Disinfectant composition

Abstract

A disinfectant composition useful in shrimp aquaculture and a process for using such a disinfectant, in which there is a composition containing a) 25 to 87 wt. % of KHSO.sub.5 containing triple salt b) 1 to 20 wt. % of at least one H.sub.2N(CH.sub.2).sub.nSO.sub.3H compound with n being 0, 1, 2 or 3, c) 1 to 8 wt. % of a surfactant d) 1 to 60 wt. % of a C.sub.2-C.sub.10 aliphatic dicarboxylic acid, optionally substituted with a hydroxyl and/or carboxylic group and e) 10 to 30 wt. % alkali metal phosphate.

Claims

1. A process for disinfecting water to be used for cultivation or containing aquatic shrimp cultures comprising adding a composition to the water, wherein said composition comprises a) 25 to 87 wt. % of KHSO.sub.5 containing triple salt; b) 1 to 20 wt. % of at least one H.sub.2N(CH.sub.2).sub.nSO.sub.3H compound with n being 0, 1, 2 or 3; c) 1 to 8 wt. % of a surfactant; d) 1 to 60 wt. % of a C.sub.2-C.sub.10 aliphatic dicarboxylic acid, optionally substituted with a hydroxyl and/or carboxylic group; and e) 10 to 30 wt. % alkali metal phosphate.

2. The process for disinfecting water according to claim 1, wherein the shrimps are aged 1 to 27 days after hatching.

3. A process for manufacturing a composition, said composition comprising a) 25 to 87 wt. % of KHSO.sub.5 containing triple salt; b) 1 to 20 wt. % of at least one H.sub.2N(CH.sub.2).sub.nSO.sub.3H compound with n being 0, 1, 2 or 3; c) 1 to 8 wt. % of a surfactant; d) 1 to 60 wt. % of a C.sub.2-C.sub.10 aliphatic dicarboxylic acid, optionally substituted with a hydroxyl and/or carboxylic group; and e) 10 to 30 wt. % alkali metal phosphate, wherein the process comprises blending together components a) to e).

4. A composition comprising a) 25 to 87 wt. % of KHSO.sub.5 containing triple salt; b) 1 to 20 wt. % of at least one H.sub.2N(CH.sub.2).sub.nSO.sub.3H compound with n being 0, 1, 2 or 3; c) 1 to 8 wt. % of a surfactant; d) 1 to 60 wt. % of a C.sub.2-C.sub.10 aliphatic dicarboxylic acid, optionally substituted with a hydroxyl and/or carboxylic group; and e) 10 to 30 wt. % alkali metal phosphate.

5. The Composition composition according to claim 4, wherein component b) comprises sulphamic acid.

6. The composition according to claim 4, wherein component d) comprises C.sub.4-C.sub.6-aliphatic dicarboxylic acid that is unsubstituted or substituted by one or more hydroxyl and/or carboxylic group.

7. The composition according to claim 4, wherein component d) comprises a aliphatic dicarboxylic acid selected from the group consisting of malic acid, citric acid, succinic acid, adipic acid, maleic acid and tartaric acid.

8. The composition according to claim 4, wherein component c) comprises an alkali metal C.sub.4-C.sub.20-alkylarylsulphonate.

9. The composition according to claim 4, wherein the composition contains a) 35 to 72 wt. % of KHSO.sub.5 containing triple salt, b) 2 to 10 wt. %, of at least one H.sub.2N(CH.sub.2).sub.nSO.sub.3H compound with n being 0, 1, 2 or 3, c) 1 to 6 wt. % of a surfactant, d) 15 to 30 wt. % of a C.sub.2-C.sub.10 aliphatic dicarboxylic acid, optionally substituted with a hydroxyl and/or carboxylic group and e) 10 to 20 wt. % alkali metal phosphate.

10. The composition according to claim 4, wherein the composition contains a) 40 to 67 wt.% of KHSO.sub.5 containing triple salt, b) 2 to 10 wt. %, of at least one H.sub.2N(CH.sub.2).sub.nSO.sub.3H compound with n being 0, 1, 2 or 3, c) 1 to 6 wt. % of a surfactant, d) 20 to 30 wt. % of a C.sub.2-C.sub.10 aliphatic dicarboxylic acid, optionally substituted with a hydroxyl and/or carboxylic group and e) 10 to 20 wt. % alkali metal phosphate.

