RU2580655C2 - Method for larval nematode ascaridata culture - Google Patents

Abstract

FIELD: medicine.

SUBSTANCE: invention refers to veterinary science and aims at preventing helminthic infections in animals and human. The method consists in the fact that running out the larvae from chorions is initiated by processing with DP-2T disinfectant.

EFFECT: declared method is effective to produce larval nematodes of various representatives of the suborder Ascarides.

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Description

The invention relates to veterinary medicine, as well as medicine, and can be used in the field of prevention of helminthiases in animals and humans, as well as in laboratory diagnostics for the creation of enzyme-linked immunosorbent assay systems based on the antigen of nematodes of the ASCARIDATA suborder derived from larvae.

Intestinal helminthiases are the most economically significant (with the exception of malaria) parasitoses of humans, animals and birds. But until now, improving their diagnosis remains an urgent problem. Classical coproscopic methods for the diagnosis of intestinal helminthiases have a number of irreparable disadvantages: dependence on the professional qualities of the researcher, low diagnostic efficiency with a weak degree of invasion.

Obviously, the most promising diagnostic method is serological based on enzyme-linked immunosorbent assay (ELISA).

However, to obtain the antigens necessary for many serological reactions, a number of methodological difficulties arise. In particular, to obtain antigen from the larvae of representatives of the Ascaridata suborder, it is necessary to stimulate the exit of nematode larvae from the egg membranes using mixtures of complex composition, in addition, having low efficiency.

The closest analogue for the patented method is the method of culturing the larvae of P. Tverdokhlebov, in which an artificial gastric juice consisting of trypsin and hydrochloric acid is used to destroy the egg membranes.

A significant disadvantage of this method is the low efficiency of the larvae.

The aim of the invention is to increase the percentage yield of larvae from the egg membranes.

The method is as follows: fertilized Ascaris suum eggs were incubated for 24 hours at 27 ° C in physiological saline with the addition of 0.2% DP-2T. For this, 5 grams of this drug was diluted in 1 liter of distilled water.

Then the eggs were washed from the preparation DP-2T by centrifugation for 10 minutes at 1000 vol. / min and incubated in penicillin vials with saline in a volume of 2 ml for 30 days.

The incubation process was monitored every two days by microscopy.

After a month of incubation, from approximately 20% of the fertilized eggs, mobile larvae entered the solution (Table 1).

In control, the release of larvae from eggs was not observed.

When using artificial gastric juice, the percentage of larvae emerging from the egg shells in our experiment was much lower (less than 5%).

Similar results were obtained on other test organisms from the Ascaridata suborder.

When processing DP-2T in the same concentration for an hour, similar results were obtained using Arcadia galli (Table 2) and Ascaris lumbricoides (Table 3) as test objects for exposure to DP-2T.

Table 1 The results of a study of the exit of Ascaris suum larvae from egg shells No. The percentage of released Ascaris suum larvae (%) samples Under the influence of DP-2T Under the influence of gastric juice (control) one. fifteen 0 2. eighteen one 3. twenty four four. 23 0 5. twenty 3 6. 19 2

table 2 Results of a study of the exit of Arcadia galli larvae from egg shells No. The percentage of released Ascaris suum larvae (%) samples Under the influence of DP-2T Under the influence of gastric juice (control) one. twenty 0 2. 25 one 3. 16 four four. eighteen 0 5. 19 3 6. twenty 2

Table 3 The results of a study of the exit of Ascaris lumbricoides larvae from egg shells No. The percentage of released Ascaris suum larvae (%) samples Under the influence of DP-2T Under the influence of gastric juice (control) one. twenty 0 2. 25 one 3. 26 four four. fourteen 0 5. 13 3 6. eighteen 2

Thus, the patented method for culturing larvae can be used to obtain larvae of nematodes of various representatives of the ascaris suborder.

Literature

1) Sergiev V.P., Lobzin Yu.V. and others. Human parasitic diseases (protozoa and helminth infections). - SPb., "Tome", 2006. - C.2 - 444.

2) Sorokina T.S. History of Medicine. - M .: “From. Center "Academy". - 2006. - P.62-81.

3) Safiullin R.T. Comparative efficiency of coproscopic methods for the diagnosis of pig helminthiases and their improvement based on standardization. / R.T. Safiullin // Tr. VIGIS. - 2001. - T.37. - S.149-159.

4) Dolbin D.A., Lutfullin M.X., Latypov D.G. Comparative effectiveness of various helminthovoscopic methods for the diagnosis of calves neoascaridosis. // Tr. VIGIS. - T.41. - M. - 2005. - P.2005. - S.154-160.

5) Dolbin D.A., Agafonova E.V., Smirnova L.R., Khazieva S.M. Ways to improve the quality of laboratory diagnosis of helminthiasis. // Kazmed. - 2007. - T. LXXXVIII. - No. 4. - S.398-402.

Claims (1)

The method of culturing the larvae of nematodes of the suborder Ascaridata, which consists in the fact that to initiate the exit of the larvae from the egg membranes, the treatment with disinfectant "DP-2T."