RU2787391C2 - Method for disinvasion against exogenous stage of development of ascarid eggs in poultry - Google Patents

Abstract

FIELD: veterinary parasitology.

SUBSTANCE: invention relates to the field of veterinary parasitology; it can be used in poultry farms of different specialization in diseases of young chickens and turkeys with ascarids. A method for disinvasion against an exogenous stage of development of ascarid eggs in poultry includes treatment, at which a complex agent is used as a disinfectant, containing, wt.%: parachlorometaximenol – 34, sulfonic acid – 26, lactic acid – 5, auxiliary components, as which surfactants are used, – 23, the rest is water up to 100. In this case, treatment of poultry houses is carried out with the complex agent in a concentration of 1.0%, at a dose of 0.2 l per m², with exposure for 2 h.

EFFECT: invention provides creation of a method for disinvasion against an exogenous stage of development of ascarid eggs in poultry, with an outdoor content technology, with available means suitable for use by both manual and mechanical methods, without damage to treated technological equipment.

1 cl, 1 tbl, 4 ex

Description

The invention relates to the field of veterinary parasitology and can be used in poultry farms of various specializations in cases of diseases of young chickens and turkeys with ascaris.

Poultry farming plays an important role in the production of animal protein, the most important component of human nutrition. Today, our country is one of the five largest countries in the world in the production of poultry meat and eggs: moreover, the main producers of poultry meat in our country are poultry farms, their share in the total volume is more than 92%.

The conducted studies have established the influence of bird keeping technology on the epizootic state of the economy.

The works of scientists have proved that every poultry farm that practices floor keeping of birds is unfavorable for parasitic diseases, and most often, helminths and especially roundworms.

Despite the implementation of anti-epizootic measures, complete prevention of parasitosis among the livestock in poultry farms is not achieved. This is evidenced by the works of domestic and foreign veterinary parasitologists [1-5].

Ascaridiosis most often and severely affects chickens and young animals up to 8-10 months of age. Adult chickens are mostly ill in a subclinical form and are carriers of invasion. Among different breeds of chickens, Russian white and jubilee chickens are most susceptible to invasion, and Rhode Island chickens are more resistant to it. At the same time, not only infected chickens, but also overwintered ascaridium eggs can serve as a source of infection of chickens with ascaridiosis. It should be noted that Ascarid females are very prolific: during her life, one female is able to release up to 50 million eggs, which are scattered everywhere in the house and in the external environment. Hence, the most dangerous foci of invasion are poultry houses, walking areas, where litter of chickens and young animals contaminated with invasive elements accumulates.

Invasion of young birds can occur at any time of the year, but the most intense course is noted during the wet warm season.

In dysfunctional poultry farms, the transmission of invasion occurs through litter, inventory, feed and water contaminated with ascarid eggs. Synanthropic birds, rodents, insects: flies, cockroaches, crickets, as well as attendants - on shoes, clothes and bird care items often become mechanical carriers of Ascarid eggs. The wide spread of ascaridiosis among chickens is facilitated by various violations of the technology of growing young animals: increased humidity of the air and bedding, crowding of birds in the premises, inadequate feeding, contact between chickens and birds of older age groups, and violations of veterinary and sanitary rules. In addition, ascaridia eggs are stable in the external environment, and even in areas with rainy summers and cold winters, bird ascaridia eggs are able to remain viable for more than a year.

To achieve sustainable results in the fight against ascariasis in chickens, scientists and practitioners have proposed a number of agents and preparations against both endogenous and exogenous stages. The problem of avian ascaridiosis continues to pose challenges to researchers, namely, to improve measures to combat invasion and to develop a way to deal with the exogenous stage of development of eggs of ascaridia of chickens with outdoor technology for their maintenance, especially replacement young chickens of egg breeds.

Among the modern means used against exogenous stages of development of eggs of ascaridia birds, it should be noted: 4% sodium hydroxide solution, which should have a temperature of at least 80 ° C; 7% ammonia solution; 2% orthochlorophenol emulsion; 10% solution of iodine monochloride, however, the effectiveness of the previously noted funds does not satisfy the demands of modern poultry farming, which require combined products consisting of several active components and excipients.

