GRAM NEGATIVE

BACTERIA PART 1
BRIAN SAMEER C. BAILE, RMT, MMPA, MMHA
2
 3

• Large, diverse group of non-spore-forming bacteria

• Wide range of habitats – large intestines (enteric),
zoonotic, respiratory, soil, water
• Most are not medically important; some are true
pathogens, some are opportunists.
• All have a lipopolysaccharide outer membrane of cell
wall – endotoxin.
4
Aerobic Gram-Negative Nonenteric
Bacilli 5

• Pseudomonas and Burkholderia – an opportunistic

pathogen
• Brucella and Francisella – zoonotic pathogens
• Bordetella and Legionella – mainly human pathogens
• Alcaligenes – opportunistic pathogen
Pseudomonas: The Pseudomonads 6
• Small Gram-negative rods with a single polar
flagellum
• Free living
• primarily in soil, sea water, and fresh water; also
colonize plants and animals
• Important decomposers and bioremediators
• Frequent contaminants in homes and clinical
settings
• Use aerobic respiration; do not ferment
carbohydrates
• Produce oxidase and catalase
• Many produce water soluble pigments.
Pseudomonas: The Pseudomonads 7
Pseudomonas aeruginosa 8

• Common inhabitant of soil and water

• Intestinal resident in 10% normal people
• Resistant to soaps, dyes, quaternary ammonium
disinfectants, drugs, drying
• Frequent contaminant of ventilators, IV solutions,
anesthesia equipment
• Opportunistic pathogen
Pseudomonas aeruginosa 9
• OBLIGATE AEROBE: MAINLY AN OXIDATIVE ORGANISM
• PIGMENTS: PYOCYANIN, PYOVERDIN, FLUORESCEIN
• OXIDASE POSITIVE; CATALASE POSITIVE
• REDUCE NITRATE
• BETA LACTAMASE (DRUG RESISTANT)
• BLUE GREEN PUS (CELLULITIS)
• BURN
• ECTHYMA GANGRENOSUM (SKIN LESION)
• OTITIS EXTERNA
• UTI
• CYSTIC FIBROSIS
Pseudomonas aeruginosa 10
• Common cause of nosocomial infections in hosts with
burns, neoplastic disease, cystic fibrosis
• Complications include pneumonia, UTI, abscesses, otitis,
and corneal disease
• Endocarditis, meningitis, bronchopneumonia, SEPTICIMIA
• GRAPE, CORN, TORTILLA LIKE ODOR
• Greenish-blue pigment PYOCYANIN, PYOVERDIN,
FLUORESCEIN
• Multidrug resistant
• Cephalosporins, aminoglycosides, carbenicillin,
polymixin, quinolones, and monobactams
Pseudomonas aeruginosa 11
• BAP: FLAT, BETA HEMOLYTIC COL. WITH GROUND GLASS
APPEARANCE
• SELLER’S: GREEN/GREEN OR GREEN/BLUE
• GROWTH AT 42 º C
12
13
Other Gram-Negative Aerobic Rods 14

• Genera Burkholderia, Acinetobacter,

Stenotrophomonas
• Similar to pseudomonads
• Wide variety of habitats in soil, water, and
related environments
• Obligate aerobes; do not ferment sugars
• Motile, oxidase positive
• Opportunistic
Burkholderia 15
• Burkholderia cepacia – active
in biodegradation of a variety
of substances; opportunistic
agent in respiratory tract,
urinary tract, and
occasionally skin infections;
drug resistant
• CAUSES CYSTIC FIBROSIS,
ENDOCARDITIS, SEPTICEMIA
AND WOUND INFECTION
• PIGMENT: YELLOW OR NEON
YELLOW
Burkholderia 16

• B. pseudomallei – generally
acquired through penetrating
injury or inhalation from
environmental reservoir;
wound infections, bronchitis
and pneumonia, septicemia
• MOTILE WITH LOPHOTRICHOUS
FLAGELLA
• CAUSES MELIOIDOSIS:
SEPTICIMAL GLANDER’S LIKE
DISEASE
Burkholderia 17

