normal bacterium flora of the gastrointestinal tract
TRANSCRIPT
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Enterobacteriaceae
Dr. Berek ZsuzsaDr. Berek Zsuzsa08 November 201108 November 2011
Normal bacterium Flora of the Gastrointestinal tract
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GRAM NEGATIVE RODS
AEROBBordetellaBrucellaFrancisella
PseudomonasAcinetobacterLegionella
FACULTATIVE ANAEROBHaemophilusPasteurella
Familia:EnterobacteriaceaeVibrionaceae
CardiobacteriumEikenellaKingellaActinobacillus
ANAEROBBacteroidesPrevotellaPorphyromonasFusobacterium
MICROAEROPHILCampylobacterHelicobacter
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Normal bacteriumFlora of the
Gastrointestinal tract
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Normal bacterium flora of theGastrointestinal tract
• Significance:experiments with germ-free animals
•• StomachStomach (103–106 cfu/g)– Low pH– Resident micro flora Ø– Transient flora +– Hypochlorhydria, stasis ⇒
Lactobacillus (Boas-Oppler) & Sarcinaspp.
•• Small intestineSmall intestine (105–107 cfu/g)– Digestive enzymes & bile, rapid peristaltic– Resident micro flora Ø– Transient flora +
•• Large intestineLarge intestine (109–1011 cfu/g)– Abundant resident micro flora
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Anaerobes90–95% of the species∼ 1011 cfu/g faecesResident
Bifidobacterium bifidumBacteroides fragilisEubacterium spp.Clostridium spp.
TransientAnaerobe KokkenFusobakterienLactobacillen
Facultative anaerobes5–10% of the species∼ 103–109 cfu/g faecesResident
Escherichia coliEnterococcus spp.
TransientKlebsiella spp.Enterobacter spp.Proteus spp.Providencia spp.Pseudomonas spp.Bacillus spp.yeasts, Protozoa
Normal bacterium Flora of the colon inadults (>400 Species)
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• Breast feeding– Resident
• Bifidobacteria (pH 5,5)– Colonisation of other species is
inhibited (vitamin K substitution!)
• Composite feeding– Primarily
• Facultative anaerobes– Later
• Bacteroides fragilis
Normal bacterium flora of thecolon in children
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Scanning electron micrograph of the colon mucosa of rat
⎢: bacterium layer, L: lumen, B: bacteria, T: intestinal tissue
Magnification: 262× Magnification: 2.624×
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Significance of the normal bacterial flora of the colon
• Decomposition of some nutrients, enterohepaticrecirculation of bile
• Production of vitamins: vitamin K & B-complex• Production of gases ⇒ distension ⇒ normal
peristaltic• Constant physical & chemical stimuli ⇒ constant
mucosal turnover• Biofilm formation, blocking epithelial receptors,
competition for nutrients, bacteriostatic products ⇒inhibition of the colonisation of pathogenic bacteria
• Constant antigen stimulus ⇒ extrauterindevelopment and functioning of the immune system
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Role of normal bacterial flora of the colon in pathological processes
•• Extra intestinal infectionsExtra intestinal infections
•• TranslocationTranslocationImmune Immune
compromised patientscompromised patientsObstructionsObstructionsShockShock
ConsequencesConsequencesEndotoxaemiaBacteriaemiaSepsis
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Bacterial translocation in rat after experimental intestinal obstruction
AA BB
A: A: lifting of the epithelial cells from the lifting of the epithelial cells from the
lamina lamina propriapropria & bacterial invasion& bacterial invasion
B: B: coccicocci and rods inand rods in
the the submucosalsubmucosal lymphatic vesselslymphatic vessels
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Alteration of the balance of the normal bacterial flora of the colon
•• CausesCauses– Malnutrition– Broad spectrum per os antibacterial therapy
•• ConsequencesConsequences– Maldigestion & maladsorption, vitamin deficiency– Alteration of the normal peristaltic, increased
gastrointestinal gas formation•• Antibiotic associated diarrhoeaAntibiotic associated diarrhoea
– Mild: „common” diarrhoea– Severe: pseudomembranous colitis
(Clostridium difficile)
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PseudomembranousPseudomembranous colitiscolitis
Thickened wall of the colon Thickened wall of the colon transversumtransversum
Characteristic yellow plaquesCharacteristic yellow plaquesC. difficileC. difficile & & neutrophilsneutrophils
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Gram negative facultativeanaerobic rods
Enterobacteriaceae I.
