RU2637856C2 - Molluscicidal means for combating with helminthosis pathogens and method for its production - Google Patents

Abstract

FIELD: biotechnology.

SUBSTANCE: method provides collection, drying of the London pride (Saponaria officinalis) plant, dried plant grinding into powder. The London pride powder is extracted with ethyl or ammonia alcohol in a weight ratio of 1:100 for 24 hours at a constant temperature of 28±2°C and mixing at 100±1 rpm. The obtained extract is filtered through filter paper. The filtrate is subjected to concentration in the HEIGOLPH rotary evaporator at a pressure of 4±2 kPa, temperature 45±2°C and 280 rpm with the subsequent concentrate drying during one day in a vacuum desiccator at a pressure of 10 kPa and obtaining of a molluscicidal means.

EFFECT: group of inventions allows to increase the effectiveness of molluscicides and to prolong the storage time of the preparation.

2 cl, 3 tbl, 3 ex

Description

The invention relates to the field of agriculture, namely to veterinary helminthology, and can be used as a molluscicidal agent in the fight and prevention of various animal helminthiases.

As a fight and prevention against helminth diseases of animals in a pharmacopoeial way, drug soap preparations (Saponaria officinalis) are used. The method allows to implement the specified purpose. The invention relates to a herbal remedy for the control of mollusks (prudoviks, planorbids and others) - intermediate hosts of the causative agents of helminth infections of animals, disinfestation of livestock pastures, and can be used in a system of seasonal preventive measures.

Helminth infections are common in most domestic, wild animals, birds, as well as humans, they cause significant economic damage to agriculture. Most helminthiases, especially pasture ones, have intermediate hosts in the biological cycle of development — insect larvae, crustaceans, fish, reptiles, and mollusks. Mollusks are sources of most trematode and nematode invasions of animals. In the body of these invertebrates, a number of stages of development of larval forms of helminths occur. In different seasons of the year, the number of larval stages of helminths varies, therefore, for the control and prevention of helminth infections in animals, one should select time periods when a large number of invasive pathogens are concentrated in the mollusk organism.

The resistance of mollusks to the effects of physical, chemical and biological factors depends on the thickness of the ciliated layer of the integumentary epithelium and the density of the collagen fibers forming the skin of the muscular skin bag of the invertebrate.

Molluscicidal treatment of grazing meadows, for example, in Central Russia, is carried out during the period of the mollusks' active state (spawning, development and growth) - in May-June (after melt-water flow) and in August-September at a water temperature of at least 13 ° C and pH not higher than 7.6. Soap preparations on pasture meadows in the habitats of mollusks (puddles, wetlands and coastal areas of water bodies, fire canals, ditches, pits, puddles, pits) are applied in the form of a working solution using a sprayer (spray gun), and in the form of a powder, manually from the windward side . Before disinvasion of flood meadows (floodplains), the introduction of organic and mineral fertilizers is stopped.

Various methods are known for killing mollusks. Methods of using various forms of molluscicides are selected depending on the biological characteristics of the mollusks, their ecology, conditions of use, biotopes, taking into account the remaining optimal technical preparations. The most well-known and often applicable methods are such as spraying, dusting, sieving or scattering granules, balls, etc., introducing molluscicides into a stream of water replenishing a pond, using aerosols, drip application, using aerosols, fumigation, folding attractive baits, coating surfaces (soaking) interspersed).

A number of molluscicides of synthetic and plant origin are known, differing in structure and mechanisms of action, toxicity and cost of the drug.

Synthetic preparations, compounds of copper, arsenic, zinc, barium, mercury and polychloroterpenes, alcohols, phenols, etc. are more promising, but their use on large areas is quite expensive. These drugs are limited available and are toxic to the environment. Many chemical compounds have high molluscicidal activity, however, for their practical use, it is necessary that, along with high molluscicidal activity, these compounds are safe for humans, plants and microorganisms. It is important that after a certain time after treatment with the molluscicidal preparation, the residual amount of chemicals contained in agricultural products obtained from the treated area is safe for animals and humans.

Herbal preparations are environmentally friendly and slightly toxic to mammals, fish, amphibians, aquatic organisms and vegetation. Also, this group of molluscicides is inexpensive, affordable and safe to use.