• B. mallei
• NON-MOTILE
• CAUSE GLANDER’S DISEASE
• (STRAUS TEST POSITIVE)
Acinetobacter 18

• OXIDASE NEGATIVE, CATALASE POSITIVE

• NON-MOTILE
• SACCHAROLYTIC
• Acinetobacter baumanii – nosocomial and community
acquired infections; wounds, lungs, urinary tract,
burns, blood; extremely resistant – treatment with
combination antimicrobials
• SELLER’S: B/G WITH YELLOW BAND
• O-F POSITIVE
Acinetobacter 19

• Acinetobacter lwoffi (MIMA POLYMORPHA)

• SELLER’S : B/G WITHOUT YELLOW BAND
• O-F NEGATIVE
Stenotrophomonas 20
• Stenotrophomonas maltophilia – forms biofilms;
contaminant of disinfectants dialysis equipment,
respiratory equipment, water dispensers, and
catheters; clinical isolate in respiratory soft
tissue, blood ,CSF; high resistance to multidrugs
• MOTILE WITH LOPHOTRICHOUS FLAGELLA
• OXIDASE NEGATIVE
• DNAse POSITIVE
• OXIDIZE GLUCOSE AND MALTOSE
• BAP: ROUGH, LAVENDER GREEN
• MALTOSE LOVING
• PNEUMONIA AND ENDOCARDITIS
Brucella and Brucellosis 21

• Tiny Gram-negative coccobacilli

• 2 species:
• Brucella abortus (cattle)
• Brucella suis (pigs)
• Brucellosis, malta fever, undulant fever, and Bang disease – a
zoonosis transmitted to humans from infected animals
• Fluctuating pattern of fever –weeks to a year
• Combination of tetracycline and rifampin or streptomycin
• Animal vaccine available
• Potential bioweapon
Brucella and Brucellosis 22

• 3-4 WEEKS BEOFRE DISCARDED AS NEGATIVE

• REQUIRES THIONINE AND BASIC FUCHSIN (B. melitensis)
• OBLIGATE INTRACELLULAR ORGANISM
• CAPNOPHILIC
• CAUSE ABORTION IN:
• 1. COW: B. abortus 3. DOGS: B. canis
• 2. SWINE: B. suis 4. SHEEPS AND GOATS: B. melitensis
• BLOOD CULTURE: CASTANEDA, BRUCELLA BROTH
• ABORTUS “BANG” RINGPROBE TEST-COLORED RING
• SKIN TEST: BRUCELLIN TEST
Brucella and Brucellosis 23
Francisella tularensis and Tularemia 24

• Causes tularemia, a zoonotic disease of mammals

endemic to the northern hemisphere, particularly
rabbits
• Transmitted by contact with infected animals,
water and dust or bites by vectors
• Headache, backache, fever, chills, malaise and
weakness
• 10% death rate in systemic and pulmonic forms
• Intracellular persistence can lead to relapse
• gentamicin or tetracycline
• Attenuated vaccine
• Potential bioterrorism agent
Francisella tularensis and Tularemia 25

• LYMPHADENOPATHY
• CONTACT IN ANIMAL THROUGH INHALATION
• REQUIRES CYSTEINE FOR GROWTH
• NON-MOTILE, ENCAPSULATED
• INTRACELLULAR BACTERIA (RESIDES IN R-E-C)
• MEDIA: GCBA: GLUCOSE CYSTEIN BLOOD AGAR
• PCA: PEPTONE CYSTEIN AGAR
• CHA: CYSTEINE HEART AGAR
Francisella tularensis and Tularemia 26
Bordetella pertussis 27

• Minute, encapsulated coccobacillus

• Causes pertussis or whooping cough, a
communicable childhood affliction
• Acute respiratory syndrome
• Often severe, life-threatening complications in
babies
• Reservoir – apparently healthy carriers
• Transmission by direct contact or inhalation of
aerosols
Bordetella pertussis 28

