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Appendix A

Luminescent bacteria risks

This appendix contains risk assessment information for Photobacterium fisheri

(synonym: Vibrio fischeri) manufactured by HACH-LANGE GmbH in Germany.

The Luminescent Bacteria Toxicity Test reagent contains freeze-dried or liquid-dried

Photobacterium fisheri bacteria. Photobacterium fisheri luminescent bacteria are

well known as non-pathogenic and innocuous.

The origin of Photobacterium fisheri is the strain number DSM 7151. The bacteria

are multiplied but not changed.

The bacteria are used as indicator organisms to identify the toxicity of

environmental or chemical samples. The Luminescent Bacteria Toxicity Test

procedure has been standardized by ISO (International Standard Organisation) ISO

11348-1, -2, -3.

A.1 Risk specifications

Table 5

gives the risk specifications for Photobacterium fisheri.

A.2 Biosafety Level 1 information

Biosafety Level 1 practices, safety equipment, and facility design and construction

are appropriate for undergraduate and secondary educational training and teaching

laboratories, and for other laboratories in which work is done with defined and

Table 5 Risk specifications

Strain number

DSM 7151 - Vibrio fischeri (Beijerinck

1889) Lehmann and Neumann 1896AL
(Bacteria) © by DSMZ-Deutsche

Sammlung von Mikroorganismen und

Zellkulturen GmbH, Braunschweig,
Germany
Name Vibrio fischeri (Beijerinck 1889)

Lehmann and Neumann 1896AL DSMZ

number 7151 = NRRL-B-11177 = ATCC

49387

Restrictions Risk

Group 1

(harmless

bacteria)

ATCC Number: 49387
NRRL Number: B-11177
Organism: Photobacterium phosphoreum

(Cohn) Beijerinck
Designations: NRRL B-11177
Depositors: NRRL

Biosafety Level

Biosafety Level 1
ATCC: American Type Culture Collection;

NRRL: ARS Culture Collection, Northern

Regional Research Laboratory

From

Biosafety in Microbiological and Biomedical Laboratories

, (BMBL) 4th Edition

(HHS Publication number (CDC) 93-8395. U.S. Department of Health and Human
Services, Centres for Disease Control and Prevention and National Institutes of
Health; U.S. Government Printing Office: Washington DC; 1999.

Summary of Contents for Eclox

Page 1: ...2886888 Eclox Rapid Response Test Kit User Manual 01 2018 Edition 5 ...

Page 2: ...Visit us at www hach com ...

Page 3: ...e waiting time and measuring time 20 3 4 Connect the luminometer to a printer 21 3 5 Connect the luminometer to a computer 23 3 6 Install LUMISsoft on the computer 23 Section 4 Chemiluminescence Toxicity Test 25 4 1 Overview 25 4 2 Prepare the reagents for luminometer calibration and sample testing 25 4 2 1 Prepare CT Reagent 2 26 4 2 2 Prepare CT Reagent 3 26 4 3 Calibrate the luminometer 28 4 4 ...

Page 4: ...rview 53 10 2 Accuracy 54 10 3 Reagent description 54 10 3 1 Quality assurance test 54 10 4 Reagent storage and preservation 55 10 5 Prepare the reagent 55 10 6 Prepare the stock suspension using the LCK491 reagent 55 10 7 Prepare the test suspension 58 10 8 Sample collection storage and preservation 59 10 9 Interferences 59 10 10 Prepare the sample 60 10 11 Prepare the test tubes 61 10 12 Measure...

Page 5: ... 10 3 Prepare a 9 dilution series D 1 values and higher 93 11 10 4 Prepare a 3 dilutions series D 1 values and higher 98 11 11 Measure the light intensity of the test suspension 101 11 12 Measure the light intensity of the test suspension after the sample dilutions are added 106 11 13 Show or send previous results 108 Section 12 Maintenance 111 12 1 General maintenance 111 12 1 1 Cleaning the kit ...

Page 6: ...Table of Contents Table of Contents Page 6 EcloxTOC fm ...

Page 7: ...y Graphical LCD display with backlight for low light conditions Data Logging Up to 60 test results recorded in full detail Up to 100 luminescent measurements for Luminescent Bacteria Toxicity Test Up to 100 screening results for Luminescent Bacteria Toxicity Test Download Capability RS232 Power Battery powered alkaline cell lithium cell AA CAUTION For quality and safety reasons only alkaline batte...

Page 8: ... pH 0 0 to 14 0 pH TDS Auto ranging 0 0 to 99 9 ppm 100 to 999 ppm 1 00 to 10 00 ppt Accuracy TDS 1 Operating temperature 0 to 50 C Battery life 250 hours with backlight continuously on Enclosure IP67 Rated waterproof immersible dust proof Warranty 1 year for the tester and 6 months for replacement sensor for manufacturing faults only Damage from use is not covered ...

Page 9: ...etting up or operating this equipment Pay attention to all danger and caution statements Failure to do so could result in serious injury to the operator or damage to the equipment Make sure that the protection provided by this equipment is not impaired Do not use or install this equipment in any manner other than that specified in this manual CAUTION Chemical exposure hazard Obey laboratory safety...

Page 10: ...ng water made or the water given for drinking and if applicable on the source water Other configurations are available including the Eclox toxicity only kit and the luminometer by itself 2 2 1 Test descriptions Eight tests are included in the Eclox Rapid Response Test Kit Chemiluminescence Toxicity Test shows the toxicity of the water sample This test uses a plant enzyme which creates light chemil...

Page 11: ...ter in a RO type of process Color Test a comparison test that compares the water sample with a calibrated gradient color disc The results are read in platinum cobalt Pt Co color units Color in water may be caused by the presence of natural metallic ions iron and manganese peat materials plankton weeds and industrial wastes Total Dissolved Solids TDS Test measures the level of dissolved solids in t...

