MICROBIOLOGY ASSIGNMENT

TUGUME Francis 22/U/21993/PS

DELIGHT Isabella 20/U/0348
MUGALU Joseph
ASIIMIRE Phionah
MBOWA Erismus
KAMUGISHA Moses
TUKWASIBWE Keneth

1. What are the specimen rejection criteria for blood culture specimens?
 samples received without appropriate preservative
 Insufficient sample received.
 Labeling or form issues of the patient (mislabeled or unlabeled)
 Specimen is not transported in the right containers
 When the specimen lab request form is soiled
 Clotted, hemolyzed, icteric samples,
 The sample is broken or has leaked in transit.
 Stability time has been exceeded

2. what are the additives in broth blood culture media? What's their role?

additives role
Sodium polyanethole sulphate (SPS) Anticoagulant. It also inhibits lysozyme,
inactivates clinically achievable
concentrations of some aminoglycosides and
polymyxin antibiotics
gelatin Counteracts the inhibition of bacterial species
by SPS in vitro
Yeast extract Promotes bacterial growth
Saponin Used in lysis centrifugation system
Hemin(X-factor) Promotes growth of fastidious organisms such
as Hemophilus influenza and Neisseria
species
NAD(V-factor) Promotes growth of fastidious organisms such
as Hemophilus
Pyridoxine Promotes growth of pyridoxine dependent
organisms such as Streptococcus species
Para-amino benzoic acid Inhibits the effects of sulfonamide antibiotics
Cysteine Improves recovery of anaerobic bacteria and
Streptococcus pneumoniae

3. What is the HACEK group of bacteria?

HACEK refers to a group of fastidious gram-negative Coccobacillary organisms. HACEK is:
H-Haemophilus species
A-Aggregatibacter species
C‐Cardiobacterium hominis
E-Eikenella corrodens
K-Kingella species.

HACEK Organisms are Associated with Infective Endocarditis

The HACEK group is based on the organisms’ propensity to cause infective endocarditis. All
HACEK organisms are commensals of the human oropharynx. One hypothesis is that they
spread hematogenously following introduction into the blood during tooth brushing, dental
cleanings, or subsequent to oral diseases such as periodontitis.
Most endocarditis is caused by Gram-positive bacteria (most commonly Staphylococcus or
Streptococcus) with a minority caused by Gram-negatives or fungi. .

Haemophilus parainfluenza is one of the most common HACEK organisms associated with
infective endocarditis.
H. parainfluenza was originally described in 1922 by Thomas M. Rivers, who isolated it from
influenza patients. He differentiated it from H. influenzae based on its requirement for NAD (V
factor) but not hemin (X factor) in contrast to H. influenzae which requires both. Rivers initially
hypothesized that it was a cause of influenza, although we now know that this disease is caused
by the influenza virus.
His isolation of H. parainfluenzae from influenza patients was likely incidental as it is a
commensal of the human oropharynx. It is not a cause of oral diseases such as dental caries or
periodontitis and is preferentially isolated from dental plaque from healthy teeth. Outside the oral
cavity, H. parainfluenzae is capable of causing a variety of infections such as otitis media,
abscesses, and pneumonia, although it is an uncommon cause of these infections.

Aggregatibacter actinomycetemcomitans was originally isolated by R. Klinger in 1912 from an

actinomycotic lesion and its species name means "occurring with actinomycetes". Its original
genus name, "Actinobacillus", referred to the internal star shapes formed by colonies on solid
media. In 2006, the genus was renamed Aggregatibacter, reflecting the propensity of organisms
in this genus to grow as discrete clumps (aggregates) in broth culture.
Although best known for its role in infective endocarditis, A. actinomycetemcomitans is also an
etiologic agent of periodontal disease. It is specifically associated with localized aggressive
periodontitis (LAP), a form of periodontal disease that afflicts adolescents. This form of disease
progresses from symptom onset to tooth loss in a matter of months as opposed to years as seen in
adult periodontitis.
Colonization with virulent strains of A. actinomycetemcomitans is most common in individuals
of African descent but relatively rare in most other ethnic groups. Among African-American
adolescents, colonization rates can be up to 16.7% with a subset of carriers (up to 20%)
progressing to LAP.
Cardiobacterium hominis was first described in 1962 and was isolated from patients with
endocarditis. Like other HACEK organisms, C. hominis is found in the oropharynx but rarely the
gastrointestinal tract. It is infrequently isolated from oropharyngeal specimens however,
potentially due to overgrowth by other commensals. Unlike the other members of the HACEK
group, C. hominis is very rarely implicated in diseases other than endocarditis.

Eikenella corrodens is named in honor of M. Eiken, the microbiologist who first discovered the
organism. The species name indicates that it pits (or corrodes) into the surface of solid agar
media. This pitting is associated with the presence of pilins, which may be important for
adhesion to host tissue. It is a member of the Neisseriaceae family but cannot be recovered on
selective media for Neisseria spp. such as Thayer-Martin agar.
Among the HACEK organisms, Eikenella is a rarer cause of endocarditis but has the ability to
cause disease at other sites. Along with A. actinomycetemcomitans, it is among the few
organisms linked to periodontal disease. Additionally, E. corrodens is associated with infection
following traumatic inoculation from the oral cavity and is found in up to 42% of abscesses from
human bite wounds.

Kingella kingae was discovered by and later named for Elizabeth King, a microbiologist who
characterized many novel species while working at the Centers for Disease Control and
Prevention. Like Eikenella, it is a member of the Neisseriaceae but unlike Eikenella, it can
usually be recovered on Thayer-Martin agar.
In comparison to others in the HACEK group, K. kingae is infrequently isolated from
endocarditis. However, it is a significant cause of osteomyelitis/septic arthritis in children aged 6
months-3 years. The organism may also be transmitted person to person by respiratory droplets
and caused a 2004 outbreak of septic arthritis in children attending

Clinical Identification of HACEK Organisms

Previously, HACEK organisms were implicated in endocarditis from which no pathogen could
be isolated (so called "culture-negative endocarditis"). This was attributed to slow growth in old
formulations of blood culture bottles and resulted in recommendations for extended incubation
(>5 days) when the presence of these organisms was suspected. However, modern blood culture
instruments reliably detect HACEK organisms within a 5-day incubation period.

HACEK organisms can be isolated on routinely used non-selective media, often growing better
on chocolate agar than on blood agar. They cannot be isolated from selective media designed for
enteric such as MacConkey agar. Identification can be performed by automated systems but
performance may be suboptimal. As with many other organisms, MALDI-TOF is emerging as a
potential identification technology for the HACEK group.
 4. What are the common bacteria contaminants in blood culture?
.Bacillus species
Coagulase negative species
Streptococci viridans
Micrococcus species
Neisseria species
Clostridium perfringens
Propionibacterium acnes