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PROF. KÁROLY NAGY M.D.,PHD,PROF. KÁROLY NAGY M.D.,PHD,PROF. KÁROLY NAGY M.D.,PHD,PROF. KÁROLY NAGY M.D.,PHD,

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PROF. KÁROLY NAGY M.D.,PHD,PROF. KÁROLY NAGY M.D.,PHD,PROF. KÁROLY NAGY M.D.,PHD,PROF. KÁROLY NAGY M.D.,PHD,

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Institute of Medical MicrobiologySemmelweis UniversityBudapest

Jawetz ,Melnik

MEDICALMICROBIOLOGY24TH EDITION

Objectives of Medical Microbiology and its place am ong Natural Sciences

„microbiology is a science of microbes invisible to naked eye”(Pasteur)

Viruses Prokaryotes

Bacteria*cyanobacteriaarcheobacteria

Eukaryots

algae* plantsprotozoa* animals*

funghi*

MEDICAL MICROBIOLOGY

- virology- bacteriology- mycology- parasitology (protozoa, parasites,insects)

*pathogens

���� in the material circulation of Earth: - C, N, S, P, CO2, nitrification, denitrification- consist of 90% of living material of the Earth- E.coli (20-25 min. division ) during 3 days > Earth mass

���� in industrial and agricultural utilization :fermentation,wine, beer, bread, pharmaceutical industry, biotechnology

���� in cohabitation with the host:

☺☺☺☺ usefulnormal flora, N -bound, anaerob intestinal bacteria

���� harmfulplant, animal and human pathogens

Role and significance of microorganisms

Viruses 10-300nm 5 -100 µm

200 µm +Bacteria 0.3-30 µm

PROTOZOA, FUNGHI

Parasites

SIZE OF MICROORGANISMS

Aristoteles ( 384-322 b.c.)

„spontaneous generation”

paralyticus poliomyelitis1500 b.c. (18th Dynasty Egypt)

„contagium vivum” - „living material” theory

Girolamo Fracastoro – founder of epidemiology, XVI.c.

First description of syphilis

Anton von Leeuwenhoek (1632-1723)

HYSTORY OF MICROBIOLOGY I

Anton von Leeuwenhoek (1632-1723)

MICROSCOPE, COMPOSITE LENS-SYSTEM247 types microscope, max. magnif.: 270 xBerkel-lake: a complete new world

1673 First description and illustration of microbes

Royal Society, London, 300 letters

1675 human red blood cells1677 humán spermatozoa1683 teeth tar1698 Tzar Peter the Great visit

Edward Jenner 1749-1823 Cattleman nodule

1796

Vaccination

Edward Jenner 1749-1823

cowpox

smallpox

HAND WASH WITH BLEACH POWDER

Semmelweis, Ignác Fülöp 1818-1865

Saviour of the mothers

1847 Puerperal fever (sepsis)

RECOGNITION OF ETIOLOGYThe pathogen is not known

prevention

Many years before the introduction of microbial origin of infections, he introduced the hand-washing with chlorine to prevent the puerperal fever caused by Streptococcus pyogenes„antisepsis”

Semmelweis Ignác Fülöp 1818-1865

Semmelweis Museum

Birthplace of Semmelweis

Semmelweis’ ashes

Semmelweis Ignác Fülöp 1818-1865

Semmelweis University Budapest

Louis Pasteur 1822 - 1895

RABIES

Louis Pasteur 1822 - 1895

Identification of etiologyThe pathogen is not known

prevention

VACCINATION!

Louis Pasteur 1822 - 1895

•Fermentation•Pasteurization

Robert Koch 1843-1910

1876 KOCH’S POSTULATES :

DISEASE - PATHOGEN

SOLID CULTURE MEDIA

Robert Koch 1843-1910

ANTRAX

Robert Koch 1843-1910

„Koch bacillus”

Dmitrij Ivanovszkij 1864-1920

1892 DISCOVERY OF VIRUSES

Tobacco mosaic disease

Transfer infection through bacterialfilters

Paul Ehrlich 1854-1915

1908

Diphteria antitoxin (side-chain theory)

Founder of theoretical basis ofMODERN IMMUNOLOGY

Salvarsan , organic arsene ,first effective therapyOf SYPHILIS

Experiment „606” (systematic work)

THEORETICAL BASIS OF CHEMOTHERAPY

Alexander Fleming 1870-1955

Penicillinum notatum

ANTIBIOTICS era

Discovery of Penicillin 1928

1945

• lysosim • development of penicillin and description

of its antibacterial effect (1929)

FLOREY, Sir Howard Walter(1898-1968)

CHAIN, Ernst Boris (1906-1979)Extraction, purification and crystallization of pen icillin (1940)

FLEMING, Sir Alexander (1881-1955)

1944. Nobel Prize in Physiology/Medicine

DOMAGK, Gerhard (1895-1964)

• „Red dye” (I.G. Farben)

• Prontosil, the first sulfonamide

1939. Nobel Prize in Physiology/Medicine(only in 1947 because of the 2nd World War)

Leeuwenhoek 1674 METHOD

Jenner 1796 EXPERIENCEPREVENTION

Semmelweis 1847

Pasteur 1885

Koch 1876 PATHOGENS

Ivanovszkij 1892

Ehrlich 1910 THERAPEUTICALAGENTS

Fleming 1928

HISTORY OF MICROBIOLOGY II

Recently:new antibioticsgenetical methods, etc.

