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Bacteria review 1) Bacteria structure and physiology A) Prokaryotes v Eukaryotes 1) Prokaryotes-no nuclear mem (1) 50s and 30s (2) dsDNA B) Cell wall components in a gram + and a gram neg. 1) Gram + = lipoteichoic acid, wall teichoic acid (ion bonding, charge maintenance, adherence to host 2) Gram neg-no crossbridging (1) Outer membrane (not lipid) (a) LPS (b) Lipid A (c) O antigen 2) Gram stain A) CV B) Iodine- keeps CV inside cell of gm + C) Alcohol washD) Safranin 3) Mycobacterium cell wall-LAB 4) Synthesis of Peptidoglycan A) GluNAC and MurNAC form peptide bridges by transglycosylation B) Side chains join by transpeptidase 1) Linkage is usually a lys-ala 2) An ala is cleaved when you bind 5) Process and purpose of spore formation A) Bacillus B) Clostridium 6) Mycoplasmas A) No cell wall 7) Capsule A) Polysaccharide or protein (bacillus anthracis) B) Block phagocytic receptors C) Block c3b deposition D) 8) Events in bacteria growth A) Lag-bacteria get bigger with no dividing B) Log-exponential growth C) Stationary –limit in nutrients D) Death-detergents, antibiotics etc 9) Oxidizers-LAB 10) Fermenters-LAB 11) Bacteria classification A) Phenotypic-growth requirements 1) Nutrient uptake (1) Group translocation-substrate altered (2) Substrate translocation-substrate unchanged (3) Facilitated diffusion 1 of 8 B) Analytic C) Genotypic-best way 1) MLE electrophoresis-more similar bacteria are more likely to exchange DNA 2) DNA restriction profile-RFLP analysislook at chromosomal banding for clones that have caused a disease 3) DNA base composition4) DNA sequencing-best way but not practical 5) DNA hybridization 12) Characterization of rRNA for genetic relatedness A) Useful because r-RNA is very conserved 13) Gram positive Bacteria A) Staph. aureus 1) Anterior nares 2) Beta hemolytic 3) Mannitol salt agar 4) Catalase and coagulase positive 5) Capsule 6) Teichoic acids 7) Protein A-binds fc fragments of Abs 8) Clumping factor-bound coagulase binds fibrin to mask in blood stream 9) Hemolysins-kill RBCs 10) Leukocidins-kill WBC 11) Penicillinase 12) Tunnels through tissue with: (1) Hyaluronidase (2) Staphylokinase-lyses formed fibrin (3) Lipase (4) Protease 13) Exfoliatin toxin 14) TSS 15) Scalded skin syndrome 16) Pneumonia, meningitis, endocarditis, osteomyletis, septic arthritis 17) Treat with vancomycin (binds to the dALA) B) Staph epidermidis 1) Novobiocin susceptible 2) Coagulase neg 3) Polysaccharide slime 4) Heart valve infections C) Staph saprophyticus 1) Novobiocin resistant 2) UTIs 3) No blood hemolysis 14) Streptococceae Bacteria review A) Group A beta hemolytic-(strep pyogenes) 1) Bacitracin susceptible 2) M protein=virulence factor (analogous to protein A of Staph a.) 3) Antiphagocytic capsule 4) Hyaluronidase 5) Cellulitis 6) Erythogenic toxin-scarlet fever 7) Streptokinases-clot busters digest fibrin 8) Hemolysins 9) DNAases 10) Protein F-binds fibronectin 11) Protein G-binds Ab 12) Associated with rheumatic fever and glomerulonephritis 13) Necrotizing fasciitis B) Group B beta hemolytic (s. agalactiae) 1) B for babies 2) Disease of newborns 3) Camp reaction positive-arrow head C) S. pneumoniae 1) Meningitis, ear infections, pneumonia 2) Optochin sensitive 3) Quelling reaction to see capsule 4) Alpha hemolytic 5) Capnophilic 6) Pneumonococcal surface protein A to inhibit opsinization 7) Autolysins to release cell components after grown in culture 8) Pnuemolysins-inhibit cilia function 9) Vaccine-protect against 23 capsule types D) Viridans strep. 