14 - Struktur, Morfologi, Klass & Molekuler Infeksi Bakteri

Mochammad Hatta (2006) Mochammad Hatta (2006)

MORFOLOGI, STRUKTUR DAN KLASIFIKASI BAKTERI

Prof. Dr. MOCHAMMAD HATTA, SpMK, Ph.D.

BAGIAN MIKROBIOLOGI, FAKULTAS KEDOKTERAN, UNIVERSITAS HASANUDDIN

MAKASSAR


Structure Bacteria

FLAGELLA

FIMBRIAE

CAPSULE

CELL WALL

PLASMA MEMBRANE

CYTOPLASMA

NUCLEAR BODY

CONTAINING DNA


NUCLEAR BODIES • STAINING AND UNSTAINING - SAME BY USUAL METHODS

• REACT POSITIVE TO “FEULGEN TEST” FOR DNA• GENETIC FUNCTIONS -- GENE EXPRESSIONS

CYTOPLASMA• NUMEROUS SMALL GRANULES CALLED “RIBOSOMES’• DIAMETER OF RIBOSOMES IS 10 - 30 uM• SOME BACTERIA CONTAINING INCLUSION GRANULES• CELL METABOLISM FUNCTIONS ---- NUTRIENT RESERVE ----

UNDER CONDITIONS OF ENERGY-SOURCE STARVATION

• VOLUTIN GRANULES (SYN. METACHROMATIC OR BABES

ERNEST GRANULES) -- CORYNEBACTERIUM DIPHTERIAE• SPECIAL METHODS FOR STAINING -- “NEISSER” STAINING


CYTOPLASMIC MEMBRANE• CONSIST MAINLY LIPO-PROTEIN --- 5- 10 uM THICK

• EXAMINED BY ELECTRON MICROSCOPE -ULTRATHIN

SECTION• CONTAIN RESPIRATORY ENZYMES & PIGMENTS

(CYTOCHROME SYSTEM)

• ENZYME OF TRICARBOCYLIC CYCLE & POLYMERASE ENZYME

CELL WALL• 10 - 25 uM THICK --- 1 um DIAMETER --- < 10.000 MOLECULAR

WEIGHT• LYSIS BACTERIA --- LETHAL DISINTEGRATION & DISSOLUTION• PLAYS AN IMPORTANT PART IN BACTERIAL DIVISION• BASAL COMPONENTS ; MUCOPEPTIDE, TEICHOIC ACID, LPS

(ENDOTOXIN)


CAPSULES• SOLID MATERIAL : POLYSACCHARIDE, POLYPEPTIDE, PROTEIN

• DEMONTRATION BY “NEGATIVE STAINING” --- INDIA INK• ENTEROBACTERIACEAE : CAPSULAR ANTIGEN (K ANTIGEN)• FUNCTION : PROTECTIVE AGAINST BACTERIOPHAGES, COLICINES,

LYSOZYME, LYTIC ENZYMES

FLAGELLA

• ACT AS ORGAN LOCOMOTION (MOTILITY)

• 0.02 uM THICK --- 1- 20 FLAGELLA/ BACTERIA -- PERITRICHOUS,

LATERAL, POLAR• FLAGELLINE ANTIGEN (SALMONELLA TYPHI) -- Vi ANTIGEN

FIMBRIAE• CERTAIN GRAM NEGATIIVE BACILLI --- ENTEROBCTERIACEAE

• FUNCTION AS ORGANS OF ADHESION


LOG OF COUNT

TIME

TOTAL BACTERIA

VIABLE BACTERIA

GROWTH OF BACTERIA

LAG PHASE

LOG PHASE

STATIONER PHASE

DECLINE PHASE


BIOLOGICAL CLASSIFICATION

• CLASS SCHIZOMYCETES :