As a combined agent, the following can be considered, which in its composition contains as active ingredients: parachloromethoxylenol, sulfonic acid, lactic acid, as well as auxiliary points: surfactants when used together in optimal concentrations.

The first two of the noted substances: parachloromethoxylenol and sulfonic acid are known as highly effective agents for infections of bacterial, viral and fungal etiology, as well as for the destruction of sporulated and non-sporulated forms of rickettsia and eimeria in poultry farming. The next substance that is part of the agent in question, lactic acid, is an antiseptic and has an effect on the shell of ascaridia eggs, facilitates access to active ingredients (DV). The substances considered above, when used together against the eggs of ascaridia of birds, have a synergistic effect on biological objects, and the auxiliary components used are surfactants that clean the floor, walls and process equipment from biocontamination and evenly distribute the active ingredient over the surface, providing high efficiency.

The task of our research was to develop a method of disinfestation against the exogenous stage of development of eggs of ascaridia of birds with outdoor technology of maintenance with available means suitable for use both manually and mechanized, without causing a damaging effect on the processed technological equipment.

The solution of this problem is achieved by the fact that as a method of disinfestation against exogenous stages of development of ascaridia of birds for the treatment of poultry houses, we used a complex agent compiled by us, which has the following composition: parachloromethoxylenol - 34%, sulfonic acid - 26%, lactic acid - 5%, auxiliary components - surfactants - 23%, water - up to 100% and we called it "ascaricide". From the sources of information available to the applicant, the specified set of existing features that allow obtaining the specified technical result is not known, therefore the claimed invention meets the condition of patentability "novelty".

The Applicant is not aware of means of a similar purpose, in which the distinctive features of the claimed invention were disclosed with the receipt of its use in any set of features of the specified technical result. Therefore, the claimed invention meets the condition of patentability "inventive step".

Evidence of the industrial applicability of the claimed is given in the following examples.

Example 1. Preparation of working solutions with different concentrations of disinfectants.

Research on this problem was carried out in the laboratory of the All-Russian Research Institute of Fundamental and Applied Parasitology of Animals and Plants named after K.I. Skryabin (VNIIP) - a branch of the Federal State Budget Scientific Institution Federal Scientific Center of VIEV RAS (Moscow).

Working solution with different concentrations of the combined des. means - "ascaricide" was prepared as follows. We took four clean glass flasks per liter and poured 700 ml of distilled water, and 5 were added to the first three of them; 10 and 20 ml of the product were thoroughly stirred for five minutes and the solution was ready for use. In the fourth flask, 700 ml of water, 40 g of crystalline phenol was added, stirred, the volume was brought to one liter, stirred again, allowed to settle, and the solution was ready for use.

Example 2. Preparation of a culture of helminth eggs Ascaridia galli.

The collection of ascaridia eggs from the faeces of infected birds was carried out by several methods. When they were isolated by the Fulleborn flotation method from samples of infected young chickens, the surface film was removed using a parasitological loop and collected in a Petri dish with distilled water, the eggs were washed three times from salt, their number was counted in one drop (10 μl), then in 1 ml, mixed with a preservative layer (1% hydrochloric acid solution) in distilled water and used in further work.

According to the second method, eggs were obtained from the gonads of sexually mature fertilized female Ascaris, which, after being removed from the intestine, were washed with saline. Sexual products were removed from the end sections of the uterus adjacent to the vagina no further than 1.0-1.5 cm, dissecting the helminth. Then the eggs were homogenized on a watch glass and mixed in a Petri dish with a preservative layer (1% hydrochloric acid solution), their number was counted in 1 ml and used in the work.

Example 3. Study of the ovocidal activity of different concentrations of the complex preparation "ascaricide".

Ovocidal activity of different concentrations of "ascaricide" in comparison with phenol was studied in a laboratory experiment in vitro after the cultivation of Ascaridia galli eggs in Petri dishes in a thermostat at 26-28°C in a humid chamber for 30 days. 2000 specimens of ascarid birds were placed in each chamber.

