Introduction and Historical  Development in Microbiology  Organism   -  object / structures invisible to the naked eye  ar...
<ul><li>Scope of study (order of decreasing size)   </li></ul><ul><ul><ul><ul><ul><li>a) Algae (Phycology)  </li></ul></ul...
BRIEF  HISTORY <ul><li>before microorganism were discovered, ancient  people regarded diseases as a form  </li></ul><ul><l...
<ul><li>Varro  = during the 2 nd  century B.C.– postulated the  </li></ul><ul><li>concept of contagion </li></ul><ul><li>...
<ul><li>Spontaneous Generation Theory </li></ul><ul><li>Aristotle – proposed the  </li></ul><ul><li>“ Spontaneous Generati...
Experiments to disprove  Spontaneous Generation Theory   <ul><li>Francesco Redi  1668 </li></ul><ul><ul><li>did an experim...
<ul><li>John Tyndall  </li></ul><ul><ul><li>proved that dust carried germs  </li></ul></ul><ul><ul><li>also demonstrated t...
<ul><li>Robert Hooke  1678 </li></ul><ul><ul><li>developed the compound microscope and was able to confirm Leeuwenhoek’s  ...
<ul><li>Louis Pasteur  </li></ul><ul><ul><li>published the results of an experiment he did to disprove  spontaneous genera...
<ul><li>Robert Koch  1876 </li></ul><ul><ul><li>establish proof of microbial etiology of 3 important diseases:  </li></ul>...
<ul><ul><li>Koch’s postulates : </li></ul></ul><ul><li>1. The organism must always be found in diseased  </li></ul><ul><li...
<ul><li>Exceptions to the Kock’s Postulate: </li></ul><ul><ul><li>#1) Carrier - some individual harbors the organism but d...
Classification and Identification of Bacteria  Protist   = group of single–celled microorganism which do not form highly d...
<ul><li>= Kingdom Protista is divided into 2 groups based on cellular organization:  </li></ul><ul><li>1. Procaryotes – lo...
<ul><li>B) Procaryotes – Lower protista  </li></ul><ul><ul><ul><li>Characteristic: </li></ul></ul></ul><ul><ul><ul><ul><li...
Laboratory Procedure Employed in the Identification  of Bacteria  1.  Isolation of bacteria in pure (axenic) culture. = de...
<ul><li>2.  Bacterial colony morphology (macroscopic) </li></ul><ul><ul><ul><ul><li>= colony is composed of descendant of ...
<ul><li>4 types of bacterial colony : </li></ul><ul><ul><ul><li>1)  Smooth (S-colonies)  - gives the appearance of  </li><...
<ul><ul><ul><li>3)  Mucoid (M-colonies)   - exhibits a water-like  glistening </li></ul></ul></ul><ul><ul><ul><li>confluen...
<ul><ul><ul><li>3.  Microscopic morphology and staining reaction. </li></ul></ul></ul><ul><ul><ul><ul><li>= determines sha...
<ul><ul><ul><ul><li>3 types of staining procedure : </li></ul></ul></ul></ul><ul><ul><ul><ul><li>1. Simple – use of 1 dye ...
<ul><li>4. Biochemical   </li></ul><ul><li>= detect different metabolic products and sugar  </li></ul><ul><li>fermentation...
<ul><ul><ul><li>5. Serological   – detect presence of antibodies in patient  </li></ul></ul></ul><ul><ul><ul><li>serum aga...
Specimen  smear  cultivation  (pure culture) microscopic    colonial morph.   exam.     biochemical    serologic   animal ...
Bacterial Morphology   Bacterial cell is a complete unit of any living organism. All its functions are genetically control...
 
<ul><li>Bacterial morphology includes:   </li></ul><ul><li>Form, Arrangement, Size and Staining reaction </li></ul><ul><li...
<ul><li>A. Coccus  (spherical-shaped) </li></ul><ul><li>= round-like a ball, perfect sphere, globe </li></ul><ul><li>= can...
<ul><ul><ul><li>B) Bacillus (rod-shaped)  </li></ul></ul></ul><ul><ul><ul><li>= cell appears longer than wide or cylindric...
<ul><ul><ul><li>C) Spiral </li></ul></ul></ul><ul><ul><ul><li>= bacteria w/ more than one somatic curved </li></ul></ul></...
<ul><li>Genus Borrelia -  much less tightly coiled having  </li></ul><ul><li>the appearance of an extremely long  </li></u...
<ul><li>  D) Intermediate forms   </li></ul><ul><li>  A. Coccobacillary–when a rod is short thick, wide/plump. </li></ul><...
II. Arrangement   = is the result of the number of planes in which the  cell divides and how they remain attached  afterwa...
<ul><li>A) Sphericals   </li></ul><ul><li>1. Singly  – occurs as a single spherical cell </li></ul><ul><li>2. Chain  – com...
<ul><ul><ul><ul><li>4. Cluster  – common with sphericals resulting from </li></ul></ul></ul></ul><ul><ul><ul><ul><li>many ...
<ul><ul><ul><li>B) Rods   </li></ul></ul></ul><ul><ul><ul><li>= divides only across their short axis  </li></ul></ul></ul>...
<ul><ul><ul><li>4.  Packets of cigarette  – arrangement like bundles. </li></ul></ul></ul><ul><li>Ex. Mycobacterium leprae...
<ul><li>III. Bacterial size </li></ul><ul><ul><ul><ul><li>1. Micrometer  </li></ul></ul></ul></ul><ul><ul><ul><ul><li>2. U...
IV. Staining Reaction   = putting a color to facilitate identification  = unstained bacteria are colorless and  transparen...
<ul><li>Types:   </li></ul><ul><li>1. Simple  = employs one dye only </li></ul><ul><ul><ul><ul><ul><li>  = sufficient to d...
<ul><li>2.b) Acidfast  = differentiate acidfast from  </li></ul><ul><li>non-acidfast bacteria </li></ul><ul><li>3. Special...
BACTERIAL CELL  ULTRASTRUCTURES
 
General Division: I. External   - Capsule  - Fimbriae  - Flagellum  II. Internal   - Cell Wall  - Cytoplasmic Membrane  - ...
<ul><ul><ul><li>BACTERIAL  ULTRASTRUCTURE </li></ul></ul></ul><ul><ul><ul><li>Bacterial Surface Coating:  </li></ul></ul><...
<ul><ul><ul><li>Slime layer </li></ul></ul></ul><ul><ul><ul><li>= loose and irregularly arranged meshwork of  </li></ul></...
<ul><li>Chemical Comp.:  </li></ul><ul><li>= water – main component 98-99% </li></ul><ul><li>= 1-2% of bacterial capsule a...
<ul><li>B. Appendages   Flagellum & Axial filaments  </li></ul><ul><li>  Fimbriae  </li></ul><ul><li>FLAGELLUM:   </li></u...
<ul><li>Chemical Comp.:  protein monomer - Flagellin  </li></ul><ul><li>Function  - Motility organelle of many pathogenic ...
Types and Arrangement of flagella in relation to the  distribution and number: I. Monotrichous  – single polar flagellum a...
<ul><li>Types: </li></ul><ul><ul><ul><li>A) Lophotrichous – tuft of flagella at one polar end </li></ul></ul></ul><ul><ul>...
<ul><ul><ul><li>  Axial Filaments   </li></ul></ul></ul><ul><ul><ul><li>= flagella-like sheathed filaments located in  </l...
<ul><li>2 types according to function: </li></ul><ul><li>1. Common/ordinary – for attachment or adherence  </li></ul><ul><...
<ul><li>Cell wall  (Peptidoglycan, Murien, Mucopeptide / Glycopeptide) </li></ul><ul><li>= complex, rigid, multilayered st...
<ul><ul><ul><li>Function : </li></ul></ul></ul><ul><ul><ul><li>1. Responsible for the characteristic  shape of bacterium <...
COMPARISON OF GRAM POSITIVE AND GRAM NEGATIVE CELL WALL Gram Positive Gram Negative Peptidoglycan  <ul><li>Very thick homo...
 
Gram Positive   Gram Negative
<ul><li>Damage to the cell wall may result to: </li></ul><ul><ul><ul><li>= when a gram positive bacteria is exposed to  </...
 
Protoplast:   comprises the naked cytoplasmic membrane and   its content  PERIPLASM  = space between the plasma inner memb...
<ul><li>Functions:   </li></ul><ul><ul><ul><li>1. Serve as selective permeability barrier  </li></ul></ul></ul><ul><ul><ul...
<ul><li>MESOSOME:   </li></ul><ul><li>  = usually seen as an invagination of the membrane  </li></ul><ul><li>associated cy...
<ul><li>NUCLEAR BODY:  (Nucleus / Nucleoid) </li></ul><ul><li>= genetic material of bacteria  </li></ul><ul><li>= contains...
<ul><li>PLASMID/EPISOME:   </li></ul><ul><li>= extrachromosomal genetic material capable of  </li></ul><ul><li>autonomous ...
RIBOSOME:   = histone-like particles composed of ribosomal RNA  (rRNA) and protein molecules found in the  cytoplasm of th...
<ul><li>INCLUSION / CYTOPLASMIC GRANULE:   </li></ul><ul><li>= large granules found anywhere in the cytoplasm </li></ul><u...
<ul><ul><ul><ul><li>2. Lipid granules  </li></ul></ul></ul></ul><ul><ul><ul><ul><ul><li>Chemical Composition  – polymer of...
ENDOSPORES:   = highly refractile body formed within vegetative  bacterial cells in response to adverse environmental cond...
<ul><li>= Composed of five parts: </li></ul><ul><li>  1.   Core – located at the center which is the spore  </li></ul><ul>...
<ul><li>= contains Calcium Dipicolinate which aids in heat  </li></ul><ul><li>resistance within the core by making the pro...
<ul><li>Classification accdg. to: </li></ul><ul><li>A.  Location  - Central, Subterminal, Terminal  </li></ul><ul><li>B.  ...
<ul><ul><ul><li>PHYSIOLOGY OF BACTERIAL CELL  </li></ul></ul></ul><ul><ul><ul><li>Nutrition   </li></ul></ul></ul><ul><ul>...
<ul><ul><ul><li>Bacterial growth requirement: </li></ul></ul></ul><ul><ul><ul><li>1) Water   – most important requirement ...
<ul><li>Types of bacteria acdg. to carbon requirement  </li></ul><ul><ul><ul><ul><li>1. Autotroph (Lithotroph) </li></ul><...
b) Nitrogen Requirement   = main reservoir of nitrogen is nitrogen gas (N 2 )  which make up 79% of earth atmosphere = mus...
4) Growth Substances   = organic nutrient essential to an organism metabolism that cannot be synthesized and must be provi...
<ul><li>II. Physical Requirement   </li></ul><ul><ul><ul><li>A)Temperature  </li></ul></ul></ul><ul><ul><ul><ul><ul><li>1....
3. Thermophiles  = 25-80 0 C  opt. 50-60 0 C = hotspring, tropical soil, hot water heater  4. Thermoduric  = 80-100 0 C = ...
<ul><ul><ul><li>B) Oxygen  </li></ul></ul></ul><ul><ul><ul><ul><li>= required by particular bacterium to satisfy its energ...
<ul><ul><ul><li>3) Facultative anaerobes  </li></ul></ul></ul><ul><ul><ul><li>= capable of growth under both aerobic and  ...
<ul><ul><ul><li>C) Hydrogen Ion Concentration (PH)   </li></ul></ul></ul><ul><ul><ul><ul><li>= optimum ph for pathogenic b...
<ul><li>Measurement of Bacterial Growth </li></ul><ul><ul><ul><li>1. Cell Concentration (cell number) number of cell per u...
<ul><ul><ul><ul><li>2) Indirect Viable Count  </li></ul></ul></ul></ul><ul><ul><ul><ul><ul><li>= plate sample of culture a...
<ul><li>2) Cell Density (Cell Mass) </li></ul><ul><li>= total protoplasm/dry weight of the cell per unit  </li></ul><ul><l...
