ПЕРЕКИСЬ ВОДОРОДА КАК ЗАМЕСТИТЕЛЬ КИСЛОРОДА, ОБЕСПЕЧИВАЮЩИЙ ЖИЗНЕСПОСОБНОСТЬ РЫБЫ В УСЛОВИЯХ ГИПОКСИИ

Ураков А.Л.

ORCID: 0000-0002-9829-9463, Доктор медицинских наук, Профессор, Ижевская государственная медицинская академия, Ижевск, Россия

ПЕРЕКИСЬ ВОДОРОДА КАК ЗАМЕСТИТЕЛЬ КИСЛОРОДА, ОБЕСПЕЧИВАЮЩИЙ ЖИЗНЕСПОСОБНОСТЬ РЫБЫ В УСЛОВИЯХ ГИПОКСИИ

Аннотация

Результаты экспериментальных исследований показывают, что сохранить жизнь рыбам в условиях гипоксии может перекись водорода. Показано, что рыба не погибает в воде от гипоксического повреждения головного мозга при отсутствии в воде растворенного кислорода, если вводить перекись водорода в эффективной дозе в воду и/или в желудок рыб. Эти данные доказывают, что рыбы всасывают перекись водорода из воды, в которой плавают, и из своего желудка. Затем в организме рыб перекись водорода в результате каталазной реакции выделяет кислород. Поэтому для сохранения рыбы живой в условиях гипоксии предложено использовать перекись водорода вместо  газообразного кислорода или в месте с ним.  

Ключевые слова: живая рыба, гипоксия, кислород, заменитель кислорода.

Urakov A.L.

ORCID: 0000-0002-9829-9463, MD, Professor, Izhevsk State Medical Academy, Izhevsk, Russia

HYDROGEN PEROXIDE CAN REPLACE GASEOUS OXYGEN TO KEEP FISH ALIVE IN HYPOXIA

Abstract

The results of experimental studies show that to save the life of a fish in hypoxic conditions can hydrogen peroxide. It is shown that fish does not die in the water from hypoxic brain damage in the absence of dissolved in water oxygen, if introduce hydrogen peroxide in an effective dose to water and/or in the stomach of the fish. These data prove that the fish suck the hydrogen peroxide from the water in which swims, and from stomach. Then in the fish body, the hydrogen peroxide resulted catalase reaction releases oxygen. Therefore, to preserve the fish living in hypoxia is proposed to use hydrogen peroxide instead of oxygen gas or in a place with him.

Keywords: live fish, hypoxia, oxygen, oxygen substitute.

Introduction

To save the live fish in the water traditionally used water aeration by atmospheric air. However, despite this about 60% of the grown and caught fish dies during transportation and storage prior to sale [1, P.117 ]. The fish dies due to lack of oxygen. So to prevent the death of fish and reduce spoilage of fish during storage has long offered different ways and means. One of the effective and tested ways of storing fish is hypothermia [3, P.50]. The fact that hypothermia is one of the most important factors of protection of the brain from hypoxic damage, and fish belong to cold-blooded animals, so it's easy allow a significant lowering of the temperature of the water environment, which are during hypoxia.

In addition, to preserve the fish alive has previously been proposed to use hydrogen peroxide, because the gills of fish capable of producing hydrogen peroxide catalase reactions, to extract from it the molecular oxygen and to ensure its absorption into the blood [2, P.1; 4, P. 100 ].

However, this proposal was noticed.

The aim of this study was to investigate the influence of hydrogen peroxide on the survival of fish in the absence of gaseous oxygen in water.

Material and methods

In the conditions of vivarium laboratory investigations of the dynamics of motional activity of 150 healthy adult fish to breed guppies and blue neons in modelling acute hypoxia at a temperature from +15 to +25°C. To study fish were placed inside a separate sealed transparent containers [3,P.97]. Hydrogen peroxide was introduced into the water with the in the range of doses from 0.05 to 0.3 ml/kg of fish. In the process of hypoxia were recorded the dynamics of motional activity of fish, in particular the frequency of the respiratory movements of the gill arches, the frequency of opening of the mouth, frequency and amplitude of the fins, as well as the sequence of color changes of the fins. Monitoring of the locomotor activity of fish was conducted by eye and using photography in the visible spectral range.

Statistical processing of the results was performed using the BIOSTAT program according to the standard technique.

Results

It is established that in normal after putting the fish in a sealed container with fresh water in the studied range of water temperature fish in the first ten minutes are remaining relatively calm. In the next period of hypoxia appear active movement of gill arches, fins, and torso. Increased frequency of mouth opening and passing the water through the gills, fish darting into container volume, they darken the color of the pectoral fins. Sometimes the fish rest for 5 - 10 seconds. Then about 50% of the fish sinks to the bottom of the container, flips and floats belly up. Regardless of the position of the fish in the water they still open mouth and the water passing through the gills, but it happens several times less than before.