1. ORDER PSEUDOMONADALES : FAMILY PSEUDOMONADACEAE

--- GENERA PSEUDOMONAS -- SPECIES P.AERUGINOSA

2. ORDER EUBACTERIALES : FAMILY ENTEROBACTERIACEAE

--- GENERA SALMONELLA -- SPECIES S. TYPHOSA

3. ORDER ACTINOMYCETALES : FAMILY MYCOBACTERIACEAE

--- GENERA MYCOBACTERIUM -- SPECIES M. TUBERCULOSIS

4. ORDER SPIROCHAETALES : FAMILY SPIROCHAETES

--- GENERA LEPTOSPIRA --- SPECIES L. ICTEROHAEMORRHAGIAE


5. ORDER MYCOPLASMATALES : FAMILY MYCOPLASMATALES

--- GENERA MYCOPLASMA --- SPECIES M. MYCOIDES

• CLASS MICROTATOBIOTES

1. ORDER RICKETTSIALES : FAMILY RICKETTSIALES --- GENERA RICKETTSIA -- SPECIES R. PROWAZEKII 2. ORDER VIRALES


BACTERIAL NUTRITIONBACTERIAL NUTRITION

THE CARBON SOURCE FOR GROWTHTHE CARBON SOURCE FOR GROWTH

-- CHEMO-SYNTHETIC AUTOTROPS OR PHOTOSYNTHETIC -- CHEMO-SYNTHETIC AUTOTROPS OR PHOTOSYNTHETIC

AUTOTROPSAUTOTROPS THE NITROGEN SOURCE OF GROWTHTHE NITROGEN SOURCE OF GROWTH

-- AMINO ACIDS AND NUCLEOTIDES -- AMINO ACIDS AND NUCLEOTIDES

THE REQUIREMENTS FOR GROWTH FACTORSTHE REQUIREMENTS FOR GROWTH FACTORS

-- -- THIAMINE, RIBOFLAVIN, NICOTINIC ACID, PYRIDOXINE,THIAMINE, RIBOFLAVIN, NICOTINIC ACID, PYRIDOXINE,

FOLIC ACID, BIOTIN, ETCFOLIC ACID, BIOTIN, ETC

INORGANIC SALTS FOR GROWTHINORGANIC SALTS FOR GROWTH -- POTASSIUM, MAGNESIUM, SODIUM, IRON AND CALCIUM -- POTASSIUM, MAGNESIUM, SODIUM, IRON AND CALCIUM


NUTRITIONAL EVOLUTIONNUTRITIONAL EVOLUTION

• ESCHERICHIA COLI CAN GROW ON A ‘ SIMPLE SYNTHETIC

MEDIUM’ WHICH CONTAINS ONLY GLUCOSE ( CARBON AND

ENERGY SOURCE ), AMMONIUM SULPHATE ( NITROGEN AND

SULPHUR SOURCE ) PHOSPHATE BUFFER, POTASSIUM CHLORIDE,

MAGNESIUM CHLORIDE AND TRACES OF OTHER INORGANIC

SALTS.

THE PHYSICAL CONDITIONS REQUIRED FOR GROWTH

• THE INFLUENCE OF OXYGEN AND REDOX POTENTIAL

THE MAJORITY OF BACTERIA ARE DESCRIBED AS

FACULTATIVE ANAEROBS.


• THE INFLUENCE OF CARBON DIOXIDETHE INFLUENCE OF CARBON DIOXIDE

SOME BACTERIA, HOWEVER, REQUIRED A MUCH HIGGER

CONCENTRATION OF CARBON DIOXIDE ( 5 - 10 % ).

• THE INFLUENCE OF TEMPERATURE 1. ON GROWTH --- THESE, AND MANY SAPROPHYTES OF SOIL

AND WATER , WHICH GROW BEST AT BETWEEN 25 o AND 40 o

CELCIUS.

2. ON VIABILITY --- THE THERMAL DEATH POINT OF A

PARTICULAR ORGANISM MAY BE DEFINED AS THE

LOWEST TEMPERATURE THAT KILL IT UNDER

STANDARD CONDITIONS, WITHIN A GIVEN TIME,

e.g. TEN MINUTES.

( e.g. ABOUT 105 o C . FOR CL. TETANI AND 115 o C. FOR CL.

