In situ analysis of native microbial communities in complex samples with high particulate loads.
In the present study a procedure combining a cell extraction method and Fluorescence In Situ Hybridization (FISH) for molecular monitoring and quantification of bacteria in soil and aquifer samples is presented. FISH was applied to bacterial cells extracted from the matrix by density gradient centrifugation. This separation method was applied to soil and aquifer samples and produced high cell recovery of 76.5%+/-4.4 and 78.0%+/-3.2, respectively. FISH, performed on the harvested cells, permitted a perfect visualization and quantification of bacteria. This approach is therefore promising for in situ detection of indigenous bacterial communities in complex samples.
Barra Caracciolo
Anna
A
Water Research Institute, CNR, Via Reno 1, 00198 Rome, Italy. caraccio@irsa.cnr.it
Grenni
Paola
P
Cupo
Cinzia
C
Rossetti
Simona
S
D001419:Bacteria / Q000235:genetics / Q000302:isolation & purification; D001431:Bacteriological Techniques; D002499:Centrifugation, Density Gradient; D004269:DNA, Bacterial / Q000235:genetics / Q000302:isolation & purification; D017753:Ecosystem; D005456:Fluorescent Dyes; D017404:In Situ Hybridization, Fluorescence; D007211:Indoles; D007472:Iohexol; D012988:Soil Microbiology; D014871:Water Microbiology
D004269:DNA, Bacterial; D005456:Fluorescent Dyes; D007211:Indoles; D007472:Iohexol; C007293:DAPI