11. The composition according to claim 4, wherein component d) comprises malic acid.

12. The composition according to claim 4, wherein component c) comprises alkali metal C.sub.10-C.sub.13-alkylbenzenesulphonate.

Description

EXAMPLES

(1) Composition 1 of the Present Invention Used in the Examples:

(2) TABLE-US-00001 component Amount [wt. %] a) KHSO.sub.5 containing 49.8 triple salt (Oxone ® mono- persulphate) Sodium chloride 1.5 b) Sulohamic acid 4.6 d) Malic acid 24 e) Na-hexameta- 14.9 phosphate c) Sodium C.sub.10-C.sub.13- 4.9 alkylbenzene sulphonate (Marion ® ARL) Fumed silica (Cabosil ® M5) 0.3 Total 100
Comparison Composition 1: Composition of the Prior Art (Virkon® Aquatic):

(3) TABLE-US-00002 Amount in Amount in composition composition [wt. %] according [wt. %] according Component to measurement to MSDS a) Potassium 49.75 25 to 50* peroxymonosulphate triple salt e) Na- 21.9 — hexametaphosphate c) Sodium C.sub.10-C.sub.13- 13.7 10-<25** alkylbenzenesulphonate d) Malic acid 8.8 <10 b) Sulphamic acid 4.4 ≤5 *note dipotassium peroxodisulphate and dipotassium disulphate are mentioned to be < 5 wt % each **Sodium alkylbenzenesulphonate
Comparison composition 2: Composition close to composition 1 of the present invention but with only 10 wt % surfactant as known from exp. 6 of WO2007/023481 for fish disinfection:

(4) TABLE-US-00003 component Amount [wt. %] a) KHSO.sub.5 containing 49.8 triple salt (Oxonee ® mono-persulphate) Sodium chloride 1.5 b) Sulphamic acid 4.6 d) Malic acid 24 e) Na-hexameta- 9.8 phosphate c) Sodium C.sub.10-C.sub.13- 10.0 alkylbenzene sulphonate (Marlon ® ARL) Fumed silica (Cabosil ® M5) 0.3 Total 100
Toxicity Test of composition of the present invention to Litopenaeus vannamei Shrimp

(5) These test will be divided to three steps, preliminary test to predict the upper and lower level of LC50 concentration, lethal toxicity test to determine the concentration that killed 50% shrimp population, and sublethal toxicity test to determine the toxicity level that not kill the organism. Lethal toxicity test data will be analyzed with probit analysis to determine LC50 values in 24 and 96 hours exposure period, respectively. The stage of the shrimp that will be used in this trial will be Nauplius and PL (PL-15).

(6) Objectives:

(7) 1. To study the toxicity levels of Disinfection composition of the present invention affected L. vannamei at different stages. Whereas Nauplius represent the youngest one which most sensitive to disinfectants and PL-15 which is more resistant than Nauplius. A more detailed classification of the respective stages can be found in the below table.

(8) TABLE-US-00004 Stage Detail Stage Days after hatching Nauplius N1-N6 0-2 Protozoea/zoea Z1-Z3 3-5 Zoea Mysis ZM 5-6 Mysis M1-M3  6-10 Mysis Post Larvae MPL 10-12 Post Larvae  PL1-PL15 12-27 2. To define the safety margin of Disinfection composition of the present invention affected L. vannamei at different stages (Nauplius, and PL-15).

Materials and Method

(9) a. Toxicity Trial for Shrimp Nauplius For this stage toxicity test should modified to 24 hours period, as in Nauplius only last for 24 hours then change to Zoea stage, The test will be divided into two different steps.

(10) i. Preliminary Test 1. The stage used was Nauplii N1-2. 2. Then, Nauplii were counted to have 130 animals into a well of a 20 well plates into 4 ml of sea water that the same with the salinity in hatchery (approximately 30 ppt=30 g salt/kg water, ppt is the unit for salinity, part per thousand). 3. Then, 1 ml of a 5% w/v solution of the disinfection composition 1 of the present invention was added to the well to make each well have 12 different concentration between 0 to 50 ppm (0 ppm, 0.5 ppm, 5 ppm, 10 ppm, 15 ppm, 20 ppm, 25 ppm, 30 ppm, 35 ppm, 40 ppm, 45 ppm and 50 ppm). No Replication used in this preliminary test. 4. The plates were incubated for 12 hours and shaken to provide aeration. After incubation, plates were observed under a stereoscope and dead animals were recorded. 5. Determine the highest concentration with 0% mortality and the lowest concentration with 100% mortality. The concentration will be the lower level and upper level of the test.

