Effectiveness of using electrochemically activated water with an acidic medium in breeding grain moth (cytotroga cerealella oliv) in biolaboratories

. The article presents, the harmful microorganisms in the air of the biolaboratory room and in the barley prepared for damage were determined during the reproduction of the eggs of the grain moth (Cytotroga cerealella oliv) used in the production of trichogram in biolaboratory conditions. The results of the study on the use of electrochemically activated water with an acidic environment (pH=3-3.5) to infect the identified microorganisms in the room air and to normalize grain moisture at the stage of contamination of barley grain with cytrago larva during the breeding process of grain moth eggs.


Introduction
Currently, in the field of cotton cultivation of our Republic, biological methods are mainly used to fight against pests, especially rootworm and bollworm, using Trichogramma fly.The process of reproduction of grain moth butterfly eggs mainly includes the following steps: preparation of barley grain for infestation with cytatroga larvae, infestation of barley grain with cytatroga larvae, care of cytatroga larvae and collection of eggs from grain moth butterflies [2].
The quality of the trichogram reproduced in this way, the preservation of its natural biological properties, depends on the technology of reproduction of the biological product and the existing conditions in the laboratory (room temperature, relative humidity, cleanliness of the room from harmful microorganisms, etc.).From this point of view, we used the part of the electrochemically activated water with an acidic environment (anolyte, pH=3-4) in order to clean the room air composition and the barley grain used in production from harmful microorganisms at the stage of the contamination of barley grain with cytatroga larvas during the reproduction of grain moth butterfly eggs in biofactories.
Researches were carried out in the scientific research laboratory of the "Chemical and Food Technologies" department of the Namangan Engineering and Construction Institute, in the "Water Analysis" laboratory of the Center for Sanitary Epidemiology, Peace and Public Health of the Namangan region, in the central laboratory of the Namangan "Suvokava" DUK (mutual cooperation agreement No. 2/2021 ) and Namangan "BIOSERVIS" LLC.

Methods and materials
The experiments consisted of 3 variants and were conducted in 3 replicates (volume 72 m 3 and wetting area 24 m 2 ) in a laboratory room.In this case, option 1 (control) is the current method in room 1, cleaning the room air with ventilation (ventilation) and regular tap water was used to normalize grain moisture, option 2, in room 2 cleaning the air of the room and normalizing grain moisture based on spraying electrochemically activated water with an acidic environment (рH=3-3.5),and the 3rd option consisted of disinfecting and normalizing grain moisture based on spraying room air with electrochemically activated water with an acidic environment (рH=4-4.5).
The "Scarlet" variety of barley was used to multiply the eggs of the grain moth butterfly (cytatroga) from barley grains [1].The temperature and relative humidity in the laboratory rooms were kept within the applicable standards (24-25 0 С and 80-85%).The experiments began with the study of the air of the laboratory room and put up for infestation harmful microorganisms in barely.
The results of the research proved that the air of laboratory room 1, selected for control, contains 25% gram-positive and 7% gram-negative cocci, the air of laboratory room 2, selected for the experiment, contains up to 24% gram-positive and 6% gram-negative cocci, and the air of laboratory room 3 contains 25% gram-positive and 7% gram negative cocci.Microorganisms which present in the air, conditionally also exist in the barley grains placed for damage in the laboratory.On this occasion, samples were taken from the barley poured for damage in the production process in order to determine the amount of harmful microorganisms in it, and examined in the "Bacteriology" laboratory.

