Method of collecting fertilised eggs (in vitro) from exciter fasciola hepatica intra vitam

FIELD: biotechnology.

SUBSTANCE: method comprises the steps: collecting from the bile ducts of liver of domestic and/or wild animals infected with fasciolas of only live adult F. hepatica. Placement them into individual tubes with filtered and centrifuged bile diluted with isotonic solution of sodium chloride 1:1. Exposure of tubes at t = 38-39°C if F. hepatica is from cattle, and t = 39-40°C if F. hepatica is from sheep and/or goats, under conditions of thermostat for 5 hours. Subsequent washing eggs in isotonic solution of sodium chloride.

EFFECT: invention enables to obtain up to 100 percent of fertilised eggs of Fasciola of species Fasciola hepatica and can be used for study in the laboratory or field experiments in solving fundamental and applied scientific tasks in the field of epizootiology, treatment and prevention of fascioliasis of domestic or wild animals.

2 ex

 

The invention relates to the field of veterinary medicine, and more particularly to Parasitology ( = helminthology) to "the Way we collect during the life of the parasite fertilized eggs (in vitro) from the pathogen parasitic zoonosis Fasciola hepatica", and is intended for use when a parasitological laboratory and/or field studies helminthological fees (during the life of the parasite in artificial environments and at temperatures typical of ruminants), fertilized eggs from pathogen parasitic zoonosis: Fasciola hepatica, which is necessary for solving fundamental and applied scientific problems in the field of epidemiology, therapy and prevention of human fascioliasis domestic and/or wild animals.

It is known that the causative agents of fascioliasis domestic and wild animals registered in many countries and continents of the globe, following the Fasciola species: Fasciola hepatica L., 1758; F. gigantica Cobbold, 1885; F. jacksoni Cobbold, 1869; F. halli Sinitsin, 1933; F. californica Sinitsin, 1933; F. indica Warma, 1953 [H. B. Demidov - 1, Kapape M. B., Abramova, V. F., Pasechnik, B. E. - 2, Singh K. P., Srivastava V. K., Prasad Α., Pandey A. P. - 7].

In the Russian Federation, countries of the former USSR and other countries of the world marked the first 2 species of Fasciola, past and present who were causing enormous economic losses in livestock farms (USA - only in the state of North Dakota annual losses due to fascioliasis of sheep and Krupnov� cattle was estimated at 7 million 650 thousand) [1].

It is known that in all countries of the world fascioliasis and currently is causing direct and indirect harm which consists of a sharp decline in all types of animal productivity, culling of valuable meat products and even death.

It is known that the loss allowed from Fasciola was especially great to use effective anthelmintic (20-30-ies of the last century), which dramatically reduce mortality [Demidov N. In. - 1]. However, the loss of valuable animals from Fasciola, including rare and endangered animal species (the Indian elephant), is noted and is currently [K. Singh, R., Srivastava V. K., Prasad Α., Pandey Α.Ρ. - 7].

It is known that the availability of food and, in particular, proteins of animal origin (milk, meat, etc.) of the population of the Russian Federation is one of the priorities of the government of our country in the coming years.

Therefore, for scientists of veterinary medicine, Russia continues to be a pressing issue to ensure prevention of diseases, including invasive (fascioliasis, trichuriasis, etc.) that contribute to the emergence of bacterial and viral diseases (tuberculosis (Tuberculosis), bovine leukemia, influenza and African swine fever (African fever) of swine foot and mouth disease of cattle (Aphthae epizooticae), rabies (Rabies), etc.), home (KRU�tion cattle, sheep, goats, camels, reindeer, yaks, domesticated elk, horses) and wild animals (boars, deer; rare and endangered species: bison, bison, etc.).

It is known that to obtain the fees of eggs, researchers received helminth eggs in the following ways:

1) destroying the female genital organs of worms [Pasechnik V. E. - 4,5];

2) flotation, sedimentation, combined [Kotelnikov G. A. - 3].

These methods have the following disadvantages:

culture of eggs the 1st way is unacceptable, as in this method of collecting eggs Fasciola hepatica:

to receive up to 100% of the fertilized eggs is impossible, since along with the fertilized eggs of Fasciola hepatica are unfertilized, which creates great difficulties in laboratory and field studies;

b) the resulting collection is very very dirty remnants of destroyed tissue of the female genital organs Fasciola hepatica that during the experiment causes the development of bacterial microflora, and putrefaction of eggs of Fasciola hepatica.

