i: TOPNOTCH MEDICAL BOARD PREP MICROBIOLOGY HANDOUT BY PACIFICO ERIC E. CALDERON, M.D. ae For inquiries, visit www.topnotchboardprep.com.ph or https://www.facebook.com/topnotchmedicalboardprep/ [eeneiniies infection filagy.—— [Diagnostic Microbiology [Parasitology Mycology Bacteriology eit MUST-KNOW TOPICS « Differences among prokaryotes and eukaryotes + Differences among bacteria, viruses, fungi, and parasites + Important nermal flora fajor mechanisms of pathogenicity # Laboratory diagnosis, WHAT ARE MICROORGANISMS? + Minute organisms += Too smail tobe seen by the unaided eye + Iodividually Wily STUDY MICROBIOLOGY? + Causative agents of infectious diseases ‘= Normal fora «+ Environmental importance ‘© Decompasers « Produce oxygen ‘2 Food chain 0 Sewage treatment (bioremediation) + lodustrial importance ‘9 Fond industry © Brewing industry ‘e Pharmaceutical industry Genetic engincering, «Research > es « Simple cell structure » Rapid rateof growth co Inexpensive to culture ties and metabolis MICROBIAL GROUPS * Microorganisms ‘9 Bacteria, protozoa, microscopic algae and fungi «+ Aceltular infectious agents ‘Viruses, vitoids andl p ic elem iophagas, plasmids and transposons: + Animal parasites ‘© Nematodes iroundworms), trematodes (flukes) and cestodes (tapeworms) 7“ ARCHARBACTERA oe rae ‘TOPNOTCH MEDICAL BOARD PREP MICROBIOLOGY HANDOUT BY PACIFICO ERIC E. CALDERON, M.D. (Characteristic Prokaryotas | Eukaryotes JONA within 2 nuclear membrane No | _ves patote division Ne ver Mare than one. site of nosome emesis | (505+ 408) [cit wai containing peptidoglycan | Ne Prokaryotic ribosomes: 50 + 30 = 70 S stands for svedberg unit cooley “ HEB + > Ge SVEDBERG UNIT ‘ Non-SL unit for sedimentation rate + Sedimentation rate s the rate at which particles ofa given size and shape travel to the bottom of the tube under centifygal force * This reflects the rate at which # molecule sediments under the centrifugal force ofa centrifuge + Thesvedberg is technically a measure of time, and is defined as exactly 1059 § (100 6) Page 1 of 76 For inquiries, visit www.topnotchboardprep. com. ph or https://www.facebook.com/topnotchmedicalboardprep/TOPNOTCH MEDICAL BOARD PREP MICROBIOLOGY HANDOUT BY PACIFICO ERIC E. CALDERON, M.D. a For inquires, visit wwew.topnotchboardprep.com.ph or Rtps://www.facebook.com/topnotchmedicalboardprep/ io ad it) ie aut Taciaieaas) {5-15 ephones) oth Ba ‘was ane we (Soh DN an “Tyne of mucous [eukaryote eukaryotic omnes ose cree eg ere acc lons income | “eset ‘neeert eset oe detest et eesceseseeesct a setae aoe sala Protencapad ana toopraten envelop] Ripa wal eatarangSepbdoghan ig wal contabwg eget erae ta 7 rene ‘fatica a vsicaton wera fas "fens Few Saag or ma? Which microscopic characteristic primarily distinguishes ‘Which ofthe following infectious agents lacicnucleic acids? | prokaryotes from eukaryotes? Viruses ‘A. Relatively small size and absence ofa nuclear membrane B. Bacteria | B. Presence of histone proteins in the nuclear area C.Viroids © Naked, cireslar chromasomes D.Prions D. Presence of subcellular structures ‘Which ofthe following organisms lack membrane sterols? | PRIONS ae [A Yeasts * Noncellular infectious proteirs B. Mycoplasmas ‘+ Naked proteins that have the sume amino acid sequence as | © Protozoa certain normal human cell surtace p teins But have folded D. Staphylococci differently — ~ + Resistant to nucleases, proteases, mary chemicals, and normat autoclaving «+ Associated with spongiform encephalopatiies, eg. Creutafeldt Jakob disease, kuru, fata familia insomnia viroiws * Obligate