• Virulence factors
• receptors that recognize and bind to ciliated respiratory
epithelial cells
• toxins that destroy and dislodge ciliated cells
• Loss of ciliary mechanism leads to buildup of mucus
and blockage of the airways.
• Vaccine – DTaP- acellular vaccine contains toxoid and
other Ags
Bordetella pertussis 29
• OXIDASE POSITIVE; UREASE NEGATIVE
• BIPOLAR METACHROMATIC GRANULES (TOLUIDINE BLUE)
• WHOOPING COUGH (PERTUSIS)
• SPECIMEN: NASOPHARYNGEAL SWAB
• DIAGNOSIS: PCR (BEST) CULTURE
• CULTURE MEDIUM:
• BORDET-GENGOU(POTATO BLOOD GLYCEROL AGAR) “MERCURY
DROPS”
• REGAN LOWE
• JONES KENDRICK CHARCOAL AGAR
• STEINER-SCHOLTE AGAR
• NASOPHARYNGEAL ASPIRATE
• TRANSPORT MEDIUM: CASEIN MEDIUM AND CAUSAMINO ACID
BROTH
30
Bordetella spp. 31
• Bordetella parapertusis
• OXIDASE NEGATIVE; UREASE POSITIVE

• Bordetella bronchiseptica
• MOTILE, OXIDASE POSITIVE, UREASE POSITIVE
Alcaligenes 32
• Live primarily in soil and water
• May become normal flora
• A. faecalis (ODORANS) – most common clinical species
• isolated from feces, sputum, and urine
• occasionally associated with opportunistic infections –
pneumonia, septicemia, and meningitis
• “FRESH APPLE ODOR”
• OXIDASE POSITIVE, CATALASE POSITIVE
• MOTILE WITH PERITRICHOUS FLAGELLA
• BAP: GREEN ZONE
• SELLERS: B/B
Legionella pneumophila and 33
Legionellosis
• Widely distributed in water
• Live in close association with amebas
• 1976 epidemic of pneumonia afflicted 200
American Legion members attending a
convention in Philadelphia and killed 29
• Legionnaires disease and Pontiac fever
• Prevalent in males over 50
• Nosocomial disease in elderly patients
• Fever, cough, diarrhea, abdominal pain,
pneumonia fatality rate of 3-30%
• Azithromycin
Legionella pneumophila and 34
Legionellosis
• PLEOMORPHIC, WEAKLY GRAM NEGATIVE ROD
• REQUIRES “CYSTEINE” AND IRON FOR GROWTH
• AIR CONDITIONING (AEROSOL), WATER COOLER TANKS, RIVERS
• FACULTATIVE INTRACELLULAR PATHOGEN
• DISEASE: LEGIONNAIRE’S DISEASE(ATYPICAL PNEUMONIA)
• “CUT GLASS” COLONY
• BROAD STREET PNEUMONIA
• PONTIAC FEVER
• DIAGNOSIS: DIETERLE STAIN: POSITIVE
• CULTURE: BUFFER CHARCOAL YEAST EXTRACT (BCYE) WITH ALPHA
KETO GLUTARATE
• FEELY GORMAN MEDIUM
35
Enterobacteriaceae Family 36

• Enterics
• Large family of small, non-spore-forming Gram-negative
rods
• Many members inhabit soil, water, decaying matter, and
are common occupants of large bowel of animals
including humans.
• Most frequent cause of diarrhea through enterotoxins
• Enterics, along with Pseudomonas sp., account for
almost 50% of nosocomial infections.
Enterobacteriaceae Family 37

• OXIDASE NEGATIVE; CATALASE POSITIVE

• REDUCE NITRATE (NO3) TO NITRITE (NO2)
• GRAM NEGATIVE RODS IN SINGLY
• MOTILE: PERITRICHOUS FLAGELLA (EXCEPT: K. pneumonia
AND SHIGELLA SPP)
• NON ENCAPSULATED (EXCEPT: K. pneumonia)
• FERMENTS GLUCOSE
38
Enterobacteriaceae Family 39