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Enterobacteriaceae
Morphology: - Gram negative rods- pili (ex.: Klebsiella, Shigella)
Cultivation:easy, simple (Agar, Blood-agar) MediaDifferentiation: pathogenic-facultative pathogenic(Biochemical reactions)
a) Selective-Mediab) Differential-Mediac) Indicator-Media
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Antigens and Virulence factors:O (cell-wall)H (Flagella)K (capsule)surface ProteinsPiliExotoxinsEndotoxin
Enterobacteriaceae
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Enterobacteriaceae
Facultative pathogensgenusEscherichia
Klebsiella GroupEnterobacterEdwardsiellaCitrobacter
Proteus GroupSerratiaProvidenciaMorganella
Obligate pathogens (genera)E. Coli
ETEC (enterotoxic)EPEC (enteropathogen)EIEC (enteroinvasive)EHEC (enterohemorrhagic)EAggEC (enteroaggregativ)
ShigellaS. dysenteriaeS. flexneriS. boydiiS. sonnei
SalmonellaS. typhiS. paratyphi
YersiniaY. pestisY. pseudotuberculosisY. enterocolitica
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Enterobacteriaceae - Facultative PathogensKlebsiella, Proteus, Escherichia etc. Extraintestinal clinical findings – ALL purulent infections:
a) urinary tract infectionsb) cholecystitis, cholangitisc) peritonitis (local, diffuse)d) pneumoniae) meningitisf) wound infectionsg) sepsish) iatrogen/nosokomial infections
Diagnosis: isolation, identificationTreatment: antibiogram (ESBL)
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Serratia marcescens - Serratia marcescens Durch Prodigiosin rotgefärbte Kolonien von Serratia marcescens auf Agargel.
www.biologie.de
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pathmicro.med.sc.edu
TSI Medium Identification
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Top row, Proteus vulgaris; second row, unidentified enteric bacterium; thirdrow, Klebsiella pneumoniae; bottom row, Vibrio alginolyticus.
helio
s.bt
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ukId
entif
icat
ion
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E. coli with Fimbriae
Urinary tract infections (UTI)Urinary tract infections (UTI)
www.spiceisle.com
•• E. coli E. coli (80%)(80%)– Predisposition– Ascending
(rarely haematogenic)– Cystitis, pyelonephritis– Virulence factors (UPEC-strains)
• Haemaggl. adhesion fimbriae– Mannose resistant (MR) fimbriae
(F-antigen), P-fimbriae (blood group P substance) ⇒pyelonephritis
– Mannose sensitive (MS) fimbriae ⇒cystitis
• Haemolysines
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• P. mirabilis, P. vulgaris– Predisposition– Ascending infection– Nosocomial
(catheter, operation)– Virulence factors
• Flagellae: motility
• Adhesion fimbriae• Urease
(pH↑, irritation, urinary stones)
P. mirabilis: Swarming
www.sciencebuddies.org
Urinary tract infections (UTI)Urinary tract infections (UTI)
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P. mirabilis
ww
w.n
rc-c
nrc.
gc.c
a
swarming
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P. mirabilis
Swarming- On cells
www.nrc-cnrc.gc.ca
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P. mirabilis
Swarming- On cells
www.nrc-cnrc.gc.ca
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biology.clc.uc.edu
Proteus vulgaris flagellar - staining
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A: Proteus, Providentia – B: Urease Test - / +
helios.bto.ed.ac.uk
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Neonatal Meningitis & Sepsis
•• E. coliE. coli K1K1(80–85%)
• K1-antigen – Identical with
Meningococcus B-ag– Tolerance ⇒
no antibody response– Translocation
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Pneumonia
• Nosokomial• Predisposition• Pathogens
E. coli,K. pneumoniae,K. oxytoca, Enterobacter spp. Bronchopneumonia Lobar pneumonia
(Friedländer)K. pneumoniae
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Klebsiella pneumoniae – Friedländer Pneumonia
ww
w.b
row
n.ed
u
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www.icbm.de
Klebsiella pneumoniae
mucoid colonies – capsule
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www.lf3.cuni.cz
Klebsiella pneumoniae
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Klebsiella pneumoniae
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Enterobacter cloacae Klebsiella pneumoniaeMucoid colonies
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Intra-abdominal Infections
Diffuse peritonitis after perforation of the colon
Free abdominal air below the diaphragm: perforation
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Enterobacteriaceae
Facultative pathogensgenusEscherichia
Klebsiella GroupEnterobacterEdwardsiellaCitrobacter
Proteus GroupSerratiaProvidentiaMorganella
Obligate pathogens (genera)Escherichia coli (species)
ETEC (enterotoxic)EPEC (enteropathogen)EIEC (enteroinvasive)EHEC (enterohemorrhagic)EAggEC (enteroaggregativ)
ShigellaS. dysenteriaeS. flexneriS. boydiiS. sonnei
SalmonellaS. typhiS. paratyphi
YersiniaY. pestisY. pseudotuberculosisY. enterocolitica
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FIGURE 25-1 Virulence mechanisms of E coli.
Medmicro
EIEC
ETEC
EAggEC
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FIGURE 25-2 Pathogenesis of E. coli diarrhoeal disease.
Medmicro
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Pathogenic E. coli strains-1
ETEC (enterotoxic)LT (heat-labile) ST (heat-stable)
ST mode of action (hyperactivation): ADP-Rybosilation GuanilcyclasecGMP water and electrolyte loss
Fimbriae (CFA = colonisation factor antigens)
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FIGURE 25-3 Cellular pathogenesis of E coli having CFA pili.
