PART 1 QUIZ- microbiology .pdf

MICROBIOLOGY, PARASITOLOGY & PUBLIC HEALTH MICROBIOLOGY o Streptococcus Aureus study of microorganisms which are large and diverse ✓ Home / Batch / Low-temp hold...

MICROBIOLOGY, PARASITOLOGY & PUBLIC HEALTH MICROBIOLOGY o Streptococcus Aureus study of microorganisms which are large and diverse ✓ Home / Batch / Low-temp hold group of microscopic organisms that can occur as either pasteurization = 63°C for 30 minutes single cell or in cluster arrangement including viruses ✓ Flash / high-temp short time (HTST) = which are microscopic but acellular 72°C for 15 seconds o "Micro” means small therefore cannot be seen by ✓ Ultra-high temperature (UHT) = 140°C the naked eye and seen using microscope for 4 seconds Parasitology – study of parasites; Most can be seen but ▪ Anaerobic Process – no oxygen some are not. ✓ Fermentation – Converting Sugar → o Organisms require direct contact by the host to Simpler substances acquire the nutrients. Not all parasites are worms o Homo – single type Public health - science and art of 3Ps o Hetero – multiple types o Preventive disease o John Tyndall o Promoting health ▪ Tyndallization – intermittent sterilization o Prolonging life ✓ AKA: Fractional Sterilization (BEQ) o Can be achieve through organized community efforts Golden age of Microbiology Benjamin Martin - publish "A new theory of CHAPTER I: BASIC CONCEPTS - INTRODUCTION consumption" HISTORY Robert Koch Early observation o discovers Bacillus anthracis – Anthrax - first Robert Hooke - discover cells, called "little boxes" or microorganism to satisfy the Koch Postulate. "prison cell" using cork cells and magnifying lens (Koch's Bacillus) (BEQ) o Cell theory: Top view it looks like a prison o Mycobacterium tuberculosis o Father of cytology o Koch's postulates Anton Van Leeuwenhoek - first person to observe live 1. a healthy person ( ) should not possess specimen under microscope microorganisms o using cheek and sperm cell, called it "animalcules" ▪ if a person has disease ( ) (+) means small animals microorganisms o Father of microbiology 2. In a person who has disease ( ), the microorganisms can be isolate and grow in Spontaneous Generation pure culture (using petri dish) out of nothing, produce new cell (BEQ) 3. Healthy persons ( ) inoculate Opponent (anti-SG) Proponent (pro-SG) microorganisms → it will have a disease Francesco Redi John Needham 4. Person with disease can re-isolate Jar with lid + meat = (-) Jar with broth + heat microorganisms and grow in pure culture maggots without lid = (+) magots o Exceptions for Koch’s Postulate Jar without Lid + meat = ▪ Normal flora - bacterial residents (reside in the (+) maggots body) of a body that will not cause disease unless inoculated elsewhere Lazzaro Spallanzani Mouth and Seen the largest normal flora Jar with broth + heat Colon population (BEQ) with lid = (-) magots Skin Staphylococcus Epidermidis Nose Staphylococcus Aureus Germ theory of disease Mouth Streptococci viridans Biogenesis theory Dental Streptococcus mutens o Pre-existing cells New cells plaque o Rudolf Virchow – anti-spontaneous generation; Colon Bacteroides > E. Coli but no proof / evidence Urethra Lactobacillus Spp. o Louis Pasteur - microorganisms are present in the Vagina Lactobacillus Vaginalis; E. Coli; air (but air is not the source itself) → SPA (BEQ) Strep. agalactiae ▪ Swan-neck / S-shape / Goose Neck flask Mouth and Candida Albicans is a normal experiment Vagina flora, a type of fungi ✓ removing of tip = (-) growth of NOTE: Considered as Sterile (no bacteria present) BLOOD microorganisms. and BRONCHI (BEQ) ✓ removing of "neck" = (+) growth in ▪ Concept of Carrier – The organism has microorganisms. disease, but no signs and symptoms present ✓ Tilt (tinagilid) (+) growth in microorganisms Example: Typhoid Mary - microorganism responsible is Salmonella typhi which has ▪ Pasteurization - aseptic technique - 2 diseases application of heat to kill microorganism o Salmonellosis – diarrhea, vomiting ✓ To avoid Milk-borne pathogens o Mycobacterium Bovis o Typhoid fever – Fever o Salmonella  MICROBIOLOGY, PARASITOLOGY & PUBLIC HEALTH Salmonella