The final stage of hypoxic damage to the fish are in a state of  belly up, about 1 minute. During this period, they observed a rare respiratory movements of the mouth, gills arches and a single twitching fins. Simultaneously, the fish periodically defecate. The total duration of a succession of intermittent jerking of the fish breed guppies at their acute hypoxia in water at 25°C lasts of 65.0 ± 0,9 (P ≤ 0.05, n = 100) seconds. Then the locomotor activity of the fish is terminated fully and finally, and the fish die. Attempts to revive them by the introduction of oxygen to water or hydrogen peroxide did not lead to success. It was found that lowering the temperature of the water with fish from 25 to 15°C extends the lifespan of the fish without air access in 2,2 – 2,3 times. Similar results were obtained also in experiments with fishes breed blue neon.

Then, experiments were conducted with hydrogen peroxide. When calculating the dose of hydrogen peroxide we was based on the assumption that 100 ml of 6% solution of hydrogen peroxide may allocate up to 1.97 l of O2 (molecular oxygen) weighing 2,816 g. Thus, we introduced hydrogen peroxide into the water with the fish in such a dose that kept the level of dissolved oxygen in the water unchanged. However, the in water appeared bound oxygen, the amount of which was similar to the amount of dissolved oxygen in the water. The results showed that the introduction of hydrogen peroxide at the indicated dose increased 2 times the lifespan of a fish in water in the absence of incoming atmospheric air and oxygen.

In particular, the results of the experiments with fish breed blue neons showed that in conditions of normal atmospheric pressure and temperature of the water in +15 - +16°C the introduction of a 6% solution of hydrogen peroxide at a dose of 0.2 ml/kg fish in the water extends the life of fish in hypoxia from 49,73 ± 2.10-99 to10 ± 4.70 minutes. Similar results were obtained in experiments with fish breed guppies and blue neons under hypoxia at normal atmospheric pressure at a water temperature of +25 - +26°C. It is shown that under these conditions the introduction of a 6% solution of hydrogen peroxide at a dose of 0.2 ml/kg fish also extends the period of preservation of live fish. So, fish breed blue neons and guppies was die in contro to 25.3 ± 1,31 and 27.4 ± 1.32 minutes (p ≤ 0.05, n = 5), and with hydrogen peroxide solution through  52,43 ± 2,43 and 57,11 ± 2.45 min (p ≤ 0.05, n = 5) (respectively).

At the same time, it was found that after 5 to 12 minutes after introduction to the water with the fish of hydrogen peroxide in a dose exceeding the therapeutic dose 4 times, all the fish die.

Then studied the effect of hydrogen peroxide on lifespan of fish in hypoxia with the introduction of hydrogen peroxide inside (with the help of a gastric tube into the stomach of the fish) and intraperitoneally (by injection). Studies have been conducted on the fish breed swordtails and rasbory trilinear.

Initially, the effect of intraperitoneal injection of 0.1 ml of 0.9% sodium chloride and 0.1% hydrogen peroxide was studied on the fish breed rasbory trilinear. The results of experiments showed that intraperitoneal injection of hydrogen peroxide in this dose accelerates the death of the fish by 7.44 ± 0.62 min (P ≤ 0.05, n = 5) compared to control (by injection of a solution of 0.9% sodium chloride).

In the next series of experiments we have investigated the influence of a solution of 0.01% and 0.05% hydrogen peroxide when introduced into the stomach of the fish. While inside the fishes breed rasbory trilinear  was introduced at 0.05 ml, and in the stomach of the fish breed rasbory trilinear was introduced 0.1 ml of a solution of 0.05% hydrogen peroxide. It turned out that after the introduction of a solution of 0.01% hydrogen peroxide in the stomach of the swordtails  and rasbory trilinear the dynamics of motional activity of fish remained the same. However, the introduction of a solution of 0.05% hydrogen peroxide has improved the dynamics of locomotor activity and survival of fish in sealed containers with water. In particular, after sealing the tank with water and fish breed swordtails or rasbory trilinear in the control series, fish were killed using 28,30 ± 0,70 and 33,90 ± 0.90 minutes (respectively). In another series of experiments after introducing into the stomach of the fishes of the solution of hydrogen peroxide in fish breed  swordtails  and rasbory trilinear died after sealing of the water container, respectively, via 33,90 ± 0.90 and 43,66 ± 0.80 minutes (P ≤ 0.05, n = 5).

In other words, enteral administration of the solution of hydrogen peroxide in a therapeutic dose is prolongs the life of aquarium fish in the conditions of sealed containers at an average 20%.

Discussion

The results of our experiments indicate that the hydrogen peroxide solution can save fish from death in conditions of hypoxia, and can kill the fish by overdose.

Hydrogen peroxide increases the resistance of fish to acute hypoxia, most likely due to the intake of oxygen into the blood via the gills and gastric wall.

Thus, the cavity of the stomach can be used for intragastric gas exchange and can become an independent method of treatment of respiratory failure and/or the preservation of life in the absence of oxygen in the inhaled air. So you can enter into the stomach not only the solution of hydrogen peroxide, and hypergazation water with gas oxygen [5, P.949; 6, P.1]. The introduction into the cavity of the stomach solution of the hydrogen peroxide, aerated with oxygen gas under excess pressure, has been called as "intragastric hyperbaric oxygenation".

