Bactiquant®-waterTotal bacterial analysis in minutes
Outline of the presentationBactiquant® waterHow it worksDocumentationApplication casesQuestions11/06/2013 Direction or Dep...
Third party verification11/06/2013 Direction or Department name 3US-EPA Verified TechnologyU.S. Environmental Protection A...
LaboriousSkilled lab personelLong analysis timeIndustry Challenges:
Problem11/06/2013 Direction or Department name 5You do not get timely results
Rapid resultsMaintain controlReduce downtimeIndustry Requirements:Save time and money
Total bacterial analysis in minutes
Bactiquant®-waterTotal bacteriaFrom days to minutes11/06/2013 Direction or Department name 8
Sampling11/06/2013 Direction or Department name 9
Enzyme targeted analysis+Bacteria-enzymeEnzyme substrate Fluorescent compound released
Enzyme targeted analysisEnzyme substrateFluorophore
Robust concept11/06/2013 Direction or Department name 12•Naturally occuring enzyme activity•High specificity for bacteria•...
Simple interpretation11/06/2013 Direction or Department name 13The more fluorescense the more bacteria ispresent in the sa...
Linearity with sample volume andreaction time11/06/2013 Direction or Department name 14y = 17,745xR² = 0,99902004006008000...
The BQ value11/06/2013 Direction or Department name 15250 ml water sample30 minutes reaction23 C (73,4 F)The Bactiquant va...
Calculating the BQ value11/06/2013 Direction or Department name 16LOD = 10 fluorescence units(Reading-blank) (vol / 250)* ...
Simple protocol•Concentrate bacteria on a sterile filter usingsyringe/vacuum manifold•Concentration from as large a volume...
Bactiquant®- AdvantagesExclusion of a cultivation stepBacteria does not have to reproduce prior to analysisReduction in pe...
Detecting all bacteriaTraditional plate counts do not measure particle bound bacteriaParticle bound bacteria are important...
Traditional plate count11/06/2013 Direction or Department name 20Plate count estimateTotal population
A comprehensive measure11/06/2013 Direction or Department name 21Organism Gram stain Phyllum/ classAthrobacter sp. Negativ...
Comparing BQ and HPC11/06/2013 Direction or Department name 22• The result does not depend on the ability ofbacteria to gr...
Comparing Bactiquant with ATP11/06/2013 Direction or Department name 23Parameter Comments Bactiquant ATPRapidity 10-30 min...
Quantification in environmentalsamples11/06/2013 Direction or Department name 24
Complex Environmental Sample11/06/2013 Direction or Department name 250123450 1 2 3 4 5 6Bactiquant valueData: Total count...
Simple samples – suspended bacteria11/06/2013 Direction or Department name 261101001000100001000001000000100000001 10 100 ...
High reproducibility11/06/2013 Direction or Department name 27Source US EPA -2011P. aeruginosa ATCC 27853Test IterationAdj...
Repeatability and inter assay reproducibility11/06/2013 Direction or Department name 28Test IterationAdjusted Fluorescence...
Excellent water quality – drinking water11/06/2013 Direction or Department name 29BQ-values < 40• 976 water samples• 200 s...
Legionella and general hygiene11/06/2013 Direction or Department name 30012345671 4 7 10 13 16 19 22 25 28 31 34 37 40 43 ...
Calibration curve – BQ Value and CFU/ml inUltra-pure Water31Courtesy of The Danish Technological InstituteSensitivity leve...
Plate counts can be predicted by the rapid methodGood accuracy with ultra-pure waterResults show plate counts of 20 CFU/ml...
Bactiquant® ApplicationsSource water/Raw water qualityWater plant (operations/process control)Distribution systems (finish...
Application Cases11/06/2013 Direction or Department name 34
Monitoring critical control points11/06/2013 Direction or Department name 35R åvand16650Tils ætning afFeCl 3 ogH2SO 45307E...
Advantages of process controlFewer resources are used on correctingproblems after they have occurredEnables the end user t...
How do we translate analyses results intooperation and optimization of processes ?Data Handling
Data handling softwareEstablish baselines based on frequent long term monitoringat Critical Control PointsEstablish operat...