typhi can be seen in poultry, hyperthermophiles - chicken and egg which means it is a food- survive in very hot borne disease - defeat Koch postulate #1 environment Mycobacterium Leprae – never grown in pure culture instead requires armadillo Kingdoms Animalia Eubacteria – true Plantae bacteria and mouse foot pad - defeat Koch's Fungi – Candida postulate #2 spp. Archaebacteria – o Causative agent of Leprosy / Protista (Protozoa, Example: Hansen disease (BEQ) Algae) Methanogen = Immunity - defeat #4 o Amoeba produce CH4 gas Edward Jenner o Dinoflagellates– from CO2 + H2O GOOD TO KNOW causes red tide poisoning Birth of chemotherapy Chemotherapy - chemicals used in therapy like cancer Biofilm cell Synthetic lab clusters of microorganisms that are attached to a surface and/or to each other o Paul Ehrlich - "magic bullet" ▪ Compound 606 / Arsphenamine / "salvarsan" e. g. dental plaque - Streptococcus mutans - offers salvation to those people who has syphilis cause by Treponema Pallidum Stages of microbial growth ▪ Arsan contains arsenic ▪ Father of Antimicrobial Chemotherapy Antibiotics o Alexander Fleming - discovered penicillin through serendipity ▪ Use 3 petri dish. the 2-petri dish grows S. Aureus, and the 3rd has Penicillium Notatum (today: Penicillium Chrysogenum) o Howard Flory, Ernst Chain & Colleague ▪ Successfully purified Penicillin from molds Generation time – time it takes to double the population (BEQ) MICROBIAL GROWTH Stage 1: Lag Phase - Metabolically active but no BEQ – High Yield replication, size, synthesis Eukaryotes Prokaryotes "Eu" means True; karyon Pro - means before; Stage 2: Log Phase/ Maximum Exponential Phase - means nucleus Karyon – nucleus replication / cell division, Nucleus + - o antibiotic susceptibility – ideal phase which DNA enclosed in a nuclear target anti-bacterial / bactericidal agent (BEQ) membrane Membrane + - Stage 3: Stationary Phase bound Mitochondria – primary site o Period of balance growth (BEQ) organelle of electron transport chain o Number of replications / divisions = Number of (BEQ) deaths (Balance or Equal) Ribosome 80s (40s and 60s) 70s (30s and 50s) Cell wall Simple Complex: Stage 4: Death Phase / Decline Phase Plants: Cellulose - Peptidoglycan o Number of deaths > Number of replications polysaccharide of glucose (murein) o Reasons for microbial death - nutrients, Fungi: Chitin - polysaccharide of NAG toxins N- acetylglucosamine (NAG) and N- Requirements acetylmuramic acid (NAM) Physical Requirement: Cell Human: cholesterol - membrane Fungi: ergosterol Temperature: Replication Mitosis - for somatic cell Binary fission temperature = metabolism = growth Meiosis - for sex cells ⬇temperature = ⬇metabolism = ⬇ growth Domains Eukarya Archae - live in Extreme temperature = protein denaturation = ❌ extreme conditions growth (no Peptidoglycan) Temperature Danger Zone for Food = 4 - 60°C e.g., extreme halophile - salt loving Psychrophiles/ Cold temperature Cryophiles Listeria monocytogenes  MICROBIOLOGY, PARASITOLOGY & PUBLIC HEALTH Mesophiles Body temperature / Moderate o Pseudomonas Growth pattern: (BEQ) temperature o Mycobacterium bottom Medically important bacteria - o Nocardia ABCEP resident Bacteria in the body o Leptospira o Actinomyces o Bacillus o Bacteroides Thermophiles Hot temperature o Corynebacterium o Clostridium Sporeformers (Bacillus, o Eubacterium clostridium) o Prevotella NOTE: Fever: Microaerophiles Aerotolerant 37.2 - Morning Require O2, Tolerates O2 but 37.7 – Evening however O2 cannot replicate in its concentration → toxic presence 37. 