Fees of eggs of Fasciola by applying flotation and combined, have the following disadvantages:

a) it is known [Kotelnikov G. A. - 3] that some saline solutions act negatively on eggs of Fasciola and cause their deformation (standardized flotation technique with a solution of lead nitrate (nitrate of lead) by G. A. Kotelnikova Fucking and V. M.; modification �kombinirovannogo method Vishnjauskas A. (1965): a solution of zinc sulfate; modification of the combined method on Skowroński, R. V. (1962): a solution of sodium thiosulfate with a density of 1.28), which is unacceptable for research in the laboratory and even in the field, as distorted outcomes: development of miracidia in the eggs of Fasciola;

b) widely used flotation method with saturated solution of sodium chloride (table salt), is not suitable for the collection of eggs of Fasciola, since the density of a solution of 1,18-1,20, and the proportion of eggs of Fasciola: 1,31-1,35;

b) unknown species, subspecies, race, strain of Fasciola.

Receiving fees of eggs of Fasciola way of sedimentation in spontaneously infected animal fascioliasis is more acceptable for conducting experiments in the field, however:

(a) in conducting research in laboratory conditions, it does not satisfy us, as the collection of eggs of Fasciola sp. this method is heavily littered with the remnants of the food of the masses, even with careful laundering of animal excrement spontaneously infected with fascioliasis;

b) in this method, unknown species (Fasciola hepatica, Fasciola gigantica) of pathogen invasion, which may be important in prevention and treatment activities in the area of distribution of other species of Fasciola (Fasciola gigantica, subspecies, strain, Rus) in the Russian Federation (this applies to the southern regions: Astrakhan, etc);

in this method of egg collection requires much time and labor in the study of a large number of faeces for the purpose of collecting a sufficiently large number of eggs of Fasciola because experimenters sometimes it takes up to several thousand eggs for laboratory and/or field research.

It is obvious that the researcher for this reason, to obtain the maximum amount (by known methods collection of eggs of Fasciola: by destroying the female genitals worms, flotation, sedimentation, combined) - up to 100% of the fertilized eggs, and it is only identified (and only known species in the way they collect eggs by destroying the female genital organs of the parasite, i.e. after the death fastsioles) pathogen parasitic zoonosis Fasciola hepatica, for experiments in field and/or laboratory conditions is extremely difficult and almost impossible.

The aim of the invention is the use for the first time developed a "Method of collecting during the life of the parasite fertilized eggs (in vitro) from the pathogen parasitic zoonosis Fasciola hepatica" to experiment with getting fees of eggs in the most sterile conditions, i.e. without contamination by tissue slices Fasciola hepatica and without entering secondary microflora from the destroyed tissues of Fasciola and external environment and, most importantly, get the f�MYM way during the life of the parasite, the maximum number is up to 100% of fertilized eggs from pathogen parasitic zoonosis: Fasciola hepatica L., 1758.

1. This object is achieved in that during the collection of eggs from Fasciola hepatica used designed us for the first time "the Way we collect during the life of the parasite fertilized eggs (in vitro) from the pathogen parasitic zoonosis Fasciola hepatica", which consists in selecting from the bile ducts of the liver, in the study of the helminthological methods at autopsy spontaneously infected with fascioliasis domestic and/or wild animal, only Mature, live helminths Fasciola hepatica in separate tubes with filtered and attentivepersonal bile 1:1 diluted in isotonic solution (0.9%) sodium chloride and exposition of tubes at t=the 38.2-39.5°C (if fastsioles from cattle) and at t=39-40,5°C (if fastsioles) from sheep and goats, for 5 hours in the condition of thermostat, and subsequent careful laundering eggs in saline solution (0.9%) sodium chloride, which differs from the known method of egg collection opening ( = destruction) of worms, that:

to collect eggs taken only Mature, live Fasciola hepatica obtained at autopsy of wild animals in national parks, hunting areas, and/or meat processing plants, slaughter houses, and in private farms (peasant, farmer, etc.), at slaughter allowed;

Fasciola hepatica are not destroyed DL� egg collection, but, during the life of the parasite, natural, active production (=lay) fertilized eggs in an artificial environment (separate for each parasite test tubes with filtered and trentepohliales bile 1:1 diluted in isotonic solution (0.9%) sodium chloride) and at a temperature characteristic of the ruminant (t=38-39°C - if Fasciola hepatica from cattle and t=39-40°C - if Fasciola hepatica from sheep and/or goats) in the face of thermostat and at the exposition time of 5 hours; in the collection virtually no: unfertilized eggs, the particles of destroyed tissue Fasciola hepatica, secondary bacterial microflora.