Page 12: ...ssary The LBT screening luminescence procedure is a simplified test procedure that uses the same reagents according to ISO 11348 but at ambient conditions LIMIT measure procedure the same as the screening luminescence procedure However the LIMIT measure procedure lets the user set a LIMIT value on the luminometer The LIMIT value is used by the luminometer to include in the test results whether the...

Page 13: ... Beaker 50 mL Color viewing tubes plastic 2x Pesticide Nerve Agent Test Strips Longpath viewing adapter Pesticide test clip Color discs 2x Luminometer EZ Arsenic Reagent 1 Powder Pillows Serial cable for luminometer EZ Arsenic Reagent 2 Powder Pillows Batteries AA Alkaline EZ Arsenic Test Strips Test record pad Arsenic reaction bottle with cap Cuvette and pipette tip set blue DPD Free Chlorine Pow...

Page 14: ...14 ...

Page 15: ...tive light units made by the reagents when exposed to samples The Eclox luminometer is made for use under extreme field conditions The Eclox luminometer components are rugged easy to use and reliable refer to Figure 1 Figure 1 Luminometer components 1 Lanyard 5 Non slip feet 2 Battery compartment 6 Cell lid 3 Battery compartment screws 7 Function keys Figure 2 4 Label 8 Display ...

Page 16: ...e is not in the cell 2 Remove the black test tube holder from the cell Make sure that the cell is clean and free from debris 3 Put the black test tube holder into the cell and close the cell lid 4 Push ON green button for several seconds to apply power to the instrument If the instrument does not energize replace the batteries refer to section 12 3 on page 112 Item number Description Function 1 So...

Page 17: ...11 Push PROCEED to do a cell zeroing test When the instrument has passed the test the Signal Level screen is shown again 12 Push and hold TEST Make sure that the signal level shown is above the minimum and below the maximum If the signal level is not above the minimum contact the manufacturer for technical support 13 Push and hold the back light button Make sure that the instrument display light c...

Page 18: ... is less than the standard deviaton of the 0 1000 range and that the precision at the 0 2000 range may be better In comparison studies of each range the phenol standardization check showed equal results to the expected 50 inhibition range Non polluted sea water samples enhance the signal give a higher light inhibition of approximately 40 As the sea water becomes more polluted the percentage inhibi...

Page 19: ... stock suspension with Diluent and do the reading again To show or change the measurement range for the Luminescent Bacteria Test 1 Push ON green button to apply power to the instrument The instrument does built in tests that make sure that the electronics and software are operating correctly 2 When the built in tests are done push PROCEED The Main Menu is shown 3 Select LUMINESCENT BACTERIA TEST ...

Page 20: ...sh DOWN or UP to change the contrast The screen shows the contrast level with a Max Min indicator bar 7 Push DOWN and UP at the same time to save the changes The Set up Menu is shown 3 3 2 Set the waiting time and measuring time This procedure only applies to the Luminescent Bacteria Test The measurement of the light intensity of luminescent bacteria is divided up into two parts Waiting time the a...

Page 21: ...ent settings are shown b To change the waiting time push CHANGE c Push STORE to save the change The Set up Menu is shown 6 To show or change the measuring time a Select Set Measuring Time and push ENTER The current settings are shown b To change the measuring time push CHANGE c Push STORE to save the change The Set up Menu is shown 3 4 Connect the luminometer to a printer Luminometer measurements ...

Page 22: ...push ON LINE SW DIP switch Switch Position Setting DIP SW1 1 Off Input Serial 2 On Printing speed High 3 On Auto loading On 4 Off Auto LF Off 5 On Setting command Enable 6 Off Printing 7 On Density 8 On 100 DIP SW2 1 On Printing columns 40 2 On User font backup On 3 On Character select Normal 4 On Zero Normal 5 On International 6 On Character 7 Off Set 8 Off England DIP SW3 1 On Data length 8 bits...

Page 23: ... on a computer by doing the instructions on the CD cover A shortcut for LUMISsoft is added to the desktop during installation In the lab LUMISsoft is used to automatically get LBT Measurement Luminescence procedure results from the luminometer during the test and put the values into LUMISsoft LUMISsoft then does calculations according to ISO 11348 LUMISsoft is also used to send previous results th...

Page 24: ...24 ...

Page 25: ...xicity Tests reagent cannot be stored under those conditions 4 2 Prepare the reagents for luminometer calibration and sample testing Prepare the chemiluminescence test CT Reagents 2 and 3 at the beginning of deployment The chemiluminescence test CT Reagents 2 and 3 are temperature sensitive and degrade at high temperatures For long term storage store the reagents in their stable forms On the first...

Page 26: ...r into the CT Reagent 2 bottle 3 Put the caps back on the bottles Shake the CT Reagent 2 bottle for 30 seconds Let dissolve for 10 minutes before use 1 Remove the CT Reagent 3 concentrate and CT Reagent 3 caps Note Make sure that the batch number for CT Reagent 3 is the same as the batch number used for CT Reagent 2 2 Push the end of the pipet into a clean 100 µL yellow pipet tip and remove from t...

Page 27: ...e the liquid by gently pushing in the operating button Put the tip into the liquid and then remove from the liquid 6 Remove the tip from the pipet and put in the waste bag Put the pipet in the storage case 7 Put the cap on CT Reagent 3 Turn over the CT Reagent 3 bottle several times to mix the solution Note An ice chest can be used in the field to extend the life of the reagent ...