intracellular para * Acellular = neha a s DYSFUNCTION DUE TO PROTEIN MISFOLDING ‘+ No human diseases known ‘TRANSPOSONS -9 “transposable” elements * Mobile geneticelements + DNA pieces that move readily from one site to another either within or between the DNA of bacteria, plasmids, and bacteriophages Ong = “Jumping genes” ® —m += Code for drug-resistant enzymes, toxins, or metabolic enzymes + Cause mutations in genes into which they insert or alter the expression of nearby genes, PATHOGENESIS OF PRION INFECTION “Thera PrP converts) Tha naw PrP la when PrP acouruttes n P16 contain to mine Onymiea oem ° Scunete ‘eo ‘TOPNOTCH MEDICAL BOARD PREP MICROBIOLOGY HANDOUT BY PACIFICO ERIC E, CALDERON, M.0. Page 2 of 76 For inquiries, visit www.topnotchboardprep.com.ph or https://www.facebook.com/topnotchmedicalboardprep/‘SPONGIFORM ENCEPHALOPATHIES + appearance of the vacuolated neurons with loss of function ‘and the lack of an immune response or inflammation. HUMAN PRION DISEASES + Creutzfeldt-jakob disease (CID) ¢ Kuru > “kurla/guria” =to shake «Variant CJD (¥C}D) + Gerstmann-Straussler-Scheinker (GSS syndrome) «¢ Fatal familial insomnia (FF!) + Sporadic fatal insomnia ‘Infection 2 Histopathol Higher Cortical Manifestation Dysfunction v v Dysdladochokinesia, Dementia ataxia, myoclonic jerks ete J + Kuru=to shake © Fatal neurodegenerative prion protein (PrP) diseases in humans Cerebellar syndrome ‘© With characteristics and relentless progression of neurological symptoms through well-defined clinical stages Inivially, cognition is fairly well-preserved. «Disease presentation is sufficiently distinctive to be easily recognized by patients, their relatives, and the local community + Clinical features © Cerebellar ataxia Voluntary tremor © Involuntary movements (choreoathetosis, myodonte jerks, fasciculations) © Euphoria, dementia emotional liability. and a loss of grasp reflexes in advanced stages © Why kuru? © Spread by the endocannibalistic funeral practices ofthe Fore ‘Family members were rituaistieally cooked and eaten following their death Brain > the most mfectious organ ** Women most commonly consumed the cooked brains ANIMAL PRION'DISEASES + Scrapie (sheep énd goats) ‘ Transmissible mink encephalopathy ‘Bovine spongiform encephalopathy (BSE) > Also known as mad cow disease «# Chronic wasting disease (mule, deer, elk) "TRANSMISSION + viainfected tissue ‘through cuts in skin ‘transplantation of contaminated tissues (cornea) ‘ouse of contaminated medical devices (brain electrodes) © ingestion of infected tissue (cannibalism) + viaioherited syndrome 1 May be sporadic SUSCEPTIBLE POPULATION ‘© women and children ofthe Fore tribe in New Guinea ‘neurosurgeons nd brain surgery patients « transplant surgeons and transplant patients ‘TOPNOTCH MEDICAL BOARD PREP MICROBIOLOGY HANDOUT BY PACIFICO ERIC E. CALDERON, M.D. ‘TOPNOTCH MEDICAL BOARD PREP MICROBIOLOGY HANDOUT BY PACIFICO ERIC E. CALDERON, M.D. For inquiries, visit www.topnotchboardprep.com.ph or https://www.facebook.com/topnotchmedicalboardprep/ ‘SPECTRUM OF DISEASE + progressive, neurodogencrative disease loss af muscle control o shivering ‘> myoclonic ferks and tremors ‘loss af coordination ‘rapidly progressive dementia ‘death ‘TREATMENT and PREVENTION ‘+ na treatment avatlable « cessation of ritual cannibalism « elimination of animal products from livestock feed « disinfection of neurosurgical tools and electrodes '5% hypochlorite solation or 1.0 M sodium hydroxide or autoclaved at 15 psi for t hour | Prionsare not normal and may cause disease because