2. Method of collecting fertilized eggs (in vitro) according to claim 1, which differs from the known methods of collecting eggs of helminths: flotation, sedimentation, combined, that:

there is no need in definitive hosts of the parasite - infected spontaneously fascioliasis by domestic and/or wild animals;

no need for different salt solutions (some of them deformed eggs of Fasciola: modification of the combined method on Vishnjauskas A. (1965): a solution of zinc sulfate; modification of the combined method on Scorescom R. V. (1962): a solution of sodium thiosulfate, which was unacceptable in conducting research in the laboratory and even in the field, as distorted results);

it is not necessary to study a large number�the number of samples from spontaneously infected with fascioliasis domestic and /or wildlife for a large number of fertilized eggs of Fasciola;

no need for the separation of Fasciola hepatica eggs from other species of helminth eggs: strongest, Capillaria sp., Trichuris sp., oocysts of Eimeria, Cryptosporidium;

there is no debris egg collection Fasciola hepatica remnants of food masses (undigested food).

Method of collecting fertilized eggs (in vitro) according to claims. 1 and 2, characterized in that: live, Mature Fasciola hepatica during the life of the parasite lay in the above solution to 100% of the fertilized eggs of Fasciola (unlike the way they collect eggs when destroy ( = kill) helminths), if necessary, you can receive up to several thousand eggs and from the known species: Fasciola hepatica, Fasciola gigantica, etc., or subspecies, race, strain (in contrast to the known methods of collecting eggs of helminths: flotation, sedimentation, combined, when a species, or subspecies, race, strain of Fasciola unknown)necessary for research in the laboratory and/or field experiments, while solving fundamental and applied problems in the field of epidemiology, therapy and prevention of human fascioliasis domestic and/or wild animals.

Comparative analysis of the claimed technical solution with the prototype (by known methods collection of eggs of Fasciola: flotation, sedimentation, combined through the destruction of helminths - female genital mutilation) shows that these differences allow to draw a conclusion on compliance for�carried out for the technical solution the criterion of "novelty."

The features distinguishing the claimed technical solution from the prototype, not identified in other technical solutions, and therefore they provide the claimed technical solution the criterion of "substantial differences"

Examples of specific performance of the proposed method.

Example 1. In the Republic of Moldova of 24 goals (12 large and 12 small cattle - sheep) spontaneously infected with fascioliasis were collected by known methods of collection: flotation - with a solution of sodium nitrate, NaNO3(sodium nitrate), sedimentation, combined-modification according to A. Vishnjauskas, [3] (control) eggs of Fasciola and at the same time proposed a "Method of gathering during the life of the parasite fertilized eggs (in vitro) from the pathogen parasitic zoonosis Fasciola hepatica" (experience): collected at the slaughterhouses in the study helminthological method by K. I. Skryabin [6] from the bile ducts of the liver 450 copies of live, Mature Fasciola hepatica in separate tubes with filtered bile 1:1 diluted in isotonic solution (0.9%) sodium chloride and exposition of tubes at t=38-39°C (unless Fasciola hepatica from cattle) and at t=39-40°C (unless Fasciola hepatica from sheep and/or goats) in a thermostat for 5 hours, and followed by thorough laundering of eggs in saline solution (0.9%) sodium chloride.

After 5 hours of working time was machined parts�Yong summary of the experiment and the results showed that what is the Method of collection during the life of the parasite fertilized eggs (in vitro) from the pathogen parasitic zoonosis Fasciola hepatica" differs from the known methods of egg collection (flotation, sedimentation, combined) that:

there is no need in definitive hosts of the parasite Fasciola hepatica - infected with fascioliasis domestic and/or wild animals;

there is no need in the preparation of various salt solutions, which are often not available in stores or have limitations (ammonium nitrate, etc.), as may be used in order to carry out terrorist acts, many of these chemicals are sold at very high prices, since the main part of these salts are produced abroad of the Russian Federation, and, consequently, the fees of eggs of Fasciola above can be very costly;

eggs of Fasciola in some salt solutions were severely deformed (method of collection of eggs of helminths: combined) that could affect the results of experiments in the field and in laboratory conditions, these deformed eggs of Fasciola to conduct our experiments were absolutely unusable;

in collecting the eggs of Fasciola known methods (flotation, sedimentation, combined) were in a very large number of helminth eggs of other genera and species: straw�gilet, trichocephalus, dicrocoelium, paramphistomum, etc., and oocysts of Eimeria, Cryptosporidium, which greatly hampered the experiment;