1) Alpha hemolytic 2) Strep mutans 3) Bacteremia 4) Endocarditis E) Enterococcus 1) Hydrolyze esculin 2) Grow in high NaCl 3) PYR hydrolysis 15) Gram positive Rods A) Listeria monocytogenes 1) Weakly beta hemolytic 2) Causes food born illnesses 3) Meningitis in neonates 4) Esculin positive 5) Catalase positive 6) MOTILE 7) Umbrella motility B) Cornybacterium 2 of 8 1) Chinese letters 2) Commensals 3) C. jeikeium-infections in immunosuppressed paitent 4) C. diphtheria-throat infection (1) Potent EXOtoxin (2) Interferes with EF-2 of the ribosome of cell and cell dies to make psuedomembrane (3) Cuases asphyxsia because plugs the trachea (4) Toxoid vaccine 5) Bacillus (1) B. cereus-food poisoning (a) Beta hemolytic and MOTILE (b) Catalase positive (c) Deposits spores in food. (2) B. anthracis (a) Endospores (b) From cattle hide (non lethal)forms lesion on skin (c) Respiratory infection-lethal (d) Has a protein capsule 16) Microbial pathogenesis A) A pathogen must: 1) Cause symptoms 2) Evade the host response, cause inflammation 3) Be acquired 4) Be able to multiply 5) Transmit to others B) Virulence factores 1) Evade complement and phagocytosis (pneumococci) 2) Eliminate T cells-(salmonella) C) Classifications of microbes 1) Non-pathogenic 2) Normal flora (1) Effect the immunocomprimised (2) Produce vitamin K in the gut 3) Opportunistic infections-attack when immune system is down 4) Disease producing organism D) Host defense mechanisms 1) Innate immunity 2) Acquired immunity 3) NK cells 4) Inflammation E) What is used to fight bacteria $$$$ 1) Extracellular bacteria-Abs, complement, phagocytes Bacteria review 2) Intracellular bacteria-Th1 cells, macrophages, and CD8 T cells 3) Viruses-Abs, interferons, NK cells 17) Neisseria A) N. meningiditis 1) Oxidase positive (enterobacteracea are oxidase neg) 2) Capnophilic 3) Fastidious 4) Polysacc. Capsule, LPS, IgA protease 5) Nasopharynx to blood to CSF 6) Oxidizes glucose and MALTOSE 7) Humans only reservoir (like shigella) 8) Asplenia and complement deficiency are at risk 9) Bacterimia 10) Purpura fulminans 11) Waterhouse-friderishsen syndromeadrenal hemorrhage 12) Empyema-exudate over brain 13) New conjugate vaccine B) N. gonorrhea 1) Nonmotile, capnophilic 2) Pili enhance attachment 3) IgA protease 4) No immunity because antigenic variation 5) Urethritis in males 6) Oxidizes GLUCOSE 7) Nucleic acid amplification and test on Thayer martin agar (chocolate + Anitbiotics) 8) Arthritis, proctitis, PID, pharyngitis, ophthalmia neonatorium (Chlamydia does this too) 18) Chlamydia A) C. trachomitis 1) Obligate intracellular 2) NO Peptidoglycan!!! Makes hard for antibiotics 3) Prevents phagolysosome fusion 4) Elementary body-infectious form 5) Reticulate-replication form 6) Not as pyogenic eye infection as the gonge (gonorrhea) 7) Most common STD 8) Lymphogranuloma venereum 9) No vaccine B) C. psittaci 1) Parrot shit 2) Diagnose with Ab titer 3) Respiratory infection 3 of 8 C) C. pneumophila 1) Atherosclerosis 2) CAD 19) Spirochetes A) Treponema pallidum 1) Primary, secondary, latent, tertiary 2) Chancre, rash, no signs, CV, gumma, CNS, Tabes dorsalis, 3) Hutchinsones incisors, mulberry molars 4) Darkfeild microscopy 5) 8th nerve deafness 6) Saber tooth tiger shins 7) Non treponemal test-RPR and VDRL can give false positives 8) Treponemal-TPPA will give positive test for life B) Borrelia 1) Stain