<ul><li>Bacterial Growth Curve  </li></ul><ul><li>(5 PHASES)  </li></ul><ul><ul><ul><li>1. Lag phase  (Phase of Rejuvenesc...
<ul><ul><ul><li>3. Stationary phase   </li></ul></ul></ul><ul><ul><ul><li>(phase of equilibrium/”plateau”) </li></ul></ul>...
 
<ul><li>Consequence of growth & multiplication is reproduction </li></ul><ul><li>3 methods of bacterial reproduction   </l...
STERILIZATION  AND   DISINFECTION
STERILIZATION  - the process of destroying all forms of microbial life in terms of their ability to reproduce and multiply...
a) Hot air oven:  = most widely used method of sterilization by dry heat  = used for sterilization of glasswares such as f...
<ul><li>2. Moist Heat  = kills bacteria faster than dry heat  </li></ul><ul><ul><ul><li>Boiling  </li></ul></ul></ul><ul><...
<ul><li>4. Tyndallization (Fractional/Intermittent sterilization)  </li></ul><ul><li>= for sterilization of liquid & semi-...
<ul><ul><ul><li>B)  Radiation </li></ul></ul></ul><ul><ul><ul><li>= produce damage to DNA resulting to killing of the orga...
<ul><ul><ul><li>D) Ultrasomic vibration   </li></ul></ul></ul><ul><li>= are sound waves with high frequency  </li></ul><ul...
Chemical agents : Disinfection  = process destroying / eliminating potentially   hazardous & pathogenic organism  = aims t...
<ul><li>Mech. of action: </li></ul><ul><ul><ul><li>1. damage cell membrane function  </li></ul></ul></ul><ul><ul><ul><li>2...
<ul><li>II – Phenolic compound   </li></ul><ul><li>= disrupt cell membrane function    leaking cell content  </li></ul><u...
III – Alcohol   = disorganize lipids    cell membrane  = denature protein  = widely used antiseptics  = has ability to re...
<ul><li>Agents that denatures protein   </li></ul><ul><ul><ul><li>1. Acid & alkalies  </li></ul></ul></ul><ul><ul><ul><li>...
<ul><ul><ul><li>3. Dyes  </li></ul></ul></ul><ul><ul><ul><ul><li>Triphenyl methane dyes (aniline acid) </li></ul></ul></ul...
<ul><ul><ul><li>Glutaraldehyde  </li></ul></ul></ul><ul><ul><ul><li>= used as cold sterilant for sterilizing medical  </li...
<ul><li>Chemotherapeutic Agent </li></ul><ul><li>= one of the most valuable method of treating infection  </li></ul><ul><l...
<ul><ul><ul><ul><ul><li>Antibiotic producers   </li></ul></ul></ul></ul></ul><ul><li>1.  Penicillium  </li></ul><ul><li>2....
<ul><li>Penicillins   </li></ul><ul><ul><ul><li>1. Natural PCN – Benzyl PCN G </li></ul></ul></ul><ul><li>  - Phenoxymethy...
Cephalosporins  = fermentation product fungus     Cephalosporium  First Generation  – bactericidal against most gram (+) ...
Second Generation  – Gram (-) bacilli – Enterobacter    - Proteus  Cefamandole - Hemophilus  Cefoxitin  –  N. gonorrhea  C...
<ul><li>II : Cell Membrane Inhibitors   </li></ul><ul><ul><ul><li>1. Polymixins – reserved for serious pseudomonas infecti...
<ul><ul><ul><ul><ul><li>IV : Inhibitors Protein Synthesis   </li></ul></ul></ul></ul></ul><ul><li>A.  Actinomycin – active...
<ul><ul><ul><ul><ul><li>V : Metabolite Analogue   </li></ul></ul></ul></ul></ul><ul><li>1.  Sulfonamide  </li></ul><ul><li...
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Micro bio dentistry.ppt

  1. 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.
  2. 2. <ul><li>Scope of study (order of decreasing size) </li></ul><ul><ul><ul><ul><ul><li>a) Algae (Phycology) </li></ul></ul></ul></ul></ul><ul><ul><ul><li>b) Fungi (Mycology) </li></ul></ul></ul><ul><ul><ul><li>c) Protozoa (Protozoology / Parasitology) </li></ul></ul></ul><ul><ul><ul><li>d) Bacteria (Bacteriology) </li></ul></ul></ul><ul><ul><ul><li>e) Virus (Virology) </li></ul></ul></ul><ul><ul><ul><li>Features of Microorganism : </li></ul></ul></ul><ul><ul><ul><li>* All microorganisms are not alike </li></ul></ul></ul><ul><ul><ul><li>* Some of them are very small while others are </li></ul></ul></ul><ul><ul><ul><li>relatively larger in size </li></ul></ul></ul><ul><ul><ul><li>* Some of them possess plant-like characteristic, </li></ul></ul></ul><ul><ul><ul><li>while others have animal-like and few of them </li></ul></ul></ul><ul><ul><ul><li>have neither plant-like nor animal-like characters </li></ul></ul></ul>
  3. 3. BRIEF HISTORY <ul><li>before microorganism were discovered, ancient people regarded diseases as a form </li></ul><ul><li>of punishment sent by GOD for the sins of men </li></ul><ul><li>disease were not new to man </li></ul><ul><li>Biblical disease - Leprosy </li></ul><ul><li>- Tuberculosis </li></ul><ul><li>- Syphilis </li></ul><ul><li>- Plague </li></ul><ul><li>treatment and prevention of these diseases were sought by sacrifices to appease the anger of God </li></ul>
  4. 4. <ul><li>Varro = during the 2 nd century B.C.– postulated the </li></ul><ul><li>concept of contagion </li></ul><ul><li> </li></ul><ul><li>diseases are transmitted </li></ul><ul><ul><ul><li>by invisible creature </li></ul></ul></ul><ul><li>Roger Bacon (13 th century) = postulated that invisible </li></ul><ul><li>living things called “germs ” produce disease </li></ul><ul><li>Fracastorius (1546) = postulated that disease is caused </li></ul><ul><li>by invisible living things and can be transmitted </li></ul><ul><li>by direct contact (person to person contact) </li></ul>
  5. 5. <ul><li>Spontaneous Generation Theory </li></ul><ul><li>Aristotle – proposed the </li></ul><ul><li>“ Spontaneous Generation Theory” </li></ul><ul><li>also called Abiogenesis </li></ul><ul><li>state that living things could develop spontaneously from non-living materials </li></ul><ul><li>supported by appearance of living creatures in decaying meat, stagnating ponds, fermenting grain, and infected wounds </li></ul>
  6. 6. Experiments to disprove Spontaneous Generation Theory <ul><li>Francesco Redi 1668 </li></ul><ul><ul><li>did an experiment with flies and wide-mouth jars containing meat </li></ul></ul><ul><ul><li>he demonstrated the appearance of maggots in decomposing meat as a result on the deposition of eggs by flie </li></ul></ul><ul><li>Rudolph Virchow 1858 </li></ul><ul><ul><li>1 st person to propose the Theory of Biogenesis </li></ul></ul><ul><ul><ul><li>‘ Cells can only arise from preexisting cells’ </li></ul></ul></ul>
  7. 7. <ul><li>John Tyndall </li></ul><ul><ul><li>proved that dust carried germs </li></ul></ul><ul><ul><li>also demonstrated the great resistance of spore to heat </li></ul></ul><ul><ul><li>introduced (Tyndallization/Fractional Sterilization) a method by which spore can be destroyed by heating using a free-flowing steam for 30-60 minutes at 100 ° C for 3 consecutive days. </li></ul></ul><ul><li>Anton Van Leeuwenhoek </li></ul><ul><ul><li>1 st to describe bacteria in 1677 with the use of a simple microscope utilizing crude lenses </li></ul></ul><ul><ul><li>considered as the Father of Bacteriology for being the first person described the 3 morphologic forms of bacteria: rod, spherical, spiral </li></ul></ul>
  8. 8. <ul><li>Robert Hooke 1678 </li></ul><ul><ul><li>developed the compound microscope and was able to confirm Leeuwenhoek’s discoveries </li></ul></ul><ul><ul><li>first person to coin the word “cell” to describe the tiniest components of the living system </li></ul></ul><ul><li>Agostino Bassi 1800 </li></ul><ul><ul><li>proved that a fungus cause a disease in “silkworm” called Muscardine </li></ul></ul><ul><li>Rayer and Davaine 1850 </li></ul><ul><ul><li>were able to observed the presence of microorganism in blood of animals dying from a disease called Anthrax. </li></ul></ul><ul><li>Edward Jenner - developed the concept of vaccination </li></ul><ul><li>Paul Erlich - developed the used of staining to demonstrate bacterial cell morphology for better visualization </li></ul>
  9. 9. <ul><li>Louis Pasteur </li></ul><ul><ul><li>published the results of an experiment he did to disprove spontaneous generation in microscopic organisms </li></ul></ul><ul><ul><li>introduced Pasteurization a method of killing organisms found in dairy products </li></ul></ul><ul><li>Joseph Lister 1860s </li></ul><ul><ul><li>demonstrated the importance of using antiseptic to control spread of disease-producing organism </li></ul></ul><ul><ul><li>uses a chemical disinfectant (aqueous phenol) to prevent surgical wound infections. </li></ul></ul><ul><ul><li>also developed the first pure culture technique using liquid medium which was the key to identification of bacteria </li></ul></ul>
  10. 10. <ul><li>Robert Koch 1876 </li></ul><ul><ul><li>establish proof of microbial etiology of 3 important diseases: </li></ul></ul><ul><ul><ul><ul><li>Cholera ( Vibrio cholerae) </li></ul></ul></ul></ul><ul><ul><ul><ul><li>Tuberculosis ( Mycobacterium tuberculosis) </li></ul></ul></ul></ul><ul><ul><ul><ul><li>Anthrax ( Bacillus anthracis) </li></ul></ul></ul></ul><ul><ul><li>formulated the Koch’s postulates which provides proof that a specific bacterium caused a particular disease. </li></ul></ul><ul><ul><li>also developed a solid culture medium for isolation of bacteria in pure culture. </li></ul></ul>
  11. 11. <ul><ul><li>Koch’s postulates : </li></ul></ul><ul><li>1. The organism must always be found in diseased </li></ul><ul><li> animals and not in healthy one </li></ul><ul><li>2. Organisms isolated from the diseased animals </li></ul><ul><li> must be grown in pure culture away from the </li></ul><ul><li> diseased animals </li></ul><ul><li> 3. Organisms grown in pure culture must initiate and </li></ul><ul><li>reproduce the disease when re-inoculated into </li></ul><ul><li>susceptible animal </li></ul><ul><li>4. Organisms must be re-isolated from experimentally infected animals </li></ul>
  12. 12. <ul><li>Exceptions to the Kock’s Postulate: </li></ul><ul><ul><li>#1) Carrier - some individual harbors the organism but do not </li></ul></ul><ul><ul><li>manifest the disease and can transmit the disease to others </li></ul></ul><ul><ul><li>#2) Some or certain organisms/bacteria cannot be cultured in </li></ul></ul><ul><ul><li> vitro (cannot grow in artificial culture media) </li></ul></ul><ul><li> Ex. Mycobacterium leprae / Treponema pallidum </li></ul><ul><ul><li>#3) Certain animals not susceptible to certain microorganism some animals are by nature immune </li></ul></ul><ul><ul><ul><li> - cannot replicate even with the same specie </li></ul></ul></ul><ul><ul><ul><li>some bacterial specie are host specific </li></ul></ul></ul><ul><ul><ul><ul><li> Ex. Vibrio cholerae cannot cause chicken cholera or vice versa </li></ul></ul></ul></ul>
  13. 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.
