Contribution of 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:[email protected] 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 , in causing chronic blepharitis Stain Droppers test and using PCR to detect the gene sequence for 16S rRNA. Nevertheless, the pathogenicity Results of emodex remains unclear and 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 , 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 shaped endospore-forming Group 1 Subgroup 1a 0.69 ± 0.24 (0.125–1) [0.750] 15 2 (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 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 Demodex-induced chronic studied patients and finally identified (PCR assay) weight standard. C(-), negative control; C(+), positive control; 1–9, 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.

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