Tracking a pollutant
Tracking a polution overview11/06/2013 Direction or Department name 40outletoutletO2Sand filterGround waterBoreholesWater ...
Tracking a pollutant
Tracking a pollution11/06/2013 Direction or Department name 42
11/06/2013 Direction or Department name 43Early warningRapid adjustmentMaintain controlValue to the customer
Cooling water11/06/2013 Direction or Department name 44
Cooling water11/06/2013 Direction or Department name 450100200300400500600700800900Leaking cansLeaking cansRepair conveyer...
11/06/2013 Direction or Department name 46Early warningRapid AdjustmentMaintain controlValue to the customer
11/06/2013 Direction or Department name 4705001000150020002500180820681270934Bactiquant®valueBottled and tap water survey,...
Optimising chlorination11/06/2013 Direction or Department name 48ReplicateRaw SampleReaction time(minutes)Volume(ml)BQV(FU...
Verifying ultrafiltration11/06/2013 Direction or Department name 498767217010002000300040005000600070008000900010000before...
Bactiquant® Value to industryTimely resultsRespond quicklyMaintain controlOptimize water treatmentsIncrease over all syste...
Return on investment11/06/2013 Direction or Department name 51Optimise operating costsReduce downtimePrevent lost producti...
QUICK FACTS: Duration: 3-5 hours. Held at the customer’s locationCOURSE OUTLINE: Proficiency in performing theanalysis...
Quality Assurance Program11/06/2013 Direction or Department name 53What is a Quality Assurance Program• The Quality Assura...
FlexibleSamplingRapidAnalysisEasyInterpretationTo wrap it up
US-EPA (Environmental Protection Agency)verified technologyRapid results – on locationRobust analysis methodReproducible a...
Whats next ?11/06/2013 Direction or Department name 56Vittorio FiguratoMarketing DepartmentELGA Process Watervittorio.figu...
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Bactiquant®-water webinar presentation

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The BactiQuant is a new patented method that provides water companies and the industry with a rapid method to estimate total numbers of bacteria in water samples. The method is well suited for HACCP and Water Safety Plans in the industry and water distribution.

With the BactiQuant, bacterial contaminants can be concentrated from large volumes of water allowing low analysis time (10-30 minutes) even at relatively low bacterial concentrations.
The use of larger sample volumes (100-5000 ml) also ensures that samples are representative for the water system to be tested.
The BactiQuant can be applied to a range of liquid samples including: Potable water, processed water, CIP (cleaning in place) and recreational water. Other liquids include: Liquid extracts of a solid material or slurries, suspended or solubilized surface samples and liquid industrial products.

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Bactiquant®-water webinar presentation

  1. 1. Bactiquant®-waterTotal bacterial analysis in minutes
  2. 2. Outline of the presentationBactiquant® waterHow it worksDocumentationApplication casesQuestions11/06/2013 Direction or Department name 2
  3. 3. Third party verification11/06/2013 Direction or Department name 3US-EPA Verified TechnologyU.S. Environmental Protection AgencyU.S. Environmental Protection Agency
  4. 4. LaboriousSkilled lab personelLong analysis timeIndustry Challenges:
  5. 5. Problem11/06/2013 Direction or Department name 5You do not get timely results
  6. 6. Rapid resultsMaintain controlReduce downtimeIndustry Requirements:Save time and money
  7. 7. Total bacterial analysis in minutes
  8. 8. Bactiquant®-waterTotal bacteriaFrom days to minutes11/06/2013 Direction or Department name 8
  9. 9. Sampling11/06/2013 Direction or Department name 9
  10. 10. Enzyme targeted analysis+Bacteria-enzymeEnzyme substrate Fluorescent compound released
  11. 11. Enzyme targeted analysisEnzyme substrateFluorophore
  12. 12. Robust concept11/06/2013 Direction or Department name 12•Naturally occuring enzyme activity•High specificity for bacteria•Amplifying signal – adjustable sensitivity