8 – Clinical Practice / can kill them LP Growth pattern: Seen o Lactobacillus pH in the middle o Propionibacterium Bacteria - 6.5 to 7.5 pH Fungi - 5.5 to 6.5 pH o Spirochetes ▪ Leprospira Acidophiles ▪ Treponema Acidic pH ▪ Borrelia Lactobacillus acidophilus, Lactobacillus casein o Campylobacter (Yakult®) o Helicobacter Neutrophiles Neutral pH (7.2-7.6) Facultative anaerobes Medically important bacteria Could thrive with or without O2 Growth pattern: seen from top to bottom Alkaliphiles/ Basophiles THE REST (E. coli.) Alkaline/ basic pH Vibrio cholera Energy source Osmotic Pressure o Phototrophs – light Osmosis - movement of water from either low solute to o Chemotrophs – redox potential (medically high solute or high solvent to low solvent (BEQ) important) Hypertonic Environment - shrinkage of cell o Halophiles – survive in hypertonic environment MICROBIAL CONTROL ▪ salt content / osmotic pressure History ▪ Vibrio parahemolyticus (seafood/ oyster Ignaz Semmelweis - proponent of handwashing poisoning) Joseph Lister - First surgeon perform surgery with the Hypotonic Environment - swelling of cell use of phenol / carbolic acid to control microorganism Isotonic – Normal – 0.9% Definition of Terms Chemical Requirement Microbial cell death - irreversible loss of all the ability to Carbon – structural backbone of living matter reproduce Heterotrophs/ Autotrophs/ Lithotrophs Sterilization - complete removal or destruction of all Organotrophs types of microorganism including spores (BEQ) organic carbon (medically inorganic carbon monoxide Disinfection – Get rid of vegetative forms but not spores important) application of agent in inanimate object. (BEQ) able to create their own food o Chlorine – DOC for municipal water supplies Example: Plants ▪ Hypochlorous acid – lead to germicidal effect Nitrogen, Sulfur, Multiple Carbon Sources o Effectiveness can be affected by: (Pussy Cat Phosphorus “Primary Natural Nutrients” Dolls) BEQ Trace element - Co, Mn, Zn, (BEQ) CHO, CHON, ▪ Population size Vitamins CHONP, Fats ▪ Concentration Oxygen ▪ Duration of exposure Antisepsis - apply the agent to living tissue or animate o Example: Iodine (considered as #1 antiseptic in Oxygen skin) o Iodine + Organic compound = Iodoform (most Obligate aerobes Obligate anaerobes common: Povidone Iodine [Betadine]) Require O2 for Grow only in the Sanitization - reduce microbial load to a level acceptable survival absence of O2 for public health purposes Growth pattern: top Die in the presence of Degerming - mechanical removal or cleansing from a only (near oxygen) oxygen local area (extracting of blood) PMNL BC  MICROBIOLOGY, PARASITOLOGY & PUBLIC HEALTH Decontamination - destruction or marked reduction in Chemical Control the number or activity of microorganism Phenol (Carbolic acid) Physical Control Previously number 1 chemical control but it is very - Heat → Denaturation of CHON irritating Moist heat Autoclave - 121°C, 15 psi, 15-20 mins "Phenol coefficient" sterilization (BEQ) o Indicator: Salmonella typhi and Staphylococcus Indicator: Bacillus sterothermophillus Aureus (BEQ) MOA: Coagulation o phenol coefficient >1 - More effective o phenol coefficient Anionic = Amphoteric > Nonionic  MICROBIOLOGY, PARASITOLOGY & PUBLIC HEALTH Ethylene Oxide Simple microscope - Single lens MOA: Alkylation o Bright vs. Dark used in gas sterilization (BEQ) ▪ Bright field: the field / background has light Use for heat-sensitive materials and microorganism is dark Indicator: Bacillus Subtilis var globigii ▪ Dark field: Dark Background, lighter color of microorganism Peroxide e.g., Spirochetes o Hydrogen peroxide - good disinfectant but poor o Light vs. Electron Microscope antiseptic ▪ Light microscope – utilizes beam of light o Benzoyl peroxide - OTC drug for acne ▪ Electron microscope – utilizes beam of electron Heavy Metals Scanning (SEM) - 3D exhibit “oligodynamic properties” (even in small quantity Transmission (TEM) - 2D it can exhibit antimicrobial properties) Compound microscope - Multiple lens Silver (silver coin, spoons, 1% AgNO3) 1% AgNO3 - prophylaxis for Ophthalmia Neonatorum STAINING Gonorrhea (not use today - obsolete) allows better visualization o replaced by Erythromycin Ointment (new DOC for NOTE: Bacteria cell wall are predominantly negatively ONG) charge that's why we use basic dye Cupric Sulfate (Blue Vitriol) – Algaecide New-Born Vaccine - Hepatitis B Vaccine, Bacillus Gram Staining: General Rules Calmette-Guerin (BCG), Vitamin K, Erythromycin All cocci are Gram-positive, All bacilli are Gram- EXCEPT NVM negative, EXCEPT Halogens Neisseria BANCEL MP Chlorine (BEQ) Veillonella Bacillus o Disinfectant of choice for municipal water supplies Moraxella Actinomyces (PD 856) Nocardia o desired