sey 7] | Ghuibtaproiem wits wich ofthe following? ‘Ah Foldingof segments polypeptide nt eomarcaly ordered units, : | fhm ac sequence na polyp chain | Protnassenbly inte thera poppeptde andite | | | zal ‘component domains | i | | D. Number and types of polypeptide units of eligomeric | “proteins and their spatial arrangement PrP knockout mice, which are engineered not to express the PrP gene, show ag obvious pathological phenotype. However, these mice have tian shown tc have abnormalities inal of the following, EXCERT which one? ‘A, Synapuic transmission BeImmunty . Cireadian rhythm Steep Which prion-related disease is caused by mutations inthe PrP gene? } A Kura |B. Sporadic c}D | & Fama cip [_b.pst Ul. BACTERIAL STRUCTURE _ BACTERIAL SHAPE AND SIZE «three shapes: ‘ccocet (spheres) ‘baci (rods) ce spirochetes (spirals) « cocel arranged in three patterns: ‘© paits (dlplococc!) ts ehains (streptocacel) clusters (staphylococet) BACTERIAL CELL WALL + All bacteria have a cell wall composed of peptidoglycan except Mycoplasma + peptidoglycan = sugar backbone (glycan) + peptide side chains (eptido) crosslinked by transpeptidase Peptidogiycan structure LED serene ert EB mene esiin see asap 1c emabpye pment Page 3 of 76 For inquiries, visit www.topnotchboardprep.com.ph or https://www.facebook.com/topnotchmedicalboardprep/For inquiries, BACTERIAL CELL WALL + Prin proteins - Facilitate passage of small, hydrophilic molecules into the cell «+ Inthe outer membrane of gram-negative bacteria > channels, allow the entry of essential substances such as sugars, amino ‘acids, vitamins, and metals as well as many antimicrobial drugs such as penicilins. ca co es pref the ca wal? ee | hata lpercofpepadogiyean ne” | cory TOPNOTCH MEDICAL, BOARD PREP MICROBIOLOGY HANDOUT BY PACIFICO ERIC E. CALDERON, M.D. it www.topnotchboardprep.com.ph or https://www-facebook.com/tapnotchmedicalboardprep/ GRAM STAINING 2 | ecole Agent ‘ Counterstan SEIS GRAM STAINING ©. B14 glycosidic bond between NAG and NAM V-I-A-S . 1 D.Tetrapepside sidechains ft Violet Acetone | ___todine Safranin GRAM-POSITIVE VS GRAM-NEGATIVE, COLOR CHANGES DURING GRAM STAIN 2 Cone =, ‘rom betes = = om map Par evivon he Pa ors Pee eran aD = Decoetamon }5—— Paneognean ——$trof ———— ~= ie = Sree CaP re poo Conran — [COMPONENT GRAM (4) CELLS GRAM (CELLS | BACTERIA NOT SEEN IN GRAM STAIN Thicker, ~ Thinner C peer + multilayer + thianer layer. NAME REASON Teichoic acids r + Too much pin cl Upopolysaccnaride | Mycobacteriae " wallsodye cannot —ACID-FAST STAIN Periplasmie space 1 penetrate HEH DARKFIEL Spirechetes + Toothinto see Pon NONE sop. very small (SEROLOGIES) [Fesoneia + Foorurake afd sven sam «Intracellular GIEMSA STAIN > Chiamydiae = very small INCLUSION BODIES, ‘ineracelular——~—~CGIEMSA/ TISSUE | eee! very small, STAINS, SAUSTINEUIGN @AC TERIA NOTSEEW INGRAM STAIN | Tisse Basse May slcroscopicaly Lak Color. | “treponema Rickersia Mycobaceria Mycoplasma Legionella | chlamydia z | SUSE eee Bates tm ome Se mews torte GRAM-NEGATIVE CELL WALLS + outer membrane of gram-negative bacteria contains endotoxia {Mpopolysaccharide} © consists of lpid A and O antigen > induction of I-1 & TNF + All gram-positive bacteria have NO endotoxin EXCEPT Listeria ‘monocytogenes ‘TOPNOTCH MEDICAL BOARD PREP MICROBIOLOGY HANDOUT BY PACIFICO ERIC E. CALDERON, M.D. ‘Which component/s of the Gram-negative cell wall is/are responsible for its nonspecific endotoxin activity? | | A. Upopolysaecharide |B Teichoicacid | . Peptidoglyean Di inthe | Which of the following is the OLDEST acid-fast