  13. 13. Simple interpretation11/06/2013 Direction or Department name 13The more fluorescense the more bacteria ispresent in the sample
  14. 14. Linearity with sample volume andreaction time11/06/2013 Direction or Department name 14y = 17,745xR² = 0,99902004006008000 10 20 30 40FlourescenceReaction time (minutes)Bactiquant®-water test on 500 ml tap water. Fluorescencemeasured as function of reaction time. Conducted at 26oCy = 0,7709xR² = 0,9959010203040506070800 20 40 60 80 100Flourescenceml tap water sampleBactiquant®-water test on tap water samples.Fluorescense measured as function of sample size.Conducted at 25oC and with 65 minutes reaction time.R2 Agar showed 28 CFU/ml in tap water.Calculation of Bactiquant Value (BQV) for this trial.1 BQV = 0.5 CFU/ml
  15. 15. The BQ value11/06/2013 Direction or Department name 15250 ml water sample30 minutes reaction23 C (73,4 F)The Bactiquant value is universal
  16. 16. Calculating the BQ value11/06/2013 Direction or Department name 16LOD = 10 fluorescence units(Reading-blank) (vol / 250)* * (Reaction time / 30) * (Temperature adjustment)
  17. 17. Simple protocol•Concentrate bacteria on a sterile filter usingsyringe/vacuum manifold•Concentration from as large a volume as possible•Can be sampled directly from sample points•Use the filter as the reaction vehicle•Saturate filter with substrate•React in filter for a set time•Typically 10 – 30 min incubation•Flush out cleaved fluorophore into cuvette•Measure using hand-held fluorometer
  18. 18. Bactiquant®- AdvantagesExclusion of a cultivation stepBacteria does not have to reproduce prior to analysisReduction in performance time from days to minutesFluorescent substrate analoguesNo permeabilization of cells necessaryRobust chemistryConcentration stepIncreases sensitivityReduction in detection timeMinimal human interventionClosed filtration unitNo handling of filter membrane necessaryRobust and repeatable
  19. 19. Detecting all bacteriaTraditional plate counts do not measure particle bound bacteriaParticle bound bacteria are important for water qualityBactiquant®-waterBactiQuant®Water detects and quantifies suspendedbacteria as well as bacteria located on particles or in aggregates.Why is this important ?
  20. 20. Traditional plate count11/06/2013 Direction or Department name 20Plate count estimateTotal population
  21. 21. A comprehensive measure11/06/2013 Direction or Department name 21Organism Gram stain Phyllum/ classAthrobacter sp. Negative Actinobacteria / grp. 20Bacillus cereus Positive Firmicutes / grp. 18Bacillus subtilis Positive Firmicutes / grp. 18Bacillus thuringiensis Positive Firmicutes / grp. 18Clostridium tetani Positive Firmicutes / clostridiaDesulfovibrio desulfouricans Positive Proteobacteria / δEnterobacter sp. Negative Proteobacteria / γEnterococcous faecalis Positive Firmicutes / bacilliEschericia coli Negative Proteobacteria / γEscherichia fergusonii Negative Proteobacteria / γFlavobacteria bacterium Negative Bacteroidetes / flavobacteriaFlavobacterium johnsoniae Negative Bacteroidetes / flavobacteriaKlebsiella pneumonia Negative Proteobacteria / γLactobacillus acidophilus Positive Firmicutes / bacilliLactobacillus reuteri Positive Firmicutes / bacilliMethanococcus aeolicus Negative Euryarchaeota / methanococciMicrococcous sodonensis Positive Actinobacteria / actinobacteriaPseudomonas aeroginosa Negative Proteobacteria / γRhodobacter sphaeroides Negative Proteobacteria / αSerratia marcescens Positive Proteobacteria / γStaphylococcous aureus Positive Firmicutes / bacilliSphingomonas Wittichii Negative Proteobacteria / αVibrio sp Negative Proteobacteria / γXanthomonas campestris Positive Proteobacteria / γSource: Brenda database
  22. 22. Comparing BQ and HPC11/06/2013 Direction or Department name 22• The result does not depend on the ability ofbacteria to grow on a specific media.• Bactiquant®-water detects and quantifies particleassociated bacteria as well as suspended bacteria.
