LABORATORY AND
LECTURE NOTES
FOR MEDICAL
TECHNOLOGISTS
, lOMoARcPSD|58847208
FASTIDIOUS GRAM NEGATIVE RODS / - Virulence Factor: Polysaccharide capsule
BACILLI (type B) which is antigenic thus, it is
General Characteristics:
important to do serotyping of capsular
- Do not grow readily on lab media antigen
- Facultative anaerobes, others are aerobic or Differentiation Tests:
microaerophilic (withstand certain amount 1. ALA / Porphyrin Test
of oxygen 10-12% O2 not 21%) -uses delta aminolevulenic acid as
- Uncommon human infection: respiratory substrate to synthesize the heme factor.
diseases Porphyrins are the by product.
- Haemophilus, Francissella, Legionella, Porphyrins will fluoresce under UV light
Bordatella and Brucella and produce pink color.
- AACEK former HACEK (Haemophilus -more accurate test to determine X factor
parainfluenza, Aggregatibacter, requirement and differentiate the types of
Cardiobactreium, Eikenella and Kingella) haemophilus
causing SBE (subacute endocarditis) -disk or liquid / tube method
-liquid / tube method: add 0.5 ml Covax
HAEMOPHILUS reagent that will confirm the presence of
General Characteristics: porphyrins if they metabolize delta
aminolevulenic acid.
- most important member of this group
-Red color presents the metabolism of
- hemo - blood, philia - loving ALA
- facultative anaerobes, require enriched
medium with fresh blood or heme/NAD 2. Horse Blood Agar / Hemolysis Test
- X factor - heme | V factor - nad -it cannot hemolyse sheep’s blood-
- multiply slowly in culture and are stabbing is recommended to readily see
capnophilic (require 5-10% CO2) the hemolysis
-V factor: staph aureus / NAD, a coenzyme
- requires CO2 incubator in the lab to culture
that transfer electrons from one to
haemophilus in order to have base
another, found in
atmosphere
RBC
- pleomorphic - exist in many forms -X factor: blood / heme, use it to
- short, cocobacillary, have filaments, non- synthesize catalase, peroxidase and ETC
motile, gram negative
Page 1 of 12
, lOMoARcPSD|58847208
-haemophilus is more abundant and bigger - can produce 6 antigenic types of capsule
near V factor / staph aureus. They satellite (A-
around staph. E). Highly virulent is the type B
capsule
-satellitism uses clear medium (TCA or
BHI)- use strips impregnated with the X - latex agglutination test - detect rapidly the
factor, with V factor and XV factor capsular antigens
Haemophilus Identification Tests Table: - do capsular or sero typing esp. for children
(<15 yrs)
- Aegypticus - negative xylose
HAEMOPHILUS INFLUENZA
- known as Pfeiffer bacillus named by
Richard Friedrich Johannes Pleiffer,
discovered it in the great influenza
Influenzae and Aegypticus pandemic in 1892
- has same reaction that is why they need to - Do not really cause influenza - Flu-like
undergo other tests like glucose or symptoms
carbohydrate fermentation tests (glucose
and xylose) - Virulence: Pilli
- Characteristics: coccobacilli, thread-like - MOT: Person to person via droplets
rods, highly pleomorphic - Diseases: most influenza causes viral
- Colonies: round, small and convex, non- diseases, but specific for h.influenza is
hemolytic pneumonia, cellulitis, epiglottitis,
- has capsule that can be lost during empyema, septic arthritis.
subculture
- Common Manifestations: bronchitis and
pneumonia esp. adults
- Influenzae
- Treatment: Beta-lactam antibiotics if the
- produces acid (glucose and xylose) and not organism do not produce B-lactamase.
lactose or sucrose
Ceftriaxone, Fluroquinolones.
Page 2 of 12
, lOMoARcPSD|58847208
-
- Vaccine: consists of type B - resembles influenza similar in microscopy
polysaccharide. Conjugated Vaccine - and colonies
vaccine added with protein to make it - Blood cultures: large clumps of
complex and be antigenic. filamentous rods
- ALA + require only NAD / V factor
- carrier rate are higher (80-90%) for non- - non-hemolytic on HBA
typable forms / do not have capsule - produce acid and gas from glucose and
- carried in the URT 2-4% healthy sucrose but not lactose or xylose
individuals
- urease and catalase positive, indole
- non-encapsulate strain are not pathogenic negative
only those who are immunocompromised - Clinical Significance: URT normal
are infected microbiota
- Diseases: endocarditis, urethritis and URT,
HAEMOPHILUS AEGYPTICUS peritonitis, cholecystitis
- Diseases: conjunctivitis, bacteremia, - not a true pathogen
septicemia, eye infections
- isolated from blood cultures HAEMOPHILUS DUCREYI
- not respiratory only in the eyes - Characteristics:
Treatment: most strains are susceptible to - small gram negative coccobacilli, short
cephalosporins (cefrotaxime and cefriaxone chains, clumps or whorls within lesions.
- 3rd generation)
- railroad track, school of fish or fingerprint
arrangements.