staining | ‘method, which requires heating the specimen during the procedure? | ‘A. Fluorochrome | B. Zicht-Neclsen | ©. Kinyoun | D. Auramine-thodamine | Page 4 of 76 For inquiries, visit www.topnotchboardprep.com.ph or https://www.facebook.com/topnotchmedicalboardprep/t rojicy TOPNOTCH MEDICAL BOARD PREP For inquiries, visit www.topnotcht 1 Which ompons fhe Gram-negative cll wall reduces | suscepubilty tlysozymes? ‘8. Telcholc acids | | | IE B. Beta-lactamases in the periplasm [ESSENTIAL COMPONENTS OF BACTERIA [RNA and protein in 50S COMPONENTS OF BAC [cel Well [As described previously ‘cytoplasmic [Upoprotein bilayer Jmembrane _withoutsterols [Ste of oxidative and cansport enzymes Protein synthesis (Genetic material lmembrane, nue na 7 invagination af plasma [Participates in cell vision | Meroveree membrane land secretion _| ‘Space between plasma [Contains many hydrolytic periplasm —|membrane and outer enzymes, including f- ilactamases, [NON-ESSENTIAL COMPONENTS OF BACTERIA [capsule — [Polysccharide _ [Protects against phagocytosis, [alycoproteim Attachment, conjugation Protein itis or Vcamaact oe lciycocaiys [Polysaccharide Fogel spore Plasmid MICROBIOLOGY HANDOUT BY PACIFICO ERIC E, CALDERON, M.D. oardprep.com.ph or https://www. facebook.con/topnotchmedicalboardprep! + Allbacterial capsiles are ‘composed of polysaccharide EXCEPT Bacillus anthracis (polypeptide of D-glutamate) «+ Spore: formed by gram-positive rods, especially Bacillus and Clostridium spp +» Extrachromosomal, double-stranded, circular DNA capable of replicating Independently ofthe bacterial chromosome. « Can sometimes be integrated into the bacterial chromosome > called episomes ‘TOPNOTCH MEDICAL BOARD PREP MICROBIOLOGY HANDO\ For inquiries, visit www.topnotchboardprep.com.ph or https: Reratnite coat © Tpesistace to heat, and chemeals |dipicotinic acic Resistance to heat, end los Genes for ato retance and ae toxins [siveoaen. ins, rane ste of mutrents in etoplsm lboiphosphats eae “SPORULATION IN BACTERIA ae a eet meen sympa bea tenet Seren cme Significance » Antibiotic resistance Resistance to heavy metals «= Resistance to UV light «Pili fimbriae) * Exotoxins and several enterotoxins. « Bacteriocins - toxic proteins produced by certain bacteria that ace lethal for other bacteria. UT BY PACIFICO ERIC E. CALDERON, M.D. Page 5 of 76 cwew facebook.com/topnotchmedicatboardprep/oma: | As part of an orientation seminar, @ medical resident received | | orientation on proper hand washing protocol while in the | hospital and treating patients. Which of the following ‘microbial characteristics necessitate the use of soap-based. | prodets over te alcohol rubs/ges commonly found in every patient room in the hospital? | A Aerobic growth D. Naked DNA virus proteins B. Cell wall teichoic acid —_E. Spore-forming i | | | a CAPS. oe | isan a snr may be cv agit ie | Fallowing organisms except which one? | A. Staphylococcus aureus eee. C.HPV 16 _D.Candida IV. BACTERIAL GROWTH + Bacterial growth is a coordinated process of acreasen Individual cell mass and size and duplication of the chromosome, followed by cell division «* Bacterial reproduction occurs via binary fission [BACTERIAL GROWTH CYCLE hae abe Sagehewe PHASE 1: LAG PHASE « Cells are depleted of metabolites as the result of unfavorable condition ‘* Adaptation to new environment ‘¢ Nutrients are incorporated «= Vigorous metabolic activity oceurs but cells do not divide «Zero growth rate PHASE 2: LOG OR EXPONENTIAL PHASE ‘= Rapid cell division occurs ‘¢ Lactam antibiotics act during this pra! + Constant