  14. 14. <ul><li>= Kingdom Protista is divided into 2 groups based on cellular organization: </li></ul><ul><li>1. Procaryotes – lower protista </li></ul><ul><li>2. Eucaryotes – higher protista </li></ul><ul><li>Eucaryotes – Higher protista </li></ul><ul><ul><ul><li>Characteristics </li></ul></ul></ul><ul><ul><ul><ul><li>1) Size greater than 5um </li></ul></ul></ul></ul><ul><ul><ul><ul><li>2) Possess a well developed true nucleus enclosed in a nuclear membrane </li></ul></ul></ul></ul><ul><ul><ul><ul><li>3) Contain sets of chromosomes </li></ul></ul></ul></ul><ul><ul><ul><ul><li>4) Cytoplasm contains mitochondria endoplasmic reticulum and vacuoles </li></ul></ul></ul></ul><ul><ul><ul><ul><li>5) Cell membrane continous with well-defined </li></ul></ul></ul></ul><ul><ul><ul><ul><li>endoplasmic reticulum and nuclear membrane </li></ul></ul></ul></ul><ul><ul><ul><ul><li>6) Ribosome (sedimentation coefficient) 80s </li></ul></ul></ul></ul><ul><ul><ul><ul><li>7) Motility organelle multistranded </li></ul></ul></ul></ul><ul><ul><ul><ul><li>Eg: Algae (Red – Brown), fungi, protozoa, slime mold </li></ul></ul></ul></ul>
  15. 15. <ul><li>B) Procaryotes – Lower protista </li></ul><ul><ul><ul><li>Characteristic: </li></ul></ul></ul><ul><ul><ul><ul><li>1) Size less than 4um </li></ul></ul></ul></ul><ul><ul><ul><ul><li>2) Nucleus primitive and is homogenous with the </li></ul></ul></ul></ul><ul><ul><ul><ul><li>cytoplasm of cell and not enclosed in a nuclear </li></ul></ul></ul></ul><ul><ul><ul><ul><li>membrane (naked) </li></ul></ul></ul></ul><ul><ul><ul><ul><li>2) Posses single chromosome (haploid) </li></ul></ul></ul></ul><ul><ul><ul><ul><li>3) Do not have mitochondria and endoplasmic reticulum </li></ul></ul></ul></ul><ul><ul><ul><ul><li>4) Cell wall made up of complex rigid layer </li></ul></ul></ul></ul><ul><ul><ul><ul><li>5) Ribosome (sedimentation coefficient) 70s </li></ul></ul></ul></ul><ul><ul><ul><ul><li>6) Motility organelle unstranded </li></ul></ul></ul></ul><ul><ul><ul><ul><li>Eg: Bacteria, Blue – Green algae </li></ul></ul></ul></ul>
  16. 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.
  17. 17. <ul><li>2. Bacterial colony morphology (macroscopic) </li></ul><ul><ul><ul><ul><li>= colony is composed of descendant of a single cell, </li></ul></ul></ul></ul><ul><ul><ul><ul><li>a clone or lump of cell </li></ul></ul></ul></ul><ul><ul><ul><ul><li>= usually the result of rapid multiplication of cell </li></ul></ul></ul></ul><ul><ul><ul><ul><li>when inoculated in a medium containing 2% </li></ul></ul></ul></ul><ul><ul><ul><ul><li>agar incubated for 18 – 24 hours in favorable </li></ul></ul></ul></ul><ul><ul><ul><ul><li>atmosphere. </li></ul></ul></ul></ul><ul><ul><ul><ul><li>= charac. microbial growth pattern on artificial media as observe when inspected with the </li></ul></ul></ul></ul><ul><ul><ul><ul><li>unaided eye </li></ul></ul></ul></ul><ul><ul><ul><ul><li>= determine size, shape, texture, presence/absence </li></ul></ul></ul></ul><ul><ul><ul><ul><li>of pigmentation, odor of the colonies </li></ul></ul></ul></ul>
  18. 18. <ul><li>4 types of bacterial colony : </li></ul><ul><ul><ul><li>1) Smooth (S-colonies) - gives the appearance of </li></ul></ul></ul><ul><ul><ul><li> homogeneity and uniform texture without </li></ul></ul></ul><ul><ul><ul><li> appearing as liquid or mucoid colonies. </li></ul></ul></ul><ul><li>= characteristic of freshly isolated wild – type </li></ul><ul><li>organism. </li></ul><ul><li>Ex. Shigella, Salmonella, Proteus and E. coli </li></ul><ul><ul><ul><li>2) Rough (R-colonies) granulated and rough in appearance. </li></ul></ul></ul><ul><ul><ul><ul><li>= produced by mutant strain of organism </li></ul></ul></ul></ul>
  19. 19. <ul><ul><ul><li>3) Mucoid (M-colonies) - exhibits a water-like glistening </li></ul></ul></ul><ul><ul><ul><li>confluent appearance. </li></ul></ul></ul><ul><ul><ul><li>= seen among organism that forms well-defined capsule or slime layer. </li></ul></ul></ul><ul><li>4) L colonies – seen among cell-walless bacteria. </li></ul><ul><ul><ul><li>Ex. Mycoplasma </li></ul></ul></ul>
  20. 20. <ul><ul><ul><li>3. Microscopic morphology and staining reaction. </li></ul></ul></ul><ul><ul><ul><ul><li>= determines shape, arrangement and staining reaction </li></ul></ul></ul></ul><ul><ul><ul><ul><li>= light microscopic examination of gram-stained preparation with the oil immersion lens and without </li></ul></ul></ul></ul><ul><ul><ul><ul><li>coverslip is routinely used to determine bacterial morphology. </li></ul></ul></ul></ul><ul><li> = staining artificially coloring the organism with dyes </li></ul><ul><li> = advantages: </li></ul><ul><li> 1) organism are made more clearly visible </li></ul><ul><li> 2) differences between all can be demonstrated </li></ul>
  21. 21. <ul><ul><ul><ul><li>3 types of staining procedure : </li></ul></ul></ul></ul><ul><ul><ul><ul><li>1. Simple – use of 1 dye only. </li></ul></ul></ul></ul><ul><ul><ul><ul><li>2. Differential – used of 2 or more dyes. </li></ul></ul></ul></ul><ul><ul><ul><ul><ul><li>A)gram – differentiates gram positive from gram </li></ul></ul></ul></ul></ul><ul><ul><ul><ul><ul><li>negative bacteria. </li></ul></ul></ul></ul></ul><ul><ul><ul><ul><ul><li>B)acidfast – differentiates acidfast from non-acidfast organism. </li></ul></ul></ul></ul></ul><ul><ul><ul><ul><li>3. Special – used to detect special bacterial structure (capsule, flagella, spores, and intracellular) </li></ul></ul></ul></ul><ul><ul><ul><ul><li> inclusion granule </li></ul></ul></ul></ul><ul><li>A) Negative </li></ul><ul><li>B) Positive </li></ul>
  22. 22. <ul><li>4. Biochemical </li></ul><ul><li>= detect different metabolic products and sugar </li></ul><ul><li>fermentation reaction of various species </li></ul><ul><li>of microorganisms. </li></ul><ul><ul><ul><li>= 60% of common pathogens are identified by metabolic test. </li></ul></ul></ul><ul><ul><ul><li>= commercial kits now available especially for identification of enterobacteria. </li></ul></ul></ul>
  23. 23. <ul><ul><ul><li>5. Serological – detect presence of antibodies in patient </li></ul></ul></ul><ul><ul><ul><li>serum against specific antigens. </li></ul></ul></ul><ul><ul><ul><ul><li>Ex. Widal test, TPI, VDRL </li></ul></ul></ul></ul><ul><ul><ul><li>6. Animal pathogenicity </li></ul></ul></ul><ul><ul><ul><li>7. Antibiotic susceptibility test – important use for the </li></ul></ul></ul><ul><ul><ul><li>purpose of treatment. </li></ul></ul></ul><ul><ul><ul><li>8. Epidemiologic test – useful in tracing source of </li></ul></ul></ul><ul><ul><ul><li>outbreak of certain bacterial diseases. </li></ul></ul></ul>
  24. 24. Specimen smear cultivation (pure culture) microscopic colonial morph. exam. biochemical serologic animal path. antimicrobial susceptibility
  25. 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.
  26. 27. <ul><li>Bacterial morphology includes: </li></ul><ul><li>Form, Arrangement, Size and Staining reaction </li></ul><ul><li>I. Form </li></ul><ul><li>= determine by heredity </li></ul><ul><li>= genetically most bacteria are monomorphic </li></ul><ul><li>(maintain a single shaped) </li></ul><ul><li>= however some environmental conditions can alter shape </li></ul><ul><li>= some are genetically pleomorphic </li></ul><ul><li>(can have many form) </li></ul><ul><li>= determined by the configuration of the cell wall </li></ul><ul><li>= detected by brightfield microscopy of stained smear </li></ul>
  27. 28. <ul><li>A. Coccus (spherical-shaped) </li></ul><ul><li>= round-like a ball, perfect sphere, globe </li></ul><ul><li>= can be oval, elongated, or flattened on one side </li></ul><ul><li>Variations : </li></ul><ul><ul><ul><li> 1. Ovoid shape- both sides rounded ends are pointed. </li></ul></ul></ul><ul><ul><ul><ul><ul><li>Ex. Streptococcus </li></ul></ul></ul></ul></ul><ul><ul><ul><ul><ul><li>2. Lancet-shape - one end is pointed, other end is flat. </li></ul></ul></ul></ul></ul><ul><ul><ul><ul><ul><li>Ex. Pneumococcus </li></ul></ul></ul></ul></ul><ul><ul><ul><ul><ul><li>3. Coffee-bean shape- flat on one side, opposite side convex or appear as letter “D” form. </li></ul></ul></ul></ul></ul><ul><ul><ul><ul><ul><li>Ex. Neisseria </li></ul></ul></ul></ul></ul>
  28. 29. <ul><ul><ul><li>B) Bacillus (rod-shaped) </li></ul></ul></ul><ul><ul><ul><li>= cell appears longer than wide or cylindrical form </li></ul></ul></ul><ul><ul><ul><li>= both sides parallel and ends are convex </li></ul></ul></ul><ul><ul><ul><li>= varies in actual form depending on the species </li></ul></ul></ul><ul><ul><ul><li>Variations: </li></ul></ul></ul><ul><ul><ul><ul><ul><li>1. Clubbed/drumstick shaped–swollen on one end. </li></ul></ul></ul></ul></ul><ul><ul><ul><ul><ul><li>Ex. C. diphtheriae / C. tetani </li></ul></ul></ul></ul></ul><ul><ul><ul><ul><ul><li>2. Corset-shaped – both sides swollen, end flat or </li></ul></ul></ul></ul></ul><ul><ul><ul><ul><ul><li>concave. Ex. Bacillus anthracis </li></ul></ul></ul></ul></ul><ul><ul><ul><ul><ul><li>3.Fusiform - both sides parallel ends pointed </li></ul></ul></ul></ul></ul><ul><ul><ul><ul><ul><li>(thick at the center and tapered) </li></ul></ul></ul></ul></ul><ul><ul><ul><ul><ul><li>towards the end </li></ul></ul></ul></ul></ul>
  29. 30. <ul><ul><ul><li>C) Spiral </li></ul></ul></ul><ul><ul><ul><li>= bacteria w/ more than one somatic curved </li></ul></ul></ul><ul><ul><ul><li>= may be regarded as bacillary form twisted to </li></ul></ul></ul><ul><ul><ul><li>form a helix. </li></ul></ul></ul><ul><ul><ul><li>2 types: </li></ul></ul></ul><ul><ul><ul><li>1. Flexible – spirals that can contract and relax </li></ul></ul></ul><ul><ul><ul><li> whose long axis bends when in motion. </li></ul></ul></ul><ul><li>Ex. Spirochetes </li></ul><ul><li> Genus Treponema - tightly coiled spiral </li></ul><ul><li>with corkscrew appearance </li></ul><ul><li>Ex. T. pallidum </li></ul>
  30. 31. <ul><li>Genus Borrelia - much less tightly coiled having </li></ul><ul><li>the appearance of an extremely long </li></ul><ul><li>undulating pole. Ex. B. recurrentis </li></ul><ul><li>Genes Leptospira - tightly coiled spiral with </li></ul><ul><li>sharp-hooked like bends. </li></ul><ul><li>Ex. L. interrogans </li></ul><ul><li>2. Rigid – spirals that cannot contract and relax and </li></ul><ul><li>move by rotation in corkscrew-like </li></ul><ul><li>(long axis remains) rigid when in motion </li></ul><ul><ul><ul><ul><li>Ex. Spirillum minor </li></ul></ul></ul></ul>
  31. 32. <ul><li> D) Intermediate forms </li></ul><ul><li> A. Coccobacillary–when a rod is short thick, wide/plump. </li></ul><ul><ul><ul><ul><ul><li>= these form is intermediate between a </li></ul></ul></ul></ul></ul><ul><ul><ul><ul><ul><li>spherical and rod. </li></ul></ul></ul></ul></ul><ul><ul><ul><ul><ul><li>Ex. Haemophilus, Brucella </li></ul></ul></ul></ul></ul><ul><ul><ul><ul><ul><li>B. Vibrio - a gently curved bacteria (comma-shaped) </li></ul></ul></ul></ul></ul><ul><ul><ul><ul><ul><li> Ex. V. cholerae </li></ul></ul></ul></ul></ul>
  32. 33. II. Arrangement = is the result of the number of planes in which the cell divides and how they remain attached afterwards.
