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General bacteriology revision notes microbiology
This document discusses foundational concepts in microbiology including key figures in the field. Louis Pasteur is identified as the father of microbiology for growing bacteria in liquid media, while Robert Koch is recognized for introducing solid media and discovering the tuberculosis and cholera bacteria. Koch's postulates are also summarized, which are the criteria used to establish a causative relationship between a microbe and a disease. The document also covers topics such as acid-fast staining, culture media, and the effect of temperature and oxygen levels on bacterial growth.
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General bacteriology revision notes microbiology
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- 3. General microbiology TONY SCARIA2010 KMC
- 4. • Father ofmicrobiology: louis Pasteur • GREW BACTERIA IN LIQUID MEDIA • Father of bacteriology : Robert Koch • INTRODUCED SOLID MEDIA • Father of chemotherapy : paul ehlrich • Father of modern antiseptic surgery : john lister • Used carbolic acid TONY SCARIA 2010 KMC
- 5. Louis Pasteur • Germcell theory • Developed anthrax rabies vaccine • Developed sterilisation technique • Sterilise,autoclave,hot air oven TONY SCARIA 2010 KMC
- 6. Robert Koch • Discoveredtubercle bacilli (kochs bacillus) and vibrio cholera TONY SCARIA 2010 KMC
- 7. Koch’s postulates • 1.The microorganism must be present in every case of the but absent from healthy host. • 2. The suspected microorganism must be isolated and grown in a pure culture from lesions of the disease. • Except : Mycobacterium leprae and Treponema pallidum,rickettsiae ,many viruses • 3. The isolated organism, in pure culture, when inoculated in suitable laboratory animals should produce a similar disease. • 4. The same microorganism must be isolated again in pure culture from the lesions produced in experimental animals. TONY SCARIA 2010 KMC
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- 10. Koch’s phenomenon • hypersensitivityreaction against tuberculosis bacilli demonstrated in guinea pigs. • guinea pigs already infected with tubercle bacillus, on challenge with tubercle bacillus or its protein, developed an exaggerated inflammatory response TONY SCARIA 2010 KMC
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- 14. • GP+VE CELLWALL IS THICKER RETAINS PRIMARY STAIN SLOW DECOLOURISATION • GP+VE CELL WALL IS ACIDIC • G N-VE HAS MORE LIPIDS GET DISSOLVED BY DECOLORISER LEAD TO FORMATION OF PORES RAPID DECOLOURISATION TONY SCARIA 2010 KMC
- 15. Acid fastness • Mycolicacid • Integrity of cell wall TONY SCARIA 2010 KMC
- 16. Acid fast staining •Ziehl neelson staining Smear stained by carbol fuchsin Decolorisation by sulphuric acid Counterstain with methylene blue Retain fuchsin (red colour) Take colour of methylene blue (blue colour) • Mycobacteria tuberculosis (25%) • Lepra (5%) • Nocardia(1%) TONY SCARIA 2010 KMC
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- 18. Acid fast organism •Bacteria • Mycobacterium • Nocardia • Rhodococcus • Legionella micdadei • Oocyst of parsite • Bacterial spores TONY SCARIA 2010 KMC
- 19. Bipolar staining • Safetypin appearance • d/t accumulation of dye at both ends TONY SCARIA 2010 KMC
- 20. Bipolar staining • Calymmobactergranulomatis • Pseudomonas mallei • Yersinia pestis • Haemophilus ducreyi TONY SCARIA 2010 KMC
- 21. GELATIN • FIRST SLDIFYINGAGENT • PROTEIN IN NATURE • REQUIRED @ CONCENTRATION 15 % • FROM ANIMAL BONE • DISADVANTAGE • PROTEOLYSED BY BACTERIA LIQUEFACTION • SOLID ONLY @ TEMP <24 *C TONY SCARIA 2010 KMC