  23. 23. Comparing Bactiquant with ATP11/06/2013 Direction or Department name 23Parameter Comments Bactiquant ATPRapidity 10-30 min 2 minEase of useModeratelyeasyVeryeasyReproducibilityPermeabilization of cellsnecessary.No YesThe robustness of assaychemistryHigh LowThe assay sensitive to waterchemistryLow HighThe overall reproducibility High LowSpecificity to Bacteria High NoUS-EPA verificationYes No
  24. 24. Quantification in environmentalsamples11/06/2013 Direction or Department name 24
  25. 25. Complex Environmental Sample11/06/2013 Direction or Department name 250123450 1 2 3 4 5 6Bactiquant valueData: Total counts 35oC, 48 hours, R2A , pour plate procedure– Middle East utility0123450 1 2 3 4 5Bactiquant valueAll data PopulationsGround water/desalinated waterTreated surface waterUntreated surface waterTVC(CFU/ml)
  26. 26. Simple samples – suspended bacteria11/06/2013 Direction or Department name 261101001000100001000001000000100000001 10 100 1000 10000 100000 1000000BactiQuant analysis result (fu/ml)HPC-DS6222(cfu/ml)
  27. 27. High reproducibility11/06/2013 Direction or Department name 27Source US EPA -2011P. aeruginosa ATCC 27853Test IterationAdjusted Fluorescence (fu)8.0 x 103 CFU/mL 4.7 x 103 CFU/mL 2.1 x 103 CFU/mL 8.7 x 102 CFU/mL1 11392 6128 2799 12492 12332 6074 2710 12473 11376 6012 2559 12974 11513 6508 2768 14375 11614 6256 2945 1375Average 11645 6196 2756 1321Standard deviation 396 196 140 83RSD (%) 3.4 3.2 5.1 6.3
  28. 28. Repeatability and inter assay reproducibility11/06/2013 Direction or Department name 28Test IterationAdjusted Fluorescence (fu)Indigenous Bacteria from Lake Water(3.7 x 102 CFU/mL)P. aeruginosa ATCC 27853(4.7 x 103 CFU/mL)Analyst 1 Analyst 2 Analyst 1 Analyst 2Average 2363 2225 6888 6691Standard deviation 152 57 333 93RSD (%) 6.4 2.6 4.8 1.4RPD (%) 6.0 2.9Source US EPA -2011
  29. 29. Excellent water quality – drinking water11/06/2013 Direction or Department name 29BQ-values < 40• 976 water samples• 200 sampling sites in Copenhagen•Plate counts (DS/EN ISO 6222) at 22 and 36oC•BQ value < 40 less than 1% probability of exceeding 200 cfu
  30. 30. Legionella and general hygiene11/06/2013 Direction or Department name 30012345671 4 7 10 13 16 19 22 25 28 31 34 37 40 43 46 49 52 55 58 61 64 67 70BQLegionella pneumophila serotype 2-14BQ value < 40Courtesy of Viborg-Kjellerup Hospital, clinical microbiologyinstitute, 2006, Denmark
  31. 31. Calibration curve – BQ Value and CFU/ml inUltra-pure Water31Courtesy of The Danish Technological InstituteSensitivity level: 1 liter sample, reaction time 5 hoursBQ- ValuesHPC(CFU/ml)
  32. 32. Plate counts can be predicted by the rapid methodGood accuracy with ultra-pure waterResults show plate counts of 20 CFU/mlcorresponds to a Bactiquant®-value of 1Ultra pure water
  33. 33. Bactiquant® ApplicationsSource water/Raw water qualityWater plant (operations/process control)Distribution systems (finished water)Industrial process waterFood and beverage productionTracking pollution eventsHACCP (hazard analysis and critical control points)Technical water (electronics, cooling towers)EffluentsRecreational waters
  34. 34. Application Cases11/06/2013 Direction or Department name 34
  35. 35. Monitoring critical control points11/06/2013 Direction or Department name 35R åvand16650Tils ætning afFeCl 3 ogH2SO 45307Efter filter209Efter ozon -Behandling70AfgangVandv ærk10GAC filter187Actifloc691SurfaceWaterIntakeSand-filtrationOzonizationUVGACAftersediment.FlocullationAl, FeOutletDistribution16650 5300690 21070190 160 BDChlorination
  36. 36. Advantages of process controlFewer resources are used on correctingproblems after they have occurredEnables the end user to be proactive