The first chamber served as a control, it remained without exposure to disinfectants, and cultivation in it took place in distilled water. A 4% aqueous solution of phenol was introduced into the second chamber with an exposure of 24 h, after which the cultivation was also carried out on distilled water. 0.5 was added to the third, fourth and fifth chamber; 1 and 2% solutions of "ascaricide", respectively, with an exposure of 24 hours when cultivated in distilled water. At the end of the exposure, Ascaridia galli eggs from chambers 2–5 were washed three times with distilled water, then they were microscopically examined to reveal morphological changes and put on cultivation. During the cultivation of eggs, aeration and control over embryogenesis were carried out once every two days.

Observations during the cultivation of eggs showed that in the first chamber (control) the development of ascaridia eggs with the formation of blastomeres was noted starting from the third day, and the formation of larvae in the eggs - 10-12 days after cultivation. The larvae in the eggs were mobile from the 14th to the 20th day, then they were at rest and became mobile only when heated. In the second chamber (phenol, 4%), an order of magnitude less blastomeres were formed during the cultivation. In the third chamber (“ascaricide” 0.5%), during the entire cultivation period, blastomeres in eggs developed only in a small part of them. In the fourth and fifth chambers (“ascaricide”, 1 and 2%, respectively), blastomeres did not develop in the eggs during the entire cultivation period.

The survival rate of eggs during cultivation was determined by counting under a microscope (100 eggs from each chamber). In the first control chamber (distilled water) there were 2% dead eggs. In the second chamber (phenol), the death toll was 88%. In the third chamber ("ascaricide", 0.5%), the dead eggs were 96%. In the fourth and fifth chambers, upon examination, all the eggs were dead.

When examining after culturing the sample from the first control chamber (distilled water), we found 98 motile larvae. In samples from the second (phenol) and third ("ascaricide", 0.5%) chambers, after centrifugation, 12 and 4 motile ascarid larvae were found, respectively. In samples from the fourth and fifth chambers, no motile larvae were found inside the eggs of ascaridia.

The following form was used to determine the intensity of effectiveness of the used disinfectants and different concentrations:

IE=(KYALK-KYALO)/KYALK×100,

where IE is the intensity of the drug (concentration),%;

NJALK - the number of eggs with larvae in the control group, ind.;

KYALO - the number of eggs with larvae in the experimental group, ind.

Using the data obtained by us in the experiment, we determined the intensity of the "ascaricide" at a concentration of 0.5%:

IE _0.5% =(98-4)/98×100=95.92%

The intensity of the tested drug "ascaricide" at concentrations of 1 and 2% was 100%, since all ascaridium eggs examined after centrifugation were dead.

It should be noted that under the action of an aqueous solution of "ascaricide" at a concentration of 1%, all laid eggs die, but their number remains (100 ind.), whereas when using a concentration of 2%, only 9 ascaridia eggs remained out of the total number of laid eggs. dead eggs that were severely deformed.

Phenol (4%), which we took as a comparison drug, showed the following effectiveness against eggs of ascaridia of birds:

IE _f =(98-12)/98×100=87.8%

Since in the fourth and fifth chambers (ascaricide concentrations of 1 and 2%) all the eggs examined after centrifugation were dead, a bioassay with material from these chambers was not carried out, and only material from the first three chambers was used.

The viability of ascaridia eggs was determined by light microscopy, by staining with vital dyes, 30 eggs from each chamber. For coloring eggs, a solution consisting of methylene blue, lactic acid and caustic alkali in a ratio of 0.05 g was used; 0.5 g and 15 ml, respectively. Live eggs from the chambers were not stained, while embryos of dead eggs were stained blue. The viability of the larvae was assessed by their mobility.

To confirm the viability of eggs after treatment with preparations, a biological test was carried out in the vivarium of the Institute on 20 chickens of the Anniversary breed, meat and egg productivity, 15 days of age, which were purchased in the poultry farm of the Moscow Region.

