# US 2017/0253902 A1 · Method for Assaying for Loss of an Organism in an Aqueous Liquid

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- Number: US 2017/0253902 A1
- Document ID: US20170253902A1
- Status: Published application
- Kind: A1
- Application number: 15/106,416
- Filed: 2014-12-15
- Published: 2017-09-07
- Earliest claimed: 2013-12-20
- Assignees: Trojan Technologies Inc.
- Applicants: Trojan Technologies
- Inventors: Hugh L. MacIntyre; John J. Cullen
- Web page: https://patentgrep.com/patent/US20170253902A1

## CPC

- C12Q 1/04: Determining presence or kind of microorganism; Use of selective media for testing antibiotics or bacteriocides; Compositions containing a chemical indicator therefor {(C12Q1/6897 takes precedence)}
- B63J 4/002: {for treating ballast water}
- C02F 1/32: with ultraviolet light
- C12Q 1/025: {for testing or evaluating the effect of chemical or biological compounds, e.g. drugs, cosmetics (antimicrobial activity C12Q1/18)}
- C12Q 1/06: Quantitative determination
- G01N 21/6428: {Measuring fluorescence of fluorescent products of reactions or of fluorochrome labelled reactive substances, e.g. measuring quenching effects, using measuring "optrodes" (in vivo A61B5/00; immunoassay G01N33/53)}
- G16H 40/63: for local operation
- C02F 1/008: {Control or steering systems not provided for elsewhere in subclass C02F}
- C02F 2103/008: {Originating from marine vessels, ships and boats, e.g. bilge water or ballast water}
- C02F 2303/04: Disinfection
- G01N 2201/12: Circuits of general importance; Signal processing
- G01N 2333/405: from algae
- G01N 2800/7004: Stress
- C12Q: MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS (immunoassay G01N33/53); COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- B63J: AUXILIARIES ON VESSELS
- C02F: TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE (separation in general B01D; special arrangements on waterborne vessels of installations for treating water, waste water or sewage, e.g. for producing fresh water, B63J; adding materials to water to prevent corrosion C23F; treating radioactively-contaminated liquids G21F9/04; regeneration of reactants for recirculation into processes, see the relevant places for the processes)
- G01N: INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES (measuring or testing apparatus or processes other than immunoassay, involving enzymes or microorganisms C12M, C12Q)
- G16H: HEALTHCARE INFORMATICS, i.e. INFORMATION AND COMMUNICATION TECHNOLOGY [ICT] SPECIALLY ADAPTED FOR THE HANDLING OR PROCESSING OF MEDICAL OR HEALTHCARE DATA

## Abstract

There is described a method for assaying for loss of viability of a photosynthetic organism (preferably a microorganism) in an aqueous liquid. In a preferred embodiment, the method comprises the step of using fluorescence to correlate the photorepair index for the organism in the aqueous liquid to survivorship of the organism (preferably a microorganism) after it is exposed to ultraviolet radiation, thereby assessing viability.

## Claims

### Claim at position 1

1-13. (canceled)

### Claim 14

A method for assaying for loss of viability of an organism in an aqueous liquid after the organism has been exposed to a stressor, the method comprising the step of assessing the ability of the organism to undergo photorepair.

### Claim 15 (depends on 14)

The method defined in claim 14, wherein the stressor is selected from the group consisting of: exposure to a chemical, exposure to mechanical energy, thermal shock, dark storage, and any combination thereof.

### Claim at position 4

16-17. (canceled)

### Claim 18 (depends on 14)

The method defined in claim 14, wherein the stressor is ultraviolet radiation having a wavelength in the range of from about 100 nm to about 280 nm.

### Claim 19

(canceled)

### Claim 20 (depends on 14)

The method defined in claim 14, wherein the assessing step comprises conducting a variable fluorescence test on the organism.

### Claim 21 (depends on 14)

The method defined in claim 14, wherein the organism is a microorganism.

### Claim at position 9

22-23. (canceled)

### Claim 24

A method for assaying for loss of viability of an organism in an aqueous liquid after the organism has been exposed to a stressor, the method comprising the step of correlating the photorepair index for the organism in the aqueous liquid to survivorship of the organism after the organism has been exposed to the stressor.