Medmicro
ETEC
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Fig.4.26 Enterotoxigenic E. coli infection. Transmission electron micrograph showing bacteria adhering to the brush-border of human intestinal mucosal cells. By courtesy of Dr. S. Knutton
ETEC
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FIGURE 25-4 Laboratory methods for isolation and identification of ETEC.
Medmicro
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Fig. 4.14 Bacterial diarrhea. Y1 adrenal cell assay for E. coli LT enterotoxin, showing normal cells (left) and cells after exposure to LT toxin (right). Note disruption of monolayer and rounding up cells. By courtesy of Dr. H.L. DuPont.
ETEC
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Pathogenic E. coli strains-2-3EPEC (enteropathogenic) O26; O55; O111; O126…
Adhaesion: Adherence Factor (EAF)Bundle Forming Pilus (BFP)Type III. Secretion-system*: Protein filaments (EspA)Tir (translocated intimin receptor)Intimin (eae*) Actin; brush-border lost cell death* Chromosome coded
EAggEC (enteroaggregativ)Fimbriae, ST-like, Haemolysin-like Toxin
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Fig.4.19 E. coli diarrhea. Electron micrograph of enteropathogenicE. coli (arrowed) attached to mucosal epithelial cells of ileum. The microvillus border of the epithelial cells has been largely destroyed by bacteria and the cells show signs of degeneration. X3000 By courtesy of Dr. J.R. Cantey
EPEC
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Fig.4.27 Enteropathogenic E. coli infection. Electron micrograph showing close, localized adherence of bacteria to human intestinal mucosal cells and localized destruction of microvilli. By courtesy of Dr. S. Knutton
EPEC
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Fig. 4.28 Enteropathogenic E. coli infection. Fluorescent actin test specific for EPEC organisms. Left: fluorescent microscopy showing aggregated actin. Right: phase contrast microscopy showing location of bacteria. By courtesy of dr. S. Knutton
EPEC
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Pathogenic E. coli strains-4
EIEC (enteroinvasive)
O28; O32; O112; O115; O124, O136; O143, O144 u.a.
Pathogenesis:s. Shigella ProteinsType III. Secretion-system
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FIGURE 25-5 Cellular pathogenesis of invasive E coli
EIEC
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Fig. 4.17 Enteroinvasive E. coli infection. Invasion of mucosal layer of the intestine by E. coli organisms. There is necrosis of the mucosal layer at the site of invasion (left). Transmission electron micrograph showing enteroinvasive E. coli organisms within HEp-2 cell (right). By courtesy of Dr. S. Knutton.
EIEC
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Fig. 4.31 Enteroinvasive E. coli infection. EIEC organisms invading HeLa cells in vitro. By courtesy of Dr. S. Knutton.
EIEC
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Pathogenic E. coli strains-5
EHEC (enterohemorrhagic) = VTEC O157:H7
SLT = Verotoxin = Shiga Toxin (stx1, 2, 2c) Mode of action: Protein synthesis inhibitor cytotoxic Haemolysin
Clinical finding, disease:HUS (haemolytic uraemic syndrome)
Haemolytic AnaemiaTrombocytopeniaacute kidney failure
Haemorrhagic colitis
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Pathogene E. coli strains
Transmission electronmicrograph of Escherichia coli O157:H7
pathmicro.med.sc.edu
EHEC(enterohaemorrhagic)= VTEC O157:H7
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EHEC (enterohemorrhagisch) = VTEC O157:H7
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Fig. 4.29 Enterohaemorrhagic E. coli infection. Assay for Shiga-like toxin (Verotoxin) produced by EHEC (Serotype O157). Left: Normal monolayer of Vero cells. Right: Destruction of Vero cells by the toxin. By courtesy of Dr. S. Knutton
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Fig. 4.30 Enterohaemorrhagic E. coli infection. Weigert stain showing fibrin ‘thrombi’ in glomerular capillaries in haemolytic uraemic syndrome. By courtesy of Dr. H.R. Powell
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Pathogen E. coli strains
DiagnosisDirect detectionSerotyping
ProphylaxisExpositions prophylaxis
TherapyWater and electrolyte substitutionAntibiogram
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Gram negative facultativeanaerobic rods
Enterobacteriaceae II.