Page 28: ...pen the luminometer lid and make sure a sample is not in the cell Then close the lid 7 Push PROCEED The test status is shown The test may go a few minutes before it is done 8 When all the cell tests are done push PROCEED 9 Open the lid of the Cuvette and 1000 µL Pipette Tip Set 10 Put one cuvette in the black cuvette holder 11 Place a blue pipette tip on the blue pipette 12 Completely push in the ...

Page 29: ...e the cuvette from the luminometer 25 Put the solution in the cuvette into the waste bottle 26 Put the cuvette into the waste bag 27 If the reference is between 300 and 900 the calibration is complete 28 If the reference is not between 300 and 900 push PROCEED and do the calibration procedure again Note New reagents may give a signal over 900 If the signal is 900 or over change the measurement ran...

Page 30: ...ralize the sample by adding two drops of pre conditioner reagent to the sample beaker Note Two drops of pre conditioner reagent can neutralize up to 15 mg L of chlorine 3 Push ON green button for several seconds to apply power to the luminometer When the built in tests are done push PROCEED The Main Menu is shown 4 Select ECLOX and push ENTER Select Measure and push ENTER Select Measure Sample and...

Page 31: ...e 11 Put the tip into the cuvette and dispense the liquid by gently pushing in the operating button 12 Remove the tip from the pipet and put in the waste bag Put the pipet in the storage case 13 Put a yellow pipet tip on the yellow pipet Use a new pipet tip for each reagent 14 Do steps 9 to 12 to put 100 µL of CT Reagents 1 2 and 3 into the cuvette 15 Open the luminometer lid Remove the cuvette fr...

Page 32: ...st Record Sheet Note For sea water samples the graph may appear higher than the reference and the Inhib may be negative 18 Push PROCEED to go back to the Measure Menu 19 Remove the cuvette from the luminometer cell Put the solution into the waste bottle Put the cuvette into the waste bag 20 Sign the Test Record Sheet Put the sample from the beaker in the wastewater drain using local operating proc...

Page 33: ...nu is shown 3 Select ECLOX and push ENTER 4 Select Previous Results and push ENTER The Previous Results Menu is shown 5 To show previous results a Select Recall Results and push ENTER b Push MORE to show more results c Push QUIT to go back to the Previous Results Menu 6 To show previous graphs a Select Recall Graphs and push ENTER b Push UP to move through the saved samples c Push SELECT when the ...

Page 34: ...built in tests that make sure that the electronics and software are operating correctly 5 When the built in tests are done push PROCEED The Main Menu is shown 6 Select ECLOX and push ENTER 7 Select Previous Results and push ENTER The Previous Results Menu is shown 8 Select Download to PC and push ENTER ...

Page 35: ... products are photosensitive and may turn dark Do not allow test strips to touch the reaction vessel solution Test strips react with gases not solution Orient the test strip pad paper side down and centered over the hole in the black cap so the generated gases can make good contact with the pad Two samples may be analyzed simultaneously with this kit 5 2 EZ Arsenic 0 500 ppb 0 10 25 50 100 250 500...

Page 36: ... for 60 seconds Do not shake or invert or allow sample to get on the strip Wait 20 minutes Swirl twice during the reaction period 5 Remove the test strip and immediately compare the developed color to the chart on the test strip bottle 0 500 ppb row Read strips in the shade ...

Page 37: ...nd one Reagent 2 powder pillow to the sample See Interferences on page 37 4 Immediately attach the cap to the reaction bottle Swirl continuously for 60 seconds Do not shake or invert or allow sample to get on the strip Wait 20 minutes Swirl twice during the reaction period 5 Remove the test strip and immediately compare the developed color to the chart on the test strip bottle 0 4000 ppb row Read ...

Page 38: ...500 or 0 4000 ppb test procedure and continue with the test Note The lead acetate must not touch the test strip Always wear gloves or wash hands thoroughly after handling lead acetate Antimony 250 ppb Tellurium Likely to interfere but not tested Acidity pH 5 Do not acid preserve samples If samples are below pH 5 adjust pH to between 5 and 6 before beginning test Nitric acid Interferes with the red...

Page 39: ...nd Fold back the protective film covering the white disc only 3 Put the white disc in the beaker that contains the sample water for at least one minute 4 Remove the pesticide strip from the sample beaker 5 Remove the protective film covering the pink disc Fold the strip in half along the perforations and push the white disc against the pink disc 6 Put the strip in the pesticide clip and put the st...

Page 40: ...asier to see 8 There are two possible results A white disc indicates POSTIVE pesticides or nerve agent are present A blue disc matching blue or darker blue than larger disc indicates NEGATIVE no pesticide or nerve agent present 9 Do the test again if a positive result is seen or compare with a test from a known clean water sample 10 Record the results ...

Page 41: ...s use the value that is in the middle of the two segments If the color disc becomes wet internally pull apart the flat plastic sides to open the color disc Remove the thin inner disc Dry all parts with a soft cloth Assemble when fully dry Undissolved reagent does not have an effect on test accuracy For free chlorine read the result immediately after the reagent is added to prevent interference fro...

Page 42: ... free chlorine read the result within 1 minute For total chlorine wait 3 minutes Read the result within 6 minutes 6 Put the second tube into the color comparator box 7 Hold the color comparator box in front of a light source Turn the color disc to find the color match 8 Read the result in mg L in the scale window ...

Page 43: ...wo segments If the color disc becomes wet internally pull apart the flat plastic sides to open the color disc Remove the thin inner disc Dry all parts with a soft cloth Assemble when fully dry The long path adapter for the low range test shows the color in the tubes from top to bottom Make sure the light source is above the tubes during the color match 8 2 Color apparent 0 100 APHA platinum cobalt...

Page 44: ...p line with sample Put the second tube into the color comparator box 5 Hold the color comparator box below a light source Turn the color disc to find the color match 6 Read the results in platinum cobalt color units in the scale window ...

