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