1. Introduction and Historical Development in Microbiology Organism - object / structures invisible to the naked eye are called microorganism = human eye cannot see object smaller than 30u (1/1000 of an inch in dia.) Microbiology – science that deals with the study of microorganisms and their activities Medical Microbiology – deals with the study of disease producing organism affecting human = also concerned with prevention and control of disease.

13. Classification and Identification of Bacteria Protist = group of single–celled microorganism which do not form highly differentiated tissue and organ system. In the early history living organism were classified into 2 kingdoms: 1. Plant 2. Animal = after the discovery of microorganism, it was found that many microorganism possess both plant and animal characteristic or neither plant or animal characteristic. So a new kingdom was proposed for microorganism (Kingdom Protista) = all bacteria, fungi, algae and protozoa are group in kingdom protista = viruses are not included because they do not have cellular organization.

16. Laboratory Procedure Employed in the Identification of Bacteria 1. Isolation of bacteria in pure (axenic) culture. = depends on source of clinical specimen = blood, spinal fluid and closed abscesses yield pure bacterial culture of microorganism = sputum, skin and body fluids, stool usually contains mixed organism.

24. Specimen smear cultivation (pure culture) microscopic colonial morph. exam. biochemical serologic animal path. antimicrobial susceptibility

25. Bacterial Morphology Bacterial cell is a complete unit of any living organism. All its functions are genetically controlled and performed by that particular cell structure whether it be physiologic or biochemical.

33. II. Arrangement = is the result of the number of planes in which the cell divides and how they remain attached afterwards.

39. IV. Staining Reaction = putting a color to facilitate identification = unstained bacteria are colorless and transparent = can be determined through the used of basic aniline dyes

42. BACTERIAL CELL ULTRASTRUCTURES

44. General Division: I. External - Capsule - Fimbriae - Flagellum II. Internal - Cell Wall - Cytoplasmic Membrane - Cytoplasm - Mesosome - Nuclear Body - Ribosome - Plasmid - Inclusion Granule - Endospore

50. Types and Arrangement of flagella in relation to the distribution and number: I. Monotrichous – single polar flagellum at one end II. Multitrichous – more than one flagella

58. Gram Positive Gram Negative

61. Protoplast: comprises the naked cytoplasmic membrane and its content PERIPLASM = space between the plasma inner membrane and the outer membrane layer = readily observe among gram negative bacteria = consist of a gell-like substances which help secure nutrients from the environment CYTOPLASMIC/PLASMA/CELL MEMBRANE: = thin elastic bilayered semi-permeable membrane lying underneath the cell wall enclosing the cytoplasm of the cell. Chemical Composition - Lipoprotein - (composed of phospholipids and protein molecules)

66. RIBOSOME: = histone-like particles composed of ribosomal RNA (rRNA) and protein molecules found in the cytoplasm of the cell. = site of action for many antibiotics that inhibit protein synthesis. = have a sedimentation coefficient of 70S and are composed of 50 S and 30 S subunits containing 16 S, 23 S and 5 S RNA respectively = procaryotic ribosome - 70s = eucaryotic ribosome - 80s Function – Site for protein synthesis Detection – Feulgin staining (-), Electron microscopy

69. ENDOSPORES: = highly refractile body formed within vegetative bacterial cells in response to adverse environmental condition and due to deficient nutrient supply = known as endospore, because it is formed inside the bacteria cell and is genetically controlled = are metabolically inactive bacterial cell that are highly resistant to desiccation, heat and various chemical agents = found in the cytoplasm of rod-shaped sporeforming bacteria of the genus Bacillus & Clostridium

76. b) Nitrogen Requirement = main reservoir of nitrogen is nitrogen gas (N 2 ) which make up 79% of earth atmosphere = must be degraded into their basic building block (Protein  Amino acid; Nucleic acid  Nucleotides) 3) Inorganic Ions = small amount needed ex. Sulfur, Phosphorous, Magnesium, Calcium, Manganese, Zinc, Cobalt, Copper

77. 4) Growth Substances = organic nutrient essential to an organism metabolism that cannot be synthesized and must be provided in the culture medium Ex. yeast extract, whole blood, serum, B-complex vitamins, amino acids, purins and pyrimidines Prototrophic = bacteria that do not require exogenous source of growth factor because they are capable of synthesizing their own Auxotrophic = bacteria that requires additional growth factor in the culture medium for growth to occur

79. 3. Thermophiles = 25-80 0 C opt. 50-60 0 C = hotspring, tropical soil, hot water heater 4. Thermoduric = 80-100 0 C = resist high temperature but cannot grow and multiply

90. STERILIZATION AND DISINFECTION

91. STERILIZATION - the process of destroying all forms of microbial life in terms of their ability to reproduce and multiply including spores PHYSICAL AGENTS FOR STERILIZATION A. HEAT: = most reliable method of sterilization = method of choice for material which are not damage by heat = denature proteins and damage membrane 2 types of heat: 1. DRY HEAT: = preferred method for sterilization of glasswares, such as glass syringes, test tube, petri plates and material such as oil, jellies and powders = less effective than moist heat as it requires longer time of heating and higher temperature = kills the organism by oxidizing their chemical constituent

92. a) Hot air oven: = most widely used method of sterilization by dry heat = used for sterilization of glasswares such as flask, pipettes, test tube, swab = principle of killin g: heating at 160 O C or 180 O C for 30 minutes b) Direct Flamming/Heating = direct heating of instrument or object in a flame till it becomes red hot = easy way of sterilization but has limited application = useful method of sterilization for non-inflamable material such as bacteriological loop or needle, forceps, scissors c) Incineration = direct burning of contaminated materials = efficient method for sterilization and disposal of soiled dressing, bedding of patient and animal carcasses

97. Chemical agents : Disinfection = process destroying / eliminating potentially hazardous & pathogenic organism = aims to destroy the disease-producing organisms Disinfectant = agents that kills growing (vegetative) form of organism but not necessarily sporeformer = applied to non-living/inanimate object Bactericidal / Germicidal = implies the killing action of chemical substances Bacteriostatic = substances that inhibit / prevent growth & multiplication of bacteria Antiseptics = subs. that either kill / prevent growth = applied to living tissue (animate object) found in the surface of skin and mucous membrane

100. III – Alcohol = disorganize lipids  cell membrane = denature protein = widely used antiseptics = has ability to remove lipids from skin surfaces Ethanol / Ethyl alcohol = skin disinfectant = disinfection clinical thermometer = effective against gram (+) , gram (-) , AF bacteria Isopropyl alcohol = most effective, less volatile = bactericidal property greater than ethanol = used as skin antiseptic = has a better fat solvent property and more bactericidal = effective at a concentration of 50-70%

107. Cephalosporins = fermentation product fungus  Cephalosporium First Generation – bactericidal against most gram (+) and many gram (-) bacteria , Except : Enterobacter Proteus ( Indole + ) Pseudomonas Cephalothin Cephapirin Cefazolin Cephradine Cephalexin Cefadroxil

108. Second Generation – Gram (-) bacilli – Enterobacter - Proteus Cefamandole - Hemophilus Cefoxitin – N. gonorrhea Cefaclor Serratia Cefuroxime Bacteroides Third Generation – less active against gram (+) but more active against gram (-) bacteria Cefoperazone Ceftazidine more active against Cefpiramide Pseudomonas Cefpirome