• Facultative anaerobes, grow best in air

• All ferment glucose, reduce nitrates to nitrites, oxidase
negative, and catalase positive.
• Divided into coliforms (lactose fermenters) and non-
coliforms (non-lactose fermenters)
• Enrichment, selective and differential media utilized for
screening samples for pathogens
Enterobacteriaceae Family 40

• OXIDASE NEGATIVE; CATALASE POSITIVE

• REDUCE NITRATE (NO3) TO NITRITE (NO2)
• GRAM NEGATIVE RODS IN SINGLY
• MOTILE: PERITRICHOUS FLAGELLA (EXCEPT: K. pneumonia
AND SHIGELLA SPP)
• NON ENCAPSULATED (EXCEPT: K. pneumonia)
• FERMENTS GLUCOSE
Enterobacteriaceae Family 41

• MAJOR GROUPS:
• LACTOSE FERMENTERS:
• RAPID LACTOSE FERMENTERS: DUE TO BETA
GALACTOSIDASE AND PERMEASE
• E. coli
• KLEBSIELLA
• ENTEROBACTER
• SLOW LACTOSE FERMENTERS/LATE LACTOSE FERMENTERS
(LLF) ONLY BETA GALACTOSIDASE IS PRESENT
• ARIZONA SP., CITROBACTER, SERRATIA AND S. sonnei
Enterobacteriaceae Family 42

• SPECIMEN: STOOL/RECTAL SWAB

• CULTURE: BILE LACTOSE MEDIUM (EMB, MacCONKEY,
ENDO AGAR)
• BIOCHEMICAL MEDIA: TSI, LIA, UREA, MRVP, SIM)
• AUTOMATED: CRYSTAL ID, API 20E, VITEK
• SEROLOGICAL ID: DETECTS “O” Ag (SOMATIC)
• “Vi” Ag (VIRULENCE)
• “H” Ag (FLAGELLAR)
• “K” Ag (CAPSULAR)
43
44
Antigenic Structures and Virulence
45
Factors

Complex surface antigens contribute to pathogenicity

and trigger immune response:
• H – flagellar Ag
• K – capsule and/or fimbrial Ag
• O – somatic or cell wall Ag – all have
• Endotoxin
• Exotoxins
46
Coliform Organisms and 47
Diseases
 Escherichia coli 48

• Pathogenic strains frequent agents of infantile diarrhea –

greatest cause of mortality among babies
• Causes ~70% of traveler’s diarrhea
• Causes 50-80% UTI
• Coliform count – indicator of fecal contamination in water
Escherichia coli: The Most Prevalent
Enteric Bacillus 49

• Most common aerobic and non-fastidious bacterium in

gut
• 150 strains
• Some have developed virulence through plasmid
transfer, others are opportunists.
Escherichia coli: The Most Prevalent
Enteric Bacillus 50

• GREENISH-METALLIC SHEEN ON EMB & ENDO AGAR

• #1 FOR UTI/CYSTITIS
• PILI, ADHESINS (VIRULENCE)
• NOSOCOMIAL INFECTION
• NEONATAL MENINGITIS
• SEPTICEMIA
Pathogenic Strains of E. coli 51
• Enterotoxigenic E. coli causes severe diarrhea due to
heat-labile toxin and heat-stable toxin – stimulate
secretion and fluid loss; also has fimbriae;
TRAVELLER’S DIARRHEA
• Enteroinvasive E. coli causes inflammatory disease of
the large intestine. SERENY TEST POSITIVE
• Enteropathogenic E. coli linked to wasting form
infantile diarrhea
• Enterohemorrhagic E. coli, O157:H7 strain, causes
hemorrhagic syndrome and kidney damage; ID 100
cells
Escherichia coli 52

• Pathogenic strains frequent agents of infantile diarrhea –

greatest cause of mortality among babies
• Causes ~70% of traveler’s diarrhea
• Causes 50-80% UTI
• Coliform count - indicator of fecal contamination in water
 Other Coliforms 53