HAEMOPHILUS
PARAHAEMOLYTICUS - - bipolar staining: both ends are stained
normal microbiota thicker
- immunocompromised individuals - Difficult to culture bec. the incubation day
- Diseases: endocarditis, pharyngitis is 10 days 33 degrees in 10% CO2, need
additional nutrients like isovitalex and Va
HAEMOPHILUS
PARAINFLUENZA - Chocolate Agar: yellow to gray dome-
shaped smooth colonies
Page 3 of 12
LECTURE NOTES
FOR MEDICAL
TECHNOLOGISTS
, lOMoARcPSD|58847208
FASTIDIOUS GRAM NEGATIVE RODS / - Virulence Factor: Polysaccharide capsule
BACILLI (type B) which is antigenic thus, it is
General Characteristics:
important to do serotyping of capsular
- Do not grow readily on lab media antigen
- Facultative anaerobes, others are aerobic or Differentiation Tests:
microaerophilic (withstand certain amount 1. ALA / Porphyrin Test
of oxygen 10-12% O2 not 21%) -uses delta aminolevulenic acid as
- Uncommon human infection: respiratory substrate to synthesize the heme factor.
diseases Porphyrins are the by product.
- Haemophilus, Francissella, Legionella, Porphyrins will fluoresce under UV light
Bordatella and Brucella and produce pink color.
- AACEK former HACEK (Haemophilus -more accurate test to determine X factor
parainfluenza, Aggregatibacter, requirement and differentiate the types of
Cardiobactreium, Eikenella and Kingella) haemophilus
causing SBE (subacute endocarditis) -disk or liquid / tube method
-liquid / tube method: add 0.5 ml Covax
HAEMOPHILUS reagent that will confirm the presence of
General Characteristics: porphyrins if they metabolize delta
aminolevulenic acid.
- most important member of this group
-Red color presents the metabolism of
- hemo - blood, philia - loving ALA
- facultative anaerobes, require enriched
medium with fresh blood or heme/NAD 2. Horse Blood Agar / Hemolysis Test
- X factor - heme | V factor - nad -it cannot hemolyse sheep’s blood-
- multiply slowly in culture and are stabbing is recommended to readily see
capnophilic (require 5-10% CO2) the hemolysis
-V factor: staph aureus / NAD, a coenzyme
- requires CO2 incubator in the lab to culture
that transfer electrons from one to
haemophilus in order to have base
another, found in
atmosphere
RBC
- pleomorphic - exist in many forms -X factor: blood / heme, use it to
- short, cocobacillary, have filaments, non- synthesize catalase, peroxidase and ETC
motile, gram negative
Page 1 of 12
, lOMoARcPSD|58847208
-haemophilus is more abundant and bigger - can produce 6 antigenic types of capsule
near V factor / staph aureus. They satellite (A-
around staph. E). Highly virulent is the type B
capsule
-satellitism uses clear medium (TCA or
BHI)- use strips impregnated with the X - latex agglutination test - detect rapidly the
factor, with V factor and XV factor capsular antigens
Haemophilus Identification Tests Table: - do capsular or sero typing esp. for children
(<15 yrs)
- Aegypticus - negative xylose
HAEMOPHILUS INFLUENZA
- known as Pfeiffer bacillus named by
Richard Friedrich Johannes Pleiffer,
discovered it in the great influenza
Influenzae and Aegypticus pandemic in 1892
- has same reaction that is why they need to - Do not really cause influenza - Flu-like
undergo other tests like glucose or symptoms
carbohydrate fermentation tests (glucose
and xylose) - Virulence: Pilli
- Characteristics: coccobacilli, thread-like - MOT: Person to person via droplets
rods, highly pleomorphic - Diseases: most influenza causes viral
- Colonies: round, small and convex, non- diseases, but specific for h.influenza is
hemolytic pneumonia, cellulitis, epiglottitis,
- has capsule that can be lost during empyema, septic arthritis.
subculture
- Common Manifestations: bronchitis and
pneumonia esp. adults
- Influenzae
- Treatment: Beta-lactam antibiotics if the
- produces acid (glucose and xylose) and not organism do not produce B-lactamase.
lactose or sucrose
Ceftriaxone, Fluroquinolones.
Page 2 of 12
, lOMoARcPSD|58847208
-
- Vaccine: consists of type B - resembles influenza similar in microscopy
polysaccharide. Conjugated Vaccine - and colonies
vaccine added with protein to make it - Blood cultures: large clumps of
complex and be antigenic. filamentous rods
- ALA + require only NAD / V factor
- carrier rate are higher (80-90%) for non- - non-hemolytic on HBA
typable forms / do not have capsule - produce acid and gas from glucose and
- carried in the URT 2-4% healthy sucrose but not lactose or xylose
individuals
- urease and catalase positive, indole
- non-encapsulate strain are not pathogenic negative
only those who are immunocompromised - Clinical Significance: URT normal
are infected microbiota
- Diseases: endocarditis, urethritis and URT,
HAEMOPHILUS AEGYPTICUS peritonitis, cholecystitis
- Diseases: conjunctivitis, bacteremia, - not a true pathogen
septicemia, eye infections
- isolated from blood cultures HAEMOPHILUS DUCREYI
- not respiratory only in the eyes - Characteristics:
Treatment: most strains are susceptible to - small gram negative coccobacilli, short
cephalosporins (cefrotaxime and cefriaxone chains, clumps or whorls within lesions.
- 3rd generation)
- railroad track, school of fish or fingerprint
arrangements.
HAEMOPHILUS
PARAHAEMOLYTICUS - - bipolar staining: both ends are stained
normal microbiota thicker
- immunocompromised individuals - Difficult to culture bec. the incubation day
- Diseases: endocarditis, pharyngitis is 10 days 33 degrees in 10% CO2, need
additional nutrients like isovitalex and Va
HAEMOPHILUS
PARAINFLUENZA - Chocolate Agar: yellow to gray dome-
shaped smooth colonies
Page 3 of 12