New diseases USA HungaryLegionellosis 1976 1980Lyme disease 1979 1981

AIDS 1981 1986

New pathogensprion, SARS

New problemsnozokomial infections

immunsuppressed patients

ANTIBIOTICS

LEGIONELLOsISLegionella pneumophilia

GASTRIC ULCERHelicobacter pylori

ARTERIOSCLEROSISChlamydia pneumoniae

NEW PATHOGENS - NEW DISEASES

B a c t e r i a

LYME DISEASEBorelia burgdorferi

NEW PATHOGENS - NEW DISEASES

V i r u s e s

AIDSHIV (retrovírus)

Hemorrhagic feverEbola vIrus(filovírus)

Kaposi’s sarcomaHHV-8 (herpesvírus)

Hemorrhagic fever,LUJO virus (Lusaka-Jo’burg)arenavirus

NEW PATHIGENS - NEW DISEASES

V i r u s e s

SARSCoronavirus

Avian influenzaInfluenza virus

NEW PATHOGENS - NEW DISEASES

P R I O N ( p r o t e i n o n l y )

Scrapie in sheepBovine Spongiform Encephalitis (cattle)Creutzfeld – Jacob disease in human

Infective proteins, no nucleic acid, reproduction in living cell

Morbidity by some acute infections in Hungary

1932 1952 1982 1992 2004VA Typhus abdominalis 22.200 1.611 10 1 1

hastifusz 2.700 62 1 0 0

VA Diphteria 19.475 2.073 0 0 0torokgyík 1.522 58 0 0 0

A Scarlatina 16.399 19.882 13.149 4.196 3.355vörheny 249 29 0 0 0

A Dysenteria 5.404 8.771 7.243 5.224 239vérhas 944 265 5 4 1

V Poliomyelitis 391 500 0 0 0gyermekbénulás 71 29 0 0 0

V Morbilli 29.592 23.747 2.521 14 6kanyaró 320 180 0 0 0

A? Iatrogenic infections 5.826 4.860 >1.000yatros = doctor 83 25 12

V vaccine A antibiotics death

CLINICAL MICROBIOLOGY

⇒⇒⇒⇒ microbiological diagnosis

⇒⇒⇒⇒ epidemiology

⇒⇒⇒⇒ pathogenesis

⇒⇒⇒⇒ therapy and prevention/control

⇒⇒⇒⇒ scientific research

IMPORTANCE OF CLINICAL MICROBIOLOGY⇒⇒⇒⇒ frequency of infectious diseases

⇒⇒⇒⇒ high mortality of infectious diseases⇒⇒⇒⇒ high number of nosocomial infections ⇒⇒⇒⇒ polyresistant pathogenes

CRITERIA DETERMING BACTERIA

���� morphology ���� staining properties ���� endospore formation���� antigenic structure���� biochemical activity���� motility

MORPHOLOGY OF BACTERIA

STRUCTURE OF BACTERIAL CELL

(no Golgi)

*

* - Nucleoid: circular single-stranded DNA

Antigen specificityAntifagociterendospore

Sexfimbrias Simple fimbrias(adhesive)

Essential components

STRUCTURE OF THE BACTERIAL CELL WALL

BLUE RED

LPS

Crystal violet – basic anylin dyeDiluted iodine sol.Alcohol/aceton differentiationFucsin or safranin after-staining

GRAM POSITIVE GRAM NEGATIVE

Pos/Peptidoglycan/BLUE Negative - RED

GRAM STAINING

- nuclear material (nucleoid),

- plasmids,- mesosome, - polyamides,- inclusion bodies, - 80% water, - protein/enzymes, carbohydrates,

lipids, anorganic ions

CYTOPLASMA

NUCLEOID (no membrane)• single stranded, circular DNA, mw: 3x 10 9

• single haploid chromosome• 1-2-4 copy (1mm, bacterium: 1-2 µµµµm) • gyrase enzyme: superhelix

GENETIC MATERIAL

• plasmides

ENDOSPOREUnfavorable nutritional and environmental condition s

FUNCTION: resistance, diagnosis

sporulation endospore (roundish or oval, central, terminal,or free ) Memb-

rane

extracellular polymer (polysaccharide) layer

- macrocapsule : thick, negative staining

Function: nutrition, antigenic property, antiphagoc ytic effect, diagnosis

THE CAPSULE

Klebsiella pneumoniae

- microcapsule : electron microscopy,serology (Quellung reaction)