### Claim at position 11

25-33. (canceled)

### Claim 34

A method for assaying for loss of viability of an organism comprised in an aqueous liquid after the organism has been exposed to a stressor, the method comprising the steps of:
(a) measuring the variable fluorescence (Fv) of an untreated sample of the organism prior to exposure to the stressor;
(b) measuring the variable fluorescence (Fv) of a treated sample of the organism after exposure to the stressor;
(c) calculating a photorepair index using the measurements obtained in Steps (a) and (b); and
(d) correlating the photorepair index calculated in Step (c) to a normalized viability for the organism.

### Claim 35 (depends on 34)

The method defined in claim 34, wherein Step (a) comprises one or more of the following:
measuring the variable fluorescence (Fv) of the untreated sample of the organism prior to exposure to radiation to obtain Fv(0)Untreated;
(ii) incubating the untreated sample in the presence of radiation at a first intensity to induce a prescribed reduction in Fv from Fv(0)Untreated;
(iii) measuring the variable fluorescence (Fv) of the untreated sample after Step (ii) to obtain Fv(1)Untreated;
(iv) incubating the untreated sample in presence of radiation at a second intensity to induce photorepair and an increase in Fv from Fv(1)Untreated; and
(v) measuring the variable fluorescence of the untreated sample after Step (iv) to obtain Fv(2)Untreated.

### Claim 36 (depends on 35)

The method defined in claim 35, wherein Step (ii) is conducted for a period of from about 10 minutes to about 120 minutes.

### Claim 37 (depends on 35)

The method defined in claim 35, wherein the first intensity is in the range of from about 2 μmol photons m−2s−1 to about 4000 μmol photons m−2s−1.

### Claim 38 (depends on 35)

The method defined in claim 35, wherein Step (ii) comprises incubating the untreated sample in presence of photosynthetically active radiation.

### Claim 39 (depends on 35)

The method defined in claim 35, wherein Step (ii) comprises incubating the untreated sample in presence of photosynthetically active radiation having one or more wavelengths substantially within the range of from about 400 nm to about 700 nm.

### Claim 40

(canceled)

### Claim 41 (depends on 35)

The method defined in claim 35, wherein the prescribed reduction in Fv in Step (ii) is in the range of from about 30% to 80%.

### Claim 42

(canceled)

### Claim 43 (depends on 35)

The method defined in claim 35, wherein Step (iv) is conducted for a period of from about 30 minutes to about 240 minutes.

### Claim 44 (depends on 35)

The method defined in claim 35, wherein the second intensity is in the range of from about 1 μmol photons m−2s−1 to about 50 μmol photons m−2s−1.

### Claim at position 23

45-46. (canceled)

### Claim 47 (depends on 35)

The method defined in claim 35, wherein Step (iv) comprises incubating the untreated sample in presence of photosynthetically active radiation having one or more wavelengths substantially within the range of from about 400 nm to about 700 nm.

### Claim 48

(canceled)

### Claim 49 (depends on 35)

The method defined in claim 35, wherein Step (b) comprises one or more of the following:
(i) incubating the treated sample in presence of photosynthetically active radiation at a first intensity to induce a prescribed reduction in Fv from Fv(0)Untreated;
(ii) measuring the variable fluorescence of the treated sample after Step (ii) to obtain Fv(1)Treated;
(iii) incubating the untreated sample in presence of radiation at a second intensity to induce photorepair and an increase in Fv from Fv(1)Treated; and
(iv) measuring the variable fluorescence of the treated sample after Step (iii) to obtain Fv(2)Treated.

### Claim at position 27

50-52. (canceled)

### Claim 53 (depends on 49)

The method defined in claim 49, wherein Step (i) comprises incubating the untreated sample in presence of photosynthetically active radiation having one or more wavelengths substantially within the range of from about 400 nm to about 700 nm.

### Claim at position 29

54-58. (canceled)

### Claim 59 (depends on 49)

The method defined in claim 49, wherein substantially the same radiation is used Step (i) and Step (iii).

### Claim 60 (depends on 35)

The method defined in claim 35, wherein Step (c) comprises calculating the photorepair index using one of the following equations:
PRI
=
[
F
v

(
2
)
-
F
v

(
1
)
]
Treated
[
F
v

(
0
)
]
Untreated
(
Eq
.

1
)
PRI
=
[
F
v

(
2
)
-
F
v

(
1
)
]
Treated
[
F
v

(
2
)
-
F
v

(
1
)
]
Untreated
(
Eq
.

2
)
PRI
=
[
F
v

(
2
)
-
F
v

(
1
)
]
Treated
F
v

(
1
)
Untreated
.
(
Eq
.

3
)

### Claim at position 32

61-65. (canceled)