CLINICAL BACTERIOLOGY – LECTURE Batch 2024

NON-FERMENTATIVE GRAM-NEGATIVE BACILLI MODE OF ACQUISITION:

GENERAL CHARACTERISTICS  Ingestion of contaminated food or water, use of contaminated medical devices,
 Opportunistic pathogens not usually found as indigenous microbiota of humans. and open skin wounds.
 Commonly found in the environment (water, soil, and vegetables), and they act as  Has been isolated from contact lenses that are rinsed with a contaminated solution
contaminants in food and hospital devices.  Also found in hot tubs and whirlpool baths.
 Can be isolated from nebulizers, ventilators, catheters, dialysate fluid, and other  TAP WATER is the reservoir of P. aeruginosa.
hospital instruments and equipment.
VIRULENCE FACTORS:
 Able to resist treatments with disinfectants such as chlorhexidine and quaternary
ammonium compounds. Exotoxin A Cytotoxicity, blocks protein synthesis
 Utilize carbohydrates by oxidation – metabolism is respiratory, not fermentative. Lipopolysaccharide Antiphagocytic, cytotoxicity
 Colorless (NLF) on MAC; K/K on TSI; fail to acidify OF medium with mineral oil. Pili Adhesion
 Most are OXIDASE-POSITIVE, have a characteristic pigmentation & color, and are Capsule Anthiphagocytic
MULTI-DRUG RESISTANT. Flagella Adhesion
Phospholipases Cytotoxicity
PSEUDOMONAS Type III secretion system Cytotoxic activity
 Belongs to the family Pseudomonadaceae. Protease Cytotoxicity
 The most commonly isolated non-fermentative bacilli.
 Obligate aerobes and their metabolism is strictly aerobic; members are usually DISEASE ASSOCIATION:
oxidizers with few asaccharolytic.  Leading cause of nosocomial infections.
 Third most common cause of gram-negative bacillary bacteremia.
 Usually grow on BAP & MAC; can survive with few nutrients (glucose & acetate).
 Causes infection of burns, giving rise to BLUE-GREEN PUS
 Can be found in cosmetics, swimming pools, hot tubs, clothing and foot wear.
 Common cause of complication in patients with CYSTIC FIBROSIS and chronic
 Catalase and Oxidase positive; K/K, gas(-), H2S(-) on TSI.
obstructive pulmonary disease.
GROUPS OF PSEUDOMONAS:  Causative agent of SWIMMER’S EARS (otitis media/externa), ECTHYMA
1. Flourescent Pseudomonas: P.aeruginosa, P.fluorescens, P.putida, P.veronii, GANGRENOSUM/ Pseudomonas dermatitis – skin lesion or wound infection,
P.monteilii, and P. mosselii WHIRLPOOL DERMATITIS/“JACUZZI” HOT TUB SYNDROME/BUSINESSMAN’S
2. Non-Fluorescent Pseudomonas: P.stutzeri, P.alcaligenes, P.pseudoalcaligenes, SYNDROME (necrotizing skin rash) - acquired from hot tub.
and P.luteola  Can cause nosocomial UTIs, endocarditis, bone infections, eye infections, infections
following burns or trauma, meningitis (rare), and infections of the nail beds.
Pseudomonas aeruginosa (Agent of Blue Pus)
 Causes hospitalised infections that are related to antibiotic resistance.
 Former name: Burkholderia pyocyaneus
 The most commonly isolated species of the genus; the most commonly encountered PIGMENTS PRODUCED BY P. aeruginosa
Gram-negative bacterium that is not a member of the Enterobacteriaceae.  PYOVERDIN – yellow-green or yellow-brown pigment.
 One of the top three (3) causes of nosocomial infection:  PYOCYANIN – blue pigment (only produced by P. aeruginosa)
 P.aeruginosa, Acinetobacter baumannii. Stenotrophomonas maltophilia
 Obligate aerobe; oxidase(+); nitrate reduction (+); motile w/ monotrichous flagella. Note: Pyocyanin combining with pyoverdin produces the green color characteristic of
P. aeruginosa colonies
 Grows well at 37°C but its good growth at 42°C differentiate it from other
Pseudomonas spp.  PYURUBIN – red pigment.