with giemsa 2) Lyme disease 3) Ixodes tick-not in AL 4) ECM rash C) Rickettsiae 1) Vectors 2) Have DNA AND RNA 3) R. Ricketsii-Vasculitis=RMSF 4) Monocyte ehrlichiosis-ehrlichia chafeensis 5) Typhus fever-R. typhi 20) Anerobes A) Bacterioides fragilis 1) Necrotizing fascitis 2) Grows on bile B) Prevotella melaninogenica 1) In the mouth C) Fusobacterium 1) Nonspore forming 2) URT and colon 3) Pulmonary abscess from vomit 4) Mixed flora 5) Pointed ends D) Actinomyces israelii 1) Sulfur granules 2) Dacyrocystitus-blockage of tear ducts 3) Actinomycosis-lumpy jaw E) Propionobacterium 1) In normal skin flora F) Lactobacillus 1) Normal flora of vagina G) Clostridium 1) Tetani 2) Botulinum Bacteria review 3) Perferingins-gas gangrene (1) Box car (2) PLC (3) Nagler test (4) Double zone of hemolysis 4) Dificile-resistant to broad spectrum antibiotics 21) Heamophilus A) X and V factors B) Person to person C) IgA protese D) 5-10% CO2 E) No hemolysis F) Capsule G) Chocolate agar H) Satellism with s. aureus I) Otits media, Sinusitis, bacterimia J) Capsule b, polysaccharide,protein carrier vaccine K) Pink eye and community aquired pnemonia L) Ducreyi (1) STD M) Aegyptis 1) Conjunctivitis 22) Bordatella A) Gram – B) Capsule C) Strict aerobe D) Oxidize but don’t ferment E) Pertussis 1) 3 stages (1) Catarrhal-1-2 weeks (2) Paroxysmal-1-6 weeks (3) Convalescent-months 2) No reservoir 3) Produces exotoxins after it adheres 4) Pertussis Toxin-adherance (1) Increase cAMP (2) Inhibit phagocytosis 5) Lethal toxin 6) Tracheal toxin 7) Endotoxin (LPS) F) DaPT - acellular vaccine, not whole bugs like DPT 23) Legionella pneumophila A) Legionarres-old men with COPD and emphysema B) In air conditioner C) Fastidious D) Facultative intracellular 4 of 8 E) No ferment, No person to person transmission (inhaled), NO vaccine F) Prevent phagolysosome fusion in macrophage G) BCYE agar, silver staining, FAb, urine polysaccharide Ag, to detect H) Cell mediated response most important 24) Moraxella catarrhalis A) URT B) Gram neg coccus 25) Mycoplasma A) Very small B) Smallest free living C) No cell wall-susceptible to drying out D) Extracellular growth on mucosal membranes E) Pneumoniae 1) Cilia 2) Person to person transmission 3) Cytotoxic after attachment 4) Walking pneumonia 5) Look for IgM (children) or seroconversion with IgG and IgM (adults) to detect 6) Role in asthma 7) Spread through households F) Hominis-opportunistic infection 1) Postpartum endometriitis G) Genitalium H) Ureaplasma species 1) Thought ot cause non chlamydial urethritits 26) DNA structure 27) DNA replication A) Initiated at OriC B) Helicase unwinds C) DNA polymerase adds bases to 3’ end D) Leading strand, lagging strand (goes 5’ to 3’) E) Okazaki fragments 28) PCR A) Denature DNA B) Add complementary oligosaccharides C) Add nucleotides D) Synthesize strand with Taq DNA polymerase 29) Prokarytotic gene organization A) Operon-group of genes. Usually gives you a long mRNA. PCR will tell you if you have an operon. B) Promoter region 1) Operator 2) -10 (TATATT) and -35 region (TTGACA) Bacteria review 3) TSS at +1 4) RNA polymerase binds the -10 and -35 C) Transcriptional apparatus 1) Beta, Beta prime, 2 alphas, omega, and sigma factor (active) are in the RNAp 2) RNAp=molecular engine of cell 3) Termination sequence-G-C rich hairpin loop D) Transcriptional sequence 1) Initiation 2) Elongation 3) Continued elongation 4) Termination 30) Lac operon (E. Coli) A) Negative regulation-(must be