growth rate ‘© Continues until either one or moré,nutriefts in the medium ‘become exhausted, or toxic metabolites accumulate and inhibit growth, PHASE 3: STATIONARYPHASE. «Exhaustion of nutri or the accumulation of toxic products cause growth togeatiscompletely (zero growth rate) + Spores are formed PHASE 4: GEGUNE OR DEATH PHASE + Most ofthe ces die because nutrients have been exhausted + Negative growth rate EFFECT OF BACTERICIDAL & BACTERIOSTATIC ANTIMICROBIALS fe Lane peer ‘TOPNOTCH MEDICAL BOARD PREP MICROBIOLOGY HANDOUT BY PACIFICO ERIC E. CALDERON, M.D. TOPNOTCH MEDICAL BOARD PREP MICROBIOLOGY HANDOUT BY PACIFICO ERIC E. CALDERON, M.D. For inquiries, visit www.topnotchboardprep.com.ph or https://www.facebook.com/topnotchmedicalboardprep/ «oxygen metabolic generates toxic products such as superoxide and hydrogen peroxide + superoxide dismutase, peroxidase and catalase are nceded to survive in aerobic environments ‘AEROBIC METABOLISM. ‘ Obligate Aerobes ‘9 completely dependent on oxygen for ATP-generation ‘+ Microaerophiles ‘ouse fermentation but can tolerate tow amounts of oxygen because they kave SOD ANAEROBIC METABOLISM + Facultative Anaerabes utilize vygan itis present, but can use fermentation ints Rbsence + Aerotolerant Aagiro exclusively anaardbe bu insenstive to the presence of oxygen + Obligate Anaerobes cannot grdw in the presence of eaygen because they lack SOD, peronidave and catalase OXYORN AEQUIREMENT ce ef | obtgaie ive | Cotas | j Sanka Steed | ancettl | Meroe { re Gi BACTERIAL OXYGEN METABOLISM ERVRONRENT EFFECT OF OXYGEN ‘GROUP Jovigae serve | Growen | No Growth IRequres 1 "Required but at iow Microseropite Ltevest¢0 ? atm) orly Growth? | No Growth z Not requred for! poner nat] lanaerobe 4 | loblgate | Microaerophiles |Facultative Anaerobes (Aerotoierant |Anaerobes Obligate anaerobes For inquiries, visit www.topnotchboardprep.com.ph or https://www.facebook.com/topnotchmedicalboardprep/‘foros: Torvoven meoca,soano pte wcroBioLocY HANDOUT BY PACIFICO ERE. CALDERON, HO. Be For inquiries, visit www.topnatchboardprep.com.ph or https://www.facebook.com/tepnotchmedicalboardprep/ Bacterial jetabolism —_ 0 ocean mre ‘OBLIGATE AEROBES neo Orman ‘osy and Nagging Bests Must Breathe Lots of oxygen. eran ig Bee ‘Nocardia Neisseria | = eae ate Bordetella/Brucella/B. cereus Legionelts | OBLIGATE ANAEROBES | ABC of Anaerobes | “Actinomyces | Bacteroides VI BACTERIAL GENETICS Barre reece aren ‘DNA TRANSFER BETWEEN BACTERIAL CELIS. * Tensfer Type of Cells acetone Procedure, Involved a | conjugation Prokaryorlc IDNA transferred by | Transduction |virustrom onecellta _|Prokaratic | another \ ri T Jamormanion [Purfed BNA ken up By [ Protarytie or [ Raesformation [cet J : can,” Transformation PROPHAGE CODED BACTERIA : ty ins of ABCDE, ] * » shiigA - like toxin: EHEC 1 Sex pilus ‘Transduction Booitnum | ‘Gnolera ! Diphtheria i Erythrogeniestrain | “GRIFFITH EXPERIMENT - TRANSWORMATION (lournai of iygiene 27 (2): 113-159, 1978) © wocoterten wes tnctates © coterie of encapesied Soomoose' ‘Semen nous —— ‘eonanea mous ‘oerored, ‘VII. NORMAL FLORA. res ~ aleroorgnsnsthararetie permanent edvats te oay [Sz > Suphwsenn cider | orl fora ue lowetrleneorganons her sul [Bete Saptecer anatomic site = + colonization resistance occurs when normal flora occupy Dental plague receptor sites preventing pathogens from binding [Colon s Bacteroides, Escherichia Lectbcs vat Vagim —— Echenchacae, 1 Seepoenccs galcin | TOPNOTCH MEDICAL BOARD PREP MICROBIOLOGY HANDOUT BY PACIFICO ERIC E. CALDERON, MO. age 7 of 76 For