The development of a safe, cheap and highly effective method of killing mollusks is an urgent task today as a fight and prevention of helminth infections in animals with their livestock (pasture) content.

The closest method for the destruction of mollusks is the pharmacopeia method for producing preparations (powder, water, alcohol, ammonia extracts) based on European spruce (Picea abies), frankincense (Bergenia crassifolia), ordinary oak (Quercus robur) [1, 2, 3]. The herbaceous plant of badass thick-leaved is territorially available only in Transbaikalia, Yakutia, Altai, Tuva. The short growing period, high productivity and wide availability of the proposed raw materials from soap dish compared with spruce and oak proves the promise of using molluscicidal properties of the plant. In order to expand the range of herbal molluscicidal preparations and expand the raw material base, it is proposed to use preparations from the roots, stems, leaves, flowers and fruits of the soapwort (S. officinalis), which is widespread. According to the well-known inventions for the destruction of mollusks, it was not known the active substance having a molluscicidal property obtained from plants. In our case, we purposefully used soap preparations, which contain surface-active substances (surfactants) - triterpene saponins with molluscicidal properties [4]. Roots and rhizomes are especially rich in saponins, up to 20-25% of these substances are found in them, of which saponaroside, hypsogenin (C ₃₀ H ₄₆ O ₄ ), saporubrin and saporubric acid are isolated. Flavonoic glycoside saponarin (C ₂₇ H ₃₂ O ₁₆ ), vitexin (C ₂₁ H ₂₀ O ₁₀ ), saponaretin were found in the leaves. Plant saponins are environmentally friendly and weakly and very slightly toxic to mammals, fish, amphibians, aquatic organisms and vegetation.

The closest analogue of the drug is a decoction of soap dish. Prior to the present invention, the authors of [4] proposed a prototype - a decoction of the roots, stems and leaves of a plant at a concentration of 1000 mg / l (0.1%). Molluscicidal effect was observed by the 6th day. We have proposed a pharmacopeia form of soap dish in the form of alcohol and ammonia extracts, which are inferior in terms of the effectiveness of the molluscicidal action to the decoction. In the proposed 0.1% concentration by the authors [4], the decoction of the plant shows poor efficiency and short shelf life. When preparing a decoction in a regime of more than 100 ° C, organic substances are partially destroyed, including saponins with molluscicidal action.

The preparations we offer (alcohol and ammonia extracts) have a longer shelf life compared to decoction and are more effective.

The purpose of the present invention is the development of an environmentally friendly, fast, affordable, cheap and effective drug with a molluscicidal effect. A chemically active group of substances (saponins) from the plant was obtained in the form of a powder, by alcohol and ammonia extraction.

Mylnianka, or Saponaria (lat. Sapondrid) - a genus of annual, biennial and perennial herbaceous plants of the clove family (Caryophyllaceae). The name of the genus comes from lat. sapo - "soap", the ability of the roots to form foam. On the territory of Russia and the CIS countries, 10 species of plants of this genus grow everywhere [5].

Saponaria officinalis is a strong expectorant and antitussive drug used to thin thick bronchial mucus in bronchitis, pneumonia, whooping cough, excruciating cough, and it also exhibits antimicrobial, diuretic, choleretic, anti-inflammatory, wound healing and anti-rheumatic effects [7]. Soap preparations increase the excretion of sweat and urine, stop heartburn and nausea. Locally (in the form of baths, lotions, gruel from powder, ointment) heals scabies, eczema, scaly lichen, purulent wounds, furunculosis, scrofula, various skin rashes, dermatitis. It is noticed that a decoction of a soap dish against scaly lichen is very effective. Gruel (ground roots with a small amount of hot boiled water) is locally used to treat purulent wounds, erysipelas, and eczema. Soapwort saponins have a local irritant effect with high hemolytic activity.

The effectiveness of the proposed method of combating pasture mollusks is reflected in the following examples.

Examples of specific performance.