 Inherently resistant to a number of antimicrobial agents.  PYOMELANIN – brown or black pigment.
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CLINICAL BACTERIOLOGY – LECTURE Batch 2024
DISTINGUISHING CHARACTERISTICS:  Pink colonies on MAC (lactose oxidizer)
 (+) Growth at 42°C  Related infection: Pneumonia in patients with cystic fibrosis,
 (+) Acetamide and citrate utilization septic arthritis, and endocarditis among users of prohibited
 (+) Gluconate production drugs
 (+) ADH; (-) LDH Burkholderia mallei  Agent of GLANDERS/FARCY DISEASE - severe infection that
 GREEN METALLIC SHEEN on BAP affects horses and donkeys and transmissible to humans
 Produce a fruity, grape-like odor or “corn tortilla-like” odor from the o Horses – pulmonary involvement
production of 2-aminoacetophenone. o Humans – fatal, begin as ulcer of skin and mucous
 Serrated confluent growth on CETRIMIDE AGAR – can grow in the presence of membrane followed by lymphadenitis and sepsis.
cetrimide (cetyltrimethylammonium bromide), a toxic substance that inhibits the  Potential bioterrorism agent.
growth of many bacteria.  NON-MOTILE; non-pigmented colonies on BAP
Note:
Burkholderia  Common name: WHITTMORE’S BACILLUS
 Cetrimide agar is a differential and selective medium that also enhances the pigment production
pseudomallei  Causes MELLIOIDOSIS/VIETNAMESE TIME BOMB DISEASE or
(pyoverdin and pyocyanin) of P. aeruginosa creating yellow-green colonies to blue-green colonies. Glander’s like infection.
 Pseudomonas species produce serum or a “floating layer” on the uppermost layer and sides of the  Motile with a polar tuft of flagella, with optimum growth at
thioglycolate tube. 42°C
 Some isolates of have wrinkled-looking colonies.  Isolated from muddy soil and rice paddies
 SELLERS AGAR promotes pigment production.  Microscopy shows presence of BIPOLAR BODIES
Pseudomonas fluorescens & Pseudomonas putida  BAP: smooth, orange colonies with mucoid-like appearance
 Isolated from contaminated blood products, cosmetics, hospital equipment, urine, on young growth.
and respiratory specimens; have been linked to transfusion-associated septicemia.  ASHDOWN medium with colistin: dry, wrinkled, deep pink or
 Produce pyoverdin, but NEITHER produces pyocyanin nor grows at 42° C. purple colonies with "earthy or soil-like odor.”
 Can produce acid from xylose: (+) xylose fermentation
 Differential test: (+) Gelatin hydrolysis for P. fluorescens ACINETOBACTER
 A member of the family Moraxellaceae.
BURKHOLDERIA  SECOND most commonly isolated, non-fermentative.Gram-negative bacilli.
 Belongs to the family Burkholderiacea.  Obligate aerobe and non-motile organism.
 Obligate aerobe and is motile by polar flagella EXCEPT B. mallei.  May be mistaken microscopically as a member of the genus Neisseria
 Generally non-pathogenic to healty individuals but may contribute to the severity of  Oxidase (-), catalase (+), LDC (+), growth may also be observed at 42°C.
infections when persistently exposed especially to the immunocompromised  Exhibits PURPLE colony on MAC; BLUE-GRAY (cornflower blue) colonies on EMB
individuals.  Related infections: Ventilator-associated pneumonia, endocarditis, meningitis,
 Acquired through contact with heavily contaminated medicai devices. nosocomial septicaemia, cellulitis, and UTI.
Burkholderia  Oxidase and LDC (+); Motile (lophotrichous flagella) Acinetobacter  Common isolate; glucose oxidizing, non-hemolytic
cepacia complex  YELLOW colonies on OFPBL (Oxidative Fermentative baumannii  Alarming cause of nosocomial infection globally due to its multi-
Polymixin B Bacitracin Lactose) antibiotic resistant property.