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GRAM NEGATIVE RODS
AEROBBordetellaBrucellaFrancisella
PseudomonasAcinetobacterLegionella
FACULTATIV ANAEROBHaemophilusPasteurella
Familia:EnterobacteriaceaeVibrionaceae
CardiobacteriumEikenellaKingellaActinobacillus
ANAEROBBacteroidesPrevotellaPorphyromonasFusobacterium
MICROAEROPHILCampylobacterHelicobacter
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Enterobacteriaceae
Morphology: - Gram negative rods- pili(ex.: Klebsiella, Shigella)
Cultivation:easy, simple (Agar, Bloodagar) MediaDifferentiation: pathogenic-facultative pathogenic(Biochemical reactions)
a) Selectiv-Mediab) Differential-Mediac) Indicator-Media
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Antigens and Virulence factors:O (cell-wall)H (Flagella)K (capsule)surface ProteinsPiliExotoxinsEndotoxin
Enterobacteriaceae
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BACTERIAL GI TRACT INFECTIONSI. Type EnterotoxinHypersecretionSmall intestine
watery diarrhea
Vibrio choleraeEscherichia coli(ETEC)
II. TypeInflammationInvasion in MucosaLarge intestine
pus, blood, mucus in stool
ShigellaE. coli (EIEC) (EPEC, EHEC)SalmonellaYersinia enterocoliticaCampylobacter jejuniAeromonas sp.Vibrio parahaemolyticus
Clostridium difficileClostridium perfringens
III. TypePenetration,GeneralisationBacterium intracellularIleum
Typhus, Sepsis
Salmonella typhiS. paratyphi A, BYersinia enterocoliticaY. pseudotuberculosisCampylobacter fetus
Exogen, peroral Infection, faecal–oral transmission route
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Enterobacteriaceae
Facultative pathogenesgenusEscherichia
Klebsiella GruppeEnterobacterEdwardsiellaCitrobacter
Proteus GruppeSerratiaProvidenciaMorganella
Obligate pathogens (genera)Escherichia coli
ETEC (enterotoxic)EPEC (enteropathogenic)EIEC (enteroinvasive)EHEC (enterohemorrhagic)EAggEC (enteroaggregativ)
ShigellaS. dysenteriaeS. flexneriS. boydiiS. sonnei
SalmonellaS. typhiS. paratyphi
YersiniaY. pestisY. pseudotuberculosisY. enterocolitica
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Salmonellae
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www.ltsa.frwww.about-salmonella.com
Salmonella sp.
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Salmonella typhi, S. paratyphi A, B, C
Antigen structure of Salmonella typhigripsdb.dimdi.de
Obligate human pathogens
AntigensOH (flagella)superficial Vi Ag = capsule
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Salmonella typhi, S. paratyphi A, B, C
PathogenesisSource of infectionsick, carriers; Contaminated food, drinking waterPortal of entrymouth gut blood Organs: spleen, liver, bile, bone-marrrow, kidneys, brainIleum: ulceration (bleeding, Perforation)
Clinical findings:Typhus abdominalisParatyphus
Please, learn the latin terms!The english enteric fever or typhoidfever does not tell, which bacteriumcaused the disease!
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Salmonella typhi, S. paratyphi A, B, C
Figure 2. Flagellar stain of a Salmonellatyphi. Like E. coli, Salmonella are motile by means of peritrichous flagella. A close relative that causes enteric infections is the bacterium Shigella. Shigella is not motile, and therefore it can be differentiated from Salmonella on the basis of a motility test or a flagellar stain. (CDC)
Figure 1. Salmonella typhi, the agent of typhoid. Gram stain. (CDC)
www.textbookofbacteriology.net
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Typhus abdominalis
Roseolenartiges, makulopapulöses Exanthem bei Typhus abdominalis gripsdb.dimdi.de
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www.wrongdiagnosis.comRose spots on abdomen of a patient with typhoid fever due to the bacterium Salmonella typhi.
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Rose spots on the chest of a patient with typhoid fever due to the bacterium Salmonella typhi. www.wrongdiagnosis.com
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Fig. 4.37 Typhoid fever. Numerous ulcers of the small intestine overlying hyperplastic lymphoid follicles (Peyer’s patches). By courtesy of Dr. J. Newman.
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Fig. 4.39 Typhoid fever. Mononuclear cells and red blood cells in the stool. Trichrome stain. By courtesy of Dr. H.L. DuPont.
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Typhus abdominalisDiagnosisDirect detection (blood, stool, Urine)Selective mediaAntibody detection
(tube-Agglutination)
ProphylaxisExpositions prophylaxisImmune prophylaxis:1) Active oral immunisation with
Ty21, an apathogenic, attenuated strain
2) Parenteral injection with Vi capsule polysaccharideS. typhi-strain Type-2
TherapyAmpicillinTrimethoprimChloramphenicol
Carriers!!
„Typhoid Mary”
www.spiceisle.com
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Salmonella - SalmonellosisUbiquitous: S. typhimurium, S. enteritidis etc.
Pathogen for birds, eggs, swine, cattle, mouse, rat and human
PathogenesisSource of infection: from infected animals, contaminated foodBacteria multiply in the food! Endotoxin free
Clinical finding:FOOD POISONING – Endotoxin effectNo bacterium multiplication in guts, self limited(Gastro)enteritis – bacteria can multiply in guts, type II. diarrhoea, not always self limited
Diagnosis: direct detection (stool, food)Prophylaxis: food and kitchen hygiene
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Figure 21-3 Invasion of intestinal mucosa by Salmonella.
medmicrotyphus
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www.idph.state.il.us
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spacescience.comSalmonella sp.