Example 1. Obtaining powder extracts of soapwort officinalis. The plant is harvested during the seasonal period from July to October, upon reaching the maximum yield of the vegetative mass. At this time, in the physiology of the plant, flowering, seed ripening and the accumulation of surface-active substances (surfactants) occur. Plants are dug up with roots, washed in tap water from the ground and dried in the shade without access to sunlight to a moisture content of 15-18%. Next, molluscicidal preparations are prepared.

The powder of the preparation from the roots, leaves, stems, flowers and seeds of the plant is prepared by grinding the dried collection in a mortar and pestle to a size of 1-3 mm particles.

The prepared drug (powder) of the soap dish is placed in a conical flask and filled with 96% ethyl or ammonia per 1 g of 100 ml of liquid. The flask is mounted on an orbital rocking chair in a thermostatic environment at a temperature of 28 ± 2 ° С. Stirring is carried out at 100 rpm. Extraction is carried out within 24 hours. The resulting extract is filtered through filter paper, and then through a 0.2 nm pore membrane filter. The filtrate is concentrated in a HEIGOLPH rotary evaporator (Germany) at a temperature of 45 ± 2 ° C and 280 ± 5 rpm, as well as an air pressure of 4 kPa. The concentrate is dried in a vacuum desiccator at room temperature and a pressure of 10 kPa for 24 hours.

The resulting product is an amorphous gel-like mass of dark green with a specific delicate odor, it is soluble in water and alcohol.

Example 2. Experience in a laboratory study of the molluscicidal properties of preparations of soapwort (Sapondria officinalis).

The work was carried out in the laboratory of epizootology of parasitic animal diseases of VNIIP, while creating artificial biotopes for mollusks.

A culture of invertebrates, intermediate hosts of helminth infections, was collected from natural biotopes of the Moscow Region. Mollusks were kept in molds with a volume of 3-5 liters of 30 pieces, plastic photocuvettes of 30-50 liters of 250 pcs., Aquariums of 60-80 liters of 300 pieces. with the presence of clay, expanded clay, settling water, field and aquatic plants. Soil and plants were collected from the territory of the research and testing department of VNIIP. Phytolamps of the Fluora type were used in the laboratory to illuminate biotopes, and aquarium compressors were used for aeration of soil and water. The viability of the mollusks was revealed by microscopy under a Levenhuk 3ST binocular microscope while heating invertebrates on the Morozov table for 5-10 minutes according to locomotor activity. The studies were carried out 3 months after the removal of all components from natural conditions. As a control, a 0.1% decoction of soap dish was used. Analysis and observation were carried out within 30 days after exposure to drugs.

The soap dish powder in the laboratory showed a molluscicidal effect on two species of ponds, small (Lymneae truncatula) and marsh (Lymneae palustris). Powder treatment of mollusk cultures was carried out by hand scattering at a dose of 10 g over an area of 1 m ^{2 of} biotope. Starting from the 17th day of the experiment of 30, invertebrates died. The mollusk viability index by the 30th day of the experiment ranged from 0% to 3.3% (Table 1). The control revealed a single death on days 27-30 in a small pond. The swamp pond was resistant to decoction of soapwort.

The soap extract of alcohol soap was used in laboratory conditions at 0.01, 0.1, 1, and 10% concentrations (Table 2). The diluted preparation was dispersed with a 1000 ml Grida hand spray at the rate of 100 ml per 1 m ^{2 of} the artificial biotope area. In aquariums with a volume of 80 l with the presence of swamp water of 50 l and cultures of planorbid (Planorbis planorbis) and an ordinary pond (Lymneae stagnalis), the drug was added in the same volume. A working solution of the drug was prepared from tap water with bringing it to the desired concentration.

Extracts of 1 and 10% concentration had a 100% molluscicidal effect. The death of invertebrates was observed in the period from 3 to 36 hours. A 0.1% solution of alcoholic extract had a 100% molluscicidal effect in the small pond, the death of invertebrates was observed in the period from 5 to 16 days. The death of 53.6 to 61.7% of invertebrates (swamp and common ponds, planorbid coils) was observed from 10 to 30 days. A solution with a concentration of alcohol extract of 0.01% practically had no molluscicidal effect. A single death of invertebrates during the month was observed in a small pond and planorbid coils (Planorbis planorbis). In the control, a single death of mollusks from 20 to 30 days was noted.