Clinically important mainly as opportunists

• Klebsiella pneumoniae– normal inhabitant of respiratory
tract, has large capsule, cause of nosocomial pneumonia,
meningitis, bacteremia, wound infections and UTIs
• ENCAPSULATED
• COMMUNITY ACQUIRED PNEUMONIA
• UTI
• WOUND INFECTION
• INFANTILE ENTERITIS
 Other Coliforms 54

• Klebsiella oxytoca:
• INDOLE TEST POSITIVE

• Klebsiella ozonae
• CAUSE DISEASE OF NASAL MUCOSA “CHRONIC ATROPHIC
RHINITIS”

• Klebsiella rhinoscleromatis
• NOSE & PHARYNX “RHINOSCLEROMA DISEASE”
 KLEBSIELLA 55

• CULTURE: EMB, MacCONKEY, XLD

• BIOCHEMICAL TEST: TSI: A/A+GAS;
• IMViC: -,-,+,+
• UREASE: +
• SIM: -,-,-
• KCN: +
• NEUFELD QUELLUNG REACTION: POSITIVE
• STRING’S TEST: POSITIVE +3% KOH
 Other Coliforms 56

Clinically important mainly as opportunists

• Enterobacter sp. – UTIs, surgical wounds
• AEROBACTER
• TSI: A/A+GAS; IMViC: +,+,-,-; UREASE :(-) SIM: -,-,+; KCN:+
• E. aerogenes: LDC(+); UREASE(-)
• E. sakazakii: YELLOW COLONIES; LDC: (+)
• E. agglomerans: YELLOW COLONIES; LDC (-)
• E. gergoviae: UREASE: (+)
• E. alvei (Haffnia alvei): CITRATE: (-), NLF
 Other Coliforms 57

• SERRATIA
• DNAse, LIPASE, GELATINASE
• RESISTANT TO COLISTIN AND CEPHALOTIN
• Serratia marcescens – produces a red pigment; causes
pneumonia, burn and wound infections, septicemia and
meningitis
• UREASE PRODUCER (K-E-S) GROUP
• NON LACTOSE FERMENTER (TSI: K/A)
• PRODIGIOSIN: (RED PIGMENT)
• ARABINOSE NEGATIVE
 Other Coliforms 58

• Serritia liquifaciens
• PINK TO RED PIGMENT
• KCN, GELATIN, ARABINOSE POSITIVE
Other Coliforms 59
• CITROBACTER: ASSOC. W/GASTROENTERITIS, UTI,
NEONATAL MENINGITIS, BRAIN ABSCESS, DIARRHEA
• LATE LACTOSE FERMENTER: ONPG POSITIVE (YELLOW)
• TSI: A/A +GAS+H2S
• LYSINE DECARBOXYLASE: (LDC) NEGATIVE
• KCN: POSITIVE
• BIOCHEMICAL SAME AS SALMONELLA, MORPHOLOGY SAME
AS E. coli
• C. freundii: INDOLE (-); H2S(+); KCN(+); MALONATE (+)
• C. diversus: INDOLE (+); H2S(-) KCN(-); MALONATE (+/-)
• C. amalonaticus: INDOLE (+); H2S(-), KCN(+), MALONATE(-)
Citrobacter 60
Other Coliforms 61
• Arizona hinshawii
• LATE LACTOSE FERMENTER: ONPG POSITIVE (YELLOW
• GROWS ON NA MALONATE
• GELATIN LIQUIFACTION (+)
• LYSINE DECARBOXYLASE
• CAUSE GASTROENTERITIS
• INTESTINAL CONTENT (REPTILES)
Other Coliforms 62
• Edwardsiella tarda
• NON LACTOSE FERMENTER: TSI: K/A+GAS+H2S
• LYSINE DECARBOXYLASE POSITIVE
• IMViC: +,+,-,-
ENTEROBACTERIACEAE 63
Noncoliform Lactose-Negative 64
Enterics
• Proteus, Morganella, Providencia
• Salmonella and Shigella
Opportunists: Proteus and Its
65
Relatives
Proteus, Morganella, Providencia – ordinarily
harmless saprobes in soil, manure, sewage,
polluted water, commensals of humans and
animals
• Proteus sp. - swarm on surface of moist agar in a
concentric pattern
• involved in UTI, wound infections, pneumonia,
septicemia, and infant diarrhea
• Morganella morganii and Providencia sp. involved in
similar infections
• All demonstrate resistance to several
antimicrobials.
Opportunists: Proteus and Its
66
Relatives
• PROTEAE: PROTEUS
• UTI, SEPTICIMIA, STAG-HORN CALCULI
• UREASE (FUSCHIA PINK) AND PAD (GREEN) POSITIVE
• DEAMINATES LYSINE: LIA: R/A
• NON LACTOSE FERMENTERS: TSI: K/A+H2S+GAS
• BURNT GUN POWDER
• SWARMING MOTILITY
• DIENES PHENOMENON
• GELATINASE (+)
Opportunists: Proteus and Its
Relatives 67