Page 45: ...lor 3 Put the tube into the left opening of the color comparator box 4 Fill a second tube to the first line 5 mL with sample Put the second tube into the color comparator box 5 Hold the color comparator box in front of a light source Turn the color disc to find the color match 6 Read the value in the scale window Multiply the value by 5 to get the result in platinum cobalt color units ...

Page 46: ...46 ...

Page 47: ... Lock Parameter key to select pH or TDS as the measurement mode 4 Add the sample to the fill line of the sensor cap 5 Put the sensor fully into the sensor cap The tester reads the value Air bubbles under the probe tip when submerged can cause slow stabilization or error in measurement Shake the tester from side to side to remove air bubbles 6 When the display is stable read the value To stabilize ...

Page 48: ...nger test life put several drops of deionized water in the sensor cap This keeps the glass bulb from becoming dry Put the sensor cap on the sensor Note To extend electrode life soak the electrode tip in tap water for a few minutes each week Note Refer to the documentation supplied with the Pocket Pro Multi 2 Tester to do conductivity salinity and temperature procedures ...

Page 49: ... Parameter key until pH shows as the measurement mode 4 Pour a pH 7 00 buffer to the fill line of the sensor cap 5 Put the sensor fully into the sensor cap The tester reads the buffer value Air bubbles under the probe tip when submerged can cause slow stabilization or error in measurement Shake the tester from side to side to remove air bubbles 6 Push the Calibration Settings key to start the cali...

Page 50: ...2 through 7 againto measure a pH 4 00 and or a pH 10 00 buffer 9 Push and hold the Calibration Settings key to exit 1 Push the Power Backlight key to set the power to on Remove the sensor cap 2 Rinse the sensor and the sensor cap with deionized water Dry with a no lint cloth 3 Push and hold the Lock Parameter key until Cond shows as the measurement mode ...

Page 51: ...d can cause slow stabilization or error in measurement Shake the tester from side to side to remove air bubbles 6 Push the Calibration Settings key to start the calibration 7 Wait for the value to stabilize then push the Calibration Settings key to keep the standard value 8 The display shows END when the calibration is complete 9 To measure a second standard 147 µS cm do step 2 then steps 4 throug...

Page 52: ...52 ...

Page 53: ...e with two exceptions Different options on the luminometer are selected to measure the sample dilutions Different options on the luminometer are selected to show or send results A LIMIT value is set by the user using the luminometer for the LIMIT measure procedure A column is added to the LIMIT measure test results that shows whether the percent inhibition measured for each sample dilution is abov...

Page 54: ...he LBT measurement luminescence procedure under more controlled conditions in a lab using additional accessories like a LUMIStherm temperature controlled incubator LTV053 10 3 Reagent description The Luminescent Bacteria Toxicity Test reagent contains living luminescent bacteria that have been grown under optimal conditions harvested and lyophilized freeze dried The reagent is a freeze dried prepa...

Page 55: ...at no more than 25 C 10 5 Prepare the reagent Prepare the Luminescent Bacteria Toxicity Test reagent in the field using the procedure in this section The amount of light made by the luminescent bacteria is affected by the temperature at which the reagent is reconstituted The luminescent bacteria and reconstitution solution must be mixed as cold as possible at refrigerator temperatures 3 to 8 C If ...

Page 56: ...onstitution solution and Diluent from refrigerator 2 Put the frozen luminescent bacteria reagent refrigerated reconstitution solution and Diluent in a cool box that contains thermal packs if possible 3 In the field remove the cap from the reconstitution solution bottle 4 Remove the foil seal and rubber stopper from the reagent bottle 5 Set the 1 0 5 0 mL pipette to 1 0 mL 6 Put the end of the 1 0 ...

Page 57: ...owly pull in 1 0 mL 8 Put the tip of the pipette into the luminescent bacteria reagent bottle and slowly dispense the solution into the reagent 9 Put the rubber stopper in the reagent bottle Swirl the reagent bottle to mix 10 Cool the reagent for 5 minutes in the cool box ...

Page 58: ...e potassium and magnesium in the Diluent stabilize the light made over time This stabilization helps keep high negative inhibitions from getting with samples that contain potassium and magnesium ions Do not make Diluent or use substitutes 1 Remove the Diluent from the cool box Remove the cap from the Diluent bottle 2 Put 14 0 mL of Diluent at refrigerator temperature in the reaction vessel using t...

Page 59: ...ferences Samples interferences can inhibit the light made by luminescent bacteria 7 Wait 15 minutes 8 Remove the pipette tip from the pipette and put in the waste bag Put the pipette in the storage case Interfering substances Interference levels and treatments Chlorine Changes the viability of the bacterial reagent Chlorine is toxic to the bacteria To remove chlorine from a sample add one powder p...

Page 60: ...to the sample if the salt concentration of the sample is more than 20 g L guide value conductivity of 35 mS cm Note The salt content of the sample should not exceed 50 g L This corresponds to a conductivity of about 70 mS cm without taking other conductive compounds into account Sodium chloride A sodium chlorine NaCl concentrations of less than 15 g L or more than 50 g L or their osmolarity equiva...

Page 61: ...levels 1 2 1 4 1 8 1 16 etc to make sure of a continuous dilution series using the dilution procedure of the manufacturer 10 11 Prepare the test tubes At the end of this procedure the test tubes contain the percent sample dilutions shown in Figure 3 Figure 3 Screening dilution series 1 Test suspension 3 Sample 2 2 NaCl solution 1 Put four test tubes in the test tube stand 2 Set the 0 2 1 0 mL pipe...