when collecting eggs by the proposed method known species Fasciola hepatica, and the application of the methods fees, egg flotation, sedimentation, combined, the species is not known, since the animals in the southern regions of the Russian Federation (Astrakhan region, etc.) can be infected and other Fasciola species: Fasciola gigantica, or subspecies: races, strains, which is of great importance when conducting preventive and therapeutic measures;

when using the known method of egg collection (sedimentation) is a standardized method of successive washes (the most appropriate way), it turned out that 5 hours working time:

a) we have managed to collect 940 eggs (control), and applying the Method of collection during the life of the parasite fertilized eggs (in vitro) from the pathogen parasitic zoonosis Fasciola hepatica" - 5990 eggs (experiment);

b) collecting eggs was a lot of eggs of other species of helminths: Trichuris sp., suborder Strongylata: Nematodirus sp., Oesophagostomum sp., Dicrocoelium lanceatum, Moniezia sp., Strongyloides papillosus, Paramphistomum sp., and oocysts of Eimeria and Cryptosporidium, which significantly reduces the performance of the experiment, as much time was spent on the separation of duties of eggs of Fasciola these inclusions;

b) collecting eggs were a numberof undigested food, which had to be washed during the entire period of the experiment - 5 hours;

g) the method is time-consuming: it's hard to find eggs of Fasciola in the sediment;

e) unknown species of Fasciola (Fasciola hepatica, Fasciola gigantica, etc.).

Summarizing the results, we found that the known methods of collection of eggs of Fasciola (flotation, sedimentation, combined) can be used to eliminate above-mentioned disadvantages only in the field, as for laboratory experiments they have fully neudovletvoreny.

Therefore, this example clearly shows the advantage of the proposed "Method of collection during the life of the parasite fertilized eggs (in vitro) from the pathogen parasitic zoonosis Fasciola hepatica" in comparison with known methods of collection of eggs of Fasciola: flotation, sedimentation, combined.

Example 2. In the study at autopsy in meat processing plants, farm, farms, of the bile ducts of the liver of domestic animals (goats, sheep, cattle), in Russia (Moscow region), in the Republic of Moldova, were collected in the amount of 600 copies Fasciola hepatica, which were used for carrying out collecting eggs in two ways.

1st (experience) by the proposed method (300 copies), i.e. by the individual (in a separate vial) egg collection from every living, sexually Mature parasite� (live, Mature Fasciola hepatica course, arbitrarily lay ( = produce) eggs in test tubes with filtered and attentivepersonal bile 1:1 diluted in isotonic solution (0.9%) sodium chloride and exhibit them in a thermostat at t=38-39°C (if F. hepatica from cattle) and at t=39-40°C (if F. hepatica from sheep and goats) within 5 hours, and followed by thorough laundering of eggs in saline solution (0.9%) sodium chloride;

and the 2nd (control) in a known manner, the collection of eggs by partitioning ( = destroy) parasites (300 copies): Fasciola hepatica, which were collected in succession (Mature and immature F. hepatica) in the helminthological dissection of the bile ducts of the liver.

In follow-up were selected in a row for 1500 eggs of Fasciola in the experience and control and were cultivated in a thermostat at t=25-30°C (the optimum temperature for the development of miracidia) and viewed under the microscope to compare our method of collecting eggs (1 - experience) and the known method is through the destruction of the parasite: Fasciola hepatica (2nd control).

The results showed that in the culture of eggs by a known Method (2nd control) developed in 12 days (up to the stage miracidia) only 49% (735 out of 1500) eggs, and when we proposed Method (experience) collection during the life of the parasite fertilized eggs (in vitro) from the pathogen parasitic zoonosis Fasciola hepatica", eggs in the amount of 1500 to 100% sluchainymi alive and developed 12 days before the stage miracidia.

Thus, the results of culturing eggs collected on our way (1 - experience) confirmed the data obtained using the proposed "method of collecting during the life of the parasite fertilized eggs (in vitro) from the pathogen parasitic zoonosis Fasciola hepatica" from domestic and/or wild animals that have been identified for the first time.

