Title Micro-NOTES-2024-EDITED (1).pdf ✅

Miguel Carlos G. Arada, RMT, ASCP(IMT), DTA, MSMT BACTERIOLOGY PROKARYOTE EUKARYOTE Nuclear body Cell division Cell wall Cytoplasmic membrane Cell organelles Site of energy production Site of protein synthesis BACTERIAL CYTOLOGY Cell wall -Defines the shape of the bacteria -Peptidoglycan consists of glycan chains of alternating: a. NAG b. NAM -Function: 1. Protects internal structure 2. Provides shape to bacteria 3. Some components responsible for pathogenicity a. M protein b. Mycolic acid Stains Simple stain  Colors the forms and shapes Differential stain  Coloring specific components GRAM STAIN REACTION Reagents Gram + Gram - P. Stain Mordant Decolorizer S. Stain GRAM POSITIVE vs GRAM NEGATIVE _______________________________________________ _______________________________________________ _______________________________________________ _______________________________________________ _______________________________________________ _______________________________________________ _______________________________________________ _______________________________________________ _______________________________________________
Miguel Carlos G. Arada, RMT, ASCP(IMT), DTA, MSMT ACID FAST STAIN Cytoplasmic Membrane - Selectively permeable membrane - Site of energy production Ribosomes - Site of protein synthesis Mesosomes - Point of attachment for chromosome - Extension of cytoplasmic membrane Inclusions - Much granules _______________________________________ _________________________ - Babes-Ernst/Metachromatic/volutin granules _______________________________________ _________________________ - Bipolar bodies _______________________________________ _________________________ Endospores - Resting cell, highly resistant to dessication, heat and chemical agents - Composition:_____________________ - Bacterial genera w/ spores 1. ______________________________ 2. ______________________________ STAINS: _____________________________________________ _____________________________________________ _____________________________________________ _____________________________________________ _____________________________________________ _______________________________________________ Capsule - Increase virulence by preventing Phagocytosis - Antigenic on the basis of serotyping by Quellung Reaction- ______________ - Colonies often slimy - Stains: __________________________ ___________________________ S. pneumoniae K. pneumoniae H. influenzae P. aeruginosa N. meningitidis C. neoformans Pili - Ordinary pili- adherence of bacteria to host cell - Sex pili- bacterial conjugation Flagella Atrichous-___________________________ Monotrichous-________________________ Amphitrichous-_______________________ Lophotrichous-_______________________ Periplasmic flagella-___________________ -Motility best seen at __________________ -Tests for motility: 1.Hanging drop method ___________________________________________ ___________________________________________ ___________________________________________ 2. Semi-solid medium Kinyoun Acid Fast Organism Non Acid-Fast Organism P. Stain Mordant Decolorizer S. Stain Ziehl- Neelsen Acid Fast Organism Non Acid Fast Organism P. Stain Mordant Decolorizer S. Stain
Miguel Carlos G. Arada, RMT, ASCP(IMT), DTA, MSMT ___________________________________________ ___________________________________________ ___________________________________________ -Stains for flagella: ___________________________________________ ___________________________________________ ___________________________________________ ---------------------------------------------------------------- BACTERIAL GROWTH FACTORS 1. Nutrients - Carbon- for synthesis of cell component a. Lithotroph- from inorganic substance Ex. CO2 b. Heterotroph- from organic substance Ex. Water, minerals, salts _______________________________________________ _______________________________________________ _______________________ -Nitrogen -Minerals- Ex. Sulfur, magnesium -Salt -Others 2. Oxygen and Carbon dioxide availability A. Obligate/strict aerobe_________________________________________ _______________________________________________ _______________________________________________ B. Obligate/strict anaerobe- _______________________________________________ _______________________________________________ _______________________________________________ C. Facultative anaerobe- _______________________________________________ _______________________________________________ _______________________ D. Aerotolerant anaerobe- _______________________________________________ _______________________________________________ _______________________________________________ E. Microaerophilic- _______________________________________________ _______________________________________________ _______________________________________________ F. Capnophilic- _______________________________________________ _______________________________________________ _______________________________________________ NOTES: _______________________________________________ _______________________________________________ _______________________________________________ _______________________________________________ _______________________________________________ 3. Temperature A. Psychrophilic/cryophilic- _______________________________________________ _______________________________ B. Mesophilic- _______________________________________________ _______________________________ C. Thermophilic- _______________________________________________ _______________________________ 4. pH A. Acidophile-____________________________ B. Alkaliphile-____________________________ ---------------------------------------------------------------- Growth cycle ---------------------------------------------------------------- 1. Lag phase- little or no multiplication 2. Log or exponential phase- organisms grow at maximum rate 3. Stationary or plateau phase- growth ceases because nutrients are exhausted or toxic metabolic products have accumulated 4. Decline phase- Viable count decreases ---------------------------------------------------------------- STERILIZATION- process that kills all forms of microbial life, including bacterial spores ________________ PHYSICAL METHOD I- Heat
Miguel Carlos G. Arada, RMT, ASCP(IMT), DTA, MSMT A. Moist heat- coagulates protein 1. AUTOCLAVE- most effective method of sterilization QC:__________________________ ________________________________ _________________________________ _________________________________ 2. FRACTIONAL- alternate heating(kills vegetative cells), incubation (spores germinate), heating (kills remaining vegetative cells and spores) a. Tyndallization-________________ _____________________________ b. Inspissation-__________________ _____________________________ B. Dry Heat- kills by oxidation 1. Flame- to sterilize inoculating loops and needles 2. Oven- for glasswares, certain metals, oils, petroleum or powders. QC:___________________________ ________________________________ 3. Incineration- most common method of treating infectious waste. _______________________________ II- FILTRATION -filtration of air is accomplished using HEPA __________________________ filters designed to remove organisms larger than 0.3 um from isolation rooms, operating rooms, and biologic safety cabinets. -Ex. Asbestos filter, Membrane filter III- IONIZING RADIATION -Plastic syringes, catheter or gloves -Short wavelength, high energy gamma rays __________________ CHEMICAL METHODS I- ETO_______________________________ - Most common chemical sterilant - Cold sterilization - Used for materials which cannot be autoclaved - Strict requirements of concentration - QC:__________________________ II-Formaldehyde vapour and vapour phase hydrogen peroxide - Sterilize HEPA filters in BSCs III- Glutaraldehyde- - Cold sterilization - Sporicidal in 3 to 10 hours - Used in medical equipment such as bronchoscopes because it does not corrode lenses, metal or rubber. IV-Peracetic Acid - Cold sterilization - Effective in the presence of organic material - Used in surface sterilization of surgical instruments DISINFECION- process that destroys pathogenic organisms,but not necessarily all organisms or spores _________________ PHYSICAL METHODS I. Boiling_______________________________ II. Pasteurization -used mostly in the food industry, eliminates food-borne pathogens and organisms responsible for food spoilage. A. Batch Method_____________________ B. Flash Method______________________ III. Non-ionizing radiation -long wavelength and low energy -do not penetrate well and organisms must have direct surface exposure, such as the working surface of a BSC for this form of disinfection to work. __________________ CHEMICAL METHODS 1. Disinfectant___________________________ 2. Antiseptic_____________________________ I.Alcohol -ethyl or isopropyl alcohol is nonsporicidal and evaporates quickly -70% vs 95% ___________________________________ ___________________________________ II. Aldehyde -toxic to humans -___________formaldehyde ___________glutaraldehyde III. Halogen -Iodine and chlorine -70% alcohol followed by an iodophor is the most common compound used for skin disinfection before drawing blood specimens for culture and surgery. -Chlorine is most often used in the form of ______________________-household bleach IV. Heavy metals