Practical and low cost tests for biological, microbiological and toxicological analysis of water contamination Prof.em.Dr.G.

Transcription

1 Practical and low cost tests for biological, microbiological and toxicological analysis of water contamination Prof.em.Dr.G.Persoone -Past Director of the Laboratory for Environmental Toxicology and Aquatic Ecology, Ghent University, Belgium -Scientific Advisor MicroBioTests Inc.

2 WATER CONTAMINATION Toxicological analyses Microbiological analyses Biological analyses

3 Toxicological analyses Short-term «conventional» aquatic toxicity tests : - 72 h Algal growth inhibition test h Daphnia immobilisation test - 96 h Fish mortality test

4 Culturing of micro-algae Infrastructure Space Labour

5 Inoculation of test vessels with micro-algae Incubation of test vessels ALGAL TEST PROCEDURE Daily counting of micro-algae

6 The waterflea Daphnia magna

7 Culturing of Daphnias Daily maintenance Daily feeding with micro-algae Infrastructure Space Labour

8 Fish toxicity test

9 Fish culturing/maintenance facilities Infrastructure Space Labour

10 Sediment contact tests Stock culturing of the amphipod crustacean Hyalella azteca

11 Equipment and space Manipulations Infrastructure Space Labour

12 Burden of most toxicity tests : Dependence of the (continuous) culturing/maintenance of live stocks of test species Infrastructure Space Time Costs

13 MICROBIOTESTS As alternatives to conventional toxicity tests

14 CONVENTIONAL TOXICITY TESTS Dependent of the (year-round) culturing of live stocks of test species - Specialised - Time consuming - Costly TOXKIT MICROBIOTESTS Culture/maintenance free - Small-scale - User-friendly - Cost-effective - Suited for routine analyses

15 Development of a battery of acute and short-chronic TOXKIT MICROBIOTESTS Worldwide application for toxicity screening and biomonitoring More than Toxkit tests have already been performed in 40 countries worldwide

16 DAPHTOXKIT magna Hatched neonates Dormant egg

17 ALGALTOXKIT Micro-algae immobilised in algal beads Algal beads stored in tube De-immobilised micro-algae Long cells as test containers

18 OSTRACODTOXKIT F 6 days direct contact test for sediments and soils (250 µm) Larvae of the crustacean Heterocypris incongruens, hatched from cysts

19 PHYTOTOXKIT A seed germination and early growth test with higher plants Direct measurement of root and shoot length in transparent test containers by image analysis

20 WATER CONTAMINATION EMERGENCIES Need for very rapid detection of toxicity CHEMICAL ANALYSES What to search for? Time needed? TOXICITY TESTS Signal for the impact of all the chemicals present

21 The RAPIDTOXKIT with larvae of the crustacean Thamnocephalus platyurus A very simple and practical test for detection of toxic hazard in water in minutes

22 The RAPIDTOXKIT microbiotest in a nutshell Suspected water Control water minutes exposure minutes exposure Addition of red microspheres (15-30 minutes) No particle uptake Uptake of red particles YES NO Toxic hazard

23 Field (in situ) analyses Toxicity of water Contamination of water by bacteria and biological residues URGENT NEED FOR FIELD TESTS

24 Development of two low cost and user-friendly kits for ultra-rapid field analyses Toxi-Screening Kit Bacterial Contamination Screening Kit

25 Basic principle of the new field kits Luminescence measurement in a portable low cost luminometer Toxi-Screening Kit : bacterial bioluminescence Bacterial Contamination Screening Kit : ATP-chemical luminescence

26 Incorporation of the equipment and materials in a handy lightweight Luminescence Measurements Case

27 Toxi-Screening Kit 1 box with 10 vials of freeze-dried luminescent bacteria 10 Unit boxes each for analysis of 2 water samples

28 Toxicity Screening 1.Test preparation Transfer 1 ml rehydration medium into vial with lyophilised bacteria 30 min rehydration 2. Sampling Take a 1 ml water sample and transfer it in the sample tube

29 3. Transfer of luminescent bacteria in control and sample tube : Transfer of 200 µl bacterial suspension + Vial with rehydrated bacteria Control tube Sample tube 4. Exposure : 30 minutes

30 5. Scoring (at start and after 30 minutes exposure) Adjust the holder to the tube Insert the tube + holder in the luminometer and close Score the luminescence (in Relative Light Units = RLU) 6. Data evaluation : % toxicity = RLU (T0-T30) sample RLU (T0-T30) control X 10 TOTAL TIME NEEDED FOR THE TEST : 1 hour