inquiries, visit www.topnotchboardprep.com.ph or https://www.facebook.com/topnatchmedicalboardprep/PNT oe AN {BACTERIAL PATHOGENESIS ‘+ pathogens are microbes that can cause disease; opportunistic pathogens cause disease only in immunacompromised people + virulence isa measure of a microbe's ability to cause disease + determined by virulence factors, such as capsules. exotoxins, fr endotoxins + IDsois the number of organisms required to cause disease in $5096 of the population BACTERIAL INFECTION ‘infection has two meanings: (1) the presence of microbes in the body and (2) the symptoms of disease ++ presence of microbes in the body does not always result in symptoms of disease (MECHANISMS OF BACTERIAL DISEASE ‘* production of toxins (both exotoxins and endotoxins) «Induction of inflammation MODES OF TRANSMISSION ‘¢ Human to Human co direct (sexual, transvaginal), fecaloral, inhalation, transplacental, blood-borne ‘* Nonhuman to Human ‘sil, water, direct animal source, vector-borne, animal excreta, fomites [BACTERIAL ADHERENCE « pill mediate attachment of bacteria + glycocalyx mediates strong adherence to surface of human cells «surface proteins called curli mediate binding to endothelium and to extracellular proteins such as fibronectin, EX | Salma Hayek and her ‘curlt hair (Salmonella) [ENZYMES IN BACTERIAL INVASION + collagenase and hyaluronidase ‘ spread through subcutaneous tissue + coagulase ‘accelerates formation ofa fibrin clot coating the organisms with alayer of fibrin + immunoglobulin A (IgA) protease allows adherence to mucous membranes + leukocidin ‘o destroys both neutrophilic leukocytes and macrophages | Mich oftefoiowing maybe cridredprinariy | responsible fr the development of elutisfrom a small | Huruncte? [A lipases Hyaluronidases Pantor-Valentineleukecidins _Hemoly page (RAUEXTUNI Bacteria with igs Protease ‘SHINe My Gong |” Streptococcus pneumoniae | Haemophilus influenzae | Neisseria Meningtidis Neisseria Gonorrhoeae VIRULENCE FACTORS + Polysaccharide Capsule © protect against phagocytosis © anticapsular antibodies allow more effective phagocytosis to occur (OPSONIZATION) © Cell Wall Proteins ‘OM protein of S. pyogenes of antiphagocytic 9 Protein A of S. aureus prevents complement activation TOPNOTCH MEDICAL BOARD PREP MICROBIOLOGY HANDOUT BY PACIFICO ERIC E. CALDERON, M.D. ‘TOPNOTCH MEDICAL BOARD PREP MICROBIOLOGY HANDOUT BY PACIFICO ERIC E. CALDERON, M.D. For inquiries, visit www. topnotchboardprep.com.ph or https://www.facebook.com/topnotchmedicalboardprep/ ‘TOXIN PRODUCTION froperty | Satan fndotosin | source iGamaentnd om canaaqunabacene lca tes Ichemistry _ Peypepnde ‘popstar ‘location of genet Plasmon baceip ewicty [igs [antigenicity ‘Waceines. |Meatatbiy [Typical diseases Ci) Pat >t Yeon a int sh (sean hee Fre a ber ements Seite Reese EXOTOXINS ‘polypeptides secreted by certain bacteria that alter specific cell functions resulting inthe symptoms of disease « havean A-B subunit structure: ‘© A subunit is the active (toxic) subunit, © Bsubunit is the binding subunic [MECHANISMS OF ACTION OF EXOTOXINS MOR ‘ADPribosyiation [Diphtheria toxin, cholera ton, Escherichia |colineat fable toxin, and pertussis toxin ‘Toxic shock syndrome toxin, taphylococeal serotonin, and erythrogeni toxin “Tetanus toxin, botulinum tox, lethal factor | |ofanthrax toxin, and scalded stn toxin {clostridium perfringens aiph ny | When an exotoxin binds to a receptor, itactivateT~ binding SIMULTANFOUSLY to a T-cell receptor and MHC Il molecule on an antigen-presenting cell WITHOUT requring an | antigen. It