Ветеринария/1 Ветеринарная медицина

COMPARATIVE METHODS OF DIAGNOSIS OF CARNIVORES NODULAR WORM DISEASE

Davletkalieva B.S. – Master student, 6M120100 - Veterinary Medicine major,A.Baitursynov Kostanai State University

Suleimanova K.U.–Candidate of science (Biology), Associate Professor at the Department of Veterinary Medicine,A.BaitursynovKostanai State University

Nowadays helminthiasesof dogs of urban and rural populations are one of the most studied and troublesome problems of veterinary medicine. It is known that many worms, parasites in dogs and cats are pathogens of farm animals and humans. Helminthiasis is not only veterinary and medical, but also a social, environmental and economic problem. Therefore, there are many works inveterinary helminthologydevoted to the diagnosis of nodular worm diseases. Speaking about the importance of diagnosis, G.A.Kotelnikov noted that in order to prevent the infection of humans and animals, it is necessary to properly applythe research methods of helminthiasis. Most of the helminth release egg cells, larvae and body fragments (segments), so accurate diagnosis of helminthiasis is the beginning of the whole chain of health measures [1, p.34; 2, p.11].

Currently there are a number of methods of diagnosis of helminthiasis, especially in the field of molecular biology, biochemistry and immunology, such as PCR, enzyme immunoassay, and others.

The ELISA method supplements well and in some cases completely replaces the direct parasitological diagnostic methods based on the detection of adult parasites or their egg cells in excreta of patients or larvae in biopsies of body and tissues [3, p.9]. The use of such methods of diagnosis is a relevant and necessary direction.

In this regard, the purpose of our research was to investigate the classical biochemical and diagnostic methods in a comparative aspect.

The material and methods of research

The work was done in 2013 - 2015 at the Department of Veterinary Medicine, A.BaitursynovKostanai State University.

The scientific and production research was carried out in the veterinary clinics of the university and "Aitar", in the Innovative research and education center and Smolinlaboratory.

There was feces and serologyanalysis ontoxocariasis and other helminth infections of dogs and cats.

Fyulleborn fecesanalysiswas carried outin accordance with standardmethods.

The immunosorbent assay was done on polystyrene plates, divided into strips. Antigens (1:50 - 1: 100) were dissolved in phosphate buffered liquor. Buffer solution was used for adsorption of excretory-secretory antigens second stage larvae of Toxocaracanis (ESAg-Tox). The same phosphate buffered saline with the addition of 0.05-0.1% Tween-20 (FSBT) was used to dilute conjugate and the sample of blood serum [4, с.15-18].

Intensity of invasion (like a relative indicator) is set with the counting chamber of Migacheva and Kotelnikov.

98 animals were examined, including 44 samples of feces of dogs aged from two weeks to 12 years old, 44 cats - 1 month - 9 years old from different administrative districts of Kostanai.

The experiments were carried outto study the diagnostic value of an enzyme immunoassay for 1-3 months old23 puppies. In addition, during an experimental study of toxocariasisof dogs (period «larva migrans») 1.5-5 months old5 puppies were examined by enzyme immunoassay.

Forexperimental infection of puppiesfeces were extracted fromanimals infested with toxocariasis. To isolate the helminth eggs from feces we used Berman-Orlov method. These helminth eggs were placed in an incubator for 5 days at t 30 ° C with relative humidity of 85%. After thatthe egg cells were given to puppies with a small amount of boiled rice for 50 larvae per puppy.Infestation of puppies were examined for toxocariasis every 15 days for 90 days.

The bloodserum of 22 dogs of different ageswas screened in ELISA for spontaneous infection of Toxocaracanis larvae.

Theresultsanddiscussion

For comparative studies with standard flotation method and enzyme immunoassay, theblood and feces samples of 98 carnivores were takenfor coprological studyto identify nodular worm diseases.

While studying 98 dogs and cats of urban population we found 42 animals infected with nodular worm diseases, which is 42.85%, including 3 animalsinfected with Dipylidiumcaninum, 2 - Diphyllobothriumlatum, 22 animals of Toxocaracanis, 14 -Toxascarisleonina, 1 - Uncynariastenocephala.