  33. 34. <ul><li>A) Sphericals </li></ul><ul><li>1. Singly – occurs as a single spherical cell </li></ul><ul><li>2. Chain – common among ovoid-form resulting </li></ul><ul><li>in one plane division with daughter cells </li></ul><ul><li>remained attached to one another </li></ul><ul><li>(Streptococci) </li></ul><ul><li>Ex. Streptococcus pyogenes </li></ul><ul><ul><ul><li>3. Pair –common with lancet-shaped and coffee- </li></ul></ul></ul><ul><ul><ul><li>bean shaped form resulting from one </li></ul></ul></ul><ul><ul><ul><li>plane division with daughter remain in </li></ul></ul></ul><ul><ul><ul><li>pair after dividing (Diplococci) </li></ul></ul></ul><ul><ul><ul><ul><li>Ex. Streptococcus pneumoniae </li></ul></ul></ul></ul><ul><ul><ul><ul><li> Neisseria gonorrheae </li></ul></ul></ul></ul>
  34. 35. <ul><ul><ul><ul><li>4. Cluster – common with sphericals resulting from </li></ul></ul></ul></ul><ul><ul><ul><ul><li>many plane division with daughter cell in </li></ul></ul></ul></ul><ul><ul><ul><ul><li>grape-like agglomeration bunch. </li></ul></ul></ul></ul><ul><ul><ul><ul><li>(Staphylococci) Ex. Staphylococcus aureus </li></ul></ul></ul></ul><ul><ul><ul><ul><li>5. Tetrads – (Packets of 4) – result from 2 plane </li></ul></ul></ul></ul><ul><ul><ul><ul><li>division with daughter cell separating from </li></ul></ul></ul></ul><ul><ul><ul><ul><li>one another to form group of 4 cells. </li></ul></ul></ul></ul><ul><ul><ul><ul><li>Ex. Graffya tetragena </li></ul></ul></ul></ul><ul><ul><ul><ul><li>6. Sarcinae – (Packets of 8) – results in 3 planes </li></ul></ul></ul></ul><ul><ul><ul><ul><li>division with cells remain attached in cube- </li></ul></ul></ul></ul><ul><ul><ul><ul><li>like group of 8. </li></ul></ul></ul></ul><ul><ul><ul><ul><li>Ex. Sarcina lutea </li></ul></ul></ul></ul>
  35. 36. <ul><ul><ul><li>B) Rods </li></ul></ul></ul><ul><ul><ul><li>= divides only across their short axis </li></ul></ul></ul><ul><ul><ul><ul><li>= fewer arrangement of bacilli than cocci </li></ul></ul></ul></ul><ul><ul><ul><ul><li>= most bacilli appears as single rods </li></ul></ul></ul></ul><ul><ul><ul><ul><li>1. Chain – result in one plane division with daughter </li></ul></ul></ul></ul><ul><ul><ul><ul><li>cell remain attached to one another. </li></ul></ul></ul></ul><ul><ul><ul><li>Ex. Bacillus anthracis </li></ul></ul></ul><ul><ul><ul><li>2. Palisade – arrangement like fence due to slipping </li></ul></ul></ul><ul><ul><ul><li>movement of daughter cells. </li></ul></ul></ul><ul><ul><ul><li>Ex. Diptheroids </li></ul></ul></ul><ul><ul><ul><li>3. Chinese-letter arrangement – common with clubbed- </li></ul></ul></ul><ul><ul><ul><li>shaped rods resulting from a snapping post </li></ul></ul></ul><ul><ul><ul><li>division movement of the daughter cells. </li></ul></ul></ul><ul><ul><ul><li>Ex. Corynebacterium diptheriae </li></ul></ul></ul>
  36. 37. <ul><ul><ul><li>4. Packets of cigarette – arrangement like bundles. </li></ul></ul></ul><ul><li>Ex. Mycobacterium leprae </li></ul><ul><li>5. Serpentine – chain arrangement commonly seen </li></ul><ul><li>among virulent strain of Myco. tuberculosis </li></ul><ul><li>C) Spirals </li></ul><ul><li>= no characteristic cell arrangement, most occur </li></ul><ul><li>singly </li></ul><ul><li>= different species vary in size, length, rigidity and </li></ul><ul><li>amplitude of their coils </li></ul>
  37. 38. <ul><li>III. Bacterial size </li></ul><ul><ul><ul><ul><li>1. Micrometer </li></ul></ul></ul></ul><ul><ul><ul><ul><li>2. Ultrafiltration </li></ul></ul></ul></ul><ul><ul><ul><ul><li>3. Ultracentrifugation </li></ul></ul></ul></ul><ul><ul><ul><ul><li>Cocci – 0.5 – 3.0um dia. </li></ul></ul></ul></ul><ul><ul><ul><ul><li>Bacilli – 0.2 – 2.0um dia. X 0.5 – 20um in length </li></ul></ul></ul></ul><ul><ul><ul><ul><li>Vibrio and Spirilla – 0.2-2.0um dia. X 0.5-100um L </li></ul></ul></ul></ul><ul><ul><ul><ul><li>Spirochetes – 0.1 – 3.0um dia. X 0.5 – 250um L </li></ul></ul></ul></ul>
  38. 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
  39. 40. <ul><li>Types: </li></ul><ul><li>1. Simple = employs one dye only </li></ul><ul><ul><ul><ul><ul><li> = sufficient to det. shaped and arrangement </li></ul></ul></ul></ul></ul><ul><ul><ul><ul><ul><li>2. Differential = employs the use of more than one dye </li></ul></ul></ul></ul></ul><ul><ul><ul><ul><ul><li> = enables microbiologist to differentiate one </li></ul></ul></ul></ul></ul><ul><ul><ul><ul><ul><li>group from the other by the color as well </li></ul></ul></ul></ul></ul><ul><ul><ul><ul><ul><li>as shape </li></ul></ul></ul></ul></ul><ul><ul><ul><ul><ul><li> = it is based on relative affinity of diff. bacterial </li></ul></ul></ul></ul></ul><ul><ul><ul><ul><ul><li>cell for the stained used </li></ul></ul></ul></ul></ul><ul><li>2.a) Gram = diff. gram (+) from gram (-) bacteria </li></ul><ul><li> = gram (+) those that retain the primary </li></ul><ul><li>stain and deep violet or purple color </li></ul><ul><li> = gram (-) those decolorize with ethyl </li></ul><ul><li> alcohol and stained by the counter </li></ul><ul><li> stain appearing pink or red in color </li></ul>
  40. 41. <ul><li>2.b) Acidfast = differentiate acidfast from </li></ul><ul><li>non-acidfast bacteria </li></ul><ul><li>3. Special </li></ul><ul><li>= use to color and determined bacterial structures like capsule, flagella, </li></ul><ul><li>inclusion granule, endospore and etc. </li></ul><ul><ul><ul><ul><ul><li>3.a) Negative staining </li></ul></ul></ul></ul></ul><ul><ul><ul><ul><ul><li>3.b) Positive staining </li></ul></ul></ul></ul></ul>
  41. 42. BACTERIAL CELL ULTRASTRUCTURES
  42. 44. General Division: I. External - Capsule - Fimbriae - Flagellum II. Internal - Cell Wall - Cytoplasmic Membrane - Cytoplasm - Mesosome - Nuclear Body - Ribosome - Plasmid - Inclusion Granule - Endospore
  43. 45. <ul><ul><ul><li>BACTERIAL ULTRASTRUCTURE </li></ul></ul></ul><ul><ul><ul><li>Bacterial Surface Coating: </li></ul></ul></ul><ul><ul><ul><li>= are extracellular polysaccharide polymers which </li></ul></ul></ul><ul><ul><ul><li>surrounds the bacterial cell </li></ul></ul></ul><ul><li>Capsule </li></ul><ul><li> = well-formed thick viscous jelly-like structure firmly </li></ul><ul><li>attached to the cell wall surrounding the cell </li></ul><ul><li> = not readily removed. </li></ul><ul><li> = easily visualized by negative staining using India </li></ul><ul><li>ink method </li></ul>
  44. 46. <ul><ul><ul><li>Slime layer </li></ul></ul></ul><ul><ul><ul><li>= loose and irregularly arranged meshwork of </li></ul></ul></ul><ul><ul><ul><li>fibrils totally detached from the cell but </li></ul></ul></ul><ul><ul><ul><li>still surrounds the cell </li></ul></ul></ul><ul><ul><ul><li>= when present are more easily washed off </li></ul></ul></ul>
  45. 47. <ul><li>Chemical Comp.: </li></ul><ul><li>= water – main component 98-99% </li></ul><ul><li>= 1-2% of bacterial capsule are chemically </li></ul><ul><li>polysaccharide, Except capsule of: </li></ul><ul><li> B. anthracis-polypeptide (D-glutamic acid) </li></ul><ul><li> S. pyogenes-Hyaluronic acid </li></ul><ul><li>Function: </li></ul><ul><li>= protection from phagocytosis </li></ul><ul><li> = correlates with virulence </li></ul><ul><li>= act as antigen (used in the identification) </li></ul><ul><li> and typing of capsulated bacteria </li></ul><ul><li> = allows bacteria to adhere/attach to various </li></ul><ul><li> surfaces in its natural environment in </li></ul><ul><li> order to survive </li></ul><ul><li>Detection: </li></ul><ul><ul><ul><ul><ul><li>1. Negative staining (India Ink) </li></ul></ul></ul></ul></ul><ul><ul><ul><ul><ul><li>2. Special staining (Capsular method) </li></ul></ul></ul></ul></ul><ul><ul><ul><ul><ul><li>3. Serological (Quellung reaction) </li></ul></ul></ul></ul></ul>
  46. 48. <ul><li>B. Appendages Flagellum & Axial filaments </li></ul><ul><li> Fimbriae </li></ul><ul><li>FLAGELLUM: </li></ul><ul><li>= long, thick, helical protein filament of uniform length </li></ul><ul><li>and diameter </li></ul><ul><li>= commonly seen among free-swimming bacteria </li></ul><ul><li>= originates in cytoplasmic membrane </li></ul><ul><li>= composed of 3 parts: </li></ul><ul><ul><ul><li>1. Basal body – anchors the flagellum to the </li></ul></ul></ul><ul><ul><ul><li>cell wall and plasma membrane </li></ul></ul></ul><ul><ul><ul><li>2. Hook – attached to the basal body and </li></ul></ul></ul><ul><ul><ul><li>connects to the filament </li></ul></ul></ul><ul><ul><ul><li>3. Filament – external to the cell and contains </li></ul></ul></ul><ul><ul><ul><li>the flagellar protein </li></ul></ul></ul>
  47. 49. <ul><li>Chemical Comp.: protein monomer - Flagellin </li></ul><ul><li>Function - Motility organelle of many pathogenic bacteria </li></ul><ul><li>Detection: </li></ul><ul><ul><ul><ul><ul><li>1. Darkfield / Phase contrast microscopy </li></ul></ul></ul></ul></ul><ul><ul><ul><ul><ul><li>(Wet mount / Unstained smear) </li></ul></ul></ul></ul></ul><ul><ul><ul><ul><ul><li>2. Brightfield/Electron Microscopy – using stained </li></ul></ul></ul></ul></ul><ul><ul><ul><ul><ul><li>specimen </li></ul></ul></ul></ul></ul><ul><ul><ul><ul><ul><li>3. Serological – use of specific antisera against </li></ul></ul></ul></ul></ul><ul><ul><ul><ul><ul><li>flagellar (H) antigen </li></ul></ul></ul></ul></ul><ul><ul><ul><ul><ul><li>4. Swarming phenomenon </li></ul></ul></ul></ul></ul><ul><ul><ul><ul><ul><li>5. Motility medium </li></ul></ul></ul></ul></ul><ul><ul><ul><ul><ul><li>6. Hanging drop preparation </li></ul></ul></ul></ul></ul>
  48. 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