activates large number of T cellsto cause acytokine | storm. What isthe nature ofthis exotoxin? A. Superantigen B, Protein synthesis inhibitor ©. ADP-ribosylator D. Protease ‘Lecithinase_ Which ofthe following kills ells by cleaving 60S ribosomal subunts? A. Bxotoxin 4 B. Shiga toxin Amikacin D. Clindamycin A. Listeriolysin O B, Anthrax toxin | | ‘Which of the foliowiog bacterial toxinsis neurotoxic? | C. Shiga-like toxin | Page 8 of 76 For inquiries, visit www.topnotchboardprep.com.ph or https://www.facebook.com/topnotchmedicalboardprep/ENDOTOXINS * lipopolysaccharides (LPS) located inthe outer membrane OF gram-negative bacteria + Lipid A's the toxic component of LPS ‘o induces the overproduction of cytokines, such as TNE & IL-1 ‘activates the complement cascade activates the coagulation cascade, resulting in disseminated intravascular coogulation ERMAN 2201 | _evaonnisaninera pare ofgram-ertive celal | ‘TOPNOTCH MEDICAL BOARD PREP MICROBIOLOGY HANDOUT BY PACIFICO ERIC E. CALDERON, M.D. For inquiries, visit www.topnotchboardprep.com.ph or https://www.facebook.com/topnotchmedicatboardprep/ ‘SPECIALIZED MEDIA FOR BACTERIAL GROWTH Bacteria Clostridium perfiingens Jeroup D streprococe! staphylococci Ww. meningitidis, |W. gonorrhoeae from sterie sites GENERALITIES ON BACTERIAL STRUCTURE ‘Ni bacteria have cell walls composed | yeoplasma ‘of peptidogivcan EXCEPT pneumoniae All gram-positive bacteria have NO Listerio ‘endotoxin EXCEPT smonoenogenes | = Sere ‘All bacterial capsules are composed | Bacifus ‘of polysaccharide EXCEPT | anthracis 4 | staphylococ: ‘All exotoxins are heat-labile ExcEPT 3 aa | enterotoxin OREN kes ‘A45/F consulted _ neck masses accompanied by fever, chills, weight ass and malaise. Fine needle aspiration was done to rule outa malignancy If the physician suspects TB lymphadenitis, ‘which ofthe following should he request fo RAPIDLY identify | the mycobacterial species? | a pcr | B. Culture on blood agar plate ©. Zichl-Neelsen staining _D, Culture on Lowensteif-fensen medium. MICROBIOLOGIC EXAMINATION + Direct Examinationand Techniques + Culture + Microbial Identification: Colony and cellular morphology: srowth charateristics under various ronditions,uillzaton of carbohyarates and other substrates; enzymatic activity, immunoassays, and genetic probes + Serodiagnosis: high or rising titer of specific IgG antibodies orthe presence of specific IgM antibodies may suggest or confirma dagnosis + Ancimicrobia! Susceptibility: Microorganisms, particularly bacteria, are tested in vitro to determine whether susceptible to antimicrobial agents nedia cond to their growth « Culture medium - a nutritive substance in which cultures of ‘microorganisms are grown ‘© May be a gel or liguid medium ‘TOPNOTCH MEDICAL BOARD PREP MICROBIOLOGY HANDOUT BY PACIFICO ERIC E, CALDERON, M. i cea paar ch Heine fesepie esaheate (G5) Semon, sete Elinghousen-MeCullough-iohnson | cen pata Feu | Leptospira interrogont Culture on Lowenstein-Jensen ‘medhum revealed typical dry, hheaped-up yellow to bufl- colored colonies of ‘Mycobacterium tuberculosis. MOLECULAR TESTS if * nucleic acid amplification tosis, nuclele acid probes, and mucele acid sequence analysis, + highly specitic, quite sensitive and mucin faster than culture + especially useful for those bacteria thatare difficult to culture such as Chlamydia and Mycobacterium species IMMUNODIAGNOSTICS ‘Diagnostic methodology that uses an ant as the primary means of detection ‘Takes advantage ofthe specificity affinity of the antibodies to Ins corresponding antigen According