Bacillus cereus Endophthalmitis

-- Michelle Callegan, Ph.D.

© MPEIR Center, 1998

Epidemiology of bacterial endophthalmitis
Clinical aspects of Bacillus cereus endophthalmitis
The contribution of Bacillus cereus proteins to virulence
Molecular pathogenesis of Bacillus cereus endophthalmitis
Relevant citations

Epidemiology of bacterial endophthalmitis

Bacterial endophthalmitis is a potentially blinding disease that can result from introduction of organisms into the eye following one of two events: 1) intraocular surgery or penetrating ocular trauma (post-traumatic endophthalmitis), or 2) hematogenous spread to the eye from a distant infectious site (endogenous endophthalmitis). Retrospective surveys of bacterial species isolated from postoperative endophthalmitis vitreous specimens report that causative agents include normal flora from the oral and nasal cavities and the skin (staphylococci, viridans streptococci, and Enterococcus faecalis). Such organisms commonly cause indolent endophthalmitis and treatment is usually successful with recovery of partial or full vision. In contrast, post-traumatic endophthalmitis-derived isolates include a variety of environmental organisms, in addition to isolated normal flora. Of such isolates, B. cereus is ranked second behind staphylococci, with estimates ranging from 27 to 46%. B. cereus has emerged as a significant cause of endogenous endophthalmitis, usually as a complication of high-grade bacteremia in patients with prolonged indwelling devices or intravenous drug abusers. The reason for its predilection for the eye in cases of endogenous endophthalmitis is not known.

Clinical aspects of Bacillus cereus endophthalmitis

Recent attention has focused on B. cereus endophthalmitis because of its extremely rapid and severe course, its refractory nature, and its invariably poor visual and anatomic outcome.
Rapid destruction of the posterior segment of the eye following penetrating trauma typifies B. cereus endophthalmitis. Penetrating trauma with introduction of organisms or metastasis of organisms from a distant infection site results in seeding of the vitreous and infection. Severe pain accompanies a rapid decline in visual acuity, exacerbated by corneal ring abscess formation, periorbital edema, and proptosis. Systemic symptoms unique to B. cereus endophthalmitis include fever, leukocytosis, and general malaise. Several clinical studies have described dismal visual and anatomic outcomes of B. cereus endophthalmitis, including the loss of all light perception and the need for enucleation or evisceration.

The recommended management of B. cereus endophthalmitis includes vitreal aspiration or vitrectomy with intravitreal injection of antibiotics (clindamycin and aminoglycosides) and intravenous antibiotic supplements. Intravitreal vancomycin has been suggested for use in all cases of B. cereus endophthalmitis, primarily because of its broad spectrum for other Gram-positive organisms. Systemic antibiotics (vancomycin, cephalosporins, ciprofloxacin) have been used concurrently for B. cereus endophthalmitis; however, some potentially effective antibiotics (aminoglycosides and vancomycin) do not penetrate readily into the vitreous. The majority of clinical reports on the use of intravitreal or intravenous antibiotics are based on the preferences of the treating physicians and, therefore, give conflicting results about the optimal antibiotic combinations, routes, and duration of therapy for B. cereus endophthalmitis. Recent reports have indicated that the outcome of therapy could be due to several factors, including the age of the patient, the duration between injury and treatment, the antibiotics chosen for therapy, and the condition of the eye upon presentation (e.g., corneal damage, retinal detachment, etc.). Despite aggressive drug and/or surgical intervention, B. cereus endophthalmitis typically results in loss of the eye within 48 hours. This outcome suggests that even if the infected eye is rendered sterile by antibiotics, ocular damage continues to occur, possibly due to toxin production by a uniquely virulent strain. This could be the case for any toxigenic infectious agent; however, because of the rapid and devastating nature of B. cereus endophthalmitis despite drastic therapeutic measures, understanding the relationship of toxin production to virulence is critical.

The contribution of Bacillus cereus proteins to virulence

The correlation of toxin production with fulminant B. cereus endophthalmitis has been hypothesized since its recognition as an unusually virulent intraocular pathogen. However, surprisingly little data exist to confirm this hypothesis. B. cereus produces a number of cytotoxins and enzymes that could contribute to the rapid course and severity of endophthalmitis, including hemolysins, lipases, enterotoxins, and proteases. This particular project focuses on four potential virulence factors: cereolysin O, cereolysin AB, hemolysin BL, and collagenase. These proteins were chosen for study because of their implied contributions to the virulence of other Bacillus infections and because of their potentially damaging activites in the eye.