Results and discussion
The results proved that: laboratory room 1 selected for control contains 26% gram-positive and 8% gram negative cocci in barley grain, laboratory 2 selected for experiment contains up to 25% gram positive and 7% gram-negative cocci in barley grain, and laboratory room 3 contains up to 26% gram positive and 7% gram negative cocci in barley grain.It can be seen from these results that the amount of microorganisms in the barley grain prepared for damage in the rooms of all options selected for the experiment is almost the same.
After the detection of harmful microorganisms in the air of the laboratory room and in the barley put for damage during the production process, we continued research on the effect of electrochemically purified water with an acidic environment on the existing microorganisms.Electrochemical activation of the water used for the research was carried out via the device MELESTA (TU 5156-002-32064511-07, certificate No. ROSS RU.AYa36.V29156) manufactured in the Russian Federation.The pH value of parts of water with acidic and alkaline environment obtained on the basis of electrochemical activation was determined with the help of universal indicator paper, the total hardness, the amount of chloride and sulfate ions was determined based on the current State Standards for "Hygienic Requirements for Drinking Water".
According to the results of the analysis, the hydrogen indicator of the water in option 1 (control) was pH=7.8, the total hardness was 5.8 mg/dm 3 , the amount of chloride ion Cl-was 53.2 mg/dm 3 , and the amount of sulfate ion SO4 -2 was 158.5 mg/dm 3 .
In option 2 (experiment), the hydrogen indicator of electrochemically activated water in an acidic environment was pH=3-3.5, the total hardness was 4.3 mg/dm 3  chloride ion Cl-was 46.4 mg/dm 3 , and the amount of sulfate ion SO4 -2 was 132.96 mg/dm 3 .In option 3, it was determined that the hydrogen index of electrochemically activated water in an acidic medium is pH=4-4.5, the total hardness is 3.6 mg/dm 3 , the amount of chloride ion Cl-was 42.3 mg/dm 3 , and the amount of sulfate ion SO4 -2 is 130.9 mg/dm 3 (Table 1).In the experiments, option 1, which was selected as a control option based on the current method, was ventilated with the air of the laboratory room with the help of ventilation, and normalization of grain moisture was carried out in ordinary tap water.
Option 2 with the portion of electrochemically activated water with a strong acidic environment (pH=3-3.5) in order to disinfect the air of laboratory room 2 and normalize grain moisture and in option 3 with the part of electrochemically activated water with a weak acidic environment in order to disinfect the air of laboratory room 3 and normalize grain moisture with (pH=4-4.5) was being processed.
The processing was carried out in two stages, and in the first stage it was carried out on the basis of spraying electrochemically activated water without touching the barley grain for 10-12 days from the day when the room air was placed to damage the barley grain.10 ml per 1 m 3 volume for one treatment, 720 ml per 72 dm 3 laboratory room for one treatment and 3600-4320 ml of electrochemically activated water were used for one production cycle.
In the second stage, after hatching of the worms from the laid eggs, the barley grains were moistened with a portion of the electrochemically activated water with an acidic environment once every 4-5 days (300 ml per cuvette).
The results obtained on the basis of the experiments carried out on the first stage showed that the air of the 1st laboratory room selected for control contained 25% gram-positive and 6% gram-negative cocci, the air of the 2nd laboratory room selected for the experiment contained 14% gram-positive and 4% gram-negative cocci, and the 3rd showed that laboratory room air contained 16% gram positive and 4% gram negative cocci.It can be seen that laboratory room 2 treated with electrochemically activated water (pH=3-3.5)with a strong acidic environment is better cleaned from harmful microorganisms compared to other options.
In the second stage, barley grains that used for production for fight against its microorganisms and for normalizing of grain moisture, the bareley grains begin heating at the day when the bareley has been damage, each cuvette was treated with 300 ml of normal tap water in the control option and with the acidic part of the electrochemically activated water in the experimental options.In option 1, 1-labaratory room , which was selected as a control option based on the current method, in order to normalize the moisture of the grain and to decontaminate the grain put into damage in 1-labaratory room used regular tap water.
In option 2, the grain put for damage in the 2-laboratory room so as to normalize the moisture and disinfect the grain we used electrochemically activated water with strong acidic environment (рН=3-3.5)and in option 3 the grain put for damage in the 3-laboratory room so as to normalize the moisture and disinfect the grain we used electrochemically activated water with strong acidic environment (рН=4-4.5).
After hatching of maggots from the eggs laid for infestation, based on grain moisture measurement (once in 4-5 days), each cuvette was moistened with 300 ml until the period of primary flight of butterflies from the grain.After notice the flight of primary butterflies from the damaged barley grain, 100 grams of samples were taken from each room and analyzed in the bacteriological laboratory to determine the amount of harmful microorganisms in it.
Analysis results: the barley sample in laboratory room 1 selected for review contains up to 26% gram-positive and 8% gram-negative cocci, the barley grain in laboratory room 2 selected for the experiment contains up to 16% gram-positive and 4% gram-negative cocci, and the barley in laboratory room 3 It was found that grain contains up to 20% gram-positive and 5% gram-negative cocci.It can be seen that the barley grain used for gilding in laboratory room 2, treated with electrochemically activated water (pH=3-3.5)with a strong acidic environment, was better cleaned from harmful microorganisms compared to other options.
During the experiment, 500 grains were taken as a sample from the cuvettes of each option, split with a knife, and the grain damage index was determined.In this case, the damage in the 1st control option was 68%, in the 2nd option 82%, and in the 3rd option 76%.From 16.03.2017until the first butterflies fly away, grain moisture was regulated in the order of options.
On 03/05/2021, the first flight of butterflies was observed in the 2nd experimental option, where the room air was sprayed with a part of electrochemically activated water that has a strong acidic environment, and it was used to regulate grain moisture.The third and first control options were observed on March 6, 2021 and March 8, 2021, respectively.Infected barley grains of each variant were collected in boxes two days after the first butterflies emerged.
The process of collecting butterflies from boxed barley and obtaining eggs from them was carried out according to the current method, observing the hydrometric mode.The results of the experiment show that 845 grams of boxed barley in option 1 (control), 920 grams of barley in option 2, and 902 grams of cytatroga eggs were obtained from barley in option 3. The results proved that 75 grams (value 112,500 sums) and 57 grams (value 85,500 sums) of 130 kg of barley in option 2 cytatroga eggs were obtained compared to the control option.
From the data presented in the table above, it can be seen that the most optimal option is to spray the room air with a part of electrochemically activated water that has a strong acidic environment (рH=3-3.5)and to normalize grain moisture, in which the damage rate of barley is 82%.Also, in this option, the initial flight of butterflies is accelerated by 3 days, saving a working day.

Conclusion
-The use of the part of the electrochemically activated water with a strong acidic environment (pH=3-3.5) in the cleaning of laboratory rooms from harmful microorganisms is effective because it is carried out directly in the production process, unlike chemical treatment; -the part of the electrochemically activated water with a strongly acidic environment (pH=3-3.5) is stable, unlike the part in the alkaline environment, and can be used for up to 15 days from the day of receipt; -in the reproduction of the eggs of the grain moth butterfly (cytatroga) from barley grain, the part of electrochemically activated water with a strong acidic environment (anolyte) can be used 10-12 days after the date of damage to the barley grain until the day when the first butterflies fly; -the most optimal option is the one-time application of electrochemically activated water in an average volume of 300 ml per 13 kg of grain in the process of reproduction of the eggs of the grain moth butterfly (cytatroga) from barley grain, which cleans the barley grain from 40-50% of microorganisms and at the same time reduces the moisture content of the grain (16% ) is ensured to be within the norm; -treatment of room air and barley grain with a part of electrochemically activated water with a strongly acidic medium (pH=3-3.5) in the breeding of grain moth butterfly eggs from barley increases production efficiency by 12-15% and saves working days.

Table 1 .
Some physical and chemical parameters of ordinary tap water and electrochemically activated water.