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www2.nphs.wales.nhs.uk
Salmonella enterica
www.textbookofbacteriology.net
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Figure 3. Salmonella sp. after 24 hours growth on XLD agar.
www.textbookofbacteriology.net
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www.textbookofbacteriology.net
Figure 4. Colonial growth Salmonella choleraesuissubsp. arizonae bacteria grown on a blood agar culture plate. Also known as Salmonella arizonae, it is a zoonotic bacterium that can infect humans, birds, reptiles, and other animals. (CDC)
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www.chromagar.com
Rambach™ Agar For detection of Salmonella spp. •Salmonella - red•other bacteria - blue, violet, colourless, or inhibited.
wqc
.ariz
ona.
edu
Isolation of Salmonellafrom Environmental Samples
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pathmicro.med.sc.edu
TSI Medium Identification
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www.textbookofbacteriology.netSalmonella typhimurium HE agar
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Enterobacteriaceae
Facultative pathogenesgenusEscherichia
Klebsiella groupEnterobacterEdwardsiellaCitrobacter
Proteus GroupSerratiaProvidenciaMorganella
Obligate pathogens (genera)Escherichia coli
ETEC (enterotoxisch)EPEC (enteropathogene)EIEC (enteroinvasive)EHEC (enterohemorrhagisch)EAggEC (enteroaggregativ)
ShigellaS. dysenteriaeS. flexneriS. boydiiS. sonnei
SalmonellaS. typhiS. paratyphi
YersiniaY. pestisY. pseudotuberculosisY. enterocolitica
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Shigella
S. dysenteriae* 10 SerotypesS. flexneri* 6 SerotypesS. boydii 15 SerotypesS. sonnei
AntigenO
* Toxin
Shigella sonnei
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Shigella
Virulence factorsExotoxincytotoxic (cell lysis!);Enterotoxic;Paralytic – letal activity
Fragment A, B (5) – Glycolipid
Inhibition of Proteinsynthesis – Binding andInactivation of 60s Ribosom subunit cell death
OMP (Ipa, Ics)Endotoxin - LPS
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Pathogenicity – ID50: 100-200 BacteriaEntry and multiplication in Epithel cells:Invasion Plasmid Antigens – IpaIntercellular Spread – Ics
Pathogenesis, clinical findingsLocal InfectionEpithel necrosis, ulcerationinhibiting AbsorptionHUS!
Shigella
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Medmicro
Shigella
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mgc.ac.cn/VFs/Figures/Shigella
Shigella
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Fig. 2: Shigella Passing Through the Mucous Membrane and …
… Invading Mucosal Epithelial Cells
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cbcm
d.ed
u
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Fig. 4.33 Shigellosis. Sigmoidoscopic view of colonic mucosa in a mild case of infection due to S. flexneri. Note the thin whitish exsudate, which is made up of fibrin and polymorphonuclearleucocytes. By courtesy of Dr. R.H. Gilman.
Fig. 4.34 Shigellosis. Sigmoidoscopic view of colonic mucosa in a fatal case of infection with S. dysenteriaetype 1 showing extensive pseudomembranous colitis. By courtesy of Dr. R.H. Gilman and Dr. F. Koster.
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Fig. 4.18 Positive Serény test. Keratoconjunctivitis in the rabbit produced by the instillation of shigella microorganism. By courtesy of Dr. H.L. DuPont.
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www.textbookofbacteriology.net
Shigella
DiagnosisDirect detection(from stool)
Differentiating –SelectiveMedia
Identification Serotyping
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pathmicro.med.sc.edu
TSI Medium Identification
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Shigella boydii colonies on blood agar
www.biologie.de
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Appearance of Colonies on Salmonella-Shigella Agar
ww
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gers
.edu
A. Klebsiella pneumoniae B. Escherichia coliKlebsiella pneumoniae & Escherichia coli are positive for acid production from fermentation of the carbohydrate(s) present.
C: Salmonella sp.
D: Proteus mirabilis
Both Salmonella sp. & Proteus mirabilis product hydrogen sulfide. E: Pseudomona aeruginosa
The
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ss.
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Shigella
ProphylaxisExpositions prophylaxis – Hygienic rules
TherapyAntibiogramTetracylin, Ampicillin, Chloramphenicol, Sumetrolim
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Enterobacteriaceae
Facultative pathogenesgenusEscherichia
Klebsiella groupEnterobacterEdwardsiellaCitrobacter
Proteus GroupSerratiaProvidenciaMorganella
Obligate pathogens (genera)Escherichia coli
ETEC (enterotoxic)EPEC (enteropathogenic)EIEC (enteroinvasive)EHEC (enterohemorrhagic)EAggEC (enteroaggregativ)
ShigellaS. dysenteriaeS. flexneriS. boydiiS. sonnei
SalmonellaS. typhiS. paratyphi
YersiniaY. pestisY. pseudotuberculosisY. enterocolitica
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Yersinia enterocolitica
Morphology Gram negative,bipolar rods
flagellae
ww
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adsw
orth
.org
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Yersinia enterocolitica www.ktl.fi
cultivationoptimum 28°C, motility 28°C
Y. enterocoliticaO:5,27 CIN-ag
www.szu.czBlood agar
AntigensO and H
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Y. enterocoliticade.wikipedia.org
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Figure 29-7 Pathogenesis of Y. enterocolitica.
