Application of PCR-SSCP for molecular epidemiological studies on the exposure of farm children to bacteria in environmental dust

Korthals, Melanie; Ege, Markus; Tebbe, Christoph C.; Mutius, Erika von; Bauer, Johann

doi:10.1016/j.mimet.2008.01.010

Cited by 37 publications

(33 citation statements)

References 33 publications

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“…31,32 As a result, outdoor microorganisms or microorganisms that can be found in farmhouses are found in air, floor dust, and even in mattress dust in homes. [31][32][33] At higher age, the child will play in the farmyard and might enter stables and have direct contact with animals that will lead to oral uptake and respiratory exposure.…”

Section: Mamp Exposure and Exposure Routesmentioning

confidence: 99%

Does diversity of environmental microbial exposure matter for the occurrence of allergy and asthma?

Heederik

Mutius²

2012

Journal of Allergy and Clinical Immunology

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161

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Section: Mamp Exposure and Exposure Routesmentioning

confidence: 99%

Does diversity of environmental microbial exposure matter for the occurrence of allergy and asthma?

Heederik

Mutius²

2012

Journal of Allergy and Clinical Immunology

Self Cite

161

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“…Agarose gel (1%) electrophoresis of DNA extracted from kefir and whey using two methods. Lanes 1 CTAB, from kefir; 2, CTAB, from whey; 3, sonication, from kefir; 4, sonication, from whey SSCP profile Generally, the SSCP method includes a conversion of the double stranded PCR products into single strand by enzymatic removal of one DNA strand, which is generated during PCR using a phosphorylated primer (Korthals et al 2008). The PCR products were detected in the agarose gel electrophoresis (Fig.…”

Section: Effects Of Samples On Dna Extraction Resultsmentioning

confidence: 99%

Comparison of DNA extraction methods for the PCR-single-strand-conformation polymorphism analysis of kefir

Chen

Wang

Han

et al. 2010

World J Microbiol Biotechnol

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Experiments were performed to determine the influence of three DNA extraction methods (i.e. lysozyme, sonication and CTAB methods) from kefir on the microbial diversity analysis by PCR-single strand conformation polymorphism (PCR-SSCP). The results showed that the band of DNA extracted using CTAB was clearer than that using other methods. In addition, the yield and purity of DNA extracted using CTAB were the highest and reached, respectively, 915 lg/ml and 1.694.The results from the experiments indicated that the CTAB-based DNA extraction method was the most efficient method for DNA extraction from kefir. The heterogeneity of PCR products, amplified from community DNA with universal primers spanning the V3 region of 16S rRNA genes, was analysed by using SSCP. The results showed that the SSCP profile based on the sonication method gave the highest microbial diversity of kefir. One conclusion from these results was that the DNA extraction method was an important factor affecting the SSCP-based microbial diversity analysis of kefir.

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“…Interestingly, studies have shown that exposure to diverse microbial populations reduce incidences of allergies and asthma in children raised in rural and agricultural/farming areas compared to reference groups (Ege et al, 2011). Analysis of dust DNA samples--isolated from matress dust samples collected from the bedrooms of children living on farms and in rural and suburban communities--via PCR-Single-Strand Conformation Polymorphism (SSCP) gel electrophoresis and sequencing (Korthals et al, 2008) revealed inverse relationship between exposure to certain micriboal groups and asthma, hay fever and atopy (Ege et al, 2011;. Bacterial species identified to be associated with this inverse relationship are Staphylococcus sp., Bacillus sp., Acinetobacter sp., Lactobacillus spp., Neisseria spp.…”

Section: Discussionmentioning

confidence: 99%

A Biophysiochemical Analysis of Settled Livestock and Poultry Housing Dusts

Gerald

McPherson

McDaniel

et al. 2014

American Journal of Agricultural and Biological Sciences

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The levels and composition of agricultural dusts are influenced by animal species, production strategy, housing type and ventilation efficiency. Agricultural dust within animal houses is complex and consists of feed particles, microbes and their products, dander, fecal matter, gases, metals and other organic and inorganic components. Livestock and poultry production facilities may be categorized as confinement, semi-confinement or pasturebased. Characterization of animal husbandry building dust will provide insight into understanding exposures experienced by animals, workers and farm visitors. The goal was to characterize biophysiochemical features of livestock dusts from swine, small ruminant, equine, poultry and cattle husbandry units. Settled dust samples were collected from livestock and poultry housing units at the University Farm and other livestock farms across the state. Morphological features were determined by electron microscopy and gravimetry. Biochemical evaluation consisted of pH determination and trace metal detection via mass spectrometry. Biological assessment centered on bacterial characterization via selective media, DNA analysis and endotoxin quantitation. Morphological analyses revealed higher levels of respirable and thoracic particles in poultry, swine, small ruminant and equine units compared to the dairy unit (p<0.01). Dusts were slightly acidic with the exception of the NCAT small ruminant unit (p<0.05). Dust endotoxin levels were consistent and bacterial species detected include Listeria and Escherichia coli. These findings suggest animal husbandry buildings harbor higher levels of smaller respirable and thoracic dust particles compared to inhalable particles. This information may be helpful in understanding dust exposures experienced by animals, farmers and agricultural workers.

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Application of PCR-SSCP for molecular epidemiological studies on the exposure of farm children to bacteria in environmental dust

Cited by 37 publications

References 33 publications

Does diversity of environmental microbial exposure matter for the occurrence of allergy and asthma?

Does diversity of environmental microbial exposure matter for the occurrence of allergy and asthma?

Comparison of DNA extraction methods for the PCR-single-strand-conformation polymorphism analysis of kefir

A Biophysiochemical Analysis of Settled Livestock and Poultry Housing Dusts

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