- 22. AGAR • MOST COMMONLYUSED • POLYSACCHARIDE (AGAROSE (70%)+ AGAROPECTIN (30 %)+INORGANIC PHOSPHATE • 1.2- 2 % IS NEEDED 2 % CONCENTRATION IN SOLID MEDIA • FROM SEA WEEDS & RED ALGAE GELIDIUM • INERT NOT HYDROLYSED BY BACTERIA • NO NUTRITION TO BACTERIA • NO GROWTH PROMOTERS OR INHIBITORS • SOLID @ TEMP <42 *C • MELTS @ TEMP > 98 *C TONY SCARIA 2010 KMC
- 23. SOFT AGAR •0.2 –0.5 % •SEMISOLID •FOR BACTERIAL MOTILITY USUAL SOLID MEDIA •1.2- 2 % 5-6 % AGAR •FORM AGAR •FOR INHIBITION OF SWARMING TONY SCARIA 2010 KMC
- 24. SWARMING GRAM POSITIVE • CLOSTRIDIUMTETANIN • BACILLUS CEREUS GRAM NEGATIVE • PROTEUS VAULGARIS • PROTEUS MIRABILIS • VIBRIO ALGINOLYTICUS • VIBRIO PARAHEMOLYTICUS TONY SCARIA 2010 KMC
- 25. OTHER SOLDIFYING AGENTS •EGG • SERUM TONY SCARIA 2010 KMC
- 26. TYPES OF CULTUREMEDIA SIMPLE MEDIA • FOR GROWING NON FASTIDIOUS BACTERIA • ONLY SOURCE OF CARBON & NITRIGEN ENRICHED MEDIUM • MEDIUM WITH EGG BLOOD SERUM SELECTIVE MEDIUM • SELECTIVE AGENT ADDED TO INHIBIT OTHER BACTERIA & PROMOTE WANTED BACTERIA Enrichment media • Liquid selective media Differential media • Medium to differentiate b/w bacteria • BA • MacConkey medium Indicator media • Indicates bacterial growth • Tellurite media TONY SCARIA 2010 KMC
- 27. SIMPLE MEDIA • PEPTONE+ H20 PEPTONE BROTH • PEPTONE BROTH + MEAT EXTRACT NUTRIENT BROTH • NUTRIENT BROTH + AGAROSE NUTRIENT AGAR TONY SCARIA 2010 KMC
- 28. ENRICHED MEDIA • BLOOD •5% BA FOR HEMOLYSIS DEMONSTRATION • 10 % BA • CHOCOLATE AGAR • EGG CONTAINING • LOWEN STEIN JENSON MEDIUM • DORSET EGG MEDIUM • SERUM • LOEFFLERS SERUM SLOPE • PPLO MEDIUM TONY SCARIA 2010 KMC
- 29. SELECTIVE MEDIA • ChangepH • Only for vibrios (pH 8.2 -8.4) • Bile salt agar • TCBS medium • Adding abx • Thayer martin medium vancomycin +colistin +nystatin • Modified Thayer martin TM medium + trimethoprim • New York city medium vancomycin + colistin + trimethoprim + amphotericin B • PPLO with penicillin • Adding chemical agents • Bile salts • MacConkey gram negative • XLD selective for salmonella & shigella • DCA selective for salmonella & shigella • Tellurite corynebacterial macLeod • 10 % salt staphylococci salt agar saltmilk agar Neisseria TONY SCARIA 2010 KMC
- 30. • By addingdyes • methylene blue agar gram negative bacteria • LJ media malachite TONY SCARIA 2010 KMC
- 31. Liquid selective broth •Alakaline peptone for vibrios • Tetrathionate broth only for salmonella • Selenite F broth for salmonella & shigella TONY SCARIA 2010 KMC
- 32. Differential media • TCBS •Thiosulfate • Citrate • Bile salt • Sucrose TONY SCARIA 2010 KMC
- 33. Differential media • Mannitolsalt agar • Only mannitol fermenting staph aureus TONY SCARIA 2010 KMC
- 34. Hugh leifson OFmediumdifferential media TONY SCARIA 2010 KMC
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- 36. Petroleum jelly tomake medium anaerobic TONY SCARIA 2010 KMC
- 37. CLED media for urinary pathogens TONY SCARIA 2010 KMC
- 38. Indicator media • Telluritemedia for Corynebacterium • Wilson blair media for salmonella TONY SCARIA 2010 KMC
- 39. Transport media • Donotallow growth of commensals • Maintain pathogens • Pikes medium streptococcus pyogenes • Thioglycolate broth for anaerobes • Cary blair broth universal stool transport media • Stuarts media & amies media for Neisseria TONY SCARIA 2010 KMC
- 40. • Salmonella, Shigella fromfaecal specimens TONY SCARIA 2010 KMC
- 41. ANEROBIC MEDIA • ROBERTSONSCOOKED MEAT MEDIA • SMITH NOGUCHI MEDIUM • HEATING & SUBSEQUENT PLATING MEDIA TONY SCARIA 2010 KMC
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- 47. Antibiotic susceptibility test TONYSCARIA 2010 KMC
- 48. MUELLER HILTON AGAR •STARCH + CASEIN HYDROLYSATE + AGAR • ANTIBIOTIC SENSITIVITY TESTING TONY SCARIA 2010 KMC
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- 50. Standard inoculum TONY SCARIA2010 KMC
- 51. 0.5 Mcfarland 1.5 * 10 ^ 8 bacteria from 4- 5 colonies TONY SCARIA 2010 KMC