Using the proposed "Method of collection during the life of the parasite fertilized eggs (in vitro) from the pathogen parasitic zoonosis Fasciola hepatica" from domestic and/or wild animals, in comparison with known methods: sedimentation, flotation, combined; egg collection through the destruction of worms, has the following advantages:

there is no need in definitive hosts of the parasite - infected with fascioliasis domestic (cattle, sheep, goats, camels, horses, etc.) or/and wild (deer, wild boars, mountain goats, elephants, etc.) animals, without which (and preferably in large quantities and with great intensity of invasion!) no researcher can conduct experiments;

known species of Fasciola: Fasciola hepatica (unlike the known methods egg collection: sedimentation, flotation, combined, when an unknown species of Fasciola: Fasciola hepatica L., 1758, F. gigantica Cobbold, 1885, F. jacksoni Cobbold, 1869, F. indica Warma, 1953, or race, strain);

shortens the time and reduces the complexity of investigations for �odd exception: the required separation of Fasciola hepatica eggs from other species of eggs of the genera and species of helminths: strongest: Nematodirus sp., Oesophagostomum sp.; Capillaria sp., trichocephalus: Trichuris ( = Trichocephalus) sp., and oocysts of Eimeria, Cryptosporidium (in contrast to the known methods of collection of eggs of Fasciola: flotation, sedimentation, combined, in the application that need to clear the charges from a large number of other species of helminth eggs and protozoan oocysts);

the necessity of opening ( = destruction) of the pathogen parasitic zoonosis Fasciola hepatica (unlike the method of collection of eggs destruction of helminths);

the need for multiple data collection of eggs of Fasciola (especially because the researcher cannot be determined using known methods egg gatherings: flotation, sedimentation, combined with what type, or race, strain operates!) sedimentation and combined methods of research of a large number of faeces of the definitive hosts of Fasciola spontaneously infected animals (cattle, sheep, goats, camels, horses, reindeer, domesticated elk, etc.) or/and wild (deer, wild boars, mountain goats, bison, bison, etc.) animals, which complicates and lengthens the work of technicians and researchers, especially in light of the intensity of infection and gives more economic benefit of the rational employment of academic staff;

excludes: contamination of egg collection scraps of tissue particles from the destroyed Fasciola hepatica,�nivana and development of secondary bacterial infections in collection of eggs offered by us is obtained.

"The way we collect during the life of the parasite fertilized eggs (in vitro) from the pathogen parasitic zoonosis Fasciola hepatica" from domestic and/or wild animals easily implemented in the laboratory;

the proposed method provides up to 100% of fertilized eggs obtained during the life of the parasite, naturally, for an arbitrary egg-laying live, Mature causative agents of parasitic zoonoses Fasciola hepatica, in an artificial environment and at a temperature characteristic of the ruminant, in tubes with filtered and attentivepersonal bile 1:1 diluted in isotonic solution (0.9%) sodium chloride (solutio Natrii chlorati isotonica) and exposition of tubes at t=38-39°C (unless Fasciola hepatica from cattle) and at t=39-40°C (unless Fasciola hepatica from sheep and/or goats) in a thermostat for 5 hours, and followed by thorough laundering of eggs in saline solution (0.9%) sodium chloride that is essential for research in the laboratory and/or field experiments, the solution of fundamental and applied scientific problems in the field of epidemiology, therapy and prevention of human fascioliasis domestic and/or wild animals.

Sources of information

1. Demidov N. In. The trematode. Fasciolata // In kN.: Helminth infections of ruminants. Under the editorship of prof, E. E. H.. M. Ed. "Spike". - 1968. Pp. 48-79.

2. Carre M. V., Abramov, V. F., Pasechnik V. E. Fast�Ales sheep and the measures against it in Moldavia // In kN.: Improving the productivity of animals in conditions of intensification of production. - Chisinau. - Ed. "Stiinta". - 1982. Pp. 105-110.

3. Kotelnikov G. A. Helminthological research animals and the environment // Directory. - M.: Kolos, 1984. - 208 p.

4. Pasechnik V. E. Cross-contamination trihotsefalami prietary dogs and sheep in Moldova, Byull. All-Union Institute of helminthology. - M. - 1984. - Vol. - 37. - P. 57.

5. Pasechnik V. E. Diss. ... candidate of veterinary science. - M. - VYGIS. - 2000. - P. 124-125.

6. Skryabin, K. I. Method complete helminthological dissection of vertebrates, including man // Ed. 1-St Moscow state University. - M. - 1928. - 36 p.

7. Singh K. R., Srivastava V. K., Prasad Α., Pandey A. P. Pathology due to Fasciola jacksoni in Indian elephants (Elephas indicus) // Indian Journal of Animal Sciences. - 1994. - 64 (8). - P. 802-804.