switched off) 1) repressor binds to the operator and transcription is blocked 2) Repressor competes with RNAp B) Induction of the Lac operon 1) Lactose binds to the active repressor and transcription still occurs C) Positive control-(must be switched on) 1) Lactose blocks active repressor 2) CRP (CAP)binds to the RNAp and transcription occurs 3) Need an inducer (CRP) and a repressor (lactose) 31) Tryptophan operon A) Have an inactive repressor B) Trp is the corepressor C) If Trp is around the bacteria wont produce more Trp. D) If Trp is low transcription will occur 32) Attenuation-Antitermination-leader sequence A) High Trp-DNA forms a hairpin terminator between 3 and 4---no transcription B) Low Trp-anti-terminator hairpin between 2 and 3 ---- transcription C) No protein synthesis-terminator between 1 and 2 and between 3 and 4---no polycistronic Trp mRNA 33) Two component system A) Sensing domain-(kdpD)relays signal to C terminus, histadine kinase is phosphorylated and then it activates the RR B) Response regulator (KdpE)-once activated binds to RNAp and activates gene expression 34) Mutation, Repair and Recombination A) Transition-purine to purine (A to G) B) Transversion-purine to pyrimidine 5 of 8 C) UV light will cause a thymine dimmer in DNA D) Repair 1) Direct-enzymatic removal 2) Excision 3) Post-replication 4) SOS-recA gene encodes for recombination proteins 5) Error prone repair-last resort 35) Gene exchange A) Transformation-naked DNA B) Transduction-virus (bacteriophage) mediated C) Conjugation-tubes between bacteria 36) Separate DNA via agarose gel electrophoresis 37) Plasmids A) Contribute to antibiotic resistance 38) Chromosomal DNA A) Can be taken up by other bacteria. For example the DNA to make a capsule can be taken up by a bacteria without a capsule. 39) Bacteriophage A) Lytic life cycle-phage particles eventually lyse B) Lysogenic life cycle-phage DNA gets cut out of host DNA and some host DNA comes with it. 40) Gene transfer A) Generalized transduction-Virus carries random DNA B) Specialized transduction-virus carries specific genes from donor to recipient C) Conjugation-plasmid or chromosomal DNA can be transferred D) Transposons-Mobile genetic elements that can transfer DNA within a cell from one position to another in the genome or between different molecules of DNA (e.g., plasmid to plasmid or plasmid to chromosome) 41) Recombinant DNA technology A) Transposon mutagenesis to ID virulence factors B) Can produce 5-10 pounds of E. Coli with recombinant DNA tech C) REstiction enzymes-read palindromic sequences (EcoR1) D) Chakrobarty can degrade oil 42) Enterobacteriaceae-all can cause UTI, wound infection, and pneumonia A) Gram neg-LPS, lipid A, O antigen 1) H antigen-flagell 2) K antigen-capsule Bacteria review (1) O 157:H7-E coli strain found in spinach B) Facultative anaerobes C) Ferment glucose D) Oxidase neg.-(oxidizers don’t ferment but fermenters can oxidize) pseudomonas is oxidase positive so good to distinguish in normal flora from EB E) Catalase posterior F) Nitrate posterior G) Peritrichous flagella H) ID with genetic relatedness of the highly conserved 16s RNA I) Normal Flora 1) Immunological stimulation 2) Opportunistic infection 3) Compete with other bacteria in gut J) E. Coli 1) Lactose (MacConkey) and indole positive 2) Shigella and salmonella are lactose neg! 3) Cause UTIs from anatomy of women,catheterization etc 4) Peritonitis-form ruptured gut. 5) Septicemia 6) Wound infections 7) Diarrhea 8) EPAC(infant diarrhea), EAEC and ETEC (travelers), EHEC (hemolytic uremic syndrome O157:H7), Enteroinvasive E. Coli (bacillary dysentery which is a little blood in the stool with watery diarrhea) K) Klebsiella pnuemoniae 1) Lactose pos. 2) Capsule 3) B-lactamase-resistent to penecillins 4) Non-motile 5) Cuased by vomiting, nosocomial, diabetes, respirator L) E. cloacae and aerogenes 1) Lactose pos. 2) Non-motile 3) UTIs 4) Cephalosporinase M) Protease mirabilis, Protease vulgaris 1) Urease positive!!!!!! 