• CULTURE: EMB: COLORLESS COL.

• MAC: COLORLESS
• VLD: COLORLESS
• BAP: SWARMING PHENOMENON,
GRAYISH COLONY
68
 Opportunists: Proteus and Its
Relatives 69
• SPECIES:
• Proteus vulgaris:
• INDOLE POSITIVE (SOURCE OF WEIL FELIX
ANTIGEN)
• Proteus mirabilis:
• INDOLE NEGATIVE
• Providencia rettgeri: TSI: K/A+GAS;
CITRATE POSITIVE
• Providencia stuartii: UREASE NEGATIVE
• Morganella morgani: TSI: K/A+GAS;
CITRATE NEGATIVE
Salmonella and Shigella 70

• Well-developed virulence factors, primary

pathogens, not normal human flora
• Salmonelloses and Shigelloses
• some gastrointestinal involvement and diarrhea but often
affect other systems
Salmonella 71

• “Vi”, “O”, “H” ANTIGENS

• NON LACTOSE FERMENTERS (TSI: K/A+H2S+GAS)
• MOTILE: EXCEPT S. pullorum & S. gallinarum
• AEROGENIC: EXCEPT S. gallinarum & S. typhi
• OSTEOMYELITIS IN SICKLE CELL DISEASE
Salmonella: SICKLE CELL ANEMIA 72
Salmonella 73

• MEMBERS:
• S. typhi (S. typhosa): “Vi”Ag is exclusive
• EBERTH’S BACILLUS
• S. cholera suis
• S. enteritidis
• BIOTYPE: A. S. paratyphi A,B,C D. S. gallinarum
• B. S. sendari E. S. pullorum
• C. S. typhimurium F. S. darby
Typhoid Fever and Other Salmonelloses74

• Salmonella typhi – most serious pathogen of the genus;

cause of TYPHOID FEVER; human host
• S. cholerae-suis – zoonosis of swine; SEPTICEMIA
• S. enteritidis – includes 1,700 different serotypes based
on variation on O, H, and capsular antigen, STOMACH
FLU; GASTROENTERITIS
• S. paratyphi A,B,C: paratyphoid fever
• Flagellated; ferments glucose
• Resistant to chemicals –bile and dyes
Typhoid Fever and Other Salmonelloses75

• SPECIMEN: BLOOD: 1ST WEEK OF INFECTION

• URINE & STOOL: 2ND TO 3RD WEEK ONWARDS
• CULTURE: MacCONKEY, EMB, SSA, HEA, XLD
• SEROTYPING: (SPECIES IDENTIFICATION)
Typhoid Fever 76

• Bacillus enters with ingestion of fecally

contaminated food or water; occasionally spread
by close personal contact; ID 1,000-10,000 cells
• Asymptomatic carriers; some chronic carriers
shed bacilli from gallbladder
• Bacilli adhere to small intestine, cause invasive
diarrhea that leads to septicemia
• Treat chronic infections with chloramphenicol or
sulfa-trimethoprim
• 2 vaccines for temporary protection
77
 Animal Salmonelloses 78