  49. 51. <ul><li>Types: </li></ul><ul><ul><ul><li>A) Lophotrichous – tuft of flagella at one polar end </li></ul></ul></ul><ul><ul><ul><li>B) Amphitrichous – single flagellum or tulf of </li></ul></ul></ul><ul><ul><ul><li>flagella at both polar end </li></ul></ul></ul><ul><ul><ul><li>C) Peritrichous – flagella distributed all over the </li></ul></ul></ul><ul><ul><ul><li>body of the bacteria </li></ul></ul></ul>
  50. 52. <ul><ul><ul><li> Axial Filaments </li></ul></ul></ul><ul><ul><ul><li>= flagella-like sheathed filaments located in </li></ul></ul></ul><ul><ul><ul><li>the periplasmic space (between the inner) </li></ul></ul></ul><ul><ul><ul><li> and outer membrane of the cell </li></ul></ul></ul><ul><ul><ul><li>= move by traveling helical wave on opposite </li></ul></ul></ul><ul><ul><ul><li>direction </li></ul></ul></ul><ul><li>= motility organelle of spirochete </li></ul><ul><li>Fimbriae / Pili / Microfibrils </li></ul><ul><li>= short, straight, thin hair-like filaments usually </li></ul><ul><li>distributed around the body of bacteria. </li></ul><ul><li>= originates in the cytoplasmic membrane </li></ul><ul><li>= found virtually among all gram (-) bacteria but not in </li></ul><ul><li>gram (+) bacteria </li></ul>
  51. 53. <ul><li>2 types according to function: </li></ul><ul><li>1. Common/ordinary – for attachment or adherence </li></ul><ul><li>to mucosal surfaces of host cell during </li></ul><ul><li>colonization and infection. </li></ul><ul><li>2. Sex pili – responsible for attachment of donor and </li></ul><ul><li>recipient cell during bacterial conjugation </li></ul><ul><li>whereby genetic material (DNA) from one cell </li></ul><ul><li>is transferred to another (reproduction). </li></ul><ul><li>Chem. Comp. - Protein (Pilin) </li></ul><ul><ul><ul><li>Detection: Electron Microscopy </li></ul></ul></ul>
  52. 54. <ul><li>Cell wall (Peptidoglycan, Murien, Mucopeptide / Glycopeptide) </li></ul><ul><li>= complex, rigid, multilayered structure that protects the </li></ul><ul><li>protoplast and the underlying fragile plasma </li></ul><ul><li>membrane </li></ul><ul><li>= found in all pathogenic free-living bacteria except </li></ul><ul><li>Mycoplasma (cell-walless bacteria) </li></ul><ul><li>Chemical Comp.: </li></ul><ul><ul><ul><li>1. Protein (Mesodiaminopimelic acid,Isomers of D-glutamic) </li></ul></ul></ul><ul><ul><ul><li>acid and D-alanine </li></ul></ul></ul><ul><ul><ul><li>2. Polysaccharide </li></ul></ul></ul><ul><ul><ul><li>(N-acetylmuramic acid and N-acetylglucosamine) </li></ul></ul></ul><ul><ul><ul><li>= responsible for rigidity of cell wall </li></ul></ul></ul><ul><ul><ul><li>3. Lipids </li></ul></ul></ul>
  53. 55. <ul><ul><ul><li>Function : </li></ul></ul></ul><ul><ul><ul><li>1. Responsible for the characteristic shape of bacterium </li></ul></ul></ul><ul><ul><ul><li>2. Provides strong structural support necessary to keep </li></ul></ul></ul><ul><ul><ul><li>bacterial cell from rupturing due to changes in </li></ul></ul></ul><ul><ul><ul><li>the environmental osmotic pressure </li></ul></ul></ul><ul><ul><ul><li>3. Contain somatic O antigen that can serologically </li></ul></ul></ul><ul><ul><ul><li>identify particular bacteria </li></ul></ul></ul><ul><ul><ul><li>4. Site of action of some antibiotics </li></ul></ul></ul><ul><ul><ul><li>5. Determines differences in gram staining reaction </li></ul></ul></ul><ul><li>Detection : </li></ul><ul><ul><ul><li>1. Microscopy of smear prepared from special </li></ul></ul></ul><ul><ul><ul><li>staining method. </li></ul></ul></ul><ul><ul><ul><li>2. Electron microscopy. </li></ul></ul></ul><ul><ul><ul><li>3. By chemical methods using lysozyme. </li></ul></ul></ul>
  54. 56. COMPARISON OF GRAM POSITIVE AND GRAM NEGATIVE CELL WALL Gram Positive Gram Negative Peptidoglycan <ul><li>Very thick homogenous single layer </li></ul><ul><li>extensively crossed-linked. </li></ul><ul><li>Constitute 50-90% of the cell wall </li></ul><ul><li>material. </li></ul><ul><li>(NAM, NAG, D-alanine) </li></ul><ul><li>Diaminophemilic acid </li></ul><ul><li>More complex layer composed </li></ul><ul><li>of thin peptidoglycan with few </li></ul><ul><li>crossed-linked and an outer </li></ul><ul><li>membrane layer. </li></ul><ul><li>Constitute 5-10% of the cell </li></ul><ul><li>wall material. </li></ul>Auxiliary compounds <ul><li>Teichoic acid (Polymer of ribitol and) </li></ul><ul><li>glycerol phosphate </li></ul><ul><li>Lipoteichoic acid </li></ul><ul><li>Surface protein </li></ul><ul><li>No teichoic acid </li></ul><ul><li>Outer membrane contains: </li></ul><ul><li>Phospholipids-located in the </li></ul><ul><li>inner membrane </li></ul><ul><li>Lipoprotien – connects OM to </li></ul><ul><li>the peptidoglycan </li></ul><ul><li>Lipopolysaccharide </li></ul><ul><li>(LPS/Endotoxin layer) </li></ul><ul><li>– located in the outer </li></ul><ul><li>layer of the OM </li></ul><ul><li>- contains the lipid A </li></ul>Penicillin sensitivity <ul><li>Sensitive </li></ul><ul><li>Resistant </li></ul>Response to lysozyme <ul><li>Digest/ Degrade </li></ul><ul><li>Resistant </li></ul>
  55. 58. Gram Positive Gram Negative
  56. 59. <ul><li>Damage to the cell wall may result to: </li></ul><ul><ul><ul><li>= when a gram positive bacteria is exposed to </li></ul></ul></ul><ul><ul><ul><li>lysosyme will degrade the peptidoglycan </li></ul></ul></ul><ul><ul><ul><li>layer resulting to complete removal of the cell </li></ul></ul></ul><ul><ul><ul><li>wall producing a osmotically fragile wall-less </li></ul></ul></ul><ul><ul><ul><li>spherical body called Protoplast </li></ul></ul></ul><ul><ul><ul><li>= since all cell wall components is removed, </li></ul></ul></ul><ul><ul><ul><li>therefore incapable of regeneration </li></ul></ul></ul><ul><li>Spheroplast </li></ul><ul><ul><ul><li>= when a gram negative bacteria is exposed to </li></ul></ul></ul><ul><ul><ul><li>lysozyme it looses the peptidoglycan layer </li></ul></ul></ul><ul><ul><ul><li>but retain the outer membrane leaving a </li></ul></ul></ul><ul><ul><ul><li>less fragile spherical body capable </li></ul></ul></ul><ul><ul><ul><li>of regenerating the cell wall. </li></ul></ul></ul>
  57. 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)
  58. 62. <ul><li>Functions: </li></ul><ul><ul><ul><li>1. Serve as selective permeability barrier </li></ul></ul></ul><ul><ul><ul><li>(transport of nutrient material in & out of cell) </li></ul></ul></ul><ul><ul><ul><li>2. Site of electron transport and oxidative phosphorylation of substances involve in the </li></ul></ul></ul><ul><ul><ul><li>generation of chemical energy (ATP) </li></ul></ul></ul><ul><ul><ul><li>3. Helps in the excretion of metabolic waste product </li></ul></ul></ul><ul><ul><ul><li>4. Site for excretion of enzyme involved in OM </li></ul></ul></ul><ul><ul><ul><li>synthesis, CW synthesis and in the secretion </li></ul></ul></ul><ul><ul><ul><li>of extracytoplasmic & extracellular substances: </li></ul></ul></ul><ul><ul><ul><ul><li>A. Permease for active uptake of nutrient </li></ul></ul></ul></ul><ul><ul><ul><ul><li>B. Respiratory </li></ul></ul></ul></ul><ul><ul><ul><ul><li>C. Polymerase that manufacture substances </li></ul></ul></ul></ul><ul><ul><ul><ul><li>of the cell wall </li></ul></ul></ul></ul><ul><ul><ul><ul><li>D. Hydrolytic </li></ul></ul></ul></ul><ul><li>Detection - Electron microscopy </li></ul>
  59. 63. <ul><li>MESOSOME: </li></ul><ul><li> = usually seen as an invagination of the membrane </li></ul><ul><li>associated cytoplasmic sac seen in Gram (+) cell </li></ul><ul><li> = contains lamellar, tubular and vesicular structure </li></ul><ul><li> = often associated with division septa </li></ul><ul><li> Chemical Composition - Lipoprotein </li></ul><ul><li> Functions: </li></ul><ul><ul><ul><li>1. Site for the synthesis of cytochrome oxidase </li></ul></ul></ul><ul><ul><ul><li>and reductase enzyme </li></ul></ul></ul><ul><ul><ul><li>2. Provide support for enzymes present in the cell </li></ul></ul></ul><ul><ul><ul><li>membrane </li></ul></ul></ul><ul><ul><ul><li>3. Responsible for compartmenting DNA during </li></ul></ul></ul><ul><ul><ul><li>cell division and sporulation </li></ul></ul></ul><ul><li> Detection - Electron microscopy </li></ul>Cytoplasm = refers to everything that is enclosed by the cytoplasmic membrane = site for most bacterial metabolism = 80% is composed of water
  60. 64. <ul><li>NUCLEAR BODY: (Nucleus / Nucleoid) </li></ul><ul><li>= genetic material of bacteria </li></ul><ul><li>= contains a single circular molecule of double </li></ul><ul><li>stranded DNA network which runs parallel to the </li></ul><ul><li>long axis of the cell </li></ul><ul><li>= found homogenous within the cytoplasm not </li></ul><ul><li>enclosed in a nuclear membrane. </li></ul><ul><li>= constitute 2-3% of the cell weight. </li></ul><ul><li>= no definite form and seen at all stages of growth </li></ul><ul><li>cycle of the bacteria. </li></ul><ul><li>Chemical Composition – DNA </li></ul><ul><li>Function: </li></ul><ul><ul><ul><ul><li>1. Controls the growth and metabolic activity of </li></ul></ul></ul></ul><ul><ul><ul><ul><li>the bacteria </li></ul></ul></ul></ul><ul><ul><ul><ul><li>2. Responsible for hereditary characteristics of the cell. </li></ul></ul></ul></ul><ul><li>Detection - Electron microscopy, Feulgin staining (+) </li></ul>
  61. 65. <ul><li>PLASMID/EPISOME: </li></ul><ul><li>= extrachromosomal genetic material capable of </li></ul><ul><li>autonomous replication usually located near </li></ul><ul><li>the chromatin body. </li></ul><ul><li>Chemical Composition – DNA </li></ul><ul><li>Function: </li></ul><ul><ul><ul><li>1. Transfer of genetic material from one cell to another </li></ul></ul></ul><ul><ul><ul><li>by conjugation (Transmissible Plasmid) </li></ul></ul></ul><ul><li>2. Carry genes for activities like: </li></ul><ul><ul><ul><ul><ul><li>a) Antibiotic resistance </li></ul></ul></ul></ul></ul><ul><ul><ul><ul><ul><li>b) Toxin production </li></ul></ul></ul></ul></ul><ul><ul><ul><ul><ul><li>c) Synthesis of enzyme </li></ul></ul></ul></ul></ul><ul><ul><ul><ul><ul><li>d) Tolerance to toxic metals </li></ul></ul></ul></ul></ul><ul><li>Detection – Feulgin staining (+), Electron microscopy </li></ul>