- „slime layer” S (smooth) - R (rough) variation

⇒⇒⇒⇒ 12-30 nm diameter, basal body, cylindric structure

FUNCTION: motility, „H” antigen

FLAGELLA

Proteus vulgaris

⇒⇒⇒⇒ Gram negative bacteria (plasmid or chromosome coded)

⇒⇒⇒⇒ short, thin (diameter: 2-10 nm)

⇒⇒⇒⇒ sex fimbriae: conjugation (donor / recipiens )

FUNCTION: colonization, virulence factor, phage rece ptor

FIMBRIAE (Pili)

⇒⇒⇒⇒ adhesion

PHYSIOLOGY OF BACTERIABINARY FISSION

1. Chromosome replication and separation 2. Cell elongation and septum formation3. Binary fisson(assymmetric division: minicells)

FACTORS AFFECTING MULTIPLICATION temperature (10 C o - 20-40 Co - 50-60 Co)pH, osmotic pressure, NaCl content (6%)

CULTURE MEDIA

S.aureus

E.coli

OBLIGATE AEROBIC (respiration): requires oxygenMICROAEROPHILIC FACULTATIVE ANAEROBIC (respiration-fermentation)OBLIGATE ANAEROBIC (fermentation): no oxygen neededAEROTOLERANT ANAEROBIC (fermentation)

NUTRITION Carbon source (autotroph, chemolithotroph, heterotroph)Nitrogen source (nitrate, nitrite reduction:assimilation, denitrifi cation:

dissimilation, nitrogen fixation) Sulfur source (sulfate reduction to hydrogen sulfide) Phosphorous source, mineral sources (Mg 2+ , FE 2+ , K + , Ca2 + etc. Growth factors (vitamins, etc.)

NUTRITION Carbon source (autotroph, chemolithotroph, heterotroph)Nitrogen source (nitrate, nitrite reduction:assimilation, denitrifi cation:

dissimilation, nitrogen fixation) Sulfur source (sulfate reduction to hydrogen sulfide) Phosphorous source, mineral sources (Mg 2+ , FE 2+ , K + , Ca2 + etc. Growth factors (vitamins, etc.)

FERMENTATION (anaerobic degradation)

RESPIRATION (aerobic degradation)

SOURCES OF METABOLIC ENERGY

Million years old microbes found under earth

Ancient bacteriain a metorite from Mars

CHANGEBILITY

ADAPTATION

Morphology of bacteriacoccae

• coccae

• in pairs

• in chain

• in groups

• in tetra forms

• coccobacillae

rods

• spindle rods

• rods with rounded edges

• rods with cut edge

• fusiform rods

• vibrios

• spirillums

spiochaets

• Borellia

• Treponema

• Leptospira

www.uccs.edu

STRUCTURE OF THE BACTERIAL CELL

Procariots: no nucleolus , no nuclear membrane

Essential components Accessory components

CapsuleAntigen specificityAntifagociterendospore

FimbriasSexfimbrias ( pilus)Simple fimbrias(adhesive)

Flagellum

Cilia

Cell wall

Membrane

Cytoplasmano Golgi

Nuclear materialcircularDuble stranded DNA(haploid)

STRUCTURE OF BACTERIAL CELL WALL

CILIACILIA

fimbria

CAPSULELPS

Gram + Gram -

(Sensitivity for antibiotics and dyes)

CAPSULE

Flagella

„Microbiologists are also „Microbiologists are also „Microbiologists are also „Microbiologists are also

with monopolarwith monopolarwith monopolarwith monopolar---- or or or or

diffuse flagellae diffuse flagellae diffuse flagellae diffuse flagellae

equipped…”equipped…”equipped…”equipped…”

www.uccs.edu

Légy

Szúnyog

Poloska

• Trypanosoma gambiense (Tsetse)

• Trypanosoma rhodesiense (Tsetse)

•Plasmodium (Anopheles)

• Leishmania (Phlebotomus, and others)

• Trypanosoma cruzi

• Bunyaviruses (sandfly Phlebotomus)

• Togaviruses

• Flaviviruses (Aedes, Culex)

• Bunyaviruses (Aedes)

• Loa loa (filariae)

• Wuchereria (filariae)

• Oncocerca (filariae)

Entomológia

Kullancs

Tetü

Bolha

Atka

• Babesia

• Flaviviruses

• Reo/Coltiviruses

• Borellia burgdorferi

• Francisella tularensis

• Ricketssia rickettsii

• Ricketssia prowa zekii

• Borellia recurrensis

• Yersinia pestis

• Rickettsia typhi

• Rickettsia akari

• Rickettsia tsutsugamushi

• (allergia, rühesség)