 BAP: Non-wrinkled yellow or yellow-green color with A. Iwoffii  Non-glucose oxidizing/asaccharolytic, non-hemolyic
“EARTHY/SOIL/DIRT-LIKE" odor. A.haemolyticus  Non-glucose oxidizing, BETA-HEMOLYIC
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CLINICAL BACTERIOLOGY – LECTURE Batch 2024
Stenotrophomonas maltophilia  Non-lactose fermenter on MAC and can grow at 42°C
 Belongs to the family Xanthomonadaceae.  Catalase (+); variable oxidase reaction since violet
 THIRD most commonly isolated non-fermentative, Gram-negative bacillus. pigment, violacein, interfere with oxidase test.
 Causes wound infection (farming equipment); contaminants in phlebotomy equipment  Gold standard for Quorum sensing study
 Strictly aerobic and motile, and it can grow at 42°C Psychrobacter  Have diplococcus-like morphology
 Oxidase (-); catalase, esculin, gelatinase, LDC and DNAse (+)  Have been isolated from processed meat, poultry products,
 Weak oxidizer of glucose; STRONG OXIDIZER OF MALTOSE and seafoods
 Colonies on BAP exhibit LAVENDER-GREEN to light purple pigmentation and distinct  Causes nosocomial eye infection among neonates.
"AMMONIACAL ODOR."; blue colored colonies on MAC  Aerobic, nonmotile, asaccharolytic, grow best between 5°
to 25° C ("cold-loving" bacteria)
Differential Characteristics of Pseudomonas, Acinetobacter, S. maltophilia
 Oxidase and nitrate reduction (+)
ORGANISM MOTILITY OXIDASE CATALASE LDC
 Can grow on gonococcal media such as TMA
Pseudomonas + + + -
 Characteristic “ROSE-PETAL” odor on BAP
Acinetobacter - - + - o P.immobilis: isolated from immunocompromised
S. maltophilia + - + + patients (AIDS) and those with fatal sepsis.
OTHER NON-FERMENTATIVE GRAM-NEGATIVE BACILLI Shewanella  Aerobic, motile, nonhalophilic; and it is a strong H2S
Alacaligenes faecalis  Oxidase and catalase (+); grows in 6.5% NaCl; nitrate putrefaciens producer
reduction and urease (-)  TSI: K/K, H2S (+); Oxidase (+)
 Asaccharolytic: O/F Test: (-/-)  Colonies on BAP appear mucoid and tan producing
 Grows well on MAC and motile by peritrichous flagella. greenish discoloration of the medium.
 With a "fruity odor" similar to APPLE or STRAWBERRY  Associated with food spoilage, mostly marine products with
 Related infections: UTI, wound, diarrhea a characteristic "sulfur-like odor or foul odor of fish."
Oligella  Aerobic rod that may appear coccoid in shape  Isolated from humans with ocular infections, otitis media,
 May colonize the distal urethra and cause a serious and and septicemia.
active infection. Elizabethkingia  Environmental organism isolated from soil, water, saline
 Catalase, oxidase, and PAD (+); gas producer; and meningoseptica solution, as well as from medical equipment.
assaccharolytic; MacConkey (-)  Aerobic, non-motile, and it ferments glucose, maltose, and
o O. urethralis - non-motile; oxidase (+) and urease (-) mannitol.
o O. ureolytica - motile; oxidase and urease (+)  Associated with meningitis and septicemia in premature
Moraxella lacunata  Agent of BLEPHAROCONJUNCTIVITIS or ANGULAR neonates and also in adults.
(Morax-Axenfeld CONJUNCTIVITIS.  Primary risk factor: Prematurity
bacillus)  Catalase, oxidase, nitrate reduction (+)  Has been isolated as a "contaminant" in tissue allograft
 Assacharolytic; MacConkey (-)  Colonies on BAP are typically circular with shiny
Chromobacterium  Opportunistic pathogen that causes neutrophil deficiency to appearance; No growth on MAC.
violaceum immunocompromised patients.  Oxidase, indole, gelatin hydrolysis, DNAse, esculin
 Colonies on BAP appear very dark purple or violet hydrolysis (+)
pigmentation (violacein) with AMMONIUM CYANIDE odor.  Produces YELLOW pigment - flavin

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