• Salmonelloses other than typhoid fever are

called enteric fevers, Salmonella food
poisoning, and gastroenteritis.
• Usually less severe than typhoid fever but
more prevalent
• Caused by one of many serotypes of
Salmonella enteritidis; all zoonotic in origin
but humans can become carriers
• cattle, poultry, rodents, reptiles, animal and
dairy products
• fomites contaminated with animal intestinal flora
Shigella and Bacillary Dysentery 79

• Shigellosis – incapacitating dysentery

• S. dysenteriae, S. sonnei, S. flexneri and S.
boydii
• Human parasites
• Invades villus of large intestine, can perforate
intestine or invade blood
• Enters Peyer’s patches instigate inflammatory
response; endotoxin and exotoxins
• Treatment – fluid replacement and ciprofloxacin
and sulfa-trimethoprim
Shigella and Bacillary Dysentery 80

• NON MOTILE
• AEROGENIC
• CAUSE BACILLARY DYSENTERY (INVASIVE)
• NON LACTOSE FERMENTERS (EXCEPT S.sonnei)
• TSI: K/A
• LYSINE DECARBOXYLASE NEGATIVE: LIA: K/A
• IMViC VARIABLE (SEROTYPING)
Shigella and Bacillary Dysentery 81

• SPECIES:
• S. dysenteriae (S. shiga): GROUP A
• S. flexnerri (STRONG’S BACILLUS): GROUP B
• S. boydii (S. ambigua): GROUP C
• S. sonnei: (COMMON ISOLATES) GROUP D
82
 The Enteric Yersinia Pathogens 83

• Yersinia enterocolitica – domestic and wild animals, fish,

fruits, vegetables, and water
• bacteria enter small intestinal mucosa, some enter lymphatic and
survive in phagocytes; inflammation of ileum can mimic
appendicitis
• MOTILE AT 22 º C (COLD GROWTH)
• ZOONOTIC, UNPASTEURIZED MILK
• “BULL’S EYE” COLONIES ON CIN (CEFSULODIN IRGASAN
NOVOBIOCIN)
• ENTEROCOLITIS
• Y. pseudotuberculosis – infection similar to
Y. enterocolitica, more lymph node
inflammation
ENTEROCOLITIS 84
Nonenteric Yersinia pestis and
Plague 85
• MICROAEROPHILIC
• NON MOTILE, GRAM (-) COCCOBACILLI
• SAFETY PIN APPEARANCE “WAYSON” STAIN
• V&W ANTIGENS
• FACULTATIVE INTRACELLULAR PARASITE
• WILD RODENTS FLEA BITE: SYLVATIC PLAGUE
• RESPIRATORY DROPLETS (HUMAN)
• DISEASE: PNEUMONIC PLAGUE, BUBONIC PLAGUE, BLACK DEATH
• STALACTITE STREAMERS IN OLD BROTH
• Tiny, Gram-negative rod, unusual bipolar staining and capsules
• Virulence factors – capsular and envelope proteins protect against
phagocytosis and foster intracellular growth
• coagulase, endotoxin, murine toxin
86
Yersinia pestis 87

• Humans develop plague through contact with

wild animals (sylvatic plague) or domestic or
semidomestic animals (urban plague) or
infected humans.
• Found in 200 species of mammals – rodents,
without causing disease
• Flea vectors – bacteria replicates in gut,
coagulase causes blood clotting that blocks
the esophagus; flea becomes ravenous
88
Pathology of Plague 89

• ID 3-50 bacilli
• Bubonic – bacillus multiplies in flea bite, enters
lymph, causes necrosis and swelling called a bubo
in groin or axilla
• Septicemic – progression to massive bacterial
growth; virulence factors cause intravascular
coagulation subcutaneous hemorrhage and purpura
– black plague
• Pneumonic – infection localized to lungs, highly
contagious; fatal without treatment
 90

• Diagnosis depends on history, symptoms, and lab

findings from aspiration of buboes.
• Treatment: streptomycin, tetracycline or
chloramphenicol
• Killed or attenuated vaccine available
• Prevention by quarantine and control of rodent
population in human habitats
BUBONIC PLAGUE 91
Oxidase-Positive Nonenteric
Pathogens 92