Clinical findingEnterocolitisLymphadenitis mesenterica
DiagnosisDirect detectionSerologySerotyping
TherapyTetracyclinChloramphenicolSumetrolim
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Yersinia pseudotuberculosis
MorphologyGram negative, bipolar rodsflagellae
Cultureeasy, grow 37°C and 20°Cmotility 20°C
PathogenicityPseudotuberculosis ofrodents
Source of infection:Sick animalsPortal of entry: mouth, mucosa Y. pseudotuberculosis
www.microbes-edu.org
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Yersinia pseudotuberculosis
Clinical findingsLymphadenitis mesentericaSeptic-typhoid FormEnteritis
DiagnosisDirect detectionSerotyping – AgglutinationSerology
TherapyTetracyclin
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Fig. 4.50 Yersinia infection. Gross specimen of ileum, showing superficial necrosis of the intestinal mucosa with several well-defined deep and superficial ulcers.
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www.llnl.gov
Y. pseudotuberculosis
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Gram negative facultativeanaerobic rods
Vibrionaceae
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GRAM NEGATIVE RODS
AEROBBordetellaBrucellaFrancisella
PseudomonasAcinetobacterLegionella
FACULTATIVE ANAEROBHaemophilusPasteurella
Familia:EnterobacteriaceaeVibrionaceae
CardiobacteriumEikenellaKingellaActinobacillus
ANAEROBBacteroidesPrevotellaPorphyromonasFusobacterium
MICROAEROPHILCampylobacterHelicobacter
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BACTERIAL GI TRACT INFECTIONSI. Type EnterotoxinHypersecretionSmall intestine
watery diarrhea
Vibrio choleraeEscherichia coli(ETEC)
II. TypeInflammationInvasion in MucosaLarge intestine
pus, blood, mucus in stool
ShigellaE. coli (EIEC) (EPEC, EHEC)SalmonellaYersinia enterocoliticaCampylobacter jejuniAeromonas sp.Vibrio parahaemolyticus
Clostridium difficileClostridium perfringens
III. TypePenetration,GeneralisationBacterium intracellularIleum
Typhus, Sepsis
Salmonella typhiS. paratyphi A, BYersinia enterocoliticaY. pseudotuberculosisCampylobacter fetus
Exogen, peroral Infection, faecal–oral transmission route
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Gram negative facultative anaerobic rods(Positive Glucose fermentation)
Oxidase positive Oxidase negative
Vibrionaceae Aeromonadaceae
Vibrio Aeromonas
Enterobacteriaceae
EscherichiaKlebsiella
EnterobacterProteusSerratia
ProvidenciaMorganella
EdwardsiellaCitrobacter
Hafnia
SalmonellaShigellaYersinia
Facultativepathogenic
Obligatepathogenic
Plesiomonas
V. choleraeV. parahaemolyticusV. vulnificus
P. shigelloides A. hydrophila
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VibrionaceaeSpecies Associated diseases
V. cholerae O1klassische & El Tor Cholera
V. cholerae O139 Cholera
V. parahaemolyticus Gastroenteritis
V. vulnificus Wound infection, sepsis
Non-agglutinable(NAG) vibrios Gastroenteritis (rarely)
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R. Koch, 1883Morphology• Gr–, curved rods• Facultative anaerobesBiochemical reactionsGlukose OF ‡Motility +Catalase +Oxidase +Nitrate Reduction +
Vibrio cholerae
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bepast.org
V. cholerae
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Antigen structure, virulence factorsO (cell wall) - 138O1 & O139H flagellar (common)Fimbriae: A, B, CO1: Bio and Serotypes
V. cholerae
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Vibrio cholerae. Leifson flagella stain (digitally colorized). CDC/Dr. William A. Clark
pathmicro.med.sc.edu
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Bio- & Serotypes of V. cholerae O1
Medmicro
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Virulence factors
Virulence factor Biologic effect
Cholera toxin Fluid & electrolytehypersecretion
Fimbriae Adhesion to mucus membrane
Accessory colonisation factor Adhesion to mucus membrane
Haemagglutination protease (mucinase)
Enteric inflammationdegradation of tight junctions
Siderophores Iron sequestration
Neuraminidase Toxin receptor up-regulation
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V. cholerae
Pathogenesis – Obligate human pathogenSource of infection: 1. humans (sick) and carriers(incubation, constant carriers and reconvalescent carriers – via stool! – flies!)2. Contaminated food (sea food!) and drinking waterReservoir: Algae, Mussel, PlanktonsTransmission: Per oralPortal of entry: GI
Toxin building in small intestineNO DISSEMINATION!Clinical finding, disease: Cholera – due to Cholera toxin
Immunity: local IgA (IgG in blood!)
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Adhaesion of V. cholerae to intestinal epithelium
TEM, ×16.000SEM, ×4.000
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Medmicro ch 24
FIGURE 24-1 Pathophysiology of cholera.
