scholarly journals Evaluation of an MPN test for the rapid enumeration of Pseudomonas aeruginosa in hospital waters

2014 ◽  
Vol 13 (2) ◽  
pp. 427-436 ◽  
Author(s):  
David P. Sartory ◽  
Danièle Pauly ◽  
Nathalie Garrec ◽  
Lucia Bonadonna ◽  
Maurizio Semproni ◽  
...  
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Membrane Filtration ◽  
Most Probable Number ◽  
Insufficient Data ◽  
Culture Methods ◽  
Probable Number ◽  
Rapid Enumeration ◽  
Relative Differences ◽  
Mpn Test ◽  
Atypical Strains

In this study, the performance of a new most probable number (MPN) test (Pseudalert®/Quanti-Tray®) for the enumeration of Pseudomonas aeruginosa from hospital waters was compared with both international and national membrane filtration-based culture methods for P. aeruginosa: ISO 16266:2006 and UK The Microbiology of Drinking Water – Part 8 (MoDW Part 8), which both use Pseudomonas CN agar. The comparison based on the calculation of mean relative differences between the two methods was conducted according to ISO 17994:2014. Using both routine hospital water samples (80 from six laboratories) and artificially contaminated samples (192 from five laboratories), paired counts from each sample and the enumeration method were analysed. For routine samples, there were insufficient data for a conclusive assessment, but the data do indicate at least equivalent performance of Pseudalert®/Quanti-Tray®. For the artificially contaminated samples, the data revealed higher counts of P. aeruginosa being recorded by Pseudalert®/Quanti-Tray®. The Pseudalert®/Quanti-Tray® method does not require confirmation testing for atypical strains of P. aeruginosa, saving up to 6 days of additional analysis, and has the added advantage of providing confirmed counts within 24–28 hours incubation compared to 40–48 hours or longer for the ISO 16266 and MoDW Part 8 methods.

Comparative study of membrane filtration and enrichment media for the isolation and enumeration of Pseudomonas aeruginosa from sewage, surface water, and swimming pools

1985 ◽  
Vol 31 (8) ◽  
pp. 686-692 ◽  
Author(s):  
A. H. Havelaar ◽  
M. During ◽  
E. H. M. Delfgou-Van Asch
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Membrane Filtration ◽  
Culture Media ◽  
Polluted Water ◽  
Most Probable Number ◽  
Cellulose Ester ◽  
Swimming Pools ◽  
Probable Number ◽  
Membrane Filters ◽  
Swimming Water

The recovery of Pseudomonas aeruginosa on several selective culture media was tested using raw sewage and secondary sewage effluent samples as well as spiked chlorinated imitation swimming water and samples from whirlpools. mPA-medium B gave good recovery of both vital and chlorine-injured P. aeruginosa and selectivity was greater than 90% when analysing whirlpool samples. It is therefore the medium recommended for examination of chlorinated swimming pools. When analysing sewage polluted water with the mPA-B medium, reduced selectivity was noted from low verification rates and from overgrowth by competitive flora. A modified medium (mPA-D; addition of cetrimide, omission of sulphapyridine and actidione) was more selective and sufficiently recovered noninjured cells. Chlorine-injured cells were completely inhibited, however. C-390 (9-chloro-9-(4-diethylaminophenyl)-10-phenylacridan) was confirmed to be highly selective for P. aeruginosa when used in spread plates at a concentration of 30 μg/mL; P. aeruginosa was slightly inhibited. However, the medium could not be used with conventional membrane filtration techniques, because cellulose ester filters interfered with the selective action of C-390. Selectivity could be improved by using Gelman Tuffryn (polysulphone) filters and increasing the C-390 concentration to 120 μg/mL. At this concentration, however, the medium was strongly inhibitory to P. aeruginosa; resuscitation only partially improved recovery. Two other membrane filtration media were tested. Both cetrimide – nalidixic acid agar and Drake's medium No. 19 were inhibitory to chlorine-injured cells. Several types of membrane filters were tested and there was little difference between them. In the most-probable-number technique, recovery of P. aeruginosa was shown to be excellent when using asparagine broth. Malachite green broth was strongly inhibitory to chlorine-injured P. aeruginosa.

scholarly journals Correlation between Quantitative PCR and Culture-Based Methods for Measuring Enterococcus spp. over Various Temporal Scales at Three California Marine Beaches