2) Highly motile, swarm 3) H2S positive 4) Tetracycline, ampicillin and cephalosporin resisntant 6 of 8 43) Diarrhea A) ETEC-watery secretory travelers diarrhea (malfunction of the Na absorption mechanisms) 1) Labile toxin 2) Stable toxin B) O157:H7-bloody diarrhea 44) Salmonella enterica A) Lactose neg B) H2S positive C) Causes 1) Typohiod fever 2) Enterocolitis-Food poisoning (typhimuriam, Enteritiditis, Newport) 3) Bacteremia D) Takes 10^5 salmonella to cause infection E) S. enterica-in normal flora of chickens. Invades epithelial cells of humans F) XLD-shows H2S as black colonies G) S. typhi (typhoid fever) 1) Vaccine-killed whole S. typhi. LPS present can give side effects. OR a live ty21a vaccine administered in capsules 2) Pathogenesis: mucosa to blood stream to spleen, liver, to secondary bacteremia H) EIEC-looks like shigella I) EHEC-bloody stool 1) HUS (hemolytic uremic syndrome)binds Gb3 or Pk antigen which is high in renal glomeruli of kids and in platelets in adults 2) O157:H7-sorbital neg. J) Shigella 1) Non-motile 2) Lactose neg 3) Very infectious-takes 200 organisms to get infection 4) 4 species 5) Transmitted by feces, fingers, food, flies, and water K) Yersinia entercolitica 1) Ulcerations in appendic 2) Normal in pigs and animals 3) Transmitted by contaminated food L) Campylobacter jejuni and C. coli 1) Gram neg in chickens, cattle and pets 2) Bloody diarrhea 3) Transmitted by food or water M) Vibrio Cholerae 1) Gram neg. 2) In oysters and seawater Bacteria review 3) Watery diarrhea 4) High dose to infect 45) Mycobacterium Tuberculosis A) Acid fast-stain with carbolfuschin B) Cause disease in AIDS patients C) Two obligate pathogens: M. tuberculosis and M. leprae-hansens disease D) Mycolic acids, abinoglactan, acyl lipids, PG, orins in cell wall E) Obligate aerobes F) Humans are the only host G) Deacreasing in the U.S. since 80s because of AIDS epidemic H) Rough appearance on a slide I) Survives drying but not UV rays, chlorine, phenols, and pasteurization J) Transmitted by droplets K) 95% of people become latent once exposed and 90% never reactivate L) Synergistic with HIV M) Types 1) Adult-upper lobes of lung 2) Childhood-middle lobes 3) Acute Tb pneumoniae-AIDS 4) Miliary-tubercles all over their body 5) Cold abscess-in a localized area (needle stick) 6) Addisons-adrenal insufficiency N) Diagnosis 1) AFB smear-quickest 2) Tuberculin skin test-15mm=infection, 10mm=at risk, 5mm=early, immunosuppressed at risk 3) Chest radiograph 4) AFB culture-way you really tell if someone has Tb 5) Digestion with NaOH (Tb survives but nothing else does) or N-acetyl-Lcysteine 6) Centrifuge O) Prevention 1) BCG vaccine 2) INH (isoniazid), prophylaxis 3) Chemotherapy 46) M. avium A) Soil, hot water like showers B) Transmit through respiratory or GI C) Resistant to many antimycobacterial drugs 47) M. kansasii A) Photochromogen 48) M. marinum-photochromogen 7 of 8 A) In water and fish tanks B) lymphocutaneous 