Contribution of Bacillus oleronius

to Demodex chronic blepharitis

A. Szkaradkiewicz1, I. Chudzicka-Strugała1, T. M. Karpiński1, O. Goślińska-Pawłowska1, T. Tułecka1,
W. Chudzicki2, A. K. Szkaradkiewicz1, R. Żaba1
1 University of Medical Sciences in Poznan, Wieniawskiego Street 3, 61-712 Poznań, Poland, tel +48618546138, e-mail:ichudzicka@vp.pl
2Military Hospital Grunwaldzka16/18, Poznań

Introduction Patients and Methods

Blepharitis is an inflammation of The studies were conducted on 68 adult patients, among whom ophthalmological and
the eyelid margins, resulting in parasitological tests permitted to distinguish a group of 38 patients with diagnosis of
extremely irritating ocular Demodex-related chronic blepharitis (group 1, including a subgroup 1a with moderate
discomfort in patients, pruritus, blepharitis and a subgroup 1b with severe blepharitis) and a group of 30 healthy
palpebral reddening and a individuals (group 2). In every studied person 6 eyelashes were epilated from each eye
decline in visual function and a number of Demodex per eyelash was scored. Demodex was detected under a
persisting in the patients. An
light microscope using 10% solution of KOH. In parallel, cultures of eyelashes was
aetiological role of Demodex
performed in nutrient broth (Difco), tryptic soy agar and sheep blood agar, at the
mites is recognized, particularly
temperature of 37˚C for 24 hours. Bacillus oleronius was identified conducting Flagella
Demodex folliculorum, in
causing chronic blepharitis
Stain Droppers test and using PCR to detect the gene sequence for 16S rRNA.
Nevertheless, the pathogenicity
of emodex remains unclear and
Results
Demodex infestations also be In the studies intensity of Demodex infestation manifested no significant differences
found in asymptomatic between subgroups 1a and 1b. From the epilated eyelashes 23 bacterial isolates were
individuals. Studies indicate that obtained , identified as belonging to B. oleronius species. (Fig.1, Fig. 2). Using the
Demodex is a non-pathogenic epilated eyelashes, culture permitted to isolate 23 strains of bacteria, originating from 18
parasite and induces morbid patients of group 1 (2 strains were isolated from patients of subgroup 1a and 16 strains
signs/symptoms only in were isolated from patients of subgroup 1b) and from 5 patients of group 2 (Table 1.).
individuals with immune deficits. Difference in the frequency of B. oleronius detection in patients of subgroup 1b and
New data related to the healthy persons (group 2) was significant (p=0.0190).
pathogenicity of the parasite
were introduced by Lacey et al.
demonstrating the presence of TABLE 1. Intensity of Demodex infestation and detection of Bacillus oleronius in the groups
Bacillus oleronius inside of patients examined
Demodex mites from patient
with papulopustular rosacea. Examined group Subgroups Mean ± SD number of Demodex No. of Bacillus Significant
Bacillus oleronius is a non- per eyelash (range in parentheses) patients oleronius
difference
motile, Gram-negative, rod- detection
Group 1 Subgroup 1a 0.69 ± 0.24 (0.125–1) [0.750] 15 2
shaped endospore-forming
(patients with (moderate blepharitis)
bacterium The pathogenic role Demodex blepharitis)
of Demodex infestation may Subgroup 1b 0.75 ± 0.25 (0.125–1) [0.8125] 18 12
16*
therefore be associated with (severe blepharitis) 1.33 ± 0.26 (1.125–1.75) [1.250] 5 4
B. oleronius. Group 2
(control group) - 0 30 5

Objective
*Significant difference as compared with the control group
Bacillus oleronius was isolated
for the first time from the
hindgut of the termite
Reticulitermes santonensis in
1995. In 2007 presence of
B. oleronius was demonstrated
inside Demodex mites. In view
of the above this study aimed at
analysis of Bacillus oleronius FIG. 1. Optical micrograph of Gram-negative, FIG. 2. Polymerase chain reaction detection of Bacillus oleronius.
involvement in pathogenesis of medium-sized rods isolated from eyelashes of Ethidium bromide stained agarose gel. M—100 bp molecular
studied patients and finally identified (PCR assay) weight standard. C(-), negative control; C(+), positive control; 1–9,
Demodex-induced chronic as Bacillus oleronius species. positive patients.
blepharitis.

Conclusion
The result indicate that B. oleronius bacteria, most probably act as a co-pathogen
in development of Demodex blepharitis severe forms.

References
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