Page 62: ...bes contain 0 2 mL of test suspension 7 Set the 0 2 1 0 mL pipette to 0 8 mL 8 Put the tip of the pipette into the 2 NaCl and slowly pull in 0 8 mL 9 Slowly dispense the 2 NaCl solution into the test tube in position 1 10 Set the 0 2 1 0 mL pipette to 0 6 mL 11 Put the tip of the pipette into the 2 NaCl and slowly pull in 0 6 mL 12 Slowly dispense the 2 NaCl solution into the test tube in position...

Page 63: ...Set the 0 2 1 0 mL pipette to 0 2 mL 17 Set the timer clock for 15 minutes contact time 18 Put the tip of the pipette into the sample and slowly pull in 0 2 mL 19 Slowly dispense the sample into the test tube in position 2 Start the timer Note No sample is put into the test tube in position 1 Test tube 1 is the non toxic reference 20 Set the 0 2 1 0 mL pipette to 0 5 mL 21 Put the tip of the pipet...

Page 64: ...me In some instances if reconstitution is done at the optimum temperature and the test is carried out at 20 C the initial light made by the bacteria can be more than 1000 Eclox light units This causes the error Detector Overload If an error occurs change the measurement range from the 0 1000 range to the 0 2000 range and do the readings again refer to Set the measurement range on page 18 22 Slowly...

Page 65: ...n select Measure Luminescence and Send to PC and push ENTER The luminometer must be connected to a computer refer to Connect the luminometer to a computer on page 23 To measure the luminescence and print the results on a printer select Screening and Send to Printer and push ENTER The luminometer must be connected to a printer refer to Connect the luminometer to a printer on page 21 5 To do the LIM...

Page 66: ...Connect the luminometer to a printer on page 21 6 Open the luminometer lid and make sure a sample is not in the cell Close the lid 7 Push PROCEED to show the test status When the cell tests are done push PROCEED again 8 Wait until the timer clock completes 15 minutes 9 Open the luminometer lid 10 Put the test tube in position 1 non toxic blank into the black test tube holder in the luminometer cel...

Page 67: ... Inhibit and rel units values for each sample dilution on the Screening Luminescence Results Sheet 14 Use the color chart on the Screening Luminescence Results Sheet to identify which sample dilutions are toxic red and which are non toxic green Note The more of your results in the red zone the stronger are the inhibitory affects of the sample the more critical is the sample 15 Put the solution in ...

Page 68: ... 3 Add one spoon of solid NaCl to 7 mL of sample 4 Fill four test tubes with the test suspension 2 NaCl and sample in the order shown in the table Start the timer after adding the first sample volume Set the timer for 15 minutes 5 Push ON green button on the luminometer Go to the Screening Luminescence Menu 6 Measure test tube 1 non toxic reference 7 Measure the sample tubes in the order 2 3 and t...

Page 69: ... 13 1 Description of screening luminescence results Reference non toxic luminescent measurements are saved as R1 to Rx The counter starts with R1 every time the storage is erased from the luminometer Sample luminescent measurements are done after a reference luminescent measurement is done Sample luminescent measurements are saved as S1 to Sx The luminometer records reference and sample measuremen...

Page 70: ...ults To show or send previous results saved on the luminometer for the screening luminescence procedure 1 Push ON to apply power to the luminometer The luminometer does built in tests that make sure that the electronics and software are operating correctly 2 When the built in tests are done push PROCEED The Main Menu is shown 3 Select LUMINESCENT BACTERIA TEST and push ENTER The Luminescent Bacter...

Page 71: ...e data to be recalled a Select the starting indicator in the From field Push SELECT to change the value Then push PROCEED b Select the ending indicator in the To field Push SELECT to change the value Then push SHOW 9 Push PROCEED to show more results 10 13 4 Show or send LIMIT measure results To show or send previous results saved on the luminometer for the LIMIT measure procedure 1 Push ON to app...

Page 72: ...nge of results saved on the luminometer select one option To show the results on the luminometer select Show selection and push ENTER To send the results to the computer select Send selection to PC and push ENTER To send the results to the printer select Send selection to Printer and push ENTER 8 If an option in step 7 was selected select the data to be recalled a Select the starting value in the ...

Page 73: ... of intensity change of control samples during the exposure time refer to the ISO standard procedure The luminometer measurements are in relative light units The luminometer measurements are used by the LUMISsoft computer program or custom made calculations to calculate percent inhibition LID EC20 and EC50 values Percent inhibition the percentage of light made by the bacteria that is inhibited by ...

Page 74: ...escent bacteria that have been grown under optimal conditions harvested and lyophilized freeze dried The reagent is a freeze dried preparation of a specially selected strain of the marine bacterium Vibrio fischeri formerly known as Photobacterium phosphoreum NRRL number B 11177 A vial of reagent contains roughly one hundred million test organisms Refer to section Appendix A Luminescent bacteria ri...

Page 75: ...ossible at refrigerator temperatures 3 to 8 C If the temperature is higher the amount of initial light made by the bacteria will be lower 11 6 1 Prepare the stock suspension using the LCK491 reagent Prepare the stock suspension by adding the reconstitution solution to the freeze fried bacteria reagent The reconstitution solution rehydrates the bacteria reagent Reconstitution solution is specially ...

Page 76: ...om the reagent bottle 4 Set the 1 0 5 0 mL pipette to 1 0 mL 5 Put the end of the pipette into a clean pipette tip 6 Put the tip of the pipette into the reconstitution solution and slowly pull in 1 0 mL 7 Put the tip of the pipette into the luminescent bacteria reagent bottle Quickly dispense the solution into the reagent 8 Put the rubber stopper in the reagent bottle Swirl the reagent bottle to m...

Page 77: ...Diluent is a specially made non toxic 2 sodium chloride NaCl solution that contains potassium and magnesia ions The marine bacterium in the reagent requires the osmotic protection that is given by the 2 NaCl in the Diluent The potassium and magnesium in the Diluent stabilize the light made over time This stabilization helps keep high negative inhibitions from getting with samples that contain pota...