form: hypochlorous acid Streptomyces Iodine Corynebacterium, o antiseptic Clostridium o Preparation: Erysipelothrix ▪ Iodine Solution Listeria, Lactobacillus ▪ Strong Iodine Solution Mycobacterium ▪ Povidone Iodine (betadine - an iodoform - Propionibacterium Iodine and organic molecule) Simple stain SUMMARY OF MOA alcoholic or aqueous solution of a basic dye; stain the Membrane Protein Oxidation whole microorganisms disruption denaturation - Halogen Component that gives the color is called Chromophore - Surfactants - Alcohol - Peroxide (seen in either cation or anion) - Phenol - Aldehyde - Preservative o Anion (- charge) → Acidic Dye - Biguanides - Ethylene oxide o Cation (+ charge) → Basic Dye - Heavy metals Negative stain - use acidic dye o e. g. India ink, nigrosine Trends of Resistance o stain the background Prions - infectious particles that contain proteins o Common in persons that eat Cadaveric Material Differential stain o resistant to heat and chemicals Use to classify organism o Examples: Gram stain - Hanz Christian Gram ▪ Creutzfeldt- Jakob Disease Acid Fast Staining ▪ Kuru o Bind to cell wall with waxy material ▪ Borna o e.g., mycobacteria / Nocardia (only acid-fast ▪ Scrapie bacteria) ▪ Bovine Spongiform Encephalopathy (BSE) - o Methods: Madcow’s disease ▪ Ziehl-Neelsen → use of heat (Sizzling) Bacteria ▪ Kinyoun → Cold o Gram (+) – “Community acquired” Acid non-acid CHAMbA o Gram (-) – “Hospital acquired”; more resistant fast fast Virus Carbol- Component: Red Red o Naked - No coating; More resistance Fucshin Fuchsia dye: Primary o Envelope - With coating; Easier to kill (pink or stain red) Carbolic acid: METHODS OF STUDY MICROORGANISMS Chemical mordant MICROSCOPY Heat Physical Mordant Red Red 1. Simple vs. Compound Microscope  MICROBIOLOGY, PARASITOLOGY & PUBLIC HEALTH Acid Decolorizer Red Colorless Gold standard in the diagnosis of most bacterial Alcohol infection (but not the first choice) → since it can Identify Methylene Secondary stain / Red green / the organism as well as antibiotic blue / Counterstain blue Turbidimetric / Tube Dilution assay - measures the Malachite absorbance and transmittance Green Cylinder plate / Disc Diffusion / Kirby Bauer Method - zone of inhibition use petri dish GOOD TO KNOW COMPOSITION Chemically Defined Medium Exact composition is known For growth of photoautotrophs and chemoautotrophs Complex/ Not Chemically Defined Medium Exact composition is not known For growth of most medically important bacteria Added nutrients Special stain Tissue Culture - For growth of viruses to visualize specific structures Capsular stain - combine negative and simple stain PHYSICAL STATE o halo appearance Liquid – no solidifying agents (agar) o Examples: Even Some Super Serial Killers Have Semi-solid – 0.5 to 1.0% agar Pretty Nice Big and Bulging Capsules Solid – 1.5-3.0% agar ▪ E. Coli (meningeal strains) ▪ S. Pneumoniae ▪ Strep. Pyogens FUNCTION / USE ▪ Salmonella Typhi ▪ Klebsiella Pneumoniae Basal Media ▪ Haemophilis influenzae Simplest ▪ Pseudomonas Aerogenosa Designed for unfastidious organisms ▪ Neisseria Meningitis Nutrient agar/ broth ▪ Bacillus Anthracis ▪ Burdotella Pertusis Enriched Media ▪ Cryptococcus Neoformans – type of fungi Fortified with added vitamins, nutrients, and other uses India ink / Nigrosin to see it’s substances needed for the growth of fastidious capsule (BEQ) organisms Endospores stains Example For organisms that o Main composition is calcium dipicolinate/ Lowenstein- Mycobacterium tuberculosis dipicolinic acid Jensen medium Fold standard in recognizing o Important for survival: resistance to heat, (contains egg) tuberculosis: Culture chemicals, dehydration o e.g., Bacillus (aerobic) and Clostridium (anaerobic). Milk agar Require high protein diet o Method: Schaeffer-Fulton Chocolate Agar Require molybdenum from diet ▪ malachite green - stains spore Plate (CAP) (present in hemoglobin) ▪ Safranin - stains cell Hemophilus influenzae o Identification: Malachite Green Staining N. meningitidis ▪ Green: Spore ▪ Red: Vegetative cell Selective Medium suppress unwanted, favors the desire Culture Media Selective For *Saboraud Dextrose