• Pasteurella multocida
• Haemophilus influenzae
• H. aegyptius
• H. ducreyi
• H. parainfluenzae
• H. aphrophilus
Pasteurella multocida 93

• Zoonotic genus; normal flora in animals

• Opportunistic infections
• Animal bites or scratches cause local abscess that
can spread to joints, bones, and lymph nodes.
• Immunocompromised are at risk for septicemia and
complications.
• Treatment: penicillin and tetracycline
Pasteurella multocida 94
• SMALL GRAM NEGATIVE RODS WITH CAPSULE
• OXIDASE & CATALASE POSITIVE
• NON-MOTILE
• ANIMAL BITES (CATS AND DOGS)
• CELLULITIS WITH LYMPHADENITIS (WOUND)
• PNEUMONIA, BRONCHITIS, SINUSITIS, PULMONARY ABSCESS
• GRAM (-) ROD THAT GROWS ON BLOOD AGAR ONLY
• INDOLE & ONPG POSITIVE
• ABLE TO REDUCE N03
• MOUSY ODOR AND GRAY COLONY; MacCONKEY (-)
• UREASE (-)
Pasteurella multocida 95
Haemophilus 96

• Tiny Gram-negative pleomorphic rods

• Fastidious, sensitive to drying, temperature
extremes, and disinfectants
• None can grow on blood agar without special
techniques – chocolate agar.
• Require hemin, NAD or NADP
• Some species are normal colonists of upper
respiratory tract or vagina (H. aegyptius, H.
parainfluenzae, H ducreyi).
• Others are virulent species responsible for
conjunctivitis, childhood meningitis, and
chancroid.
Haemophilus 97

• BLOOD LOVING
• REQUIRES X & V FACTOR EXCEPT:
• H. ducreyi: X FACTOR ONLY
• H. parainfluenza: V FACTOR ONLY
• H. paraphrophilus: V FACTOR ONLY (SBE)
• PORPHYRIN (ALA) TEST (X FACTOR): RED COLOR (=)
Haemophilus 98
• H. influenzae (PFEIFFER’S BACILLUS)
• acute bacterial meningitis, epiglottitis, otitis media, sinusitis,
pneumonia, and bronchitis
• subunit vaccine Hib
• POLYRIBITOL CAPSULE: IgA PROTEASE
• MOUSY ODOR
• X (HEMIN) & V (NICOTINAMIDE ADENINE DINUCLEOTIDE) FACTOR
• NASOPHARYNX
• OTITIS MEDIA (MAJOR CAUSE)
• BRONCHITIS
• PNEUMONIA
• EPIGLOTTIS
• B-LACTAMASE
• NO GROWTH ON MacCONKEY
Haemophilus 99
• MENINGITIS: (TYPE b H. influenza)
• LATEX AGGLUTINATION TEST (ANTIGEN SCREEN ON CSF)
• SEROTYPE BY QUELLUNG TEST
• CULTURE MEDIA: BAP WITH S. aureus “SATELLITE
PHENOMENON”
• CAP: (X&V)
• FILDES ENRICHMENT
• LEVINTHAL AGAR
Haemophilus 100

• H. ducreyi – chancroid STD

• CAUSE “SOFT CHANCRE” (CHANCROID) : SKIN LESION,
DISCHARGES, SWAB
• SCHOOL OF RED FISH
• GROWTH ON CAP + VANCOMYCIN
Haemophilus 101

• H. aegypticus –
conjunctivitis, pink eye
• “KOCH WEEK’S BACILLUS”
• REQUIRES X&V FACTOR

• H. parainfluenzae and H.
aphrophilus – normal oral
and nasopharyngeal flora;
infective endocarditis
Haemophilus 102

• H. influenzae – acute bacterial meningitis, epiglottitis,

otitis media, sinusitis, pneumonia, and bronchitis
• subunit vaccine Hib
• H. aegyptius –conjunctivitis, pink eye
• H. ducreyi – chancroid STD
• H. parainfluenzae and H. aphrophilus – normal oral and
nasopharyngeal flora; infective endocarditis
Haemophilus 103
TO BE CONTINUED 104