BOTULINUM )


THE INFLUENCE OF MOISTURE AND OF DESICCATIONTHE INFLUENCE OF MOISTURE AND OF DESICCATION

FOUR-FIFTHS BY WEIGHT OF THE BACTERIAL CELL CONSISTSFOUR-FIFTHS BY WEIGHT OF THE BACTERIAL CELL CONSISTS

OF WATER AND AS, IN THE CASE OF OTHER ORGANISM, OF WATER AND AS, IN THE CASE OF OTHER ORGANISM,

MOSITURE IS ABSOLUTELY NECESSARY FOR GROWTH MOSITURE IS ABSOLUTELY NECESSARY FOR GROWTH

THE INFLUENCE OF HIDROGEN -ION CONCENTRATIONTHE INFLUENCE OF HIDROGEN -ION CONCENTRATION

THE MAJORITY OF COMMENSAL AND PATHOGENIC BACTERIA THE MAJORITY OF COMMENSAL AND PATHOGENIC BACTERIA

GROW BEST AT A NEUTRAL OR VERY SLIGHTLY ALKALINE GROW BEST AT A NEUTRAL OR VERY SLIGHTLY ALKALINE

REACTION ( PH 7.2 TO 7.6 ) REACTION ( PH 7.2 TO 7.6 )

THE INFLUENCE OF LIGHT AND OTHER RADIATIONSTHE INFLUENCE OF LIGHT AND OTHER RADIATIONS ULTRA VIOLET RAYS RAPIDLY BACTERICIDAL, e.g. DIRECT ULTRA VIOLET RAYS RAPIDLY BACTERICIDAL, e.g. DIRECT

SUNLIGHT OR RADIATION FROM A MERCURY VAPOUR LAMP SUNLIGHT OR RADIATION FROM A MERCURY VAPOUR LAMP

THE INFLUENCE OF OSMOTIC PRESSURETHE INFLUENCE OF OSMOTIC PRESSURE

FOR MOST SPECIES THE UPPER LIMIT OF SODIUM CHLORIDEFOR MOST SPECIES THE UPPER LIMIT OF SODIUM CHLORIDE

CONCENTRATION PERMITTING GROWTH LIES BETWEEN 5 - 15 %, CONCENTRATION PERMITTING GROWTH LIES BETWEEN 5 - 15 %,

HALOPHILIC (OSMOPHILIC HALOPHILIC (OSMOPHILIC))


METABOLIC FUNCTIONS OF THE VITAMIN B GROUP

B VITAMIN ORGANISM REQUIRING

• THIAMINE STAPH. AUREUS

• NICOTINIC ACID L. ARABINOSUS

• RIBOFLAVIN L. CASEI

• PYRIDOXAL CL. WELCHII

• PANTHOTHENIC ACID BRUCELLA ABORTUS

• P AMINOBENZOIC ACID CL. ACETOBULYTICUM

• FOLIC ACID CL. TETANI

• BIOTIN CL. TETANI

• VITAMIN B 12 L. LACTIS


1. Fermentasi asam homolaktat (beberapa streptococcus dan lactobacillus

NADH NAD

asam piruvat asam laktat

2. Fermentasi beralkohol (khamir)

NADH NAD

Asam piruvat asetildehide etil alkohol

CO2

3. Fermentasi asam campuran (E.coli dan beberapa bakteri saluran pencernaan yang lain

NADH NAD

Asam piruvat Asam laktat

CO2

Asetil oksalat Asetil-CoA + Asam format

NADH NADH

NAD NAD

Asam suksinat Etil alkohol Asam asetat H2 + CO2


4. Fermentasi glikol-butelin (Enterobacter, Bacillus, Pseudomonas) Fermentasi ini menghasilkan sejumlah kecil produk akhir yang sama dengan fermentasi campuran asam, tetapi sebagai tambahan, sebagian besar asam piruvat diubah menjadi 2,3-glikol butilen seperti terlihat di bawah.

