Regrine B.

Lagarteja, RMT MICRO 203

2013-88648


IDENTIFICATION OF BACTERIAL UNKNOWNS

SPECIMEN: URINE
SPECIMEN NO.: 11

Common Isolates for Urinary Tract Infections:
Adapted from: Lippincotts Microcards: Microbiology Flash Cards, 3
rd
Edition

MORPHOLOGY
Grams Stain: Gram (+) cocci, singly

Diagram for Differential Diagnosis of Pathogenic Gram (+) cocci
Adapted from: Lippincotts Microcards: Microbiology Flash Cards, 3
rd
Edition
Regrine B. Lagarteja, RMT MICRO 203
2013-88648

CULTURAL CHARACTERISTICS

Blood Agar Plate: Pinpoint, alpha-hemolytic colonies
MacConkey Agar: no growth

BIOCHEMICAL CHARACTERISTICS

Biochemical Test Result
Taxo P Resistant
Bacitracin Resistant
Coagulase Negative
Growth in 6.5% NaCl Positive
Bile Esculin Agar Positive
Catalase Negative
Sorbitol Positive*
Litmus Milk Reduction Positive*
Gelatinase Negative*
*not performed
Colony count: >1x10
5
CFU/mL

SUSPECTED ISOLATE: Enterococcus faecalis
Pathogen? YES

Entercococcus faecalis
Enterococcus faecalis is a member of the human gut flora (Gladwin & Trattler, 2003). It is known to
cause nosocomial, life-threatening infections in humans (Ryan & Ray, 2004). It is frequently found in
root canal teeth ranging from 30-90% of the cases (Rocas, Siquierra, & Santos, 2004; Ryan & Ray,
2004).

Physiology
E. faecalis is a nonmotile, facultatively anaerobic microbe; it ferments glucose without gas production,
and does not produce a catalase reaction with hydrogen peroxide. It can produce a pseudocatalase
reaction if colony is extracted from blood agar. It produces a reduction of litmus milk, but does not
liquefy gelatin. Growth on nutrient broth is consistent with it being facultatively anaerobic. It can
catabolize carbohydrates, glycerol, lactate, malate, citrate, arginine, agmatine, and many keto acids.
Enterococci survive extremely harsh environments including extreme alkaline pH (9.6) and salt
concentrations. They resist bile salts, detergents, heavy metals, ethanol, azide, and desiccation. They
can grow in the range of 10C to 45C and survive at temperatures of 60C for 30 min. (Stuart,
Schwartz, Becson, & Owatz, 2006).

Pathogenesis
E. faecalis causes endocarditis, urinary tract infections (UTI), meningitis, and biliary tract infections
(Hidron, Edwards, & Patel, 2009) (Murray, 1990) (Harapavat & Nissim, 2012). Several factors
contribute to its pathogenicity. Cytolysin, a plasmid encoded hemolysin, and its combination with
gentamicin resistance contributes to a 5-fold increase in the risk of death among human bacteremia
patients. (Huycke, Spiegel, & Gilmore, 1991) (Chow, et al., 1993).

Regrine B. Lagarteja, RMT MICRO 203
2013-88648

E. faecalis is resistant to commonly used antimicrobial agents such as aminoglycosides, aztreonam,
cephalosporins, clindamycin, and trimethoprim-sulfomethoxazole. Vancomycin-Resistant Enterococci
(VRE) infections are also common. Treatments to VRE include linezolid and daptomycin. (Hirt,
Schlievert, & Dunny, 2002)

Virulence Factors
Endures prolonged periods of nutritional deprivation
Binds to dentin and proficiently invades dentinal tubules
Alters host responses
Suppresses the action of lymphocytes
Possesses lytic enzymes, cytolysin, aggregation substance,pheromones, and lipoteichoic acid
Utilizes serum as a nutritional source
Resists intracanal medicaments (i.e. Ca(OH)2)
Maintains pH homeostasis
Properties of dentin lessen the effect of calcium hydroxide
Competes with other cells
Forms a biofilm
(Stuart, Schwartz, Becson, & Owatz, 2006; Harapavat & Nissim, 2012).
Reference
Ryan, K., & Ray, C. (2004). Sherris Medical Microbiology. USA: McGrawHill.

Hidron, A., Edwards, J., & Patel, J. (2009). NHSN annual update: antimicrobial-resistant pathogens
associated with healthcare-associated infections: annual summary of data reported to the National
Healthcare Safety Network at the Centers for Disease Control and Prevention, 2006-2007. Infection
Control and Hospital Epidemology , 29, 996-1011.

Murray, B. (1990). Life and Times of Enterococcus. Clinical Microbiology Review , 3, 46-65.

Rocas, I., Siquierra, J., & Santos, K. (2004). Association of Enterococcus faecalis with different forms
of periradicular diseases. Journal of Endodontics , 30, 315.

Stuart, C., Schwartz, S., Becson, T., & Owatz, C. (2006). Enterococcus faecalis: Its Role in Root
Canal
Treatment Failure and Current Concepts in Retreatment. Journal of Endodontics , 32, 93.

Harapavat, S., & Nissim, S. (2012). Lippincott's Microcards: Microbiology Flash Cards, 3rd Edition.
Philadelphia, USA: Lippincott Williams & Wilkins.

Huycke, M., Spiegel, C., & Gilmore, M. (1991). "Bacteremia caused by hemolytic, high-level
gentamicin-resistant Enterococcus faecalis.". Antimicrobial Agents Chemotherapy , 35 (8), 16-24.

Chow, J., Thal, L., Perri, M., Vazquez, J., Donabedian, S., Clewell, D., et al. (1993). Plasmid-
associated hemolysin and aggregation substance production contribute to virulence in experimental
enterococcal endocarditis. Antimicrobial Agents Chemotherapy , 37 (11), 2474-7.

Regrine B. Lagarteja, RMT MICRO 203
2013-88648

Hirt, H., Schlievert, P., & Dunny, G. (2002). "In vivo induction of virulence and antibiotic resistance
transfer in Enterococcus faecalis mediated by the sex pheromone-sensing system of pCF10.".
Infectious Immunology , 70 (2), 716-23.

Gladwin, M., & Trattler, B. (2003). Clinical Microbiology Made Ridiculously Simple. USA: MedMaster
Inc.