- 52. • Inoculated into millor Hilton agar @ 37 * c • Interpreted @ 16- 18 hrs TONY SCARIA 2010 KMC
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- 54. Methods of antibioticsensitivity • Dilution best method best method / reference mthod • Exact Minimum inhibitory concentration can be determined • Broth dilution • Microbroth microtitre plate • Macrobroth testtubes • Agar dilution • More time consuming • Kirby bauer disc diffusion method TONY SCARIA 2010 KMC
- 55. Lowest concentration ofAbx just inhibit bacterium inmmedium MIC (minimum inhibitory concentration) TONY SCARIA 2010 KMC
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- 58. Kirby bauer disc diffusion method TONYSCARIA 2010 KMC
- 59. Stokes method ofdisc diffusion TONY SCARIA 2010 KMC
- 60. E STRIP METHOD EPSILOMETER E TEST (GRADED CONCENTRATION OF ABX SHEET) TONY SCARIA 2010 KMC
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- 62. Bacterial growth curve TONYSCARIA 2010 KMC
- 63. • Generation time time required for 1 binary fission • Viable count no of live bacteria • total count no live + Killed bacteria TONY SCARIA 2010 KMC
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- 66. Lag phase • Stagefor adaptation • Variable for different bacteria • Metabolically active • Size of bacteria is maxmm @ end of lag phase • No replication count (viable & total) remains constant TONY SCARIA 2010 KMC
- 67. Log phase /exponential • Stage of active replication • Viable count ↑ • Total count increase ↑ • Size decreases TONY SCARIA 2010 KMC
- 68. Stationery phase • Gradualnutrient depletion • No of bacteria multiplying= no of bacteria dying • Viable count constant • Total count increases • Sporulation toxin production occurs TONY SCARIA 2010 KMC
- 69. Declining phase /death phase • Total nutrient depletion • Most bacteria are dying • Viable count decreases • Total count is constant TONY SCARIA 2010 KMC
- 70. • Maximum effectof abx adhesion is maximum in log phase TONY SCARIA 2010 KMC
- 71. Maximum cell sizein log phase Cells are smaller stain uniformly used for microbiological stains • Spore formation • Exotoxin production • Antibiotic proudtion Involutional forms are seen TONY SCARIA 2010 KMC
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- 73. Types of culture Batchculture • With fixed amount of nutrients • Bacterial growth curve present • Closeed system Continous culture • Maintenance of log phase • In chemostat or turbidistat • Fresh nutrients added • Toxic nutrients removed TONY SCARIA 2010 KMC
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- 75. Obligate aerobes • Grow Onlyin aerobic conditions Facultative anaerobe • Prefer aerobic condition but can also grow in anaerobic condiions Microaerophilic anerobes • Prefer anaerobic conditions but can tolerate low amount of o2 Obligate anaerobe • Grow only in anerobic environment & cannot tolerate aerobic condition Capnophilic • Grows better in 5-10 % CO2 • Brucella abortus Vibrio cholera Most pathogenic bacteria Campylobacter Helicobacter Brucella abortus Clostridium Bacteriodes TONY SCARIA 2010 KMC
- 76. Temperature requirement Psychrophilic • <20*c • Bacteria responsible for Spoiling of referigertaed foods mesophilic • 25 – 40 *c • Most human pathogens Thermophilic • High temperature • Indicator of sterilization • Cause spoilage of canned foods TONY SCARIA 2010 KMC
- 77. Spore formation • NOTA METHOD OF REPRODUCTION • In unfavourable condition • SPORES ARE ACID FAST • Destroyed by autoclaving TONY SCARIA 2010 KMC
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- 80. L form /lister forms TONY SCARIA 2010 KMC
- 81. Resolution • Eye 0.2mm • Light microscope 0.2um • Electron microscope 0.2nm TONY SCARIA 2010 KMC
- 82. UREASE PRODUCING BACTERIA •PUNCH • PROTEUS MIRABILIS • UREAPLASMA UREALYTICUM • NOCARDIA • CRYPTOCOCCUS • HELOBACTER PYLORI TONY SCARIA 2010 KMC