Agar Fungi *Colistin-Nalidixic Agar or Gram-positive organisms (CAN) Thayer-Martin Neisseria gonorrhea Loeffler’s Serum Medium, Corynebacterium diphtheria Tellurite Medium Campy BAP, Skirrow’s agar Campylobacter jejuni Bordet-Gengou Medium Bordetella pertussis Flagella – Uses Carbolfuchsin Thiosulfate citrate bile salts Vibrio spp. sucrose (TCBS) CULTURE MEDIA Nutrient material for microbial growth in laboratory Bismuth Sulfate Agar Salmonella Typhi  MICROBIOLOGY, PARASITOLOGY & PUBLIC HEALTH Charcoal Yeast agar Legionella Pneumophila ▪ Normal gut flora produces Vitamin K2 (Menaquinone) help in 1972 clotting factor Differential Medium Vitamin K1 (Phytonadione) – found in Subdivided a big group of microorganisms into vegetables (Petchay) categories to classify or differentiate microorganisms ▪ Case: hemorrhagic disease of the newborn Thioglycolate ▪ Sterile intestine = decrease normal flora = o For growth of obligate anaerobes Vitamin K = clotting factor (1972) o Thioglycolate broth: the ONLY reducing medium o Simple Media using Candle Jar/ Gas Pak® → Commensalism - one organism benefits and the other provides anaerobic environment one is unharmed / unaffected Lactose Fermentation o e. g. Staphylococcus Epidermidis in human skin o MacConkey agar (MCA) and Eosin Methylene Blue (EMB) Parasitism - one organisms benefits and the other one o for Enterobacteriaceae is harmed o e.g., helminths - inside the human intestine Culture Medium Lactose Non-Lactose o Balantidium coli – protozoa that can cause Fermenters Fermenters intestinal ulcer in human ▪ Reservoir: Pigs (BEQ) Examples E. coli - green Shigella metallic sheen Salmonella Competitive Exclusion Klebsiella Yersinia o e. g., Clostridium Difficile infection - due to chronic Eenterobacter Proteus antibiotic use called Pseudomonas Colitis citrobacter Pseudomonas o Overgrowth of C. Difficile may cause Colitis in the Serratia colon MCA Pink colonies Colorless o Pseudo membrane - nagkumpol kumpol at EMB Purple black Colorless namatay na mga microorganisms colonies o Predation – one organism benefit and the other one may die Blood Agar Plate ▪ Bdellovinrio bacteriovirus (PACOP) – kill or o check hemolytic pattern cause damage against G (-) Bacteria o Streptococcus Organisms Type of Hemolysis Zone of FEATURES OF INFECTIOUS DISEASE Hemolysis BEQ α-hemolytic Partial hemolysis Green Pathogenicity Virulence β-hemolytic Complete hemolysis Clear / no change ability to cause overt degree of pathogenicity or γ-hemolytic Non-hemolytic Red disease; kaya bang disease evoking power of magcause ng sakit microorganisms; Quantitative ability to cause Selective and Differential disease; gaano kagaling Mannitol and Salt Agar - S. Aureus magcause ng sakit 75% NaCl - selective Example: Opportunistic Determinants: Mannitol - differentiation Pathogen Toxigenicity - ability of o Fermented: Changes color from red to golden do not cause disease agent to produce yellow in healthy person but toxin become virulent with Invasiveness - ability MICROBE-HOST INTERACTION immunocompromised of microorganism to DEFINITION OF TERMS and unhealthy grow, reproduce and Pathology - study of disease e.g., Streptococcus spread into the body o Etiology -cause Species which targets o Pathogenesis - how the disease develops Meningitis in Pathogen - disease causing microorganisms HIV/AIDS patient Antigenicity - ability of agent to induce antibody production Resistance - ability of agent to survive adverse Reservoir - continuous source of microorganisms / environmental conditions infection Immunogenicity - Produce specific immunity Living Reservoir Non-Living Reservoir Infection - colonization of body by pathogens Humans - common Fomites - non-living Disease - when infection results in change in the state Animals - zoonotic like soil. object that of health infection takes part in direct o Example: St. and indirect SYMBIOSIS Louis transmission Relationship between two organisms; Host-parasite Encephalitis o tetani (causative relationship Virus (Wild agent: tetanus) - Mutualism - both organisms’ benefit birds) soil is an o Normal gut flora & human o B. Coli - Pigs important reservoir