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Institut für Angewandte Mikrobiologie, Universitt Gieen, Giessen, Germany
Institut für Hygiene und ffentliche Gesundheit, Universitt Bonn, Bonn, Germany
Pneumologische Fachpraxis Dr. Rolke, Frohsinnstrae 13, 63739 Aschaffenburg, Germany
Allergologisch-immunologisches Labor Prof. J. Sennekamp, Bonn, Germany
ABSTRACT
A case of extrinsic allergic alveolitis (EAA) caused by Sphingobacterium spiritivorum is described. The symptoms were associated with the use of a steam iron. The water reservoir was heavily contaminated with S. spiritivorum (106 CFU ml–1). This is the first report of S. spiritivorum as a causative agent of EAA.
CASE REPORT
A 34-year-old woman presented with a dry cough, thorax constriction, and overall weakness. During the preceding 5 months, she had experienced these dry cough episodes twice a week. On examination, the patient was febrile (39°C), but a clinical lung examination showed no striking diagnostic findings. The high-resolution computer tomography-Roentgen image was normal; however, a lung function test showed a restriction of the vital capacity (71% of the adjusted reference value), and the diffusion capacity was also reduced (66 to 68%).
During a hospital stay of several days' duration, the patient was free of discomfort. In the bronchoalveolar lavage specimen, the lymphocytes were increased up to 41% (norm, <10%), and neutrophils were increased up to 6% (norm, <2%) with a normal total cell count. This bronchoalveolar lavage specimen cell pattern indicated an acute phase of extrinsic allergic alveolitis (EAA; hypersensitivity pneumonitis). The erythrocyte sedimentation rate was increased up to 20/35 mm. The patient's conventional chest X ray and high-resolution computer tomography images were free of any alterations, based on criteria outlined previously (11).
The patient's detailed history revealed that she had used an iron before each of the episodes of discomfort. She had used a steam iron which is additionally equipped with a liquid reservoir for wetting of the clothes.
Using the agar gel diffusion test (Ouchterlony assay described previously [13]), the water of this liquid reservoir showed a clear precipitation line with the serum of the patient, indicating the presence of precipitating immunoglobulin G (IgG) antibodies in the serum of the patient against antigens in the water.
No IgG antibodies could be detected against a wide panel of organisms which are commonly associated with EAA, including Saccharopolyspora rectivirgula, Thermoactinomyces vulgaris, Aspergillus fumigatus (also not in an IgG immunoblot), Aspergillus versicolor, Aspergillus niger, Penicillium brevicompactum, Cladosporium cladosporioides, Alternaria species, Aureobasidium pullulans, Rhizopus nigricans, Pseudomonas aeruginosa, Pseudomonas fluorescens, Fusarium culmorum, Acremonium strictum, Stenotrophomonas maltophilia, Streptomyces albus, Cephalosporium acremonium, Sporobolomyces roseus, Trichoderma viride, Ustilago species, Pseudallescheria species, Exserohilum rostratum, Phoma betae, Sporothrix schenckii, Leucogyrophana pinastri, Stachybotrys chartarum, Trichosporon cutaneum, Torulopsis glabrata, Rhodotorula species, and Geotrichum candidum, by the usual solid-phase peroxidase IgG enzyme-linked immunosorbent assay using Nunc Maxisorb microplates, as described by Tijssen (14).
The liquid originating from the water reservoir of the steam iron was investigated for the presence of microorganisms. A total of 106 CFU ml–1 were detected on tryptone soy agar (TSA; Oxoid) (48 h of incubation at 37°C), a pure culture with yellow-pigmented colonies. A clear precipitation line was again found in the Ouchterlony assay against this organism (isolate B1) (13), with the serum of the patient indicating that this organism is responsible for the EAA of the patient. According to the criteria of the "EAA working group" by Sennekamp (13), sufficient criteria are fulfilled for the diagnosis of EAA (exposure to allergens, EAA-typical symptoms, precipitating antibodies, restricted lung function, and EAA-specific blood parameters.)
The patient was free of discomfort after using an iron without a liquid reservoir. On Gram stain, isolate B1 was found to be a gram-negative rod. The 16S rRNA gene was analyzed as described by Kmpfer et al. (4), with the modification described elsewhere (5). The 16S rRNA sequence of strain B1 was a continuous stretch of 1,449 bp. Sequence similarity calculations after a neighbor-joining analysis indicated that the closest relative of strain B1 was S. spiritivorum DSM 2582T, accession no. AJ459411 (99.7% similarity). Fatty acid analysis of whole-cell hydrolysates prepared after growth on tryptone soy agar for 24 h using the Sherlock system (MIDI, Newark, DE) (6) and comparison with the Sherlock database provided the identification result of S. spiritivorum with an identity score of 0.631 (results not shown). Selected results of the physiological characterization, according to Kmpfer et al. (7), are given in Table 1 in comparison with S. spiritivorum NCTC 11386T. DNA-DNA hybridization experiments were performed with B1 and S. spiritivorum NCTC 11386T using the method described by Ziemke et al. (17) and modified as described by Kmpfer et al. (5). Strain B1 showed 98% DNA-DNA similarity (mean value of the results of four hybridizations) to S. spiritivorum NCTC 11386T, indicating that B1 is a member of this species since the similarity demonstrated was >70%.
Discussion. S. spiritivorum has been occasionally isolated from human clinical specimens (4, 12). Blood and urine have been reported to be the most common sources for the isolation of S. spiritivorum (2). Recently, Tronel et al. (15) reported a case of bacteremia caused by an S. spiritivorum-like organism, and details on the induction of apoptosis of human leukemic cells by bacterial ceramides and sphingophospholipids of S. spiritivorum have been published by Minamino et al. (9).
A second Sphingobacterium species, S. multivorum, has been isolated more frequently from clinical specimens but has only rarely been associated with serious infections (1, 3, 8, 10, 15). Interestingly, most recently a respiratory infection caused by S. multivorum was reported (16), indicating a possible clinical role of sphingobacteria in the respiratory tract.
To our knowledge, neither an allergic reaction against sphingobacteria nor the association of a hypersensitivity pneumonitis with the use of a steam iron has been described in the literature. Thus, this is the first report of a steam-iron-induced allergy presenting as EAA (hypersensitivity pneumonitis) caused by S. spiritivorum.
Nucleotide sequence accession number. The DNA sequence of the 16S rRNA gene of isolate B1 was deposited in EMBL/GenBank under accession number AJ878852.
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