2CH2COCOOH CH2COHCOOH + CO2

Asam piruvat C O

CH3

Asam asetolaktat

NAD NADH CO2

CH3CHOHCHOHCH3 CH3CHOHCOCH3

2,3-Glikol butilen Asetoin


5. Fermentasi asam propionat (Propionibacterium dan Veillonella

3-Asam piruvat Asam asetat +CO2

2-Asam oksaloasetat

2CO2

(terikat enzim) 2- Asam suksinat

2-Propionil-CoA CoA

2-Asam propionat 2-Suksinil-CoA

2- Metil malonil-CoA


6. Asam butirat, butanol, fermentasi aseton (Clostridium) 4CO2

4-Asam piruvat 4-Asetil-CoA Asam asetat

NADH

CO2 NAD

Aseton 2-Asetoasetil-CoA Etil alkohol

NADH

NAD

Isopropil alkohol 2-Krotonil-CoA

NADH

NAD

2-Butiril-CoA

NADH

NAD

Asam butirat Butanol


Klasifikasi berdasarkan energi dan sumber karbonKlasifikasi berdasarkan energi dan sumber karbon

KELOMPOKKELOMPOK SUMBER ENERGISUMBER ENERGI SUMBER SUMBER KARBONKARBON

KemoheterotrofKemoheterotrof Oksidasi senyawa Oksidasi senyawa organikorganik OrganikOrganik

KemoautotrofKemoautotrof

Oksidasi senyawa Oksidasi senyawa anorganik :anorganik :

seperti Amoniak, seperti Amoniak,

sulfida dan seyawasulfida dan seyawa

besibesi

COCO22

FotoheterotrofFotoheterotrof CahayaCahaya OrganikOrganik

FotoautotrofFotoautotrof CahayaCahaya COCO22


Transport elektron dan fosforilasi oksidatif berdasarkan model kemiosmosis Mitchell.

Membran bakteri NADH + H+

NADH dehidrogenase

H

H + H+

CoQ e

H + CoQ-H CoQ-H H+

H + CoQ-H2 H+

FeS

sit c O2 + H+

e

sit a a3

H2O ADP ATPase 2H+ + Pi ATP

Flavoprotein FeS

Saluran protein

Sit.b


Fotosintesis tumbuhan hijau

NADP

Freedoksin NADPH

Sitokrom b

ADP

ATP

Sitokrom f e- Sitokrom e- Quinom

ADP H2O 2e- + H+ +1/2O2

ATP

Cahaya Klorofil Cahaya Klorofil

Fotosistem I Fotosistem II

e-

e


Pembentukan ATP dan NADH ole Nitrosomonas yang autotrof

2NH4+ + 3O2 2NO2 - + 2H2O + 4H+ + 4e-

ATP ADP ATP ADP

2e + Cyt a1 Cit C Cit b Flavoprotein

ATP

Cyt a3

NADP + H+ ADP

ADP

ATP

H2O NADPH

Menghasilkan Memerlukan energi

energi


Jalur Embden-Meyerhof untuk disimilasi glukosa Glukosa (enam karbon) ATP ADP

Glukosa 6-fosfat

Fruktosa 6-fosfat ATP ADP

Fruktosa 1,6-difosfat Dihidroksiaseton Gliserildehide 3-fosfat (tiga karbon) fosfat 2NAD penerima H terkahir 2NADH (2) 1,3-Asam difosfogliserin

2 ADP 2 ATP

(2) 3-Asam fosfogliserin

(2) 2-Asam fosfogliserin H2O

(2) Asam fosfoenolpiruvat 2 ADP 2 ATP

(2) Asam piruvat

Produk akhir


Ikatan Peptida mengkaitkan asam amino menjadi satu untuk membentuk protein besar

CH3CHCOOH + CH2 CH COOH

NH2 OH NH2

Alanin serin

CH3CHCOOH + H2O

NH ikatan peptida

C O

CH NH2

CH2OH Dipeptida

alanin dan serin


CH2 OOC4H7

CH OOC4H7 + 3H2O

CH2 OOC4H7

CH2OH

CHOH + 3CH3(CH2) 2COOH

CH2OH asam butirat

Gliserol yang dibebeskan kemudian dapat dimetabolisme melalui jalur Embden- Meyerhof, dan asam lemaknya dapat diuraiakan memlaui asetat pada daur asam citrat.