to protein detected 41. Antigen Detection ‘ Immunofluorescent antibody tests © BIAs, including enzyme-linked immunesorbent assays (USA), and agglutination tests 2. Antibody tests Western blot immunoassays © Lepto MAT Corr SEROLOGIC TESTS ‘* Most ofthe immanodiagnastic tests would use the seram-> contains antibodies «Usually for diagnosis of viral infections with long incubation period prior to the appearance of clinical manifestations ‘Epstein-Barr virus the hepatitis viruses, and HiV + Typically, testing for antibodies to these viruses is te frst step + Followed later by nucleic acid amplification Page 9 of 76 For inquiries, visit www.topnotchboardprep.com.ph or https://www. facebook com/topnatchmedicalboardprep/IMMUNOFLUORESCENT (IF) ANTIBODY STAINING ' More specific than other staining techniques but also more cumbersome to perform + Commonly uses fluorescetn-labeled antibodies * Use of direct IF staining on specimens from patients is more dificult and less specific ‘+ Common uses: Bordetella pertussis or Legionella pneumophila in colonies isolated on culture media sar, em brit tld ee UE ak Paulie sit cupers Sst Ante trae hare Ser ia Bs Inthe indiret fluorescent antibody test, the Buorescent dye is attached to antibody made against human igG- RABIES LEAT ‘The rables antibody used for the dFA testis primarily directed against the nucleoprotein (antigen) of the virus. + Rabies virus replicates in the cytoplasm of cells, and infected cells may contain large round or oval inclusions containing collections of nucleoprotein (N) or smaller collections of antigen that appear as dust-like fluorescent particles if stained by the dFA procedure, SEROLOGIC TESTS FOR SYPHILIS + Nontreponemal -screening tests, easy to perform, low coe, * ‘oVenereal Disease Research Laboratory (VDRL}- require tmlroscopic examination to detect flocculation 6 Rapld plasma reagin (RPR) © Antigen: cardiolipin, cholesterol, and purify lectin (Others: Unheated serum reagin (USR) and tmjuldine red ‘unheated serum test (TRUST) ‘+ Treponemal antibody tests— for coffirmation co measure antibodies against T pallidum antigens ‘oT pallidum enzyme immundassey (TPEIR} ELISA or EIA 1. Coat the wells of plastic'terodilutton trays with desired antigen have metabolic processes closer to those of the human host than to prokaryotic bacterial pathogens ‘TREATMENT OF PROTOZOAN INFECTIONS. + Protoznal diseases are less easily treated than bacterial, infections, + Many of the antiprotozoal drugs cause serious toxic effects in the host 0 Calls showing high metabolic activity: neuronal, renal ‘tubular, intestinal, and bone marrow stem cells + Most have not proved tobe safe for pregnant patients ‘OVERVIEW - PROTOZOA Type and (Pls [Blood and tissue !/212P/082: | Trypanosoma sp. 7 L ENTAMOEBA HISTOLYTICA PARASITE BIOLOGY + Pseudopod-forming nonflagellated protozoa 1 Most invasive parasite among Entamoeba «Eukaryotic organism that lacks membrane- bound organelles (SSION | INFECTIVESTACE, DIAGNOSTIC STAGE] ‘trophozoites, mature ests, immature cysts. + fecalorel route | » mature cysts PATHOGENESIS: Virulence Factors + lectin mediates adherence + amebapores for penetration + eysteine proteases for cytopathic effect SPECTRUM OF DISEASE * cyst carrier state ‘+ amebic colitis ‘dysentery without fever o flask-shaped colon ulcers + ameboma ‘associated with dysentery + amebic liver abscess ‘9 most common extraintestinal form anchovy sauce-lke aspirate Page 13 of 76 For inquiries, visit www.topnotchboardprep.com. ph or https://www.facebook.com/topnotchmedicalboardprep/