  37. 37. How do we translate analyses results intooperation and optimization of processes ?Data Handling
  38. 38. Data handling softwareEstablish baselines based on frequent long term monitoringat Critical Control PointsEstablish operational limits
  39. 39. Tracking a pollutant
  40. 40. Tracking a polution overview11/06/2013 Direction or Department name 40outletoutletO2Sand filterGround waterBoreholesWater work1212800
  41. 41. Tracking a pollutant
  42. 42. Tracking a pollution11/06/2013 Direction or Department name 42
  43. 43. 11/06/2013 Direction or Department name 43Early warningRapid adjustmentMaintain controlValue to the customer
  44. 44. Cooling water11/06/2013 Direction or Department name 44
  45. 45. Cooling water11/06/2013 Direction or Department name 450100200300400500600700800900Leaking cansLeaking cansRepair conveyer beltLeaking cansDisinfection over weekendChlorine pump defect over nightBQvalueDate
  46. 46. 11/06/2013 Direction or Department name 46Early warningRapid AdjustmentMaintain controlValue to the customer
  47. 47. 11/06/2013 Direction or Department name 4705001000150020002500180820681270934Bactiquant®valueBottled and tap water survey,MalaysiaCourtesy of TECHKOR, Korea
  48. 48. Optimising chlorination11/06/2013 Direction or Department name 48ReplicateRaw SampleReaction time(minutes)Volume(ml)BQV(FU)RSD %1 15 15 107662 15 15 104953 15 15 102964 15 15 102965 15 25 10264Average 10423 2,00 %NaOCL – 15minutes1 15 15 9892 15 15 7083 15 15 6134 15 50 518Average 707 28 ,70 %Fracking data2012Courtesy of Mycometer Inc., USA
  49. 49. Verifying ultrafiltration11/06/2013 Direction or Department name 498767217010002000300040005000600070008000900010000before filtration after filtration Recirculation 10 daysBQ-valueCourtesy of BIN-X, Denmark22
  50. 50. Bactiquant® Value to industryTimely resultsRespond quicklyMaintain controlOptimize water treatmentsIncrease over all system reliability
  51. 51. Return on investment11/06/2013 Direction or Department name 51Optimise operating costsReduce downtimePrevent lost productionExtend equipment life
  52. 52. QUICK FACTS: Duration: 3-5 hours. Held at the customer’s locationCOURSE OUTLINE: Proficiency in performing theanalysis Applications, theory, andprecautions Implementation optionsCOURSE OUTCOME:This course is designed to providethe customer with the skills andexpertise to use the BactiquanttechnologyTraining and certification
  53. 53. Quality Assurance Program11/06/2013 Direction or Department name 53What is a Quality Assurance Program• The Quality Assurance Program has beendeveloped for our experienced customers• The Quality Assurance Program enables thecustomer to perform in house Proficiencytesting and verification of instrument function.How is the quality assurance carried out?• Customer measures the fluorescence signalof the samples according to the qualityassurance protocols.• The results are sent to Mycometer’s testinglaboratory who issues a quality test reportfor the proficiency test and the instrumentverification.
  54. 54. FlexibleSamplingRapidAnalysisEasyInterpretationTo wrap it up
  55. 55. US-EPA (Environmental Protection Agency)verified technologyRapid results – on locationRobust analysis methodReproducible and repeatableData handling softwareTraining and certification programComprehensive product package
  56. 56. Whats next ?11/06/2013 Direction or Department name 56Vittorio FiguratoMarketing DepartmentELGA Process Watervittorio.figurato@veoliawater.comBook a practical demonstration:

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