Identification of Enterobacteriaceae coliforming 2012274048 정소화 발 표 2012274017 정솔잎 자료조사 2012274031 정주희 자료조사 2012274051 조민정 ppt 제작 2012274018 주현우 자료조사 2010251114 김덕호 자료조사 Identification of Enterobacteriaceae coliforming organisms and related diseases Identification of Enterobacteriaceae coliforming organisms and related diseases

contents Family Enterobacteriaceae Identification Antigenic Structures & Virulence Factors Coliform organism & disease Other coliforms

Family Enterobacteriaceae

Enterobacteriaceae Characteristics : - Small and straight rods - Gram negative - Resident of Soil, water, decaying matter, large bowel of humans and animals - spore (x), - Most motile (except Shigella & Klebsiella & Yersinia ) - Facultative anaerobe - Oxidase (-), Catalase (+) - Glucose ferment, Nitrate reduction - ferment Carbohydrates (anaerobic pathway)

Enterobacteriaceae Diarrheal illnesses : two basic mechanism 1. Toxigenic infection – (a) 2. Invasive infection – (b) Enterics : more than 50% of nosocomial infection -E.coli, Klebsiella, Proteus, Enterobacter, Serratia, Citrobacter.

Enterobacteriaceae Coliforms : Rapid lactose-fermenting enteric bacteria E.coli, Klebsiella, Enterobacter, Hafnia, Citrobacter, Serratia Non-coliforms : Non-lactose-fermenting or Slow lactose-fermenting bacteria - Opportunistic, normal gut flora - Pathogenic enterics - Pathogenic nonenterics

Identification Enrichment media : selenite or GN broth Selective media -MacConkey agar -EMB agar(Eosin Methylene Blue agar) -Hektoen enteric agar TSI agar IMViC(Indole test, MR test, VP test, Citrate test) Urease test Motility test

Identification Coliform : Pink MacConkey agar EMB agar Non- coliform : 반투명 EMB agar Coliform : 검은색 Non – coliform : 옅은 연보라색 Hektoen enteric agar Coliform : orange, salmonpink Non – coliform : green, blue-green

Identification TSI agar IMViC Urease test Motility test

+ - Genus Indole Methy red Voges-Proskauer Citrate TSIA Motility Escherichia + - A/A G MOTILE Klebsiella SPECIES VARIABLE NON-MOTILE Enterobacter Citrobacter H2S Serratia K/A

Antigenic Structures Complex surface antigens H : flagellar antigen K : capsule and/or fimbrial antigen O : somatic or cell wall antigen

Antigenic Structures Variety of subspecies or serotypes Serotyping - Classifying some species - Pinpoint the source of outbreaks - Epidemiological tool

Antigenic Structures Endotoxin (lipopolysaccharide,LPS) Exotoxin Capacity to overcome host defenses Multiply in the tissues and blood

Virulence Factors Gram-negative bacteria freely transfer chromosome or plasmid - Drug resistance - Toxigenicity - Adaptive traits - Alter their pathogenicity Virulence genes : code for enterotoxins, capsules, hemolysins, fimbriae

Coliform organism & disease

Escherichia coli The most prevalent enteric basillus Colon bacillus The best known coliform facultative anaerobic & non-fastidious bacterium in the gut

Pathogenic Strains of E. coli (1) Enterotoxigenic E. coli (ETEC) severe diarrheal illness(similar to cholera) two exotoxins -LT(heat-labile toxin) -ST(heat-stable toxin) heightened secretion, fluid loss fimbriae : adhesion to the small intestine (2) Enteroinvasive E. coli (EIEC) Invasion and ulceration of the mucosa of the large intestine Inflammatory disease

Pathogenic Strains of E. coli (3)Enteropathogenic E.coli (EPEC) infantile diarrhea (4)Enterohemorrahgic E.coli (EHEC) E.coli O157:H7 hemorrhagic colitis hemolytic uremic syndrome(HUS) , severe damage to the kidney

Clinical diseases of E. coli (1) infantile diarrhea the greatest single cause of mortality among babies crowded tropical regions : poor sanitary facilities , contaminated water supplies malnourished, further loss of body fluids and electrolytes → fatal (2) traveler's diarrhea 70% exotic pathogen(ETEC) Within 7-8days : profuse, watery diarrhea, low grade fever, nausea, vomiting

Clinical diseases of E. coli (3) urinary tract infections(UTI) 50%~80% of UTI in healthy people -more common in women - short urethras →(promote) ascending infection to bladder(cystitis) & kidneys (4) other extraintestinal infections -neonatal meningitis, pneumonia, septicemia, wound infections (complicate surgery, enndoscopy, tracheostomy, catheterization, Renal dialysis, immunosurpessant therapy,)

E. coli and the Coliform Count the indicator present in larger numbers, can survive in the environment, easier & faster to detect than true pathogens if E. coli is present in a water sample → fecal pathogens Salmonella, viruses, pathogenic protozoa →judge unsafe to drink

Other coliform

Other coliform Klebsiella, Enterobacter, Serratia, Citrobacter natural host defenses fail septicemia, endotoxemia

Other coliform 1) Klebsiella pneumoniae respiratory tracts - chronic lung infections large capsule : prevents phagocytosis nosocomial pneumonia, meningitis, bacteremia, wound infections, and UTIs(urinary tract infections)

Other coliform 2) Enterobacter & Hafnia spp. 3) Citrobacter spp. inhabit : soil, sewage, dairy products surgical wounds, spinal fluid, sputum, blood Urinary tract infections(UTIs) lethal when introduced into the bloodstream. 3) Citrobacter spp. inhabit : soil, water, human colon opportunist in urinary tract infections , bacteremia in debilitated persons

Other coliform 4) Serratia marcescens inhabit : soil, water, intestine produces an intense red pigment Serratia pneumonia (prevalent in alcoholics), burn and wound infections, fatal septicemia, meningitis .

Q&A

QUIZ

QUIZ 1. What is NOT true about Enterobacteriaceae ? 1. spore-forming 2. Glucose ferment 3. Nitrate reduction 4. Oxidase (-) 5. Catalase (+)

QUIZ 2. what does the following next colifrom : Creates nuclear black, sometimes tinged with a metallic luster on the surface 1. MacConkey agar 2. EMB agar 3. Hektoen enteric agar 4. TSI agar

QUIZ 4. How can E. coli cause infantile diarrhea in infants but not in their mothers?

QUIZ 5. What are two basic mechanisms of diarrheal illness? ①toxigenic infection ②invasive infection ③aerial infection ④contact infection

QUIZ 6. Which is not surface antigen? 1.H antigen 2.K antigen 3.O antigen 4. None of above. 5. All of these.

QUIZ 7. Which strain of E.coli causes HUS? a. EHEC b. EAEC c. EPEC d. EIEC

Reference 임상미생물학 실습/전국임상병리학과 교수 협의회/ 고려의학/ 2010/ 4판 임상미생물학 실습/전국임상병리학과 교수 협의회/ 고려의학/ 2010/ 4판 미생물학/ 최호형/ 아카데미서적/ 2004/ 1판 Foundations in microiology/ Talaro Chess/ McGraw-Hill/ 2012/ 8th 아카데미 생명과학 사전/생명과학사전위원회/아카데미서적 미생물학/willey,, sherwood/7판/김영민 외 8명/라이프 사이언스 미생물학 실습/김선희 외 7명/고려의학 밀레니엄 미생물학/김관천 외 10명/ 정문각 MICROBIOLOGY/Kathleen Park Talaro/McGraw Hill/612-614p

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