2011 ◽  
Vol 78 (4) ◽  
pp. 1237-1242 ◽  
Author(s):  
Reagan R. Converse ◽  
John F. Griffith ◽  
Rachel T. Noble ◽  
Richard A. Haugland ◽  
Kenneth C. Schiff ◽  
...  
Keyword(s):  
Quantitative Pcr ◽  
Membrane Filtration ◽  
Empirical Relationship ◽  
Time Of Day ◽  
Most Probable Number ◽  
Spatial And Temporal Variability ◽  
Culture Methods ◽  
Fecal Indicator ◽  
Probable Number ◽  
Content Type

ABSTRACTSeveral studies have examined how fecal indicator bacteria (FIB) measurements compare between quantitative PCR (qPCR) and the culture methods it is intended to replace. Here, we extend those studies by examining the stability of that relationship within a beach, as affected by time of day and seasonal variations in source.Enterococcusspp. were quantified at three southern California beaches in the morning and afternoon using two qPCR assays, membrane filtration, and defined-substrate testing. While qPCR and culture-based measurements were consistently and significantly correlated, strength of the correlation varied both among and within beaches. Correlations were higher in the morning (0.45 < ρ < 0.74 [P< 0.002]) than in the afternoon (0.18 < ρ < 0.45 [P< 0.021]) and higher when the fecal contamination was concentrated (0.38 < ρ < 0.83 [P< 0.001]) than when it was diffuse (0.19 < ρ < 0.34 [P< 0.003]). The ratios of culture-based and qPCR results (CFU or most probable number [MPN] per calibrator cell equivalents [CCE]) also varied spatially and temporally. Ratios ranged between 0.04 and 0.85 CFU or MPN per CCE and were lowest at the beach affected by diffuse pollution. Patterns in the ratios over the course of the day were dissimilar across beaches, increasing with time at one beach and decreasing at another. The spatial and temporal variability we observed indicate that the empirical relationship between culture-based and qPCR results is not universal, even within a beach.

A Most Probable Number Method for the Enumeration of Legionella Bacteria in Water

1991 ◽  
Vol 24 (2) ◽  
pp. 143-147 ◽  
Author(s):  
N. A. Grabow ◽  
R. Kfir ◽  
W. O. K. Grabow
Keyword(s):  
Water Samples ◽  
Membrane Filtration ◽  
Quantitative Method ◽  
Fluorescent Antibody ◽  
Most Probable Number ◽  
Probable Number ◽  
Comparative Tests ◽  
Antibody Staining ◽  
Direct Fluorescent Antibody ◽  
Yeast Extract Agar

A new quantitative method for the enumeration of Legionella bacteria in water is described. Appropriate tenfold serial dilutions of water samples concentrated by membrane filtration are plated in triplicate on buffered charcoal yeast extract agar. After incubation for 3 days representative smears from individual plates are tested for the presence of Legionella by direct fluorescent antibody staining. The number of positive plates in each dilution is used to calculate the Legionella count by means of conventional most probable number statistics. In comparative tests on a variety of water samples this method yielded significantly higher counts than previously used procedures.

The relationship between three potential pathogens and pollution indicator organisms in Nova Scotian coastal waters

1983 ◽  
Vol 29 (10) ◽  
pp. 1261-1269 ◽  
Author(s):  
W. J. Robertson ◽  
R. S. Tobin
Keyword(s):  
Membrane Filtration ◽  
Indicator Bacteria ◽  
Fecal Coliforms ◽  
Most Probable Number ◽  
Indicator Organisms ◽  
Fecal Indicator ◽  
Filtration Method ◽  
Probable Number ◽  
Pollution Indicator ◽  
Nova Scotian

Fifteen stations, in two estuaries, along the Northumberland Strait of Nova Scotia were examined between June and September 1981 for a relationship between the concentrations of commonly monitored fecal indicator bacteria and the potential pathogens Candida albicans, Pseudomonas aeruginosa, and Vibrio parahaemolyticus. Increased densities of these three organisms were usually associated with high densities of indicator bacteria. Whereas C. albicans and P. aeruginosa occur in human fecal wastes, V. parahaemolyticus is indigenous to the marine environment and positively responds to elevated nutrient levels in sewage. There is also some evidence that these bacteria survive as long or longer in marine waters than the common indicator bacteria. While membrane-filtration techniques for the enumeration of C. albicans and P. aeruginosa proved satisfactory, a V. parahaemolyticus membrane-filtration method lacked specificity and was supplemented by a most-probable-number method. In marine recreational and shellfish waters, these three organisms could complement fecal coliforms and fecal streptococci as indicators of human fecal contamination.