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textbookofbacteriology.net
Cholera toxin: mode of action
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ww
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1. 2.
3.
4.
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Cholera Toxin and Pertussis Toxin
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Cholera Pandemies• India, estuarine of Ganges• Intercontinental travels, trading, wars ⇒
seven pandemics from 1817• At present: 7th pandemic, V. EL Tor
– 1961: Asia– 70’s–80’s: Africa, Europe, Oceania– 1991: South-America
• 1992 : V. cholerae O139 „Bengal”– Rapid spread (Asia, Europe, USA)– No cross immunity with O1-strains– 8th pandemic?
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7th Cholera Pandemy 1961
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Cholera cases, WHO: 2000–2001
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Cholera outbreaks
Cholera camp in Mozambique
Cholera clinic in Mozambique
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Cholera: clinical picture• rice- - water diarrhoea
(25 L/day)• Dehydration• Haemoconcentration• Blood pH • Serum K+ , Na+
• Serum Glucose ↑• Shock• Letality
– Untreated• Classical: 60%• El Tor: 15–30%
– Treated: 1%
Rice-water diarrhoea
Baby before and afterrehydration
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Cholera Beds
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• Patients history!• specimen: stool• Direct microscopy:
dark field microscope
Diagnosis
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Transport: Alkaline PeptonwaterCulture: TCBS medium
Diagnosis
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IdentificationBiochemical Reactions
Serotyping (O1, O139)
Antibiogram
Diagnosis
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V. cholerae
Prophylaxis of Cholera
Expositions prophylaxisClean drinking water! – and food hygiene and sewage water system – no mix with drink water!
Mass education, adhering to hygienic rulesIsolation; Quarantinetreating, disinfectingWater boilingFollow up carriersReport to WHO
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• Immune prophylaxisVaccination
– Vaccine (against O1 ony)• Killed bacterium, parenteral• Killed bacterium + B-subunit Toxoid per os• Gene manipulated, attenuated V. cholerae per os
– Protection of 3–6 month, efficacy 50–60%– Travellers to endemic areas– WHO: false feeling of security (!)
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Prevention of cholera: mass education of hygienic rules
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Therapy of Cholera
Salt-sticks
Coke
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Therapy of Cholera
• Fluid and electrolyte substitution– Intravenous– Per os Oral Rehydration Fluid
ORF• Glucose 20g/l• NaHCO3: 2,5 g/l• NaCl: 3,5 g/l• KCl: 1,5 g/l
• Antibiotic– Ciprofloxacin– Doxycycline
Peru
Bangladesh
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Vibrio parahaemolyticus
www.city.niigata.niigata.jp
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www.fehd.gov.hk
Gastroenteritis
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Fig. 10.20 Cellulitis. Severe infection with bullous lesions due to V. vulnificus infection following immersion of leg in brackish water.
V. vulnificus
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V. vulnificus
Fig. 10.21 Vibrio cellulitis. Haemorrhagic, bullouslesions of V. vulnificussepsis. By courtesy of Dr. J.R. Cantey
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A. hydrophila P. shigelloides
Cellulitis Myonecrosis
A. hydrophila infection in fish
Gastroenteritis
Sea food
Natural water sources
web.umr.edu
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Aeromonas hydrophila
www.buddycom.com
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www.buddycom.comweb.umr.edu
Aeromonas hydrophila
Plesiomonas shigelloideshttp://www.vetmed.wisc.edu
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Aeromonas hydrophila
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Aeromonas hydrophila
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To be continued…
Zakynthos, 2004
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Microaerophilic Gram negative rods –
Campylobacter andHelicobacter
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GRAM NEGATIVE RODS
AEROBBordetellaBrucellaFrancisella
PseudomonasAcinetobacterLegionella
FACULTATIVE ANAEROBHaemophilusPasteurella
Familia:EnterobacteriaceaeVibrionaceae
CardiobacteriumEikenellaKingellaActinobacillus
ANAEROBBacteroidesPrevotellaPorphyromonasFusobacterium
MICROAEROPHILCampylobacterHelicobacter
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BACTERIAL GI TRACT INFECTIONSI. Type EnterotoxinHypersecretionSmall intestine
watery diarrhea
Vibrio choleraeEscherichia coli(ETEC)
II. TypeInflammationInvasion in MucosaLarge intestine
pus, blood, mucus in stool
ShigellaE. coli (EIEC) (EPEC, EHEC)SalmonellaYersinia enterocoliticaCampylobacter jejuniAeromonas sp.Vibrio parahaemolyticus
Clostridium difficileClostridium perfringens
III. TypePenetration,GeneralisationBacterium intracellularIleum
Typhus, Sepsis
Salmonella typhiS. paratyphi A, BYersinia enterocoliticaY. pseudotuberculosisCampylobacter fetus
Exogen, peroral Infection, faecal–oral transmission route
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Spiral formed & curvedGram-negative bacteria
Spirillaceae
Campylobacter Helicobacter Spirillum
C. jejuni H. pylori S. minus
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Most important species of Campylobacter genus