Page 78: ...he LUMIStherm Note The LUMIStherm should be operating at 15 C 7 Set the 0 2 1 mL pipette to 0 5 mL 8 Put the end of the pipette into a clean pipette tip 9 Put the tip of the pipette into the reaction vessel and slowly pull in 0 5 mL of the test suspension 10 Slowly dispense the test suspension into the test tube in position B1 11 Do step 9 and 10 again until each test tube in Row B and Row C conta...

Page 79: ... D 1 13 Remove the pipette tips from the pipettes and put in the waste bag Put the pipettes in the storage case 1 Remove the Diluent from the refrigerator Remove the cap from the bottle 2 Put 20 parts Diluent solution D at refrigerator temperature into the reaction vessel using a pipette 3 Remove the stock solution rehydrated reagent from the refrigerator Remove the rubber stopper from the reagent...

Page 80: ...on D at refrigerator temperature into a clean reaction vessel using a pipette 7 Put 1 part stock suspension S at refrigerator temperature into a clean reaction vessel using a pipette For example 0 2 mL S 10 mL D 8 Put the cap on the reaction vessel and shake to mix thoroughly Put a 1 50 label on the reaction vessel 9 Put one half of the new empty test tubes in Row B and one half of the test tubes ...

Page 81: ...nto the reaction vessel that contains the 1 20 test suspension and slowly pull in 0 2 mL of the test suspension 13 Slowly dispense the test suspension into the test tube in position B1 14 Do step 12 and 13 again until the test tubes in position C1 B2 and C2 contain 0 2 mL of test suspension 15 Set the 0 2 1 mL pipette to 0 5 mL ...

Page 82: ...pense the test suspension into the test tube in position B3 18 Do step 16 and 17 again until each test tube in Row B and Row C position 3 and higher contains 0 5 mL of test suspension 19 Cool the filled test tubes in the LUMIStherm at 15 C for 15 minutes 20 Remove the pipette tips from the pipettes and put in the waste bag Put the pipettes in the storage case ...

Page 83: ... sample and wait for 10 minutes High oxygen consumption Causes light inhibition that is not caused by toxicity pH pH related light inhibition may occur if the pH is below 6 0 or above 8 0 The pH of the sample must be within 7 0 2 pH units of the standard Sodium chloride A sodium chlorine NaCl concentrations of less than 15 g L or more than 50 g L or their osmolarity equivalents in a sample will ca...

Page 84: ...e by more than 5 intotal 3 Add solid NaCl to the sample until the concentration in the sample is 2 w v For example weigh out 0 3 g of NaCl and dissolve it in 15 mL of sample or dissolve one spoon of solid NaCl LCX058 in 7 mL of sample The concentration of salt in the test should not exceed 35 g L Note Do not add NaCl to the sample if the salt concentration of the sample is more than 20 g L guide v...

Page 85: ...re This procedure makes dilutions ranging from undiluted to a dilution ratio of 1 16 in Row A This corresponds to D values of 2 to 32 in the test Figure 6 as 0 5 mL of the sample dilution is added to 0 5 mL of the test suspension during the test in Row B and C Adding test suspension to the sample dilutions increases the sample dilutions in row A by a factor of two as final test concentration Note ...

Page 86: ... Slowly dispense the 2 NaCl solution into the test tube in position A9 5 Set the 1 0 5 0 mL pipette to 1 5 mL 6 Put the tip of the pipette into the 2 NaCl solution and slowly pull in 1 5 mL 7 Slowly dispense the 2 NaCl solution into the test tube in position A8 8 Do steps 6 and 7 againto put 1 5 mL of 2 NaCl solution in each test tube in positions A7 A6 A5 A4 A3 A2 and A1 Note Do not put 2 NaCl in...

Page 87: ...sample into the test tube in position A8 15 Do steps 13 and 14 againto put 1 5 mL of the sample into the test tube in position A10 Figure 7 on page 89 shows the contents of the test tubes in Row A after this step is completed 16 Pull the solution in A9 into the pipette 2 to 3 times to mix the sample dilution 17 Start making the sample dilution series in Row A Pull in 1 5 mL of solution from A9 and...

Page 88: ...ion in A8 into the pipette 2 to 3 times to mix the sample dilution 21 Pull in 1 5 mL of solution from A8 and put it into A6 using the pipette Pull the solution in A6 into the pipette 2 to 3 times to mix the sample dilution 22 Pull in 1 5 mL of solution from A6 and put it into A4 using the pipette Pull the solution in A4 into the pipette 2 to 3 times to mix the sample dilution 23 Pull in 1 5 mL of ...

Page 89: ... to D values of 2 to 4 Figure 8 in the test as 0 5 mL of the sample dilution is added to 0 5 mL of the test suspension during the test in Row B and C Adding test suspension to the sample dilutions increases the sample dilutions in row A by a factor of two as final test concentration 24 Keep the dilution series at 15 C for at least 5 minutes to correct the temperature Figure 7 Before the dilution s...

Page 90: ... 2 2 NaCl solution 1 Put 4 empty test tubes into Row A of the LUMIStherm 2 Set the 1 0 5 0 mL pipette to 1 0 mL 3 Put the end of the pipette into a clean pipette tip 4 Put the tip of the pipette into the 2 NaCl solution and slowly pull in 1 0 mL 5 Slowly dispense the 2 NaCl solution into the test tube in position A3 6 Set the 1 0 5 0 mL pipette to 1 5 mL ...