The molluscicidal effect of small and bog ponds on the ammonia extract of soapwort is shown in Table 3. The preparation was applied by dispersion with a 1000 ml Grida hand sprayer in the same dose of 100 ml of working solution per 1 m ² of biotope area as the alcohol extract. In laboratory conditions, 0.01% and 0.1% aqueous solutions of ammonia soapwort extract showed a single death of mollusks from the drug within 18-30 days of the study. A 1.0% aqueous solution showed high efficiency (100%) when exposed to invertebrates for 4-8 hours. After 10-15 minutes, invertebrates ceased to actively move, the body was completely immersed in the shell. Microscopic examination of mollusks (magnification x2 - 20) marked a marked secretion of mucus (a protective reaction of the invertebrate organism to the contact irritation of the drug). In the control with a 0.1% decoction of Mylavochka, a single death of mollusks from 28 to 30 days was noted.

Example 3. Conducting field studies to study the molluscicidal effect of the alcoholic extract of common soapwort (S. officinalis). On the territory of walking pastures of the Moscow, Ryazan and Bryansk regions, tests were carried out on invertebrates in natural biotopes. In the spring, two places were selected with stations of lung-breathing mollusks with an area of 12 and 27 ha. In the studied territories, there is stagnant water from meltwater, which remains until June-July in the form of puddles and pits. During the spring flood season (March, April), water channels of no more than 12 cm deep can form in these places. In May-June, 5 biotopes from each place with mollusks inhabiting it were selected. The density of natural planting of invertebrates in the areas of biotopes with a hydrological regime of 11 to 48 m ² was different: L. truncatula - from 9 to 54 ind./m ² , L. palustris - from 18 to 184 ind./m ² , L. auricularia - from 0 to 18 ind./m ² , Physa fontinalis - from 17 to 82 ind./m ² . Treatment with a molluscicidal preparation based on an alcoholic extract was carried out by spraying with a ZhUK apparatus OP-209 with a volume of 9 l at the rate of 100 ml of a 1% solution per 1 m ^{2 of} biotope. The results of the study were taken into account on the 7th, 14th and 30th days of the experiment. As a result of the experiments performed on natural biotopes with mollusks, only empty shells of invertebrates were recorded during the period of taking into account the results in amounts from 2 to 18 ind./m ² .

The proposed method, which includes the use of alcohol or ammonia extract of the soapwort, provides a quick, cheap, environmentally friendly and effective fight against mollusks - intermediate hosts of animal helminths.

List of sources used

1. RU 2239321, A01N 65/00, 2004.

2. RU 2148319, A01N65 / 00, 2000.

3. RU 2141204, A01N 65/00, A01K 61/00, 1999.

4. Gorokhov V.V., Sturgeon B.C. Shellfish and their use in agriculture. - M .: Kolos. - 1987. - 224 p.

5. Drum E.I. Botany: a textbook for students. higher textbook. institutions. - M.: Publ. Center "Academy", 2006. - 448 p.

6. Danikov N.I. Healing poisonous plants. - M .: RIPOL classic, 2005 .-- S. 319-323.

7. Peshkova G.I., Schröter A.I. Plants in home cosmetics and dermatology. Directory. SMEs. 2001.668 s.

Claims (2)

1. A method of obtaining a molluscicidal agent, including collecting, drying a plant for soap soap (Saponaria officinalis), grinding with a pestle in a mortar of a dried plant for soap soap to a powder state, characterized in that the powder is poured into a conical flask for extraction with ethyl or ammonia in a weight ratio of 1 : 100 for 24 hours at a constant temperature of 28 ± 2 ° C and stirring at 100 ± 1 rpm, while the resulting extract is filtered through filter paper, and then concentrated in rotary isp at a pressure of 4 ± 2 kPa, a temperature of 45 ± 2 ° C and 280 rpm, after which it is dried for a day in a vacuum desiccator at a pressure of 10 kPa. 2. Molluscicidal agent for combating animal helminthiasis, which is a gel-like extract of soapwort (S. officinalis) obtained by the method according to claim 1.