31 Bacterial Contamination Screening Kit with 10 Unit boxes each for analysis of 2 water samples

32 Screening of Bacterial Contamination Tube with ATP reagents Compartment with liquid ATP extractant Compartment with solid ATP reagent

33 DIRECT TEST for evaluation of the degree of contamination by bacteria and biological residues 1.Test performance 1 minute Transfer 200 µl water sample in the top compartment of the tube Transfer the contents of the top compartment into the lower compartment of the tube Adjust the holder to the tube and shake

34 2. Test scoring (after 2 minutes) Insert the tube + holder in the luminometer and close Score the luminescence (in Relative Light Units = RLU) TOTAL TIME NEEDED FOR THE DIRECT TEST : 5 minutes

35 3. Evaluation of the degree of contamination by bacteria and biological residues RLU score Degree of contamination < 50 Very low < 200 Low Significant > 1000 Very high

36 MEMBRANE FILTER TEST for evaluation of the degree of contamination by bacteria 1. Elimination of interferences from dissolved chemicals 2. Elimination of dissolved extracellular ATP 3. Elimination of non-bacterial ATP 4. Possibility of application to estuarine/marine waters 5. Application to larger volumes of water samples

37 1.Test preparation : Placing of pre-filter and membrane filter in mini-filter holders Unscrew the 2 parts of One of the 2 filter holders Put a 10 µm pre-filter in the hollow part of the filter holder Reassemble the 2 parts of the filter holder Proceed the same way for plaing a 0.2 µm membrane filter in the second filter holder

38 2. Test performance Membrane filter holder Pre-filter holder Pre-filter holder Connect the 2 filter holders Take a 10 ml water sample with the syringe Membrane filter holder Put the syringe on the connected filter holders and flush the water through the fillter holders

39 Remove the sealed cover of the tube with the pincette Take the membrane filter out of the filter holder with the pincette Submerge the membrane filter for 1 minute in the ATP extracting medium Adjust the holder to the tube and shake Transfer the contents of the top chamber into the lower chamber of the tube

40 3. Test scoring Insert the holder + tube in the luminometer and close Score the luminescence TOTAL TIME NEEDED FOR THE MEMBRANE FILTER TEST <10 minutes

41 4. Evaluation of the degree of bacterial contamination - Recalculate the scored RLU value to 1 ml of filtered water sample - Estimate the degree of bacterial contamination of the water RLU/ml Degree of bacterial contamination < 200 Very low < 1000 Relatively low > 1000 Significant > 5000 High > Very high

42 + + Luminescence Measurements Case + Toxi-Screening Kit Bacterial Contamination Screening Kit A breakthrough for field testing of suspected waters for their toxicity and/or bacterial contamination Ultra-rapid User-friendly Low cost

43 Biological analyses of water contamination

44 All-In portable microscope

45 - Small size (only 28x16x10 cm) - Lightweight (only 1.1 kg) - Handy carrying case Suited for microscopic observations in the laboratory as well as in the field

46 Same characteristics as conventional laboratory microscopes Revolver with 3 objectives and 2 eyepieces for magnifications from 20X to >1000X Cross table to clamp slides and freely move them in all horizontal directions Electrical low voltage illumination with adjustable light intensity

47 Provided with various additional UNIQUE assets Incident (= top) illumination for examination of nontransparent objects

48 Microscopic view of dry brine shrimp cysts with bottom illumination Same view with top illumination

49 Microscopic view of dry yeast pellets with bottom illumination Same view with top illumination

50 Battery holder with 3 batteries (1.5V) for use of the microscope anywhere and anytime independently of the electricity means NB: This allows for up to 600 observations (of 30 minutes each) with the same batteries

51 Provided with a webcam ocular and software for direct observations on a computer screen

52 Provided with multiple accessories for microscopic observations and preparation of specimens and slides

53 Transparent chamber with 4 compartments for direct observations of organisms under the microscope Minifiltration unit for capture and concentration of planktonic organisms CD Rom with 2 programmes for image transfer to computer Nonbreakable (plastic) slides and coverslips

54 Special nonbreakable coverslips with grid for counting and direct length measurements 1 cm

55 Storage of all removable parts and accessories in specific compartments of a panel which also fits in the carrying case

56 All-In portable microscope The ultimate for microscopic analyses in the laboratory as well as in the field Price? Incredibly low!!

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