Species Reservoir Diseases Frequency
C. C. jejunijejuni Poultry, pig, cattle, rabbit
Gastroenteritis, sepsis, meningitis, Guillan-Barré
Frequent
C. coli Pig, poultry, cattle, sheep
Sepsis, gastroenteritis, meningitis
Uncommon
C. fetus Cattle, sheepSepsis, gastroenteritis, meningitis
Uncommon
C. lari Poultry, pig, cat, monkey, horse
Gastroenteritis, sepsis Rare
C. upsalensis Dog, cat Gastroenteritis, sepsis, abscess ?
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Campylobacter
MorphologyGram negative curved rods (0,3–0,6 μm)
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Campylobacter
Polar flagellum
Transmission electron micrographs of Campylobacter jejuni, negatively stained to enhance contrast.
www.wadsworth.org
www.shef.ac.uk Campylobacter jejuni
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www.indigo.com
Campylobacter jejuni
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CultureMicroaerophilic
5–7% O25–10% CO2
Thermophilic: 42ºC
Campylobacter
Campy-blood-agarmedinfo.ufl.edu
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www.biomerieux.com
Special media
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Campylobacter C. jejuni, SEM
C. jejuni, SEM
Biochemical reactionsNon-fermenting • Catalase +, Oxidase +• Nitrate reduction +• Antigen Structure
– O, H, K (Serotyping)
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Virulence factors• flagellum ⇒ motility• Adhaesion Factors• Invasion Factors (?)• Cytotoxin• C. fetus: S-Protein ⇒ inhibition of binding of C3b ⇒
antiphagocytic effect• LPS
Complication:• Guillan-Barré SyndromeSimilar structure: LPS-core and glycosphingolipids of
Gangliosids (GM1, GM2) ⇒Antibodies against GM1 ⇒Autoimmun process ⇒ Demyelinisation
Campylobacter
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C. coli & Guillan-Barré syndrome
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Epidemiology
• Zoonotic infection!• Source of infection:
contaminated food & waterInsufficient heat on meat• Human to human: Faecal-oral
route (children). Rarely• ID: 500 (low)• Very common (in developing
countries 80% of diarrhealdiseases)
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Campylobacteriosis: Clinical picture
• Incubation: 1–2 days• Bloody diarrhoea• Fever• Abdominal pain, cramps• Spontaneous healing:
within 1–7 days• Complications
– Prolonged course– Systemic infection– Reactive arthritis– Guillan-Barré syndrome
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Diagnosis of Campylobacter Infections
Campylo-agarculture
Biochemical identification
• Sample– Faeces– Blood culture, CSF– Food
• Culture (microaerophilic, thermophilic)• Identification• Antibiotic susceptibility test
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Therapy & Prevention of Campylobacter Infections
• Supportive (water, electrolytes)• Antibiotic therapy
– Gastroenteritis• Erythromycin, Doxycycline, Ciprofloxacin,
Amoxicillin/clavulanic acid– Systemic infection
• Carbapenem, Aminoglycoside, Chloramphenicol
• Prevention: food hygiene
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Helicobacter pyloriMorphology• Gram negative, spiral-
formed rod
Polar flagellae: Motility
Culture• MicroaerophilicBiochemical reactions• Non-fermenting• Catalase +• Oxidase +• Urease + (!)
H. pylori
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Virulence factors of H. pylori• Adhaesins (HOP)• Flagellae (Motility)• Mucinase, phospholipase• Urease activity• Vacuolating cytotoxin• Protein CagA
Translocation viaType IV. Secretion-systemchanging cytoskeletonIl-8, Il-1, TNFα Stimulation
Urease positive
Cytotoxic effect in HeLa cells
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Epidemiology and clinical findings ofH. pylori Infections
Epidemiology• Common worldwide• Reservoir: humans• Transmission
– Faecal-oral– Oro-oral (saliva)– Endoscope!
Clinical pictureAcute GastritisChronic-active GastritisGastroduodenal UlcusTumor
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H. pylori & peptic ulcer
Chronic gastritis
Duodenal ulcer
H. pylori on gastric mucosa, SEM
H. pylori histology, silver impregnation
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H. pylori & gastric carcinoma
Antral adenocarcinomagastroscopic finding
Gastric adenocarcinoma withliver metastasis & ascites
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Diagnosis of H. pylori
• direct detection of Helicobacter Antigen in stool sample• biopsy: Histopathology, culture, molecular biology (PCR)• radioactive Urea expiration test• serology: AB detection
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Radioactive Urea expiration test
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Microbiological Diagnosis of H. pylori
H. pylori culture
Biochemical identification
H. pylori ELISA (IgG, IgA)
H. pylori Western blot
culture –Biopsy Material
antibodies – Blood serum
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H. pylori Eradication
• Combinations - Therapy with– Proton pump inhibitors– Antibiotics
• Clarithromycin + Metronidazole• Amoxicillin + Metronidazole• Doxycycline + Metronidazole
– Eradication: 90%
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THE END
Santorini, 2009