Method of collecting fertilized eggs (in vitro) from the pathogen Fasciola hepatica in life, consisting in the selection of the bile ducts of the liver infected with Fasciola domestic and/or wild animals only live Mature F. hepatica, placing them in separate vials with filtered and attentivepersonal bile 1:1 diluted in isotonic solution of sodium chloride, and the exposure tubes at t=38-39°C, if F. hepatica from cattle, and at t=39-40°C, if F. hepatica from sheep and/or goats, in the conditions of thermostat for 5 hours, and subsequent careful laundering eggs in isotonic sodium chloride.



 

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2 cl, 2 dwg, 1 ex

FIELD: construction.

SUBSTANCE: static, dynamic or vibration sensing is carried out preliminary at the selected points to the depth from 1 m with respect to the top of the earth fill. At the same time the samples of compacted soil of undisturbed structure are selected in order to determine the moisture and density of skeleton of the specified soil from several drilled wells at points at a distance of not more than 1 metre in plan from sensing points. Laboratory researches of standard compaction with definition of compacting factor depending on the density of soil skeleton, are carried out on the selected samples of soils from the body of compacted fill. Construction of correlation dependence is performed between the specified values of compaction factor and values of the resistance to penetration of standard cone into the soil during sensing, taking into account determinations previously performed in the laboratory followed by evaluation of compaction quality of the earth fill.

EFFECT: improving the accuracy of definition and identifying the areas of non-compacted soil for its subsequent local postcompaction.

3 cl

FIELD: medicine.

SUBSTANCE: method includes the preparation of smear from peripheral blood with preliminary fixation with methyl alcohol, drying, washing with distilled water. After that, the smears are placed in a potassium chloride solution in a ratio of 0.57 g of potassium chloride per 100 ml of distilled water for 20 min and washed with distilled water. Additionally prepared is a mixture of solutions, prepared ex tempore, containing a solution "A" and "B". The solution "A" includes a 50% silver nitrate solution in an amount of 5 g of silver nitrate + 5 ml of distilled water. The solution "B" includes a 2% solution of gelatin on a 1% formic acid solution in an amount of 15.8 ml of distilled water + 0.2 ml of 100% formic acid + 4.0 ml of 10% gelatin. The solutions "A" and "B" are mixed in an amount of 5 ml of each, in darkness, with further submergence of the blood smears for 20 min in darkness in the thermostat at a temperature of 37°C with the further submergence of the smears into distilled water for 2-3 seconds. After that, they are twice subjected to a 8 min exposure in a 5% sodium thiosulphate solution in darkness in the thermostat at a temperature of 37°C. After that, they are washed successively with tap water and distilled water, after-staining is performed in the Romanovskiy dye for 30 min. After that, the smears are washed again with tap water, air-dried, placed in the Canadian balm and covered with a coverslip.

EFFECT: increased quality of smear staining and provision of a possibility to identify and further evaluate parameters of nucleolus organiser regions.

4 tbl, 6 ex

FIELD: biotechnology.

SUBSTANCE: invention is a kit for the laboratory diagnostics of infections caused by urogenital mycoplasmas Mycoplasma hominis and Ureaplasma urealyticum, containing the lyophilized transport medium for the storage of test samples, freeze-dried nutrient medium for detecting Mycoplasma hominis and/or Ureaplasma urealyticum, and strips for identification of Mycoplasma hominis and Ureaplasma urealyticum for the semi-quantitative determining the titer of the pathogen and for determining the sensitivity of Mycoplasma hominis and Ureaplasma urealyticum to antibiotics, which comprises three types of strips, the first of which is designed for identification of Mycoplasma hominis and Ureaplasma urealyticum and for semi-quantitative determining of their titre, and some wells of the first type of strips that are designed for identification and determining the titre of Mycoplasma hominis contain arginine, brilliant blue and clarithromycin, the other part of the wells of this strip that are designed for identification and determining the titre of Ureaplasma urealyticum contain urea and lincomycin, the second type of strips designed for determining the antibiotic susceptibility of Mycoplasma hominis, contains specific reagents for detection of Mycoplasma hominis - arginine, brilliant blue and clarithromycin, and also eight antibiotics - doxycycline, josamycin, midecamycin, ofloxacin, ciprofloxacin, sparfloxacin, moxifloxacin, clindamycin, adsorbed to the wells of the strips in the same concentration, the third type of the strips designed to determine the antibiotic susceptibility of Ureaplasma urealyticum contains specific reagents for detection of Ureaplasma urealyticum - urea and lincomycin, and also eight antibiotics - doxycycline, josamycin, clarithromycin, erythromycin, roxithromycin, azithromycin, ofloxacin and sparfloxacin, adsorbed to the wells of the strips in the same concentration.