  62. 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
  63. 67. <ul><li>INCLUSION / CYTOPLASMIC GRANULE: </li></ul><ul><li>= large granules found anywhere in the cytoplasm </li></ul><ul><li>= are accumulation of organic and inorganic substances </li></ul><ul><li>which serves as source of energy and nutrient </li></ul><ul><li>supply to the organism </li></ul><ul><li>Kinds: </li></ul><ul><li>1. Babes Ernst / Volutin / Metachromatic granules </li></ul><ul><ul><ul><li>Chemical Composition – polymerized </li></ul></ul></ul><ul><ul><ul><li>metaphosphate which can be used in the </li></ul></ul></ul><ul><ul><ul><li>synthesis of ATP </li></ul></ul></ul><ul><ul><ul><li>Function – reserve energy supply </li></ul></ul></ul><ul><li>Demonstration – simple staining / special </li></ul><ul><li>staining (Albert’s stain) </li></ul>
  64. 68. <ul><ul><ul><ul><li>2. Lipid granules </li></ul></ul></ul></ul><ul><ul><ul><ul><ul><li>Chemical Composition – polymer of beta- </li></ul></ul></ul></ul></ul><ul><ul><ul><ul><ul><li> hydroxybutyric acid </li></ul></ul></ul></ul></ul><ul><ul><ul><ul><ul><li>Function – reserve food supply. </li></ul></ul></ul></ul></ul><ul><ul><ul><ul><li>3. Sulfur granules – derived energy by oxidizing </li></ul></ul></ul></ul><ul><ul><ul><ul><li>sulfur and sulfur-containing </li></ul></ul></ul></ul><ul><ul><ul><ul><li>compounds. </li></ul></ul></ul></ul><ul><ul><ul><ul><ul><li>Function – reserve energy supply for sulfur- </li></ul></ul></ul></ul></ul><ul><ul><ul><ul><ul><li>containing bacteria. </li></ul></ul></ul></ul></ul><ul><ul><ul><ul><li>4. Glycogen granule – polymer of glucose. </li></ul></ul></ul></ul><ul><ul><ul><ul><ul><li>Function – reserve food supply. </li></ul></ul></ul></ul></ul>
  65. 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
  66. 70. <ul><li>= Composed of five parts: </li></ul><ul><li> 1. Core – located at the center which is the spore </li></ul><ul><ul><ul><ul><ul><li>cytoplasm/protoplast. </li></ul></ul></ul></ul></ul><ul><ul><ul><ul><ul><li>- contains the nucleus and enzyme dipicolinic acid </li></ul></ul></ul></ul></ul><ul><ul><ul><ul><li>2. Spore wall – enclosing the core. </li></ul></ul></ul></ul><ul><ul><ul><ul><li>3. Cortex – laminated structure surrounding the spore </li></ul></ul></ul></ul><ul><ul><ul><ul><li>wall </li></ul></ul></ul></ul><ul><ul><ul><ul><li>4. Spore coat – multilayered membrane enclosing the </li></ul></ul></ul></ul><ul><ul><ul><ul><li>cortex </li></ul></ul></ul></ul><ul><ul><ul><ul><li>5. Exosporium – the outermost covering of the spores </li></ul></ul></ul></ul><ul><ul><ul><ul><li>which give the spore a rigid appearance. </li></ul></ul></ul></ul>
  67. 71. <ul><li>= contains Calcium Dipicolinate which aids in heat </li></ul><ul><li>resistance within the core by making the protein </li></ul><ul><li>and nucleic acid more resistant to denaturation </li></ul><ul><li>= germinate under favorable nutritional condition after </li></ul><ul><li>an activation process that involves damage to the </li></ul><ul><li>spore coat </li></ul><ul><li>= helpful in identifying some specie of bacteria especially </li></ul><ul><li>those sporeforming bacilli (Clostridium and Bacillus) </li></ul>
  68. 72. <ul><li>Classification accdg. to: </li></ul><ul><li>A. Location - Central, Subterminal, Terminal </li></ul><ul><li>B. Shape - a) Ovoid </li></ul><ul><li> b) Circular </li></ul><ul><li>C. Swollen/not swollen </li></ul><ul><li>(swollen when the diameter is more than the of the) </li></ul><ul><li> bacterial cell, not swollen diameter the same as bacterial cell </li></ul><ul><li>Detection: </li></ul><ul><ul><ul><li>1. Microscopy of stained smear </li></ul></ul></ul><ul><ul><ul><ul><li>A) Negative staining </li></ul></ul></ul></ul><ul><ul><ul><ul><li>B) Positive staining </li></ul></ul></ul></ul><ul><ul><ul><ul><li>2. Phase contrast microscopy </li></ul></ul></ul></ul>
  69. 73. <ul><ul><ul><li>PHYSIOLOGY OF BACTERIAL CELL </li></ul></ul></ul><ul><ul><ul><li>Nutrition </li></ul></ul></ul><ul><ul><ul><li>= process by which chemical substances (nutrients) </li></ul></ul></ul><ul><ul><ul><li>either organic or inorganic are use in cellular </li></ul></ul></ul><ul><ul><ul><li>activities of microorganism for metabolism </li></ul></ul></ul><ul><ul><ul><li>and growth </li></ul></ul></ul><ul><ul><ul><li>Growth </li></ul></ul></ul><ul><li>= orderly increase of all chemical constituents of the </li></ul><ul><li>cell including size and number </li></ul><ul><li>= process entails replication of all cellular structures, </li></ul><ul><li>organelles and protoplasmic components of </li></ul><ul><li>the cell </li></ul><ul><li>Generation time </li></ul><ul><li>= measure of the growth rate of microbial species. </li></ul><ul><li>= varies in length of time accdg. to environmental </li></ul><ul><li>conditions. </li></ul>
  70. 74. <ul><ul><ul><li>Bacterial growth requirement: </li></ul></ul></ul><ul><ul><ul><li>1) Water – most important requirement </li></ul></ul></ul><ul><ul><ul><li> = vehicle for the entry of all nutrients into the </li></ul></ul></ul><ul><ul><ul><li>cell and elimination of their waste product </li></ul></ul></ul><ul><ul><ul><li> = form an integral part of the cell protoplasm </li></ul></ul></ul><ul><ul><ul><li>2) Nutrient requirement : </li></ul></ul></ul><ul><li> = source of Carbon and Nitrogen </li></ul><ul><li>a) Carbon = major building block for constructing </li></ul><ul><li>cell material </li></ul><ul><li> </li></ul>
  71. 75. <ul><li>Types of bacteria acdg. to carbon requirement </li></ul><ul><ul><ul><ul><li>1. Autotroph (Lithotroph) </li></ul></ul></ul></ul><ul><li>= req. only H 2 O, CO 2 & inorganic substances </li></ul><ul><li>= utilize CO 2 as source of carbon </li></ul><ul><li>= Photoautotroph- light as energy source = Chemoautotroph - chemical reaction as </li></ul><ul><li>energy source </li></ul><ul><li> 2. Organotroph </li></ul><ul><li>= requires organic substances for growth </li></ul><ul><li>= unable to utilize CO 2 as energy source </li></ul><ul><li> Photoorganotroph - light as energy source </li></ul><ul><li> Chemoorganotroph - chemical reaction as </li></ul><ul><li>energy source </li></ul><ul><li> 3. Heterotroph </li></ul><ul><li>= requires both organic and inorganic </li></ul><ul><li>substances for growth </li></ul>
  72. 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
  73. 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
  74. 78. <ul><li>II. Physical Requirement </li></ul><ul><ul><ul><li>A)Temperature </li></ul></ul></ul><ul><ul><ul><ul><ul><li>1. Phychrophile/Cryophile </li></ul></ul></ul></ul></ul><ul><ul><ul><ul><ul><li>= 5-30 0 C opt. 10-20 0 C </li></ul></ul></ul></ul></ul><ul><ul><ul><ul><ul><li>= grow at refrigeration temp. of 4 0 C - 8 0 C </li></ul></ul></ul></ul></ul><ul><ul><ul><ul><ul><li>= responsible spoilage food refrigeration </li></ul></ul></ul></ul></ul><ul><ul><ul><ul><ul><li>= cold –loving bacteria </li></ul></ul></ul></ul></ul><ul><ul><ul><ul><ul><li>= found normally in cold water </li></ul></ul></ul></ul></ul><ul><li>2. Mesophiles </li></ul><ul><li>= 10 – 45 0 C opt. 20-40 0 C </li></ul><ul><li>= saprophytic – 26-35 0 C </li></ul><ul><li>= parasitic – 37-45 0 C </li></ul><ul><li>= bacteria pathogenic for human </li></ul>
  75. 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
  76. 80. <ul><ul><ul><li>B) Oxygen </li></ul></ul></ul><ul><ul><ul><ul><li>= required by particular bacterium to satisfy its energy </li></ul></ul></ul></ul><ul><ul><ul><ul><li>needs 5 groups of bacteria on the basis of their O 2 </li></ul></ul></ul></ul><ul><ul><ul><ul><li>requirement </li></ul></ul></ul></ul><ul><ul><ul><ul><ul><li>1) Obligate anaerobes </li></ul></ul></ul></ul></ul><ul><ul><ul><ul><ul><li>= grows only under condition of high </li></ul></ul></ul></ul></ul><ul><ul><ul><ul><ul><li>reducing intensity and for which </li></ul></ul></ul></ul></ul><ul><ul><ul><ul><ul><li>oxygen is toxic </li></ul></ul></ul></ul></ul><ul><ul><ul><ul><ul><li>(complete absence of oxygen) </li></ul></ul></ul></ul></ul><ul><ul><ul><ul><ul><li>= requires oxygen - free environment to </li></ul></ul></ul></ul></ul><ul><ul><ul><ul><ul><li>survive </li></ul></ul></ul></ul></ul><ul><ul><ul><ul><ul><li>2) Aerotolerant anaerobes </li></ul></ul></ul></ul></ul><ul><ul><ul><ul><ul><li>= organism that are not killed by exposure </li></ul></ul></ul></ul></ul><ul><ul><ul><ul><ul><li>to oxygen </li></ul></ul></ul></ul></ul>
  77. 81. <ul><ul><ul><li>3) Facultative anaerobes </li></ul></ul></ul><ul><ul><ul><li>= capable of growth under both aerobic and </li></ul></ul></ul><ul><ul><ul><li>anaerobic condition </li></ul></ul></ul><ul><ul><ul><li>4) Obligate aerobes </li></ul></ul></ul><ul><ul><ul><li>= requires oxygen for growth </li></ul></ul></ul><ul><ul><ul><li>5) Microaerophilic </li></ul></ul></ul><ul><ul><ul><li>= organism that grows best at low oxygen tension </li></ul></ul></ul><ul><ul><ul><li>6) Capnophiles </li></ul></ul></ul><ul><ul><ul><li>= requires 5-10% carbon dioxide and oxygen on </li></ul></ul></ul><ul><ul><ul><li>primary isolation </li></ul></ul></ul>