(11) ii. Definitive Test 1. After the value of lower and upper level is found by preliminary test, divide the range between the value into 6 concentrations (for examples lower level=0 ppm and upper level=25 ppm, so the concentration will be 0 ppm, 1 ppm, 2 ppm, 4 ppm, 8 ppm, 16 ppm and 32 ppm) 2. Then, Nauplii were counted to have 130 animals into a well of a 6 well plates into 4 ml of sea water that the same with the salinity in hatchery (approximately 30 ppt). 3. Then, 1 ml of a 5% w/v solution of the disinfection composition of the present invention was added to the well to make each well have 12 different concentration between 0 to 50 ppm (0 ppm, 1 ppm, 2 ppm, 4 ppm, 7 ppm, 11 ppm, 16 ppm, 22 ppm, 29 ppm, 37 ppm, 46 ppm and 50 ppm). Each concentration is tested with 5 replications. 4. The plates were incubated for 24 hours and shaken to provide aeration. After incubation, plates were observed under a stereoscope and dead animals were recorded. 5. Analyze the result with the so called probit analysis to determined LC50 value.

(12) b. Toxicity Trial for PL-15 Shrimp PL-15 stage is selected to be tested because during this stage the shrimp gills will be developed completely usually this stage is the latter stage that send to grow out ponds. The test is be divided into two different steps.

(13) i. Preliminary Test 1. PL-12 will get from nearby hatchery with proper biosecurity measurement and will be acclimatized through three days conditioning process in the lab with giving feeding regime normally with live feed/artemia and artificial feed. During acclimatization the shrimp will be reared in 500 L container and equip with 4 aeration system from 4 sir stones. Commercial feed with 10% of the body weight was given 3 times a day and live feed (artemia 20 artNaupli) 2 times a day. 2. After two days, count ten PL-15 of L. vannamei and put it in 5 aquarium (25×35×40 cm) filled up with sea water and equipped with aeration tools throughout the period. 3. Water quality will maintain to be stable with salinity 30 ppt, and water temperature between 28-30 degree Celsius. 4. Disinfection composition of the present invention will prepare 60 minutes before use and diluted with fresh water. 5. Prepare the experimental kit with concentration level of Disinfection composition of the present invention between 0 to 1000 ppm (the concentration will be 0 ppm, 1 ppm, 10 ppm, 100 ppm, 1000 ppm). No Replication used in this preliminary test. 6. Determine the highest concentration with 0% mortality and the lowest concentration with 100% mortality after 12 hours. The concentration will be the lower level and upper level of the test.

(14) ii. Definitive Test 1. After the value of lower and upper level is found by preliminary test, divide the range between the value into 7 concentrations (for examples lower level=0 ppm and upper level=100 ppm, so the concentration will be 0 ppm, 10 ppm, 20 ppm, 40 ppm, 80 ppm, 100 ppm and 200 ppm). 2. Count Ten PL-15 of L. vannamei that already pass the 3 days conditioning phase and put it in 18 aquarium (25×35×40 cm) filled up with 10 Litres sea water and equipped with aeration tools throughout the period. Each concentration will have three replications. 3. Disinfection composition 1 of the present invention will prepare 60 minutes before use and diluted with fresh water and then apply directly to the tank to get the concentration 4. Water quality will maintain to be stable with salinity 30 ppt, and water temperature between 28-30 degree Celsius. 5. Take note of the PL rate of death hourly in the first 12 hours, then twelve hourly for 96 hours. 6. Analyze the test result to find out the concentration level that causes 50% of death rate of the PL within 96 hours. 7. Analyze the result with probit analysis to determined LC50 value.
Results:

Example 1: Present Invention

(15) Lethal toxicity test to determine the concentration that killed 50% shrimp population, and sub-lethal toxicity test to determine the toxicity level that not kill the organism. Lethal toxicity test data analyzed with probit analysis to determine LC50 values in 96 hours exposure period. The stage of the shrimp that will be used in this trial will be Nauplii and post larval white shrimps at 15 days of age (PL-15) of white shrimp (Litopenaeus vannamei, Synonym Penaeus vannamei). Shrimp were exposed to a series of disinfectant concentrations for 24 h and 96 h, respectively.

