Andersson M A, Nikulin M, Köljalg U, Andersson M C, Rainey F, Reijula K, Hintikka E L, Salkinoja-Salonen M
Department of Applied Chemistry and Microbiology, University of Helsinki, Finland.
Appl Environ Microbiol. 1997 Feb;63(2):387-93. doi: 10.1128/aem.63.2.387-393.1997.
Microbial toxins and eukaryotic cell toxicity from indoor building materials heavily colonized by fungi and bacteria were analyzed. The dominant colonizers at water-damaged sites of the building were Stachybotrys chartarum (10(3) to 10(5) visible conidia cm-2), Penicillium and Aspergillus species (10(4) CFU mg-1), gram-negative bacteria (10(4) CFU mg-1), and mycobacteria (10(3) CFU mg-1). The mycobacterial isolates were most similar to M. komossense, with 98% similarity of the complete 16S rDNA sequence. Limulus assay of water extracts prepared from a water-damaged gypsum liner revealed high contents of gram-negative endotoxin (17 ng mg-1 of E. coli lipopolysaccharide equivalents) and beta-D-glucan (210 ng mg-1 of curdlan equivalents). High-performance liquid chromatography analysis of the methanol extracts showed that the water-damaged gypsum liner also contained satratoxin (17 ng mg-1). This methanol-extracted substance was 200 times more toxic to rabbit skin and fetus feline lung cells than extract of gypsum liner sampled from a non-water-damaged site. The same extract contained toxin(s) that paralyzed the motility of boar spermatozoa at extremely low concentrations; the 50% effective concentration was 0.3 microgram of dry solids per ml. This toxicity was not explainable by the amount of bacterial endotoxin, beta-D-glucan, or satratoxin present in the same extract. The novel in vitro toxicity test that utilized boar spermatozoa as described in this article is convenient to perform and reproducible and was a useful tool for detecting toxins of microbial origin toward eukaryotic cells not detectable in building materials by the other methods.
对被真菌和细菌大量定殖的室内建筑材料中的微生物毒素和真核细胞毒性进行了分析。该建筑受水损害部位的主要定殖菌为展青霉(10³至10⁵个可见分生孢子/cm²)、青霉属和曲霉属菌种(10⁴CFU/mg)、革兰氏阴性菌(10⁴CFU/mg)和分枝杆菌(10³CFU/mg)。分枝杆菌分离株与科莫森分枝杆菌最为相似,其完整的16S rDNA序列相似度为98%。对取自受水损害石膏衬里的水提取物进行鲎试剂检测,结果显示革兰氏阴性内毒素(17 ng/mg大肠杆菌脂多糖当量)和β-D-葡聚糖(210 ng/mg短梗霉多糖当量)含量很高。对甲醇提取物进行高效液相色谱分析表明,受水损害的石膏衬里还含有葡萄穗霉毒素(17 ng/mg)。这种甲醇提取物对兔皮肤和猫胎儿肺细胞的毒性比取自未受水损害部位的石膏衬里提取物高200倍。同一提取物含有能在极低浓度下使公猪精子运动麻痹的毒素;50%有效浓度为每毫升0.3微克干物质。这种毒性无法用同一提取物中存在的细菌内毒素、β-D-葡聚糖或葡萄穗霉毒素的量来解释。本文所述的利用公猪精子的新型体外毒性试验操作简便、可重复,是检测建筑材料中其他方法无法检测到的微生物来源毒素对真核细胞毒性的有用工具。