Nonfermentative Gram-Negative Bacilli

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1

oxidative

nonfermentative of glucose or other carbs in absence of oxygen; require oxygen as final hydrogen ion acceptor; no reaction on TSI: K/K H2S negative

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nonfermentative gram negative bacilli

obligate aerobes, oxidase positive, grows better on blood than MacConkey, non spore formers

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natural water sources

location most are found

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small amount of indole produced

modified indole result of nonfermenters

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Oxidative fermentative (OF) Medium of Hugh Leifson

tests for several carbs: glucose, maltose, lactose, mannitol, and xylose

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bromthymol blue indicator

medium within OF

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yellow - positive

OF in acidic environment

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green/blue - negative

OF in alkaline environment

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oxidative

OF result positive in open tube and negative in closed tube

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fermentative

OF result positive in both open and closed tubes

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nonsaccharide

OF result negative in both open and closed tubes

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negative NO3 reduction - fermenters

red color change with zinc indicating presence of nitrates

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positive NO3 reduction - nonfermenters

no color change with addition of zinc indicating nitrates have been reduced completely to nitrogen gas (N2)

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pseudomonadaceae

nonfermentative, gram negative bacilli; motile with polar flagella; oxidize glucose and other carbohydrates; strict aerobes; usually cytochrome oxidase positive

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fluorescent pseudomonadaceae

produce pyoverdin, fluorescent pigment; includes aeruginosa, fluorescens and putida

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stutzeri pseudomonaceae

scil dentitrifiers; includes stutzeri and mendocina

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pseudomallei

resistant to polymyxin B and colistin; includes mallei, pseudomalle, cepacia, and maltophilia

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Pseudomonas aeruginosa

most frequently isolated nonfermentor (75-80%); strict aerobe; fruity overripe grape odor; grows well at 35-7 and 42 C; motile with single polar flagellum; dull spreading beta hemolytic colonies with feathery edge on blood agar; do not ferment lactose; unable to utilize carbohydrates K/K in TSI; oxidase positive; oxidizes glucose but not maltose; ADH (arginine) +; LDC (lysine) -; opportunistic and nosocomial; antibiotic resistant

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pyocyanin pigment

blue pigment that is soluble in water and chloroform; only produced in P. aeruginosa

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pyoverdin, fluorescein, pigment

yellow fluorescent pigment; found in fluorescens and putida

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bluish green

pigment pyocyanin and pyoverdin produce together on certain media

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cystic fibrosis

patients produce mucoid strain of Pseudomonas aeruginosa

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alginate

produced in cystic fibrosis patients which surrounds the bacterial cell wall and protects it from phagocytosis

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Pseudomonas aeruginosa virulence

mucoid strain (alginate) of CF, fimbriae, flagella, eslastase, collagenase, protease, pyocyanin, oxotoxin A, hemolysis, and lipopolysaccharide

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P. fluorescens

food spoilage; rare cause of UTI

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P. putida

rare causes of septicemia

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P. stutzeri and P. mendocina soil denitrifiers

use ammonium as the sole nitrogen source and acetate as the sole carbon source

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P. stutzeri

tan, wrinkled colonies

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P. mendocina

smooth, buttery colonies

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Burkholderia mallei

glanders; acquired through inhalation of organism; only nonmotile pseudomonad; smooth, creamy to white colonies on blood agar; weakly oxidase positive and fails to grow at 42C

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glanders

infectious disease of horses, goats, sheep, and donkeys

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Burkholderia pseudomallei

meliodosis; subacute disease; possible bioterrorism agent; oxidizes many carbohydrates

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meliodosis

glanders-like disease; found in soil and stagnant water in SE Asia and Australia

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subacute disease

pulmonary infection and chronic cellulitis

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Burkholderia cepacia

found in soil, IV fluids and disinfectants; onion bulb rot in plants and foot rot in humans; yellow sedated colonies, weakly oxidase positive; opportunistic colonizing skin and respiratory tract; pneumonia; isolated with cystic fibrosis and chronic granulomatous disease