The experimental chickens were numbered and individually weighed and divided according to the principle of analogues into four groups (5 animals each) and kept in mesh cages from 16 to 51 days of life. At 16 days, all chickens were subjected to coproscopic examination according to the Darling method; according to its results, they were free from helminth eggs and oocysts of eimeria. Chickens were distributed as follows: group 1 - control uninfected, they were asked 1 ml of saline solution using a micropipette at 16 days; group 2 - control infected, which received 1 ml of a suspension of ascaridia eggs (400 copies) per 1 head; group 3 - infected with material after exposure to phenol (4%) at the same concentration of ascaridia eggs (400 ind./ml); Group 4 - infected with material after exposure to "ascaricide" (0.5%) at the same concentration of ascaridia eggs.

During the entire time of the experiment, the chickens of all four groups were kept in similar conditions and received the same diet. During the entire period of the experiment (35 days, from 16 to 51 days of life), the chickens were subjected to daily clinical observations of their general condition, behavior, food and water intake, and visible physiological changes.

To determine the concentration of ascaridia eggs in the faeces, samples were taken separately from chickens of each group for 20; 25; 30 and 35 days after inoculation.

In vivo studies. According to clinical studies, chickens of different groups had only minor differences. In chickens of the 1st group (intact control), no ascaridium eggs were found at all periods of the study, while in chickens of the 2nd group (infected control), eggs were found after 25; 30 and 35 days after inoculation in the amount of 840 to 3500 ind./g of litter. In chickens of the 3rd group (phenol), eggs were found 30 and 35 days after inoculation in the amount of 460-1600 ind./g of litter. Chickens of the 4th group (“ascaricide”, 0.5%) were also isolated with feces of eggs 30 and 35 days after inoculation in the amount of 280-740 ind./g of litter (Table 1).

On the 36th day of the experiment, the chickens of all four groups were subjected to weighing, slaughter and autopsy. According to the autopsy data, chickens of the 1st control group were free from ascaris. In chickens of the 2nd control group, an average of 46 copies were found in the intestines. ascaridium. In chickens of the 3rd group (phenol), ascaridium was found in the intestines of two chickens in the amount of 7 and 16 copies. In chickens of the 4th group (“ascaricide”, 0.5%), ascaridium was found in the intestines of only one chicken in the amount of 8 copies.

The results of a bioassay with experimental infection of chickens showed total infestation of infected control chickens (extensinvasion, EI - 100%), which indicates the successful functioning of this model, with a sufficiently high intensity of ascaridiosis invasion (the intensity of invasion of AI - 46 ind.).

Chickens of the 3rd group, which received material treated with a 4% phenol solution, were infected with ascaridia by 40% (EI 40%), with an average invasion intensity of 11.5 ind. (AI - 11.5 ind./head).

The smallest infection with ascaris was in chickens of the 4th group, which received material treated with a 0.5% solution of "ascaricide".

The results of weighing the chickens showed that the uninfected control had the largest increase in relation to the initial weight, while the chickens of group 2 (infected control) had the smallest. Undoubtedly, ascaridium in the intestines of infected chickens had a negative effect on feed digestion and growth. Chickens of both groups, treated with preparations of ascaridium suspensions, were characterized by intermediate values of this indicator. The average daily gain in live weight of chickens followed a similar pattern.

The obtained data show that under the influence of intensive ascaridiosis infestation, the chickens of the infected control (group 2) during the experiment had a significantly lower increase in live weight - by 29.4% than the chickens of the uninfected control (p<0.05). In groups 3 and 4 (receiving suspensions of eggs treated with phenol and "ascaricide" (0.5%), respectively), lagged behind the uninfected control by 23.98 and 16.1%, respectively.

Example 4. Production test of the effectiveness of the complex agent "ascaricide" against eggs of ascaridia of turkey poults.

The effectiveness of the complex remedy "Ascaricide" 1% against Ascaridium eggs of turkey poults in a production test was established experimentally with the artificial laying of ascaridium eggs on control sites compared with the base preparation phenol 4% at an exposure of 2 hours. The test was carried out in the conditions of a turkey farm in the Moscow region in March-April 2021. In poultry house No. 2 with a concrete floor, where cleaning and disinfection were carried out in advance, three sites of 1 m ² were selected. Each platform was separated from the other by plastic slats and the platforms had a flat surface to prevent leakage of disinfectant solution. At the start of the test, the prepared culture of A. galli diluted with 50 ml of distilled water was uniformly applied to the sites using a small spray gun and allowed to soak for 20 minutes. 2000 infested Ascarid eggs were applied to each plot. After the expiration of time, a 1% ascaricide complex agent was applied to the experimental site No. 1, the norm was 0.2 l per 1 m ² at an exposure of 2 hours, a 4% phenol solution was applied to the experimental site No. 2 at a rate of 0.5 l per 1 m ² and exposure 2 hours. Site No. 3 was the control and the preparation for disinvasion was not applied.