Conclusion

Consequently, the hydrogen peroxide solution, that introduced into the water in which fish swim to and/or in the stomach, is able to maintain fish life in sealed containers in the absence of aeration of water by air in the absence of dissolved in water oxygen gas. Thus, the results demonstrate that the hydrogen peroxide solution is able to replace gaseous oxygen for fish.

I express my gratitude to the students of L.V. Chernova and E.L.Fisher for assistance in conducting the experiments.

Список литературы / References

1. Чернова, Л.В. Влияние температуры на динамику цвета плавников и двигательной активности взрослых аквариумных рыбок при острой гипоксии / Л.В.Чернова // Международный научно-исследовательский журнал. - 2014. - № 3-4 (22). - С. 117-118.
2. Ураков, А.Л. Способ сохранения живой рыбы при транспортировке и хранении / А.Л. Ураков, Н.А.Уракова, Р.К.Агарвал, А.П.Решетников // RUS Патент № 2563151. 2015. Бюл. №  26.
3. Ураков, А.Л., Уракова Н.А., Чернова Л.В. Влияние температуры, атмосферного давления, антигипоксантов и химического «аккумулятора кислорода» на жизнеспособность рыб в воде без доступа воздуха/ А.Л.Ураков, Н.А.Уракова, Л.В.Чернова // Международный журнал прикладных и фундаментальных исследований. - 2014. - № 8 - 2. - С. 48 – 52.
4. Ураков А.Л. Эффективный способ хранения живой рыбы в воде на основе применения перекиси водорода/ А.Л.Ураков, Л.В.Чернова, П.Б.Акм аров// Вестник Ульяновской ГСХА. - 2016. - № 2. (34). - С. 96 -100.
5. Ураков А.Л. Введение гипероксигенированного раствора перекиси водорода в желудок повышает устойчивость организма к гипоксии/ А.Л.Ураков// Успехи современного естествознания. - 2015. - № 1. - С. 946 – 950.
6. Ураков А.Л. Средство для повышения устойчивости к гипоксии/ А.Л.Ураков, Н.А.Уракова, Д.Б.Никитюк// RUS Патент № 2604129. – 2016. – Бюл. № 34.

Список литературы на английском языке / References in English

1. Chernova L.V. Vlilanie temperatury na dinamiku ctveta plavnikov i dvigatel’noj activnosti vzroslyh akvariumyh rybok vzroslyh akvariumnyh pybok pri ostroj gipoxii [The effect of temperature on the dynamics of the color of the fins and motor activity of adult aquarium fish in acute hypoxia]  / L.V.Chernova // Mezhdunarodnij nauchno-issledovatel’skij zhurnal  [International research journal]. - 2014. - № 3-4 (22). - P. 117-118. [in Russian]
2. Urakov А.L. Sposob sohranenija zhyvoj ryby pri transportirovke I hranenii [The method of keeping of live fish during transportation and storage] / А.L. Urakov, N.А.Urakova, R.K.Agarval, A.P.Reshetnikov // RUS Patent №  2563151. - Bul. - 2015. - №  26. [in Russian]
3. Urakov А.L., Urakova N.A., Chernova L.V. Vlijanie temperatury, atmosfernogo davlenija, antigipoksantov i himicheskogo “akkumuljatora kisliroda” na zhiznesposobnost’ pyb v vode bez dostupa vozduha [Influence of temperature, atmospheric pressure, antihypoxants and chemical "oxygen battery" on the viability of the fish in the water without access to air] / A.L.Urakov, N.A.Urakova, L.V.Chernova // Mezhdunarodnyj zurnal prikladnyh i fundamental’nyh issledovanij [International journal of applied and fundamental research] - 2014. - № 8 - 2. - P. 48 – 52. [in Russian]
4. Urakov A.L. Effektivnyj sposob hranenija zivoj ryby v vode na osnove primenenija perekisi vodoroda [Effective method of storage of live fish in water on the basis of application of hydrogen peroxide] / A.L.Urakov, L.V.Chernova, P.B.Akmarov// Vestnik Ulianovskoj GSHA [Bulletin of the Ulyanovsk State Agricultural Academy] - 2016. - № 2. (34). - P. 96 -100. [in Russian]
5. Urakov A.L. Vvedenie giperoxigenirovannogo rastvora perekisi vodoroda v zheludok povishaet ustoichivost’ organizma k gipoksii [Introduction hyperoxygenating solution of hydrogen peroxide in the stomach increases the body's resistance to hypoxia] / A.L.Urakov// Uspehi sovremennogo estestvoznanija [Success of Modern Science]. - 2015. - № 1. - P. 946 – 950. [in Russian]
6. Urakov A.L. Sredstvo dlja povyshenija ustoichivosti k gipoksii [Agent for improving resistance to hypoxia] / A.L.Urakov, N.A.Urakova, D.B.Nikitjuk// RUS Патент № 2604129. - Bul. - 2016. - № 34. [in Russian]