The studies showthat there are from3to 17 helminth egg cells of different species in 1 g feces, which indicates low and medium infestation of worm diseases in this area. The results are given in Table 1.

Тable 1 – The results ofdiagnostic study

№ п/п	Animal species	Numbero fanimal sexamined	Sick animals
			Coprologicalexamination	Extensive invasion, %	ELISA	Extensive invasion, %
1	Dogs	44	13	29,5	24	54,54
2	Cats	44	9	20,4	11	25

Analyzing the data in Table 1, we found 13 infected animalswhen studying 44 dogs by Fyulleborn method,which is 29.5%, ELISA identified 24 diseased animals, which is 54.54%, 44 cats by coprological method identified 9 sick animals, which is 20.4%, by ELISA - 11 sick cats, which is 25%.

Based on the epizootic monitoring of nodular worm diseases and ourresearch, we have found that toxocariasis isthe most frequent helminth infection in Kostanai. We examined three groups of dogs:

• The first group - 18 puppies, 6 months old

• The second group - 22 dogs, from 6 months to 2 years old,

• The third group - 24 dogs, 2 years and older.

The research was carried out by two ELISAin summer.

Table 2 - Efficiency of ELISA for epizootological monitoring at toxocarosis dogs

Groups of animals	Numberofanimalsexamined	Positive take	Extensive invasion	The average value of the extinction ELISAin groups of animals
				ELISА 1	ELISА 2
6 months old puppies	18	12	66,67	0,6	0,4
Male dogs	12	9	75	0,5	0,3
Female dogs	6	3	50	0,7	0,5
Dogs from 6 months to 2 years old	22	6	27,27	0,9	0,6
Dogs 2 yearsandolder	24	4	16,67	1,2	0,8

As Table 2shows, the largest extensiveness of invasion was in puppies under 6 monthsold, that is 66.67%. The invasion is common among males. The dogs from 6 months to 2 years old were infected at 27.27%, and the distribution of extensiveness of invasion remained. Males of toxocarosisinvasion reached 36.36 and females - 18.18%. The dogs of the third group had antibodies in the blood in 16.67% of the animals examined.

We have found that antibodies in blood serum were foundin a group of older femaledogs more frequently than in males. That may indicate the presence of tissue larvae in females and their more intensive accumulation with age. This biological phenomenon provides parasite circulation in the "mother - fetus" system,leads to intra-uterine infection of puppies and allows parasites to take a broader ecological niche [5. p. 319].

Figure 1 - Indicators of the diagnostic methods efficiency of nodular worm diseases.

Figure 1 shows thatimmunosorbent assay is more efficient. Its indicatorexceeds by13.27% in comparison with the coprological method.

Conclusion

Thus, nodular worm disease is one of the most common groups of diseases among dogs and cats. It should be noted that some types of helminthiasisis a threat to humans, too. Worms have harmin the body of animals: toxic (eccrisis), mechanical (injury of organs and tissues), transmissible (carry various pathogens), allergic (reaction to the presence of a foreign protein). These effects on the body of an animal can lead to serious consequences, that is why timely measures for the diagnosis, prevention and elimination of pathogenic effects on the bodyare so important. The use of immunoassay and classical methods of diagnosis of nodular worm diseases are effective, but ELISAis the most accurate and efficient method, as it allows to revealhelminthiasis in the early stages of its development in comparison with the classical method. In addition, the enzyme immunoassay allows you to quickly identify groups of infested animals for further detailed examination and to establish a final diagnosis.

Refernces:

1. «3і: intellect, idea, inovation» №1, 2014 // М.К. Мustafin, М.Zh. Аubakirov, А.Т. Zharmagambetov“The results of the application of advanced diagnostic method ofhelminthiasisofanimals”. P. 32-36;

2. Veterinary of farm animals №9, 2012 // N.I. Тumolskaya“Helminthiasisof pets”. P. 11-12;

3. Veterinary of farm animals №9, 2011 // L.М. Dedkova“Enzyme-linked immunosorbent assay in the diagnosis of helminthiasis”. P. 9-14;

4. Instructions for use D-2752 ELISA – Best // Novosibirsk, 2012. P. 15-18;

5. N.А. Romanenko,Padchenko, N.V. Chebyshev // Sanitary parasitology. Мoscow, 2000. P. 319.