Cereolysin O is a thiol-activated cytolysin related to a group of oxygen-labile enzymes produced by several Gram-positive species, including streptolysin O of Group A streptococci, L. monocytogenes listeriolysin, and pneumolysin of S. pneumoniae. These enzymes have been implicated as virulence factors in experimental models of infection. Cereolysin O binds to cholesterol, a component of all mammalian cell membranes, as an initial step prior to cell lysis. Therefore, cells of the posterior segment of the eye are prone to damage. Cereolysin O is lethal in murine models of intravenous injection; however, whether cereolysin O contributes to fulminating B. cereus endophthalmitis is not known.

Cereolysin AB has been characterized as a cytolytic unit unrelated to cereolysin O. Cell lysis occurs via sequential action on membranes by independent activities of sphingomyelinase and phosphotidylcholine-phospholipase C. These components of cereolysin AB combine synergistically to lyse erythrocytes, but each can confer resistance to phagocytosis by neutrophils. Cereolysin AB has been proposed as an evolutionary precursor of Clostridium perfringens alpha-toxin, a noted virulence factor. Because of their membrane-specific activities, either cereolysin O or AB could damage or lyse retinal cells directly, or induce the production of inflammatory mediators which could contribute to tissue damage during infection.

Initial interest in B. cereus as a diarrheal-type food poisoning agent brought about the characterization of the vascular permeability factor as a potential cause. Recent characterization of this enterotoxic factor (also known as hemolysin BL) led to the implication of its involvement in B. cereus endophthalmitis. Hemolysin BL is a tripartite toxin, with hemolytic, dermonecrotic, and emetic activities, suggesting its potential to exert multiple toxic effects upon ocular tissues during fulminant endophthalmitis. Using an in vitro retinal button toxicity assay and in vivo vitreal injections of sterile supernatants and purified hemolysin BL, Beecher et al. suggested that B. cereus exotoxins, including hemolysin BL, can cause ocular damage during fulminant endophthalmitis. In these studies, however, hemolysin BL accounted for only half of the retinal toxicity of B. cereus supernatants, suggesting that full virulence is likely a multifactorial process.

A recent review suggested that B. cereus collagenase could be involved in ocular virulence. Collagenolytic enzymes can readily damage the vitreal and retinal architecture and have been reported to contribute to the virulence of Pseudomonas aeruginosa ocular infections. Collagen is a major constituent of the vitreous, supplying a potential target for this enzyme during endophthalmitis. Collagenolytic B. cereus strains have been isolated from cases of peridontitis and endophthalmitis, further suggesting that collagenase could contribute to virulence.

Other less well characterized B. cereus factors that could contribute to virulence include emetic toxin (cereulide), hemolysin II (murine lethal toxin), and phosphotidylinositol-phospholipase C (phosphatasemic factor). Emetic toxin (cereulide) has been reported to cause emesis in rhesus monkeys and vacuole formation in HEp-2 cells. Hemolysin II is a thermolabile toxin unrelated to cereolysin, that is lethal in simian models of intravenous administration. The phosphotidylinositol-phospholipase C has been shown to cause a phosphatasemia when injected intraveneously into rabbits. What contributions these proteins make, if any, to fulminant endophthalmitis have yet to be identified.

Molecular pathogenesis of Bacillus cereus endophthalmitis

Historically, studies of the pathogenesis of bacterial infections in ocular and non-ocular models have focused on the contribution of specific proteins to disease. Ocular models employing such techniques have included experimental models of keratitis and endophthalmitis. The application of this approach for B. cereus is challenging because of the probable presence of several chromosomally-encoded toxins or enzymes that could contribute to virulence during endophthalmitis. Genetically constructed isogenic mutants defective in one specific protein have provided the most reliable data. The most plausible evidence that a bacterial gene product contributes to a disease is validated by generating isogenic mutants specifically deficient only in the virulence factor under investigation. The molecular studies employed during these particular studies are designed to identify B. cereus virulence factors following this approach.

Relevant Citations

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