Prokaryotic community analysis with CARD-FISH in comparison with FISH in ultra-oligotrophic ground- and drinking water.
OBJECTIVE
We compared the applicability of catalysed reporter deposition fluorescence in situ hybridization (CARD-FISH) and FISH to enumerate prokaryotic populations in ultra-oligotrophic alpine groundwaters and bottled mineral water


RESULTS
Fluorescent oligonucleotide probes EUB338 and EUB338mix (EUB338/EUB338-II/EUB338-III) were used to enumerate bacteria and probes EURY806 and CREN537 for Euryarchaea and Crenarchaea, respectively. Improved detection of Planctomycetales by probe EUB338-II was tested using a different permeabilization step (proteinase K instead of lysozyme). Total detection efficiency of cells in spring water of four different alpine karst aquifers was on average 83% for CARD-FISH and only 15% for FISH. Applying CARD-FISH on bottled natural mineral waters resulted in an average total hybridization efficiency of 89%, with 78% (range 77-96%) bacteria and 11% (range 3-22%) identified as Archaea.


CONCLUSIONS
CARD-FISH resulted in substantially higher recovery efficiency than FISH. Hence, CARD-FISH appears very suitable for the enumeration of specific prokaryotic groups in ground- and drinking water.


CONCLUSIONS
This study represents the first evaluation of CARD-FISH on ultra-oligotrophic ground- and drinking water. Results are relevant for basic research and drinking water distributors. Archaea can comprise a significant fraction of the prokaryotic community in bottled mineral water.
Wilhartitz
I
I
Institute of Chemical Engineering, Department for Applied Biochemistry and Gene Technology, Vienna University of Technology, Getreidemarkt, Vienna, Austria.
Mach
R L
RL
Teira
E
E
Reinthaler
T
T
Herndl
G J
GJ
Farnleitner
A H
AH
D001105:Archaea / Q000302:isolation & purification*; D001419:Bacteria / Q000302:isolation & purification*; D005618:Fresh Water / Q000382:microbiology; D017404:In Situ Hybridization, Fluorescence / Q000379:methods; D008900:Mineral Waters / Q000382:microbiology*; D015345:Oligonucleotide Probes; D014871:Water Microbiology; D014881:Water Supply
D008900:Mineral Waters; D015345:Oligonucleotide Probes