Used for anaerobiosis Centrifuges To separate supernatant and pellet Durham tubes Used to detect gas production in sugar fermentation media Haemagglutination plate For viral culture detection. Inoculation loop Used to inoculate test samples into culture media for bacterial or fungal cultures. Microtitre plates For ELISA test (Enzyme linked immunosorbent assay)  Cont…. Canister Containing a stand for Petri dishes and generally is used to carry and store plates. Washing bottles For storing distilled water for washing purposes. Cool box Carrying samples from sampling site to laboratory. Vacutainer tubes For collection of blood from animals for blood agar preparation (The one with anticoagulant). Measuring cylinder Measuring volumes of liquid reagents and distilled water. Tally counter For enumeration of bacteria and bacteria colonies Micropipette Loading bacterial, fungal or viral DNA into agarose gel for electrophoresis, preparing master mix (measuring micro volumes). Water bath Warming culture media especially which cannot be autoclaved  Cont… Weighing balance For measuring mass of media, chemicals and reagents Syringes For blood specimen collection. Bijou bottles Used for holding culture medium and biochemical tests Safety cabinet