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Ralstonia

includes pickettii and radiobacter; slow growing nonfermenters found in environmental sources; generally not pathogenic isolated from blood, urine, and sputum

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Acinetobacter

obligate aerobe, oxidase negative catalase positive, nonmotile coccibacilli that demonstrate as plump paired rods or diplococci; nosocomial and opportunistic; non pigmented, domed, mucoid colonies with smooth to pitted surfaces that are non hemolytic; cannot reduce nitrate or grow anaerobically; grow well on MacConkey; penicillin resistant

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Acinetobacter baumanni

may be confused with Enterobacteriaceae; oxidase negative and grow on MacConkey

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Acinetobacter Iwoffi

ammonia-like odor

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Acinetobacter

may mimic Neisseria in gram stains from female genital tract; differentiate through oxidase which will be negative from Neisseria +

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Stenotrophomonas maltophilia

yellow-to-tan on TSA; lavender green to pale lavender on sheep blood agar; grows at 42 C; oxidase negative oxidizing glucose weakly and maltose strongly; ammonia-like odor; found in food, water, soil, and plants; opportunistic and nosocomal

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Moraxella

nonmotile, small GN diplococci or diplobacilli; no growth on MacConkey; oxidase and catalase positive; nonsaccharolytic; differentiate from Neisseria (blood agar growth and carbohydrate test) and Acinetobacter through oxidase

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Moraxella catarrhalis

resembles Neisseria morphologically and serologically; normal flora of upper respiratory tract; increasingly opportunistic; otitis media, sinusitis, pneumonia, endocarditis, conjunctivitis, bacteremia, wound infections, and meningitis; grows on TSA at 35-37 C; gray smooth opaque non hemolytic colonies; oxidase and DNAse positive; fails to produce acid; slides like a hockey puck

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Chryseobacterium

member of Flavobactericeae family; found in environment, sinks, ice machines and water baths; nosocomial and opportunistic; lavender green on blood agar due to gelatins; poor or no growth on MacConkey

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Chryseobacterium indologenes

dark yellow colonies; occasional cause of bacteremia in immunosuppressed

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Chryseobacterium meningosepticum

most significant human pathogen; pale yellow colonies; meningitis in newborns; pneumonia, endocarditis, and bacteremia in immunosuppressed

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Kingella

require blood for growth; includes denitrificans and kingae; rare pathogens

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Pseudomonas, Burkholderia, Ralstonia, and Stenotrophomonas

motile with poor flagella

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Chryseobacterium, Flavobacterium, and Moraxella

nonmotile and oxidase positive

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Acinetobacter

nonmotile and oxidase negative

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Alcaligenes, Achromobacter, and Oligella

motile with peritrichous flagella

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fimbriae and polar flagella

pseudomonas attachment virulence factors

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pyoverdin and elastase, protease, collagenase, hemolysin, and leukocidan tissue invasion

pseudomonas tissue invasion and immune inhibition virulence factors

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alginate capsule, exotoxin A, type III secretion system and Exo S, T, U, and Y toxins

pseudomonas endotoxin virulence factors

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fimbriae

attach to host cells and activate pro inflammatory genes

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polar flagella

attach to host cells for motility and inactivation of interleukin 8

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pyoverdin

binds iron

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elastase tissue invasion

digests elastin

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protease tissue invasion

protein degradation

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collagenase tissue invasion

collagen destruction

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hemolysin tissue invasion

lyse red blood cells

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leukocidan tissue invasion

lyse white blood cells

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alginate capsule

inhibits phagocytosis and chemotaxis of neutrophils, and activation of complement

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exotoxin A

inhibits protein synthesis and facilitates infection

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Type III secretion system

injects toxins into host cells Exo S, T, U, and Y

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Exo S, T, U, and Y

toxins that have cytotoxic and other effects