49) M. scrofulaceum A) Granulomatous cervical lymphadenitis 50) M. fortuitum-M. chelonei complex A) Skin, pulmonary B) Rapid growers 51) M. leprae A) Hansens disease B) Tuberculoid leprosy 1) Hypopigmentation 2) Anesthesia 3) Cooler parts of body 4) Detected with AFB test 5) Non progressive 6) Intact CMI C) Lepromatous 1) Depressed CMI 2) Bacteremia 3) Leonine facies 52) Nocardia-strictly aerobic actinomycete A) Partially acid fast, gram positive B) N. asteroids most common C) 4 types 1) Pulmonary 2) Localized extrapulmonary and systemic 3) Cutaneous 4) Mycetomas D) Susceptible to sulfonamides E) Found in soil 53) Antibiotics A) Cell wall 1) Beta lactams-mimic D-ALA-D-ALA (1) Penicillin, cephalosporin, carbapenum 2) Non beta lactams (1) Cycloserine-cant make D-ALA (2) Glycopeptides (vancomycin)-binds d-ala (3) Bacitracin-binds lipid carrier (good against gram positive) (4) Isonazid-inhibits mycolic acid synthesis (Tb treatment) B) Ribosome 1) Tetracycline-prevent attachment of tRNA-AA to 30s (Gram + , ricketsia and Chlamydia, mycoplasmas,STDs) 2) Aminoglycosides-bind 30s and inactivate the initiation complex (gram – aerobes), degraded by stomach, take IV Bacteria review 3) Macrolides-bind 50s and prevent release of t-RNA (mycoplasmas, Chlamydia, legionella) 4) Oxazolidinones-bind 50s and prevent formation of initiation complex 5) Chloraphenicol and clindamycin-bind 50s and prevent peptide bond formation (treat anaerobes) 6) Streptogramins-bind 50s and prevent peptide elongation and premature release C) Cell membrane 1) Polymyxin 2) Daptomycin-depolarization of mem, influx of K D) DNA replication 1) Rifampin-prevents transcription, cant use alone 2) Metronidizole-only anaerobes, directly damages DNA 3) Flouroquinolones-inhibit DNA gyrase E) Antimetabolites 1) Sulfonomides-mimic PABA (precursor for folic acid) 2) Trimethoprim-inhibits dihydrofolate reductase 54) Antibiotics 2 A) MIC-minimum inhibitor concentration B) MBC- minimum bactericidal concentrationlowest # of antimicrobial to kill 99.9% of bacteria C) Bacteriostatic-inhibits growth, D) Bacteriocidal-kills bacteria, good in immunosuppressed E) Agar disc diffusion-qualitative test F) Agar gradient diffusion-quantitative test 55) Desirable properties of an antibiotic A) Selective toxicity B) Water soluble C) Bactericidal D) High conc. in serum for several hours E) Broad spectrum F) Minimal effect on normal flora G) Low potential for inducing resistance H) Minimal side effects 56) Drug resistance increasing because A) People are sicker B) New devices and procedures C) Ineffective infection control D) Increase in antimicrobial prophylaxis E) Emperic polymicrobial antimicrobial therapy 57) Characteristics favoring resistance 8 of 8 A) Ability to exchange DNA B) Ability to survive adverse environments C) Easilycolonize, infect,and transmit between people D) Reservoirs in the body 58) Types of resistance A) Innate-gm negs are resistant to vancomycin because it is too big to get inside cell, isonazids don’t work on gm – because no mycolic acids B) Acquired-clinically worrisome 59) Mechanisms of antibiotic resistance A) Enzyme inactivation-beta lactamase B) Metabolic bypass-bypass folic acid C) Decrease permeability-antibiotic cant get into cell D) Active efflux-of antibiotic E) Altered targets-mutate the DNA gyrase and quinolones cant work 60) Some drug resistant bacteria A) S. aureus, E. faecium, S. pneumoniae, ESBLproducing E. coli and Klebsiella spp., and P. aeruginosa.