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House dust is a reservoir for different kinds of biological and non-biological materials originating from building materials, furnishing, occupants, pets, 1353 bp 1078 bp 872 bp

Detection of streptomycetes in house dust--comparison of culture and PCR methods.

Indoor air, no. 2 (2004): 112-119

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摘要

Streptomycetes are gram-positive, spore producing, filamentous bacteria common in soil, but also present in indoor environments. They are potent producers of secondary metabolites and inducers of inflammatory responses in vitro. Polymerase chain reaction (PCR)- and culture-based detection methods for streptomycetes in house dust samples w...更多

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重点内容
  • Streptomycetes are gram-positive filamentous bacteria of the class Actinobacteria and the order Actinomycetales (Stackebrandt et al, 1997)
  • House dust is a reservoir for different kinds of biological and non-biological materials originating from building materials, furnishing, occupants, pets, 1353 bp 1078 bp 872 bp
  • 310 bp Fig. 2 Agarose gel electrophoresis of Polymerase chain reaction (PCR) products amplified from different amounts of Streptomyces californicus-spores
  • Pseudomonas sp. and Streptomyces sp. were found in 11 and 9%, respectively, of dust samples taken from classrooms (Smedje and Norback, 2001). 8 · 106 cfu/g mesophilic bacteria, and 2 · 104 cfu/g thermophilic bacteria, e.g. bacilli and actinomycetes, were detected from carpet dust (Macher, 2001)
  • Mesophilic actinomycetes were cultured from 36% of the dust samples
  • Diversity analysis based on the 16S rDNA of the streptomycete isolates revealed that the majority of the isolates belonged to the S. albidoflavus-cluster, which is the largest of the Streptomyces-clusters
方法
  • Dust samples

    Dust samples were collected from homes as a part of a research project monitoring the connection between microbial conditions and moisture damage in homes.
  • The homes were visited by a trained building expert who made a walk-through inspection for moisture damage according to a checklist, which has been developed earlier (Nevalainen et al, 1998), and recorded visible signs of moisture and mould.
  • According to the results of the inspection, the homes were divided into categories using a two-level classification of moisture damage (Haverinen et al, 2001).
  • Homes with no signs of moisture or mould or having, at a maximum, one small dried sign of moisture on a limited area, were considered as nondamaged, and all homes having more severe damage were considered as damaged
结果
  • Counts of cultivable actinomycetes ranged from 4.5 · 102 to 1 · 105 cfu/g.
  • Total counts of viable bacteria in house dust were determined.
  • They ranged from 1.1 · 104 to 2.1 · 107 cfu/g with a mean value of 3.4 · 106 cfu/g.
  • There was no difference in the counts of cultivable actinomycetes and total bacterial count between the moisture-damaged and non-damaged houses
结论
  • House dust is a reservoir for different kinds of biological and non-biological materials originating from building materials, furnishing, occupants, pets, 1353 bp 1078 bp 872 bp

    310 bp Fig. 2 Agarose gel electrophoresis of PCR products amplified from different amounts of Streptomyces californicus-spores.
  • 8 · 106 cfu/g mesophilic bacteria, and 2 · 104 cfu/g thermophilic bacteria, e.g. bacilli and actinomycetes, were detected from carpet dust (Macher, 2001).
  • Hameed et al (1999) found bacilli and micrococci to be the most abundant bacterial species in carpet dust, streptomycetes were isolated and identified to represent the species S.
  • The 16S rDNA sequence analysis revealed that all the 15 isolates, which were sequenced, belonged to the genus Streptomyces.
  • Other species were present, some of which did not cluster with known species
总结
  • Introduction:

    Streptomycetes are gram-positive filamentous bacteria of the class Actinobacteria and the order Actinomycetales (Stackebrandt et al, 1997).
  • They are soil bacteria, present in all kinds of soils, but found in other habitats as well (Williams et al, 1989).
  • Some of the secondary metabolites are volatile, such as geosmin, which gives samples contaminated with streptomycetes their earthy, musty odor (Gerber and Lechevalier, 1965; Sunesson et al, 1997)
  • Objectives:

    The aim of this study was to evaluate the usefulness of the previous PCR method developed by the authors (Rintala et al, 2001) for the detection of streptomycetes in house dust samples.
  • Methods:

    Dust samples

    Dust samples were collected from homes as a part of a research project monitoring the connection between microbial conditions and moisture damage in homes.
  • The homes were visited by a trained building expert who made a walk-through inspection for moisture damage according to a checklist, which has been developed earlier (Nevalainen et al, 1998), and recorded visible signs of moisture and mould.
  • According to the results of the inspection, the homes were divided into categories using a two-level classification of moisture damage (Haverinen et al, 2001).
  • Homes with no signs of moisture or mould or having, at a maximum, one small dried sign of moisture on a limited area, were considered as nondamaged, and all homes having more severe damage were considered as damaged
  • Results:

    Counts of cultivable actinomycetes ranged from 4.5 · 102 to 1 · 105 cfu/g.
  • Total counts of viable bacteria in house dust were determined.
  • They ranged from 1.1 · 104 to 2.1 · 107 cfu/g with a mean value of 3.4 · 106 cfu/g.
  • There was no difference in the counts of cultivable actinomycetes and total bacterial count between the moisture-damaged and non-damaged houses
  • Conclusion:

    House dust is a reservoir for different kinds of biological and non-biological materials originating from building materials, furnishing, occupants, pets, 1353 bp 1078 bp 872 bp

    310 bp Fig. 2 Agarose gel electrophoresis of PCR products amplified from different amounts of Streptomyces californicus-spores.
  • 8 · 106 cfu/g mesophilic bacteria, and 2 · 104 cfu/g thermophilic bacteria, e.g. bacilli and actinomycetes, were detected from carpet dust (Macher, 2001).
  • Hameed et al (1999) found bacilli and micrococci to be the most abundant bacterial species in carpet dust, streptomycetes were isolated and identified to represent the species S.
  • The 16S rDNA sequence analysis revealed that all the 15 isolates, which were sequenced, belonged to the genus Streptomyces.
  • Other species were present, some of which did not cluster with known species
表格
  • Table1: Frequencies of moisture-damaged and streptomycete positive and negative samples, determined by PCR and culture
  • Table2: Frequency distribution of the PCR and culture positive dust samples
Download tables as Excel
基金
  • The work was financially supported by the Finnish Work Environment Fund (grant 101053), the Foundation for Finnish Inventions, and the East-Finnish Foundation for High Technology
研究对象与分析
dust samples: 47
Polymerase chain reaction (PCR)- and culture-based detection methods for streptomycetes in house dust samples were compared. A total of 47 dust samples were investigated, and the presence of streptomycetes was determined by cultivation on tryptone-yeast-extract-glucose agar and PCR. The 16S rRNA gene of actinomycete isolates from house dust was partially sequenced to investigate if they belong to the genus Streptomyces

house dust samples: 47
Ó Indoor Air (2003). Practical Implications A PCR-based detection method for streptomycetes was tested with 47 house dust samples and compared with culture. PCR-detection revealed more positives than culture

culture-negative samples: 24
If using binary data (presence or absence of moisture damage or streptomycetes), and Fisher’s exact test, a correlation, however not statistically significant, between moisture damage and the occurrence of streptomycetes was observed (P 1⁄4 0.127 for culture, and P 1⁄4 0.054 for PCR). The correlation between the results obtained by the culture and PCR methods was poor; three culture-positive samples were PCR-negative, and 24 culture-negative samples were PCR-positive (Table 2). The detection limit of the culture method was calculated by dividing the lowest spore count by the sum of the volumes of the plated dilutions that were counted

strains: 4
The remaining five isolates did not show clear affiliation to any of the sequences of known streptomycetes used in the analysis. However, the sequences of these four strains had 98.8–99.6% sequence similarity to 16S rDNA sequences of different Streptomyces sp. House dust is a reservoir for different kinds of biological and non-biological materials originating from building materials, furnishing, occupants, pets, 1353 bp 1078 bp 872 bp

dust samples: 47
However, also other species were present, some of which did not cluster with known species. In this study of 47 dust samples, a positive PCR amplification was revealed in 81% of the dust samples, the reagent controls always being negative. It shows that genetic material of streptomycetes is frequently present in house dust, although they cannot always be cultivated from these samples

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A Hyvärinen
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