lipase

tributrin


Thiobacillus (oksidasi belerang)

2S + 3O2 + 2H2O 2H2SO4

Nitrosomonas (oksidasi amoniak)

2NH4Cl + 3O2 2HNO2 + 2HCl + 2H2O

Nitrobacter (oksidasi nitrit)

2NaNO3 + O2 2NaNO3

Berbagai marga (oksidasi hidrogen)

2H2 + O2 2H2O

Siderocapsa (oksidasi senyawa-senyawa besi)

4FeCO3 + O2 + 6H2O 4Fe(OH) 3 + 4CO2


Daur asam sitratDaur asam sitrat

CH3 CoA CH3

Metabolisme karbohisrat C O C O + CO2

CH3 S CoA Asam piruvat Asetil CoA

COOH COOH

COOH CH2 CH2

C O HO C COOH C COOH

CH2 CH2 CH

COOH COOH COOH

Asam oksaloasetat Asam sitrat Asam cis-akonitat

NADH + H2O NAD COOH COOH

HCOH CH2

CH2 HC COOH COOH HOCH

COOHAsam malat Asam isositrat


Asam Malat Asam isositrat

NADCO2

NADH ADP ATP

COOHCOOH COOH COOH

FADH2 FAD GTP GDP CoA CH2

CH CH2 CH2

CH2

CH CH2 CH2 CO2

CoA NADH NAD C 0 COOH COOH CO SCoA COOH

Asam fumarat Asam suksinat Suksinil-CoA Asam -keto- glutamat


MICROBIAL VIRULENCE

• LOSS OF VIRULENCE : WHEN KEPT IN ARTIFICIAL MEDIUM

--- MEASUREMENT OF VIRULENCE CONSISTS OF TESTING FOR

ABILITY TO INFECT, WITH OBSERVABLE CONSEQUENCES,

“NORMAL” ANIMALS SUCH AS A STANDARD STRAIN MICE

• THE ED50 : THE POINT ON DOSE SCALE AT WHICH THIS LINE

INTERSECT THE 50 PERCENT POINT IN THE CUMULATIVE EFFECT

IN HALF THE ANIMALS INOCULATED WITH BACTERIA.

• TEST VIRULENCE : TO KNOW TOXIN EFFECT FROM INFECTION

OF CORYBACTERIUM DIPTHERIAE


TesTes virulensivirulensi Untuk mengetahui strain toxigenikUntuk mengetahui strain toxigenik ( (menghasilkan menghasilkan

toksin) dan non-toxigenik (tidak menghasilkan toksin) dan non-toxigenik (tidak menghasilkan toksin).toksin).

Tes Tes ELEKELEK (invitro) : (invitro) :

- Kertas saring direndam antitoksin dalam- Kertas saring direndam antitoksin dalam

agar --- Diatas ditanam basil difteri – agar --- Diatas ditanam basil difteri –

inkubasi 24 jam.inkubasi 24 jam.

- Hasil : (+) hasilkan toksin : tampak garis - Hasil : (+) hasilkan toksin : tampak garis presipitasi. presipitasi.

(-) tidak hasilkan toksin : tidak ada (-) tidak hasilkan toksin : tidak ada garis presipitasi garis presipitasi..


•TesTes in-vivoin-vivo

250 ug antitoksin (s.c.)

2 jam

Marmot

Basil difteri pasien (s.c.)