EFFECT: use of the kit enables to detect simultaneously Mycoplasma hominis and Ureaplasma urealyticum, to evaluate their titre and to determine the sensitivity to different spectrum of antibiotics, and for Mycoplasma hominis and for Ureaplasma urealyticum different antibiotics are used.

2 dwg, 2 tbl, 3 ex

FIELD: biotechnology.

SUBSTANCE: method of determining sensitivity of fluorescence spectra comprises cultivation of P. aeruginosa strains on nutrient media, stimulating the synthesis of pioverdine, centrifugation and filtration of cultures for obtaining samples, sample irradiation with spontaneous ultraviolet radiation in the wavelength range of 200-300 nm, obtaining the fluorescence spectra, the resistance marker of the strain to antibiotics is the presence in a band of fluorescence of the peak in the wavelength range of 435-445 nm. The sensitivity marker of the strain to antibiotics is the presence in a band of fluorescence of the peak in the wavelength range of 455-470 nm.

EFFECT: invention enables to simplify the technology of optical methods of the study and to optimise treatment of bacterial infections.

2 dwg

FIELD: biotechnologies.

SUBSTANCE: suspension of Tetrahymena pyriformis cells and test substance is put into detector cells of device for automated calculation of live infusoria number for further detection of live cells per minute, and similar dynamics of live cell number changes in time for compared substances allow for concluding on the equal activity of substances. Calculation methods are based on determination of either infusoria incubation time with the test substance, required for death of half of infusoria, or on cell death rate constant, which can be calculated conveniently by inclination tangent of straight line characterising dependence of moving cell number logarithm on time during observed decline in live cell number. Both values depending on membrane activity characterise membrane activity of test substances and can be used for activity comparison.

EFFECT: membrane activity determination in substances with possible process standardisation.

4 cl, 3 dwg, 1 tbl, 3 ex

FIELD: biotechnology.

SUBSTANCE: nutrient medium comprises enzyme-autolyzate of cattle spleen, monosubstituted potassium phosphate, disubstituted potassium phosphate, potassium gluconate, mesoinosite, soluble starch, gelatin, L-cysteine, iron (III) sodium salt, high-grade ink and distilled water in a predetermined ratio of components.

EFFECT: invention enables to improve the accuracy of determining of antibiotic susceptibility of cultures of legionellosis causative agent.

3 tbl 8 ex

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention refers to biotechnology and concerns a method for preparing a pertussis component of complex vaccines. The presented method involves the B. pertussis culture growth on dense Bordet-Gengou and/or CCA nutrient media, the grown colonies selection according to the morphological characteristics, the selected colonies passage, the bacterial mass growth, the culture washout, the extinction reduction of the prepared pertussis suspensions to 10 international optical units, the agglutination reaction to measure agglutinogens 1, 2, 3 and the selection of pertussis suspensions, wherein the agglutinogen content is determined by type-specific serum dilution 1:3200 and more. The detected B. pertussis culture expressing agglutinogens 1, 2, 3 actively after the lyophilisation is used to prepare the pertussis component of complex vaccines.

EFFECT: characterised invention enables preparing the potent pertussis component of complex vaccines.

1 dwg, 5 tbl, 1 ex

FIELD: medicine.

SUBSTANCE: method involves the preliminary sorption of a specific ligand in polystyrene tray wells; the above ligand is specified in: haemoglobin, myoglobin, collagen, fibrinogen, fibronectin, immunoglobulin A; that is followed by adding a microorganism cell suspension into the tray wells with the sorbed ligand, incubating the cell suspension in the tray wells for 15 minutes, sampling a suspension aliquot from the well and adding it to wells of another or the same tray containing 0.5% sodium chloride or 0.2 M sodium phosphate; the bacterial cell adhesion is assessed by measuring a decrease of the optical density of the prepared diluted suspensions at wave length 600 nm as compared to the well references free from ligands.

EFFECT: invention is characterised by a high test rate of the microorganism adhesion to ligands of various nature, high reproducibility, using a minimum amount of the microorganism biomass, with no need for hazardous chemicals to be used.

7 tbl, 7 ex

FIELD: chemistry.