Page 91: ...NaCl solution into the test tube in position A1 10 Set the 1 0 5 0 mL pipette to 2 0 mL 11 Put the tip of the pipette into the sample and slowly pull in 2 0 mL 12 Slowly dispense the sample into the test tube in position A3 13 Set the 1 0 5 0 mL pipette to 1 5 mL 14 Put the tip of the pipette into the sample and slowly pull in 1 5 mL 15 Slowly dispense the sample into the test tube in position A4 ...

Page 92: ...92 16 Do steps 14 and 15 againto put 1 5 mL of the sample into the test tube in position A2 17 Keep the dilution series at 15 C for at least 5 minutes to correct the temperature ...

Page 93: ...in Row B and C Adding test suspension to the sample dilutions increases the sample dilutions in row A by a factor of two as final test concentration Note The test tubes in the higher Row A positions are more concentrated The pipette is moved from A9 to A4 higher concentration to lower concentration when making the dilution series so the pipette tip does not need to be replaced during this procedur...

Page 94: ...of the pipette into the 2 NaCl solution and slowly pull in 1 0 mL 5 Slowly dispense the 2 NaCl solution into the test tube in position A9 6 Set the 1 0 5 0 mL pipette to 2 0 mL 7 Put the tip of the pipette into the 2 NaCl solution and slowly pull in 2 0 mL 8 Slowly dispense the 2 NaCl solution into the test tube in position A1 9 Set the 1 0 5 0 mL pipette to 1 5 mL ...

Page 95: ...solution in each test tube in positions A7 A6 A5 A4 and A3 13 Set the 1 0 5 0 mL pipette to 2 0 mL 14 Put the tip of the pipette into the sample and slowly pull in 2 0 mL 15 Slowly dispense the sample into the test tube in position A9 16 Put the tip of the pipette into the sample and slowly pull in 2 0 mL 17 Slowly dispense the sample into the test tube in position A2 18 Set the 1 0 5 0 mL pipette...

Page 96: ...in Row A after this step is completed 22 Pull the solution in A9 into the pipette 2 to 3 times to mix the sample dilution 23 Start making the sample dilution series in Row A Pull in 1 5 mL of solution from A9 and put it into A7 using the pipette Pull in the solution in A7 into the pipette 2 to 3 times to mix the sample dilution 24 Pull in 1 5 mL of solution from A7 and put it into A5 using the pip...

Page 97: ...he pipette 2 to 3 times to mix the sample dilution 27 Pull in 1 5 mL of solution from A6 and put it into A4 using the pipette Pull in the solution in A4 into the pipette 2 to 3 times to mix the sample dilution 28 Keep the dilution series at 15 C for at least 5 minutes to correct the temperature Figure 10 Before the dilution series is started 9 dilutions D 1 and higher 1 Sample 2 2 NaCl solution ...

Page 98: ...ple dilution is added to 0 5 mL of the test suspension during the test in Row B and C Adding test suspension to the sample dilutions increases the sample dilutions in row A by a factor of two as final test concentration Figure 11 shows the contents of the test tubes in Row A at the end of this procedure Figure 11 Dilution series 3 dilutions D 1 values and higher 1 Sample 2 2 NaCl solution 1 Put 5 ...

Page 99: ...ip of the pipette into the 2 NaCl solution and slowly pull in 1 5 mL 8 Slowly dispense the 2 NaCl solution into the test tube in position A3 9 Do steps 7 and 8 againto put 1 5 mL of 2 NaCl solution into the test tube in position A1 10 Put the tip of the pipette into the sample and slowly pull in 1 5 mL 11 Slowly dispense the sample into the test tube in position A5 12 Set the 1 0 5 0 mL pipette to...

Page 100: ...slowly pull in 2 0 mL 14 Slowly dispense the sample into the test tube in position A4 15 Do steps 13 and 14 againto put 2 0 mL of sample into the test tube in position A2 16 Keep the dilution series at 15 C for at least 5 minutes to correct the temperature ...

Page 101: ...r several seconds to apply power to the luminometer When the built in tests are done push PROCEED The Main Menu is shown 2 Select Luminescent Bacteria Test and push ENTER 3 Select Measure and push ENTER 4 Select Measure Luminescence and push ENTER 5 Select one option that is shown To measure luminescence and save the results on the luminometer select Measure Luminescence and Save and push ENTER To...

Page 102: ...ell tests are done push PROCEED again 3 If testing D 2 values and higher set the 0 2 1 0 mL pipette to 0 5 mL If testing D 1 values and higher set the 0 2 1 0 mL pipette to 0 8 mL 4 Open the luminometer lid 5 Put the test tube in position B1 into the black test tube holder in the luminometer cell 6 Close the luminometer lid Push MEASURE ...

Page 103: ...puter or printed 9 Open the lid of the luminometer Remove the test tube from the luminometer and put it back in the LUMIStherm 10 Put the test tube in position C1 into the black test tube holder in the luminometer cell 11 Close the luminometer lid Push MEASURE 12 If testing D 2 values and higher put the pipette in test tube A1 and pull in 0 5 mL of sample dilution If testing D 1 values and higher ...

Page 104: ...older in the luminometer cell 16 Close the luminometer lid Push MEASURE 17 If testing D 2 values and higher put the pipette in test tube A1 and pull in 0 5 mL of sample dilution If testing D 1 values and higher put the pipette in test tube A1 and pull in 0 8 mL of sample dilution Put the pipette into the test suspension and slowly dispense the sample in C1 Mix with the pipette 18 When the B2 measu...

Page 105: ...m the test tube in position A2 has been added to the test tubes in position B2 and C2 If testing D 2 values and higher 0 5 mL of the sample dilution from the test tube in position A2 has been added to the test tubes in position B2 and C2 0 5 mL of the sample dilution from each test tube in Row A A3 and higher has been added to the test tubes in Row B and C that have the same position number e g fr...

