1. M Luqman M.Phil Microbiogy 0212 We make invisible world visible.

2. Bacteriology Study of bacteria Bacteria; On the basis -----how they generate energy Anaerobic Obligate Aerotolerant Aerobic Obligate Facultative Microaerophilic 3. Obligate Anaerobes Require anoxic environment (grow either by anaerobic respiration or fermentation) Can not tolerate O2 , O2 compounds & O2 radicals (O2 -, OH- , H2O2, 1O2) Grow best at low Eh They lack Cytochrome System Superoxide Dismutase O2 - + O2 - + 2H+ H2O2 + O2 Catalase 2H2O2 2H2O + O2(Bubbles) Cont 4. Peroxidase H2O2 + NADH + H+ 2H2O + NAD+ Superoxide Reductase O2- + 2H+ rubredoxin reduced H2O2 + rubredoxin oxidized Aerotolerants Strictly fermentative Can grow either in the absence or presence of O2 5. Bacilli Gram ve bacilli Bacteroides group (B fragilis) Prevotella (P melaninogenica) Porphyromonas Fusobacterium (Pleomorphic) Gram +ve, bacilli, non spore former Actinomyces (A israelli) Lactobacilus Propionibacterium, Pleomorphic (P acnes) Eubacterium, bifidobacterium and arachnia (Pleomorphic) Gram +ve, bacilli, spore former Clostridium spp. How they survive in aerobic host? Fig 1; B fragilis Fig 2; Fusobacterium spp. Cont 6. Cocci Gram ve Veillonella Gram +ve Peptostreptococcus Spirochetes Borrelia, treponema Can microbiome be considered an organ of the body? Fig 4; Peptostreptococcus spp. Fig 3; Veillonella spp. 7. Source may be endo or exogenous Exogenous (Environment) Soil Clostridium spp. Endogenous Oral anaerobes Intestinal anaerobes 8. Resist exposure to oxygen Aerotolrance Evade host defence mechanism Prevent phagocytises Inactive complement Establish anaerobic environment Mixed infection with facultative or aerobic bacteria Signal host cells to increase O2 consumption Resistance to antibiotics Intrinsic resistance to penicillin Plasmids, transposons, insertion sequences Obtain nutrients for growth Protease Glycohydrolases 9. Toxin Clostridial Toxins Endotoxin (Lipopolysaccharide) Promotes abscess formation Capsule Involve in abscess production Enzymes Nucleases lipases Proteases Hyaluronidase Colegenase lactamase 10. Brain abscesses Peptostreptococci and B fragilis Oropharyngeal infections Oropharyngeal anaerobes; actinomyces, Prevotella melaninogenica, Fusobacterium species Pleuropulmonary infections Peptostreptococci; Fusobacterium species; P melaninogenica, B fragilis in 2025% Intra-abdominal infections Liver abscess:Mixed anaerobes in 4090%; facultative organisms Abdominal abscesses B fragilis; other gastrointestinal flora How they displace from their normal area to infected area? Cont 11. Female genital tract infections Vulvar abscesses: Peptostreptococci and others Tubo-ovarian and pelvic abscesses: P bivia and P disiens; peptostreptococci; others Skin, soft tissue, and bone infections Mixed anaerobic flora Bacteremia B fragilis; peptostreptococci; clostridia; propionibacteria;`others Endocarditis B fragilis 12. Anaerobic Specimens include: blood Aspirates from body fluids Pleural Pericardial Cerebrospinal Peritoneal Joint fluids Urine collection Abscess contents Deep aspirates from wounds 13. Use a two tube system 1st tube contains swab---anaerobic environment 2nd tube contains pre-reduced anaerobically sterilized media. 14. Because of better survival of pathogen greater quantity of specimen less contamination with extraneous organism are often achieved 15. Anaerobic Transport Medium Transport Devices having anoxic environment (N2, H2, CO2) Syringes Swabs in; Tube containing CO2 Carey and Blair semisolid media Tissue Specimen in; Anaerobic Jar Petri dish in a sealed bag 16. Gram stain must be performed in laboratory....because... Organisms seen on Gram stain that cannot be grown in aerobic cultures Typical morphology for anaerobes on Gram stain 17. Anaerobic growth on proper media containing antibiotic-suppressing aerobes Growth in anaerobic zone of fluid or agar media Gas presence foul-smelling odour Characteristic colonies Colonies of pigmented Prevotella or Porphyromonus spp. may fluoresce red under ultraviolet light, and older colonies produce a typical dark pigment 18. Solid or Liquid media----must provide anaerobic environment----- indicator resazurin pink (+ O2), no colour ( - O2) Anaerobic Jar Candle Jar reduces O2 environment only CO2 tension Gas Pak Jar Palladium Catalyst O2 reduced to CO2 Sodium bicarbonate & Sodium borohydride Reacts with H2O to form H2 & CO2 19. Methylene Blue Strip Indicator Blue ( O2 present) White ( O2 not present) Anaerobic Chamber Anaerobic environment Filled with inert gases (85% N2, 10% H2, 5% CO2) Oxy-Plate Oxyrase enzyme Combines O2 with H2 20. Anaerobic Disposable Plastic Bags Anaerobic bags Generate H2 & CO2 Resazurin indicators Bag is heat sealed 21. Plates are checked at > 18-24 hours for faster growing species like Cl. Perfringens & B.fragilis & daily thereafter up to > 5-7 days for slowly growing species like Actinomyces, Eubacterium & propionibacterium Genus is determined by - gram stain, cellular morphology, Gas-liquid chromatography Species determination is based on fermentation of sugars & other biochemical determination 22. Jonathen Cohen, William G Powderly and Steven M Opal, Chapter 173; Anaerobic Bacteria, in Infectious Diseases, 3, 3rd edn, 2 vols. (Mosby Elsevier, 2013), II. David J. Hentges, Anaerobes: General Characteristics, in Medical Microbiology, ed. by Samuel Baron, 4th edn (Galveston (TX): University of Texas Medical Branch at Galveston, 1996) [accessed 15 December 2013]. Gerard J. Tortora, Berdell R. Funke and Christine L. Case, Microbial Growth, in Microbiology An Introduction, 10th edn (Benjamin Cummings: Pearson Education,, 2013). Jawet, Melnick and Adelberg, Infections Caused by Anaerobic Bacteria, in Jawetz, Melnick, & Adelbergs Medical Microbiology, 24th edn (The McGraw-Hill Companies, 2013).