After 2 hours, swabs were taken from the surface of each area using a brush and distilled water. The wash collected from the surface of each site was transferred into a plastic bottle, the volume was adjusted to 1000 ml, mixed 2-3 times and left for 24 hours at room temperature (+20±2°). After the time elapsed, the contents were drained to the mark, the remaining 30 ml of the solution was used for the bioassay.

Washouts collected from sites No. 1, No. 2 and No. 3 were used for bioassay on 15-day-old turkey poults. There were 4 chickens in each group, which were given 1 ml of suspension with A. galli eggs at a dose of 500 copies. on the head.

The assessment of the general condition of the experimental turkey poults after the appointment of a suspension of ascaridia eggs was assessed according to daily clinical observations.

To establish the infestation with ascaridia, experimental turkey poults were subjected to coproscopic studies according to Fülleborn 15, 20, 25 and 30 days after the bioassay.

According to the results of the studies, the turkey poults of the first group, which received ascaridium eggs treated with the "ascaricide" agent, were free from ascaridia. In the second group, one chicken was infested with ascaridia (11 ind.), while in the control group all chickens were infected with ascaridia, their number at autopsy showed from 27 to 45 ind. to the intestines.

In a production test, the "ascaricide" 1% showed high efficiency against eggs of ascaridia of turkeys: IE - 100%. While the intensity of the base preparation of phenol 4% was 84.7%.

The results of laboratory studies on the study of the ovocidal properties of the "ascaricide" complex agent at different concentrations in an in vitro experiment on distilled water showed that under the influence of its different concentrations, the number of dead Ascaridia galli eggs (intensity) was: for a 0.5% solution - 95 ,nine%; 1 and 2% - 100%. Phenol (4%) used as a reference drug showed an intensity of 87.8%.

Due to the fact that, according to the test results, the concentrations of "ascaricide" 1 and 2% showed the same 100% efficiency, then for wide practical use for treatment against Ascaridia eggs in poultry houses during their preparation for settling with young chickens, for economic reasons, it should be recommend a 1% concentration of the drug.

Tests have shown the high efficiency of the complex remedy "ascaricide" in 1% concentration against ascaridia of young chickens and turkeys. A production test of the "ascaricide" 1% showed high efficiency against eggs of ascaridium turkeys: IE - 100%. This concentration can be recommended for use in the poultry farms of the country when preparing poultry houses for settling with young animals against exogenous stages of ascaridia.

Sources of information

1. Akbaev M.Sh., Vasilevich F.I., Akbaev R.M. et al. Parasitology and invasion diseases of animals. - M., 2008. - 776 p.

2. Instructions on measures for the prevention and elimination of animal diseases by helminthiases. - M., 1999. - 72 p.

3. Rules for disinfection and disinfestation of objects of state veterinary supervision. - M., 2002. - 74 p.

4. Safiullin R.T. Parasitic diseases of birds, means and methods of control. - M., 2019 - 260 p.

5. Chalysheva E.I., Safiullin R.T. Epizootic situation on intestinal parasitic protozoa of young turkeys at poultry farms in Central Russia // Collection of scientific articles based on the materials of the international scientific conference "Theory and practice of combating parasitic diseases". - M., 2019. - S. 690-695.

Claims (3)

A method for disinfestation against the exogenous stage of development of eggs of ascaridia birds, including treatment, characterized in that a complex agent is used as a disinfectant, containing, wt. %: parachlormethoxymenol 34 sulfonic acid 26 lactic acid five auxiliary components: surfactants 23 the rest is water up to 100 poultry houses are treated with a complex agent at a concentration of 1.0% at a dose of 0.2 l per m ² with an exposure of 2 hours.