Page 106: ... dependent ISO 11348 states that the test must be done under temperature controlled conditions at 15 C using a thermostat LUMIStherm LTV053 If the Luminescent Bacteria Toxicity Test is done at ambient temperature record the temperature The results of tests done at different temperatures cannot be compared directly To measure the light intensity of the test suspension after the sample dilutions are...

Page 107: ...o the luminometer saved to the computer or printed 6 Open the lid of the luminometer Remove the test tube from the luminometer and put it back into the LUMIStherm 7 Do steps 3 to 6 again until all the test tubes in Row B and Row C have been measured Measure the test tubes in the same order that the light output of the test suspension was measured e g B1 C1 B2 C2 etc 8 Put the solution in the test ...

Page 108: ...t the luminometer to a computer on page 23 2 Start LUMISsoft 3 In LUMISsoft select Transfer Options Interface Protocol Connect 4 Do the procedure in this section To send previous results to a printer do the steps in Connect the luminometer to a printer on page 21 and then do the procedure in this section To show or send previous results saved on the luminometer 1 Push ON green button to apply powe...

Page 109: ...stage the results cannot be sent to the LUMISsoft 4 7 To show or send a specific range of results saved on the luminometer select one option To show the results on the luminometer select Show selection and push ENTER To send the results to the computer select Send selection to PC and push ENTER To send the results to the printer select Send selection to Printer and push ENTER 8 If an option in ste...

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Page 111: ... Cleaning the luminometer Keep the luminometer clean at all times If the surface is dirty clean the surface with a damp cloth Important Note Do not let water get into the luminometer cell If water gets into the cell remove the cell insert and remove the moisture with a clean dry cloth Replace the cell insert 12 2 Decontamination If the Eclox Rapid Response Test Kit comes into contact with any chem...

Page 112: ...nd dispose of them in accordance with local operating procedures 4 Put four new batteries AA Alkaline in the luminometer Make sure the battery polarity is correct 5 Put the battery cover on the luminometer with the Battery Cover Screw Tool 6 Push ON green button to apply power to the luminometer 7 Do the pre deployment checks refer to section 3 2 1 Test the operation on page 16 Figure 13 Replace t...

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Page 114: ...Visit us at www hach com ...

Page 115: ...ontact technical support 03 LCD display test failed Continue operation If fault occurs again contact technical support 04 Non volatile RAM test failed Memory failure Contact technical support 05 Storage measurements corrupt All saved measurements will be erased Continue operation If fault occurs again contact technical support 06 Configuration settings corrupt LCD contrast and measurement range wi...

Page 116: ...t technical support 105 Cannot recall measurements Remove power and then apply power to the luminometer If fault occurs again contact technical support 106 Cannot store usage counter Remove power and then apply power to the luminometer If fault occurs again contact technical support 107 Cannot store measurements Remove power and then apply power to the luminometer If fault occurs again contact tec...

Page 117: ...uL tip set each 2887200 Serial comms lead cable luminometer each 2888200 Test record sheets 15 pkg 2888300 Waste bottle 250 mL each 2888400 Chemiluminescence Toxicity Test Chemiluminescent Reagent Set 100 tests each 94 9004 Cuvette and 1000 µL Pipet Set 2 2887400 Cuvette and 1000 uL Pipette Tip Set each 2887400 Cuvette for Eclox 500 pack each 30 0015 100 µL Pipet Tip Set 2 2887200 Chemiluminescenc...

Page 118: ...73200 Color disc DPD chlorine 0 3 4 mg L each 990200 Color viewing tube plastic with cap 4 pkg 4660004 DPD free chlorine reagent powder pillows 100 pkg 1407799 DPD total chlorine reagent powder pillows 100 pkg 1407699 Caps plastic color viewing tubes 4660004 4 pkg 4660014 Chlorine standard solution 50 75 mg L 2 mL PourRite ampule 20 pkg 1426820 Color viewing tube glass 6 pkg 173006 Stoppers glass ...

Page 119: ...reaction vessels rack for 8 reaction vessels stand for 40 test tubes variable pipette 0 2 1 0 mL variable pipette 1 0 2 0 mL pipette tips and timer clock each LCW490 Case each 4660800 Dilution solution 1000 mL each LCX048 2 NaCl solution 250 mL each LCK481 NaCl solid in bottle with dosing spoon 25 g each LCX058 Pipette variable 0 2 1 0 mL each BBP078 Pipette variable 1 0 5 0 mL each BBP065 Pipette...

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Page 121: ...t and facility design and construction are appropriate for undergraduate and secondary educational training and teaching laboratories and for other laboratories in which work is done with defined and Table 5 Risk specifications Strain number DSM 7151 Vibrio fischeri Beijerinck 1889 Lehmann and Neumann 1896AL Bacteria by DSMZ Deutsche Sammlung von Mikroorganismen und Zellkulturen GmbH Braunschweig ...

Page 122: ... cause infection in the young the aged and immunodeficient or immunosuppressed individuals Vaccine strains that have undergone multiple in vivo passages should not be considered avirulent simply because they are vaccine strains Biosafety Level 1 represents a basic level of containment that relies on standard microbiological practices with no special primary or secondary barriers recommended other ...

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Page 124: ...O 80539 0389 U S A Tel 970 669 3050 800 227 4224 U S A only Fax 970 669 2932 orders hach com www hach com HACH LANGE GMBH Willstätterstraße 11 D 40549 Düsseldorf Germany Tel 49 0 2 11 52 88 320 Fax 49 0 2 11 52 88 210 info de hach com www de hach com HACH LANGE Sàrl 6 route de Compois 1222 Vésenaz SWITZERLAND Tel 41 22 594 6400 Fax 41 22 594 6499 ...

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