(16) A probit regression analysis was used to calculate standard LC.sub.50 values and their 95% confidence limits. a) Nauplius stage is selected because this is the first stage in hatchery process. For this stage toxicity test should modified to 24 hours period, as in Nauplius only last for 24 hours then change to Zoea stage. b) PL-15 stage is selected to be tested because usually this stage is the latter age that send to grow out ponds.

(17) Definitive test for a) Nauplii and b) PL15 respectively, the details are below:

(18) Results: the mortality rates of 24 hours period for Nauplius and of 96 hours period the mortality rates for PL-15

(19) a) Lethal concentrations for Nauplii

(20) TABLE-US-00005 LC* LC Minimum** Maximum** (%) (ppm) Dosages (ppm) Dosages (ppm) LC50 8.317 5.723 12.087 LC85 21.532 14.817 31.291 LC90 26.965 18.555 39.185 *LC is the abbreviation of lethal concentration. A LC50 is concentration of a substance which required to kill 50% of the tested population. The lower the LC50 value is the more toxic the substance is. A LC90 is concentration of a substance which required to kill 90% of the tested population. **The minimum and maximum dosage is a 95% confidence interval, meaning that with 95% confidence the true value is within this interval.

(21) b) Lethal concentrations for PL-15

(22) TABLE-US-00006 LC LC Minimum Maximum (%) (ppm) Dosages (ppm) Dosages (ppm) LC50 55.204 45.739 66.629 LC85 71.635 59.352 86.459 LC90 76.188 63.124 91.954
Comparison Composition 1:

(23) Examples 1a) and 1b) were repeated but instead comparison composition 1 was used Results:

(24) TABLE-US-00007 Example Comparison Stages of L. 1a) composition 1 vannamei LC50 (ppm) Nauplius 8.317 4.538 PL-15 55.204 28.207

(25) TABLE-US-00008 Example Comparison 1b) composition 1 Stages of L. 95% confidence interval vannamei LC50 (ppm) Nauplius 5.723 4.222 12.087 4.846 PL-15 45.739 24.398 66.629 32.099
Result:

(26) Direct comparison of the lethal concentrations for Litopenaeus vannamei Nauplii of the composition 1 of the present invention and the comparison composition 1 shows that toxicity is decreased for the composition of the present invention. Direct comparison of the lethal concentrations for Litopenaeus vannamei PL-15 of the composition 1 of the present invention and the comparison composition 1 shows also a reduced toxicity. Hence, it was proven that with the present invention a disinfectant composition with improved properties in particular for shrimps of the age of 1 to 27 days, after hatching, in particular in the stage from being Nauplii up to the PL-15 phase was provided.

(27) Comparison Composition 2:

(28) Examples 1a) and 1b) were repeated but instead comparison composition 2 was used Results:

(29) TABLE-US-00009 Example Comparison Stages of L. 1a) composition 2 vannamei LC50 (ppm) Nauplius 8.317 2.724 PL-15 55.204 27.7005

(30) TABLE-US-00010 Example Comparison 1b) composition 2 Stages of L. 95% confidence interval vannamei LC50 (ppm) Nauplius 5.723 2.5632 12.087 2.8848 PL-15 45.739 24.625 66.629 30.776
Result:

(31) Direct comparison of the lethal concentrations for Litopenaeus vannamei Nauplii of the composition 1 of the present invention and the comparison composition 2 shows that toxicity is decreased for the composition of the present invention around 3 fold.

(32) Direct comparison of the lethal concentrations for Litopenaeus vannamei PL-15 of the composition 1 of the present invention and the comparison composition 2 shows also a nearly 2 fold reduced toxicity. Hence, it was proven that with the present invention a disinfectant composition with improved properties in particular for shrimps of the age of 1 to 27 days, after hatching, in particular in the stage from being Nauplii up to the PL-15 phase was provided.

(33) Hence, even the fish disinfection composition with lower amounts of surfactant than the Virkon® Aquatic product as known from WO 2007/023481 still shows a poor performance compared to compositions of the present invention.

FINAL CONCLUSION

(34) The disinfectant presented in this invention relates to a disinfectant composition for the shrimp aquaculture and a respective process for using such a disinfectant showed that it can even be further improved with regard to toxicity over a similar commercial product available on the market as well as known fish disinfection compositions known in the prior art.