Colony-Forming Unit Enumeration by a Plate-MPN Method

1983 ◽  
Vol 46 (10) ◽  
pp. 836-841 ◽  
Author(s):  
S-T. TAN ◽  
R. B. MAXCY ◽  
W. W. STROUP
Keyword(s):  
Standard Technique ◽  
Most Probable Number ◽  
Plate Method ◽  
Probable Number ◽  
Microbial Counts ◽  
Colony Forming Unit ◽  
Pure Cultures ◽  
Petri Plate ◽  
Mpn Test ◽  
Surface Plate

Concepts of the standard surface plate method and the most probable number method (MPN) were combined to provide a new enumeration technique (plate-MPN). Three discrete 0.01-ml samples of an appropriate decimal dilution were inoculated onto each quadrant of a pre-dried petri plate. The discrete spots from the inoculum were then observed for growth after incubation. Results were interpreted analogous to a 3-tube MPN test using presently available tables. Application of the test to pure cultures and mixed flora provided no evidence to indicate the plate-MPN technique to be any less accurate than the standard technique for microbial counts. The plate-MPN technique was less precise than the standard technique. However, the plate-MPN technique has many advantages over traditional methods.

scholarly journals Rapid single-tube confirmatory test forEscherichia coli

1982 ◽  
Vol 89 (1) ◽  
pp. 149-154 ◽  
Author(s):  
J. L. Smith ◽  
S. Rockliff
Keyword(s):  
Membrane Filtration ◽  
Confirmatory Test ◽  
Most Probable Number ◽  
Standing Committee ◽  
Traditional Methods ◽  
Probable Number ◽  
Single Tube ◽  
Laboratory Service ◽  
The Public ◽  
E Coli

SUMMARYA single-tube confirmatory test that allows a result to be obtained in 4 h has been developed from the single-tube confirmatory test recommended by the Joint Committee of the Public Health Laboratory Service and the Standing Committee of Analysts (PHLS/SCA, 1980). A variety of river, lake and reservoir samples were examined for the presence ofE. coliusing either most probable number (MPN) or membrane filtration (MF) technique, and the PHLS/SCA recommended confirmatory medium (LTMB) was evaluated against traditional methods. To improve the performance of LTMB, the medium was modified and this modified medium when used in 0·1 ml volumes and incubated for 4 h at 44°C provided 99% agreement with traditional methods.

scholarly journals Evaluation of Culture Methods To Identify Bovine Feces with High Concentrations of Escherichia coli O157

2007 ◽  
Vol 73 (16) ◽  
pp. 5253-5260 ◽  
Author(s):  
J. Trent Fox ◽  
David G. Renter ◽  
Michael W. Sanderson ◽  
Daniel U. Thomson ◽  
Kelly F. Lechtenberg ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Sensitivity And Specificity ◽  
Immunomagnetic Separation ◽  
Most Probable Number ◽  
Culture Methods ◽  
Potassium Tellurite ◽  
Probable Number ◽  
E Coli ◽  
Concentration Sensitivity ◽  
High Concentrations

ABSTRACT Our objective was to evaluate methods for identifying cattle with high concentrations of Escherichia coli O157 in their feces. In two experiments, feces were collected from cattle orally inoculated with nalidixic acid (Nal)-resistant E. coli O157, and direct plating of diluted feces on sorbitol MacConkey agar with cefixime and potassium tellurite (CT-SMAC) containing Nal was considered the gold standard (GS) method. In experiment 1, methods evaluated were preenrichment direct streak, immunomagnetic separation with most probable number (MPN), and postenrichment direct streak with MPN, all using CT-SMAC. The mean concentration of Nal-resistant E. coli O157 in samples (n = 59) by use of the GS was 3.6 log10 CFU/g. The preenrichment streak detected >3.0 log10 CFU/g samples with a 74.4% sensitivity and 68.8% specificity. Postenrichment direct streak-MPN and immunomagnetic separation-MPN concentrations were correlated significantly with GS concentrations (r = 0.53 and r = 0.39, respectively). In experiment 2 (480 samples), pre- and postenrichment direct streaking performed in triplicate and spiral plating on CT-SMAC were evaluated. For preenrichment streaks, sensitivity was 79.7% and specificity was 96.7% for detecting >3.0 log10 CFU/g when the criterion was positive cultures on at least two plates. For spiral plating at that concentration, sensitivity and specificity were 83.9% and 56.3%, respectively. Postenrichment streaking performed relatively poorly. Triplicate preenrichment streaks of 1:10-diluted feces on CT-SMAC may be useful for identifying cattle shedding high concentrations of E. coli O157. Estimates of sensitivity and specificity enable appropriate application of methods and interpretation of results and may enhance applied research, surveillance, and risk assessments.

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