4 hari

4 hari

Tetap hidup

Mati

Kesimpulan :

Basil difteri penderita mengandung toksin


• TissueTissue cultureculture ((BiakanBiakan jaringanjaringan))

Jaringan dalam medium

RPMI 1640 + HBSS + PBS

( Complete medium)

Biakan

18 jamLihat dibawah mikroskop

* jaringan mati :

basil menghasilkan

toksin

* jaringan hidup :

Basil tidak menghasil

kan toksinMasukkan basil difteri pasien


Gram positive (blue) and Gram negative (red) cocciGram positive (blue) and Gram negative (red) cocci


Staphylococcus Staphylococcus epidermidis epidermidis (novobiocin (novobiocin susceptible, susceptible, upper)upper)

Staphylococcus Staphylococcus saprophyticus saprophyticus (novobiocin (novobiocin resistant, lower)resistant, lower)


Staphylococcus Staphylococcus aureus aureus culture on culture on

blood agarblood agar


Streptococcus Streptococcus pyogenespyogenes (group A) (group A) shows shows ββ-hemolysis -hemolysis on blood agar (upper on blood agar (upper left)left)

Streptococcus Streptococcus pneumoniaepneumoniae shows shows αα-hemolysis (right)-hemolysis (right)

Enterococcus not Enterococcus not produce hemolysin produce hemolysin (lower left(lower left))


Enteric bacteria Enteric bacteria on MacConkey on MacConkey agaragar

Lactose-positive Lactose-positive bacteria show bacteria show pink colonies pink colonies (upper left)(upper left)

Lactose-negative Lactose-negative bacteria have bacteria have colorless colorless colonies (lower colonies (lower right)right)


Biochemical reactions for identification of a fresh enteric Biochemical reactions for identification of a fresh enteric bacterial isolate by the API 20E procedurebacterial isolate by the API 20E procedure


Black Black colonies of colonies of Salmonella Salmonella after growth after growth on bismuth on bismuth sulfite agarsulfite agar


Clostridium Clostridium perfringensperfringens

Shows double Shows double zones of zones of hemolysis hemolysis surrounding surrounding colonies after colonies after incubation incubation anaerobically on anaerobically on blood agarblood agar


Respiratory syncytial infection of cells in nasopharyngeal aspirate is Respiratory syncytial infection of cells in nasopharyngeal aspirate is revealed by immunoflourescencerevealed by immunoflourescence


Powassan virus antigen surrounding infected cells of mouse brain is Powassan virus antigen surrounding infected cells of mouse brain is revealed by immunoperoxidase stainingrevealed by immunoperoxidase staining


Contagious Pustular Contagious Pustular Dermatitis (ORF) Dermatitis (ORF)

a. ORF lesion ona. ORF lesion on forefinger forefinger

b. Electron micrographb. Electron micrograph of ORF virion of ORF virion


Microsporum Microsporum caniscanis colony on colony on Sabouraud’s Sabouraud’s agaragar


Microsporum canis Microsporum canis macroconidia (spore)macroconidia (spore)


Dermatophyte hyphae in KOH preparation of skin Dermatophyte hyphae in KOH preparation of skin scrapingscraping


Microsporium gypseumMicrosporium gypseum macroconidia (spore) macroconidia (spore)


Histoplasma Histoplasma capsulatum capsulatum macroconidiamacroconidia


Trichopyton rubrumTrichopyton rubrum colony on Sabouraud’s agar colony on Sabouraud’s agar


Coccidioides immitisCoccidioides immitis. . Arthrospores after cultureArthrospores after culture


Coccidioidomycosis. Lung section shows sporangiosporeCoccidioidomycosis. Lung section shows sporangiospore


Aspergillus fumigatus Aspergillus fumigatus colony on Sabouraud’s agarcolony on Sabouraud’s agar


Cryptococcus neoformansCryptococcus neoformans. Budding yeast surrounded by the large . Budding yeast surrounded by the large mucoid capsule is demonstrated by negative staining with nigrosinmucoid capsule is demonstrated by negative staining with nigrosin


Aspergillus fumigatusAspergillus fumigatus conidiosphores conidiosphores


Ziehl Neelsen staining from sputa before Ziehl Neelsen staining from sputa before decontaminationdecontamination


Ziehl Neelsen staining from sputa after Ziehl Neelsen staining from sputa after decontaminationdecontamination

Documents

14 - Struktur, Morfologi, Klass & Molekuler Infeksi Bakteri