Bugcount® Wastewater (QG21W)

Key terms

• ATP: Adenosine triphosphate, energy molecule of living cells.
• tATP (Total ATP): All ATP in the sample (living + dead cells).
• dATP (Dissolved ATP): ATP released from dead or compromised cells.
• cATP (Cellular ATP): ATP inside living cells, representing active biomass.
• RLU: Relative Light Units (measurement from luminometer).
• RLU ATP1: Calibration signal from UltraCheck 1.
• fbATP (Floc-Bulking ATP): ATP from bulking floc in suspended-growth aerobic systems.
• agATP (Attached Growth ATP): ATP from microorganisms attached in fixed-film systems.
• AVSS (Activated Volatile Suspended Solids): Mass of living organisms in the sample.
• ABR (Active Biomass Ratio): Share of suspended solids that are living microorganisms.
• BSI (Biomass Stress Index): Stress level of the microbial community.
• Specific fbATP (s-fbATP): Relative quantity of bulking floc to total floc.  As this number increases, the risk of bulking conditions increases.
• Specific agATP (s-agATP): Relative quantity of attached microorganisms to total microorganisms. As this number decreases, the risk of process failure due to biomass detachment increases.

1.1 Log in to Relay™

• Log in (or create your account) at http://relay.luminultra.com.

If you are new to Relay™, follow the guides to first create a site and sample location.

• Connect to your luminometer, then select RUN TESTS to begin a new test.
• As you work through these test kit instructions, you can initiate RLU readings directly within Relay™ by selecting READ on any input. Once all inputs are entered, your results will automatically calculate.

1.2 Rehydrate the Luminase^W enzyme

• Gently mix the Buffer and Luminase^W enzyme.
• Wait 5 minutes for solution to dissolve.

After rehydration, keep Luminase refrigerated or frozen; it stays effective for 2–4 weeks at 4°C (39°F) or 3–6 months when frozen between uses. Before testing, let it thaw and reach room temperature (takes about 1 hour). Do not apply direct heat to warm it faster. For more details, visit my.luminultra.com.

1.3 Calibration using UltraCheck 1 (RLU ATP1)

• Hold the UltraCheck 1 bottle vertical and add 2 drops (100µL) of it to a 12x55mm test tube.
• Pipet 300µL of Luminase^W into the tube.
• Swirl the tube, place it in the luminometer, and take the RLU ATP1 reading (select READ) within 10 seconds.

If RLU ATP1 ≤ 500 rehydrate a new bottle of Luminase^W.

Once UltraCheck 1 is opened, it must be used within 3 months. After 3 months, discard and use a new bottle.

2. Total ATP analysis (RLU tATP)

2.1 Extraction

• Mix sample well and test before the sample settles.
• Using a wide-mouth pipet tip, add 1mL of sample to a 2mL UltraLyse 30²¹ (extraction) tube.
• Cap, mix and allow 1 minute for incubation.

2.2 Dilution

• Pour the UltraLyse 30²¹ (extraction) tube contents into a new 8mL UltraLute/Resin (dilution) tube.
• Transfer the mixture between the tubes at least 3 times. Cap, mix and allow beads to settle.

2.3 Assay

• Add 100µL of the UltraLute/Resin (dilution) solution to a 12x55mm test tube.
• Use a new pipet tip to add 300µL of Luminase^W.
• Swirl the tube, place it in the luminometer, and take the RLU tATP reading (select READ) within 10 seconds.

3. Dissolved ATP analysis (RLU dATP)

3.1 Dilution

• Gently mix the sample and test before the sample settles.
• Using a wide-mouth pipet tip, add 100µL of sample to a 10mL LumiSolve tube.
• Cap and invert at least 3 times to mix, then allow 1 minute for incubation.

3.2 Assay

• Add 100µL of the 10mL LumiSolve solution to a 12x55mm test tube.
• Use a new pipet tip to add 300µL of Luminase^W.
• Swirl the tube, place it in the luminometer, and take the RLU dATP reading (select READ) within 10 seconds.
• In Relay™, select SAVE TEST.

4. Analysis

5. Analysis

5. Analysis

Tip: Relay™ does the work For you

When all inputs are entered in Relay’s Run tests workflow, the results will calculate automatically. Select Save Test to catalog and share your results with your team.

\mathrm{tATP}\;(\mathrm{ng\ ATP}/\mathrm{mL})
=
\frac{\mathrm{RLU}_{\mathrm{tATP}}}{\mathrm{RLU}_{\mathrm{ATP1}}}
\times
11\;(\mathrm{ng\ ATP}/\mathrm{mL})

\mathrm{dATP}\;(\mathrm{ng\ ATP}/\mathrm{mL})
=
\frac{\mathrm{RLU}_{\mathrm{dATP}}}{\mathrm{RLU}_{\mathrm{ATP1}}}
\times
101\;(\mathrm{ng\ ATP}/\mathrm{mL})

\mathrm{fbATP}\;(\mathrm{ng\ ATP}/\mathrm{mL})
=
\frac{\mathrm{RLU}_{\mathrm{fbATP}}}{\mathrm{RLU}_{\mathrm{ATP1}}}
\times
36.7\;(\mathrm{ng\ ATP}/\mathrm{mL})

\mathrm{agATP}\;(\mathrm{ng\ ATP}/\mathrm{units})
=
\frac{\mathrm{RLU}_{\mathrm{agATP}}}{\mathrm{RLU}_{\mathrm{ATP1}}}
\times
\frac{36.7\;(\mathrm{ng\ ATP})}{g\ \mathrm{or}\ \#\ \mathrm{media}}

\mathrm{cATP}\;(\mathrm{ng\ ATP}/\mathrm{mL})
=
\mathrm{tATP}\;(\mathrm{ng\ ATP}/\mathrm{mL})
-
\mathrm{dATP}\;(\mathrm{ng\ ATP}/\mathrm{mL})

\mathrm{AVSS}\;(\mathrm{mg\ Biomass}/\mathrm{L})
=
\mathrm{cATP}\;(\mathrm{ng\ ATP}/\mathrm{mL})
\times
0.5

\mathrm{ABR}\;(\%)
=
\frac{\mathrm{AVSS}\;(\mathrm{mg\ Biomass}/\mathrm{L})}
     {\mathrm{TSS}\;(\mathrm{mg}/\mathrm{L})}
\times
100\%

\mathrm{BSI}\;(\%)
=
\frac{\mathrm{dATP}\;(\mathrm{ng\ ATP}/\mathrm{mL})}
     {\mathrm{tATP}\;(\mathrm{ng\ ATP}/\mathrm{mL})}
\times
100\%

\mathrm{s\text{-}fbATP}\;(\%)
=
\frac{\mathrm{fbATP}\;(\mathrm{ng\ ATP}/\mathrm{mL})}
     {\mathrm{tATP}\;(\mathrm{ng\ ATP}/\mathrm{mL})}
\times
100\%

\mathrm{s\text{-}agATP}\;(\%)
=
\frac{\mathrm{agATP}\;(\mathrm{ng\ ATP}/\mathrm{mL})}
     {\mathrm{tATP}\;(\mathrm{ng\ ATP}/\mathrm{mL})
      +\mathrm{agATP}\;(\mathrm{ng\ ATP}/\mathrm{mL})}
\times
100\%

Troubleshooting

Additional Resources

myLuminultra:
https://my.luminultra.com

Contact support:
https://my.luminultra.com/hc/en-us/requests/new