  78. 82. <ul><ul><ul><li>C) Hydrogen Ion Concentration (PH) </li></ul></ul></ul><ul><ul><ul><ul><li>= optimum ph for pathogenic bacteria 7.2 – 7.6 </li></ul></ul></ul></ul><ul><ul><ul><ul><ul><li>1. Acidophilic – below ph 6.5 – 7.6 </li></ul></ul></ul></ul></ul><ul><ul><ul><ul><ul><li>2. Basophilic – (alkalophilic) – 8.4 – 9.0 </li></ul></ul></ul></ul></ul><ul><ul><ul><ul><ul><li>3. Normophilic (neutrophilic) – ph 7.5 – 8.0 </li></ul></ul></ul></ul></ul><ul><ul><ul><li>D) Osmotic Pressure </li></ul></ul></ul><ul><ul><ul><ul><li>= direct pressure – resistant </li></ul></ul></ul></ul><ul><ul><ul><ul><li>= osmotic pressure – sensitive </li></ul></ul></ul></ul><ul><ul><ul><ul><li>= plasmolysis – shrinkage </li></ul></ul></ul></ul><ul><ul><ul><ul><li>= osmophile – bacteria that can grow on high </li></ul></ul></ul></ul><ul><ul><ul><ul><li>osmotic pressure </li></ul></ul></ul></ul><ul><ul><ul><ul><li>= Ex. Halophile – seawater bacteria grow best at </li></ul></ul></ul></ul><ul><ul><ul><ul><li>high concentration of salt </li></ul></ul></ul></ul>
  79. 83. <ul><li>Measurement of Bacterial Growth </li></ul><ul><ul><ul><li>1. Cell Concentration (cell number) number of cell per unit </li></ul></ul></ul><ul><ul><ul><li>volume of culture. Can be counted directly by a </li></ul></ul></ul><ul><ul><ul><li>microscope counting chamber. </li></ul></ul></ul><ul><ul><ul><li>DETERMINATION: </li></ul></ul></ul><ul><ul><ul><ul><li>1) Total direct plate count </li></ul></ul></ul></ul><ul><ul><ul><ul><ul><li>A) Bacterial Counting Chamber </li></ul></ul></ul></ul></ul><ul><ul><ul><ul><ul><li>(Petroff-Hauser Counter) </li></ul></ul></ul></ul></ul><ul><ul><ul><ul><ul><li>B) Coulter Counter (Electron Particle Counter) </li></ul></ul></ul></ul></ul><ul><ul><ul><ul><ul><li>= measures both distribution of size </li></ul></ul></ul></ul></ul><ul><ul><ul><ul><ul><li>and number of bacteria in </li></ul></ul></ul></ul></ul><ul><ul><ul><ul><ul><li>bacterial suspension </li></ul></ul></ul></ul></ul>
  80. 84. <ul><ul><ul><ul><li>2) Indirect Viable Count </li></ul></ul></ul></ul><ul><ul><ul><ul><ul><li>= plate sample of culture and make dilutions </li></ul></ul></ul></ul></ul><ul><ul><ul><ul><ul><li>of microbial population  inoculate </li></ul></ul></ul></ul></ul><ul><ul><ul><ul><ul><li>suitable solid medium  incubate  </li></ul></ul></ul></ul></ul><ul><ul><ul><ul><ul><li>formation of viable colony </li></ul></ul></ul></ul></ul><ul><li>USES: </li></ul><ul><ul><ul><li>A) Solving problems on bacterial cell division </li></ul></ul></ul><ul><ul><ul><li>B) Genetic </li></ul></ul></ul><ul><ul><ul><li>C) Infection </li></ul></ul></ul><ul><ul><ul><li>D) Microbial Inactivation </li></ul></ul></ul>
  81. 85. <ul><li>2) Cell Density (Cell Mass) </li></ul><ul><li>= total protoplasm/dry weight of the cell per unit </li></ul><ul><li>volume of culture </li></ul><ul><li>= include living and dead cell </li></ul><ul><ul><ul><li>DETERMINATION: </li></ul></ul></ul><ul><ul><ul><li>1. Absorbance Spectrophotometer </li></ul></ul></ul><ul><ul><ul><ul><ul><li>= measuring optical density of broth culture of </li></ul></ul></ul></ul></ul><ul><ul><ul><ul><ul><li>microorganism </li></ul></ul></ul></ul></ul><ul><ul><ul><li>2. Turbidimetric Technique </li></ul></ul></ul><ul><ul><ul><ul><ul><li>= useful in determining mass of cells during bacterial growth </li></ul></ul></ul></ul></ul><ul><ul><ul><li>3. Nitrogen Determination </li></ul></ul></ul><ul><ul><ul><li>4. Centrifugation </li></ul></ul></ul><ul><li>USES: </li></ul><ul><ul><ul><li>1. Study of bacterial nutrition </li></ul></ul></ul><ul><ul><ul><li>2. Biochemistry </li></ul></ul></ul>
  82. 86. <ul><li>Bacterial Growth Curve </li></ul><ul><li>(5 PHASES) </li></ul><ul><ul><ul><li>1. Lag phase (Phase of Rejuvenescence/Phase of) </li></ul></ul></ul><ul><ul><ul><li>Physiologic Growth </li></ul></ul></ul><ul><ul><ul><ul><li>= after inoculation  bacteria requires a period of </li></ul></ul></ul></ul><ul><ul><ul><ul><li>adjusting and adapting to new environment </li></ul></ul></ul></ul><ul><ul><ul><ul><li> marked increase macromolecular </li></ul></ul></ul></ul><ul><ul><ul><ul><li>component of the cell  increase in cell size </li></ul></ul></ul></ul><ul><ul><ul><ul><li>but no detectable increase in cell number </li></ul></ul></ul></ul><ul><li>= no cell division occur </li></ul><ul><ul><ul><li>2. Exponential phase (Logarithmic phase) </li></ul></ul></ul><ul><ul><ul><ul><li>= cell in state of balanced growth </li></ul></ul></ul></ul><ul><ul><ul><ul><li>= characterized sharp rise in growth curve indicating </li></ul></ul></ul></ul><ul><ul><ul><ul><li>rapid growth and multiplication </li></ul></ul></ul></ul><ul><li>= bacterial cell double its growth per unit of time </li></ul>
  83. 87. <ul><ul><ul><li>3. Stationary phase </li></ul></ul></ul><ul><ul><ul><li>(phase of equilibrium/”plateau”) </li></ul></ul></ul><ul><ul><ul><ul><li>= manifestation of unbalanced growth  growth rate zero </li></ul></ul></ul></ul><ul><ul><ul><ul><li>= viable count remains constant for a short period eventually gives way to decreasing population </li></ul></ul></ul></ul><ul><li> = number of living bacteria equals number of </li></ul><ul><li>bacteria dying </li></ul><ul><ul><ul><li>4. Phase of Decline (Death phase) </li></ul></ul></ul><ul><ul><ul><ul><li>= growth rate decreases  complete cessation of multiplication </li></ul></ul></ul></ul><ul><ul><ul><ul><li>= result in the decline in total viable count </li></ul></ul></ul></ul><ul><ul><ul><ul><li>= most bacteria dies due to : </li></ul></ul></ul></ul><ul><ul><ul><ul><ul><li>1. lack of nutrient material in the medium </li></ul></ul></ul></ul></ul><ul><ul><ul><ul><ul><li>2. accumulation of waste product excreted by </li></ul></ul></ul></ul></ul><ul><ul><ul><ul><ul><li>the bacteria </li></ul></ul></ul></ul></ul><ul><ul><ul><ul><ul><li>3. change in ph of the environment </li></ul></ul></ul></ul></ul>
  84. 89. <ul><li>Consequence of growth & multiplication is reproduction </li></ul><ul><li>3 methods of bacterial reproduction </li></ul><ul><ul><ul><li>1. Binary fission </li></ul></ul></ul><ul><ul><ul><li>2. Branching </li></ul></ul></ul><ul><ul><ul><li>3. Sporulation </li></ul></ul></ul><ul><li>2 methods of bacterial multiplication </li></ul><ul><ul><ul><li>1. Binary fission </li></ul></ul></ul><ul><ul><ul><li>2. Branching </li></ul></ul></ul>
  85. 90. STERILIZATION AND DISINFECTION
  86. 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
  87. 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
  88. 93. <ul><li>2. Moist Heat = kills bacteria faster than dry heat </li></ul><ul><ul><ul><li>Boiling </li></ul></ul></ul><ul><ul><ul><li>= 30 minutes boiling at 100 0 C is adequate to kill </li></ul></ul></ul><ul><li>vegetative cell of bacteria, fungi, protozoa </li></ul><ul><li>= sporeformer requires 5½ hours boiling to kill the </li></ul><ul><li>organism </li></ul><ul><ul><ul><li>2. Free – Flowing stream </li></ul></ul></ul><ul><li>= live stream  Arnold sterilizer </li></ul><ul><li>= same sterilizing action with boiling </li></ul><ul><li>3 . Steam Under Pressure </li></ul><ul><li>= heating material at 15 lbs/square inch pressure for 15 – 20 minutes at 121 0 C </li></ul><ul><li>= most efficient and reliable method of sterilization = commonly used for sterilizing of: </li></ul><ul><ul><ul><ul><li>a. Hospitals surgical & medical instrument </li></ul></ul></ul></ul><ul><ul><ul><ul><li>b. Bacteriological media not destroyed by heat </li></ul></ul></ul></ul><ul><ul><ul><ul><li>c. Hospital supplies </li></ul></ul></ul></ul>
  89. 94. <ul><li>4. Tyndallization (Fractional/Intermittent sterilization) </li></ul><ul><li>= for sterilization of liquid & semi-solid material easily </li></ul><ul><li> destroyed by heat . </li></ul><ul><li> Ex. culture medium contg. egg, serum, </li></ul><ul><li>carbohydrate </li></ul><ul><li>= heating material at 80-100 0 C 30 minutes 3 conc. days </li></ul><ul><li>= kills both sporeformer & non-sporeformer bacteria </li></ul><ul><ul><ul><li>5. Pasteurization </li></ul></ul></ul><ul><li>= consist of heating material at 62 0 C for 30 minutes </li></ul><ul><li>followed by rapid cooling </li></ul><ul><li>= use to kill only disease – producing organism found in </li></ul><ul><li>milk, beverages, milk products and in preparation </li></ul><ul><li>bacterial vaccine </li></ul><ul><li>Mechanism of moist heat: Denetuaration and coagulation of </li></ul><ul><li>protein inside bacterial cell </li></ul>
  90. 95. <ul><ul><ul><li>B) Radiation </li></ul></ul></ul><ul><ul><ul><li>= produce damage to DNA resulting to killing of the organism </li></ul></ul></ul><ul><li> = 2 types : </li></ul><ul><ul><ul><ul><li>A. Non-ionizing radiation </li></ul></ul></ul></ul><ul><ul><ul><ul><li>Ex. Ultraviolet rays – have low penetrating power </li></ul></ul></ul></ul><ul><ul><ul><ul><li>= used to destroy bacteria, fungi, viruses that </li></ul></ul></ul></ul><ul><ul><ul><ul><li>are sensitive to U-V treatment </li></ul></ul></ul></ul><ul><li>= primarily used to control airborne infections in </li></ul><ul><li>nurseries, hospital wards & OR </li></ul><ul><ul><ul><ul><li>B. Ionizing radiation </li></ul></ul></ul></ul><ul><ul><ul><ul><li>Ex: X-ray, alpha, beta and gamma rays </li></ul></ul></ul></ul><ul><li>= have greater penetrating power </li></ul><ul><li>= use extensively for sterilization of cutgut , </li></ul><ul><li> plastic syringe, catheter, prosthesis, </li></ul><ul><li>swab and dressing pack </li></ul><ul><li> C. ) Filtration = process of separating microorganism from </li></ul><ul><li>contaminated solution </li></ul><ul><li> = employs the use of filters for removal of microorganism </li></ul><ul><li>that are sensitive to heat </li></ul><ul><li> = for sterilization of heat – labile C.M containing serum, </li></ul><ul><li>plasma, urea </li></ul>
  91. 96. <ul><ul><ul><li>D) Ultrasomic vibration </li></ul></ul></ul><ul><li>= are sound waves with high frequency </li></ul><ul><li>causes desruption & disintegration of the cell </li></ul><ul><li>= uses 1. research laboratories </li></ul><ul><li> 2. treating sewage H 2 O </li></ul><ul><ul><ul><li>E) Lyophilization ( Freeze-drying ) </li></ul></ul></ul><ul><li>= not reliable </li></ul><ul><li>= causes protein denaturation & damage of the </li></ul><ul><li>membrane w/ leakage of intracellular </li></ul><ul><li>organic compounds </li></ul><ul><li>F) Laser’s </li></ul><ul><li>= sterilization of medical instrument and operating </li></ul><ul><li> room </li></ul>
  92. 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
  93. 98. <ul><li>Mech. of action: </li></ul><ul><ul><ul><li>1. damage cell membrane function </li></ul></ul></ul><ul><ul><ul><li>2. denatures protein </li></ul></ul></ul><ul><ul><ul><li>3. induce extensive nucleic acid damage </li></ul></ul></ul><ul><li>Agents that damage cell membrane function </li></ul><ul><li>I – Surface active agents : = destroys structural integrity of cell membrane </li></ul><ul><li>(protein & lipids) </li></ul><ul><li>= interferes w/ normal membrane function </li></ul><ul><li>= widely used as detergent, wetting agent and emulsifier </li></ul><ul><li> 3 types: </li></ul><ul><li> cationic = most useful </li></ul><ul><li> = includes Quarternary ammonium cpd., </li></ul><ul><li> Benzalkonium chloride ( Zephiran ) </li></ul><ul><li> anionic = soap, surfactant, sanitizer </li></ul><ul><li> = help remove microbe by rubbing </li></ul><ul><li> nonionic = not effective, non-toxic </li></ul>
  94. 99. <ul><li>II – Phenolic compound </li></ul><ul><li>= disrupt cell membrane function  leaking cell content </li></ul><ul><ul><ul><li>A) Carbolic acid (phenol) </li></ul></ul></ul><ul><li>= excellent for disinfecting feces, blood, pus, </li></ul><ul><li>sputum and other proteinous material </li></ul><ul><li>= primarily use for testing new bactericidal agent </li></ul><ul><ul><ul><li>B) Cresol / Creolin </li></ul></ul></ul><ul><li>Lysol - alkyl derivative phenol </li></ul><ul><li> = disinfect inanimate object esp. </li></ul><ul><li>contaminated with TB bacilli </li></ul><ul><li>Creolin - septic tank disinfectant </li></ul><ul><ul><ul><li>C) Diphenyl compound (Hexachlorophene) </li></ul></ul></ul><ul><li>= effective against gram + bacteria (staph & strep) </li></ul>
  95. 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%
  96. 101. <ul><li>Agents that denatures protein </li></ul><ul><ul><ul><li>1. Acid & alkalies </li></ul></ul></ul><ul><ul><ul><li>2. Alcohol </li></ul></ul></ul><ul><ul><ul><li>3. Acetone </li></ul></ul></ul><ul><li>Agents that modify functional group of protein & nucleic acid </li></ul><ul><li>1. Salts of heavy metals – mercury, silver , arsenic </li></ul><ul><li>Silver cpd.  widely used as antiseptic </li></ul><ul><li> </li></ul><ul><li> AgNO 3 (silver nitrate) – routinely used as </li></ul><ul><li>prophylaxis against opthalmia neonatorum </li></ul><ul><ul><ul><li>2. Oxidizing agents (Halogen Compound) </li></ul></ul></ul><ul><li>iodine – wound/skin disinfectant </li></ul><ul><li>- active against tubercle bacilli </li></ul><ul><li>chlorine – H 2 O disinfectant </li></ul><ul><li>Hydrogen peroxide – use for cleaning wound </li></ul>
  97. 102. <ul><ul><ul><li>3. Dyes </li></ul></ul></ul><ul><ul><ul><ul><li>Triphenyl methane dyes (aniline acid) </li></ul></ul></ul></ul><ul><ul><ul><ul><li>Acidine dyes (Flavines) – yellow color </li></ul></ul></ul></ul><ul><ul><ul><li>4. Alkalyting agent (Formaldehyde, Glutaraldehyde,) </li></ul></ul></ul><ul><ul><ul><ul><li>Ethylene oxide </li></ul></ul></ul></ul><ul><ul><ul><ul><li>= exert lethal effect on protein substances </li></ul></ul></ul></ul><ul><ul><ul><ul><li>Formaldehyde – in aqueous solution is bactericidal, </li></ul></ul></ul></ul><ul><ul><ul><ul><li>sporocidal and also effective against </li></ul></ul></ul></ul><ul><ul><ul><ul><li>viruses </li></ul></ul></ul></ul><ul><ul><ul><ul><li>= use for inactivating vaccine and </li></ul></ul></ul></ul><ul><ul><ul><ul><li> preservation of fabrics </li></ul></ul></ul></ul><ul><ul><ul><ul><li>formalin = in aqueous solution of 37% </li></ul></ul></ul></ul><ul><ul><ul><ul><li> formaldehyde is used for preserving </li></ul></ul></ul></ul><ul><ul><ul><ul><li> fresh tissue specimen </li></ul></ul></ul></ul><ul><ul><ul><ul><li>= as gas, used for fumigating operating </li></ul></ul></ul></ul><ul><ul><ul><ul><li> room, hospital ward, laboratories </li></ul></ul></ul></ul>
  98. 103. <ul><ul><ul><li>Glutaraldehyde </li></ul></ul></ul><ul><ul><ul><li>= used as cold sterilant for sterilizing medical </li></ul></ul></ul><ul><ul><ul><li>and surgical instruments </li></ul></ul></ul><ul><ul><ul><li>Ethylene Oxide </li></ul></ul></ul><ul><ul><ul><li>= used for sterilization of polyethylene tubing, </li></ul></ul></ul><ul><ul><ul><li>heart lung machine, electronic hospital </li></ul></ul></ul><ul><ul><ul><li>devices and sensitive medical instrument </li></ul></ul></ul>
  99. 104. <ul><li>Chemotherapeutic Agent </li></ul><ul><li>= one of the most valuable method of treating infection </li></ul><ul><li>Antibiotics : are naturally occurring metabolic products of </li></ul><ul><li>primarily soil bacteria & fungi </li></ul><ul><li>Desirable Properties : </li></ul><ul><ul><ul><li>1. Selective Toxicity </li></ul></ul></ul><ul><ul><ul><li>2. Bactericidal </li></ul></ul></ul><ul><ul><ul><li>3. Should not be allergenic and with no adverse reaction </li></ul></ul></ul><ul><ul><ul><li>on large doses or prolonged use </li></ul></ul></ul><ul><ul><ul><li>4. Active in the presence of plasma, body fluids &exudates </li></ul></ul></ul><ul><ul><ul><li>5. Should have broad-spectrum activity </li></ul></ul></ul><ul><ul><ul><li>6. Water soluble & stable </li></ul></ul></ul><ul><ul><ul><li>7. Susceptible organism should not become resistant </li></ul></ul></ul><ul><ul><ul><li>8. Cheap </li></ul></ul></ul>
  100. 105. <ul><ul><ul><ul><ul><li>Antibiotic producers </li></ul></ul></ul></ul></ul><ul><li>1. Penicillium </li></ul><ul><li>2. Bacillus </li></ul><ul><li>3. Streptomyces </li></ul><ul><li>4. Cephalosporium </li></ul><ul><li>Mech. of action : </li></ul><ul><li>1. Interfere with cell wall synthesis </li></ul><ul><li>2. Interfere with cell membrane function </li></ul><ul><li>3. Interfere with Protein synthesis </li></ul><ul><li>4. Interfere with nucleic acid metabolism </li></ul><ul><li>Cell Wall Inhibitors : </li></ul><ul><ul><ul><li>1. Beta – lactam antibiotics </li></ul></ul></ul><ul><ul><ul><ul><li>A. Penicillin </li></ul></ul></ul></ul><ul><ul><ul><ul><li>B. Cephalosphorins </li></ul></ul></ul></ul><ul><ul><ul><li>2. Fosfomycin </li></ul></ul></ul><ul><ul><ul><li>3. Cycloserine </li></ul></ul></ul><ul><ul><ul><li>4. Vancomycin </li></ul></ul></ul><ul><ul><ul><li>5. Bacitracin </li></ul></ul></ul>
  101. 106. <ul><li>Penicillins </li></ul><ul><ul><ul><li>1. Natural PCN – Benzyl PCN G </li></ul></ul></ul><ul><li> - Phenoxymethyl PCN V </li></ul><ul><ul><ul><li>2. Semi – synthetic PCN </li></ul></ul></ul><ul><ul><ul><ul><li>A) Penicillinase – resistant PCN’s </li></ul></ul></ul></ul><ul><li>Methicillin </li></ul><ul><li>Nefcillin </li></ul><ul><li>Isoxazolyl PCN – Cloxacillin </li></ul><ul><li>Drug of choice - Dicloxacillin </li></ul><ul><li>For PCN resistant - Oxacillin </li></ul><ul><li>S. aureus & S. epidermides </li></ul><ul><ul><ul><ul><li>B) Extended spectrum PCN’s </li></ul></ul></ul></ul><ul><li>Ampicillin </li></ul><ul><li>Amoxicillin </li></ul><ul><ul><ul><ul><li>C) Antipseudomonas </li></ul></ul></ul></ul><ul><li>Carbenicillin </li></ul><ul><li>Ticarcillin </li></ul><ul><li>Piperacillin </li></ul>
  102. 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
  103. 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
  104. 109. <ul><li>II : Cell Membrane Inhibitors </li></ul><ul><ul><ul><li>1. Polymixins – reserved for serious pseudomonas infection </li></ul></ul></ul><ul><ul><ul><li>2. Polyenes </li></ul></ul></ul><ul><li>Amphotericin B – for deep seated mycoses </li></ul><ul><li>Nystatin – seperficial & cutaneous fungus infection </li></ul><ul><ul><ul><li>3. Azoles – Imidazole antifungal agent – Ketaconazole </li></ul></ul></ul><ul><li> - Miconazole </li></ul><ul><li> - Clotrimozole </li></ul><ul><ul><ul><li>4. Triazoles – Itraconazole </li></ul></ul></ul><ul><li> - Fluoconazole </li></ul><ul><li>III : Inhibitors of DNA function </li></ul><ul><li>1. Mitomycin </li></ul><ul><li>2. Nalidixic acid & Quinolones </li></ul><ul><li>3. Norfloxacine & Ciprofloxacin </li></ul><ul><li>4. Metronidazole – for anaerobic infection </li></ul><ul><li>5. Novobiocin – gram (-) </li></ul><ul><li>6. Griseofulvin </li></ul>
  105. 110. <ul><ul><ul><ul><ul><li>IV : Inhibitors Protein Synthesis </li></ul></ul></ul></ul></ul><ul><li>A. Actinomycin – active against gram (+) & gram ( -) </li></ul><ul><li>B. Rifampicin – major drug for TB & Leprosy </li></ul><ul><li>C. Streptomycin </li></ul><ul><li> Gentamycin Aminoglycosides </li></ul><ul><li> Tobramycin </li></ul><ul><li> Amikacin </li></ul><ul><li>D. Chloramphenicol </li></ul><ul><li>E. Nitrofurans </li></ul><ul><li>F. Erythromycin </li></ul><ul><li>G. Clindamycin / Lincomycin </li></ul>
  106. 111. <ul><ul><ul><ul><ul><li>V : Metabolite Analogue </li></ul></ul></ul></ul></ul><ul><li>1. Sulfonamide </li></ul><ul><li>= gram (+) gram, (-) Nocardia, Chlamydia, Pneumococcus </li></ul><ul><li>2. Sulfone </li></ul><ul><li>= Myco. leprae </li></ul><ul><li>3. Para-aminosalicyclic acid </li></ul><ul><li>(PAS) – tuberculosis </li></ul>

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