SUBSTANCE: group of inventions relates to biotechnology and can be used for biotesting of toxicity of environmental media. The invention discloses a Vibrio aquamarinus bacterial strain, a method of determining toxicity of samples using said strain and use of the strain as a testing culture for determining chemical toxicity of samples. The method comprises measuring luminescence of the Vibrio aquamarinus VKPM V-11245 strain in the presence of at least one chemical toxicant of a sample, said toxicant being ZnSO4 or CuSO4 or K2Cr2O7 or oil or diesel fuel or bilge water or phenol or a heavy metal. Luminescence of said strain is measured without a chemical toxicant and the measured luminescence levels are compared. Change in luminescence indicates toxicity of the analysed sample.

EFFECT: invention improves reliability of detecting toxic substances in the environment.

9 cl, 4 tbl, 4 ex

FIELD: agriculture.

SUBSTANCE: invention relates to the field of quality control of disinfection. The method comprises disinfection of premises, selection of test objects which are used as a suspension Bacillus subtilis of the strain ATCC PTA-6737 (PB6) sorbed on the carrier - the filter paper with an adhesive surface part. The test objects are placed in the premises of disinfection and added to the nutrient medium. The tubes are heated to 60-80°C and cultured at 37°C for 12-24 hours followed recording the result by recording the growth of bacteria after 12 hours. Clouding of the nutrient medium to form a mucous film is indicative of inadequate quality of disinfection, absence of clouding and formation of a film after 24 hours is indicative of the satisfactory disinfection.

EFFECT: invention enables to simplify the quality control of disinfection of premises.

2 tbl

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention refers to biotechnology and microbiology and represents a method for identifying a cluster of antigens coding staphylococcal proteins that are exotoxins. The present method is implemented by performing a single multiple-primer polymerase chain reaction in two reaction mixtures, first of which contains primers to genes coding such staphylococcal proteins, as thermonuclease, beta-glucosidase in the species S.aureus, S.epidermidis, S.haemolyticus, S.lugdunensis, S.saprophyticus, and the second one - to set1, set2, set3, set4, set5 genes coding the exotoxins. That is followed by a comparative analysis of amplified gene fragments prepared in two sample aliquots and a positive control reference according to the identification table. The present invention also discloses a test system for implementing the above method, which contains DNA recovery components, PCR components, and result analysis components. The PCR components contain a 10-merous buffer solution, pH 8.4, deionised sterile water, one positive control reference, Taq polymerase, mixture of four dNTPs, mixture of primers No.1 containing epi-F, epi-R, aur-F, aur-R, hae-F, hae-R, lug-F, lug-R, sap-F, sap-R in a ratio of 1:1:1:1:1:1:1:1:1:1, and a mixture of primers No. 2 containing set1-F, set1-R, set2-F, set2-R, set3-F, set3-R, set4-F, set4-R, set5-F, set5-R in a ratio of 1:1:1:1:1:1:1:1:1:1.

EFFECT: invention enables recovering the cluster of genes coding the staphylococcal proteins, a molecular weight of which makes 25 to 35 kD consisting of almost 200 amino acid residues and having a tertiary structure high-homologous with some staphylococcus superantigens (enterotoxins, TSST-1 toxins) and with pyrogenous streptococcal exotoxin C.

2 cl, 3 tbl, 1 ex

FIELD: chemistry.

SUBSTANCE: claimed solutions deal with a method of generating a starter culture, the starter culture and a fermentation method with an application of the said starter culture. The claimed method of generating the starter culture includes impact on a parent bacterial strain, which contains, at least, a part of the locus CRISPR, with a bacteriophage to obtain a mixture of bacteria, which contains a bacteriophage-resistant variant strain, containing the modified locus CRISPR, which contains, at least, one additional spacer in the said modified locus CRISPR; independent impact on the same parent bacterial strain with the same bacteriophage to obtain the mixture of bacteria, which contains other bacteriophage-resistant variant strain, containing the modified locus CRISPR, which contains, at least, one additional spacer in the said modified locus CRISPR, different from the additional spacer in the first bacteriophage-resistant variant strain, selection of the said bacteriophage-resistant variant strains from the mixtures of bacteria and their separation.

EFFECT: claimed inventions make it possible to obtain bacteriophage-resistant cultures and can be applied in the food industry in manufacturing fermented products.

29 cl, 23 dwg, 20 tbl, 23 ex

FIELD: biology.

SUBSTANCE: invention is designated for biotesting water and aqueous extract samples. Daphnia are immersed firstly in solution or water for culturing to be tested and then daphnia are transferred into lightproof chamber with outlet hole for culturing and this chamber is immersed into water. Time passing out daphnia from the testing chamber to light is measured and toxicity of analyzed solution is estimated by difference time in daphnia passing out. Method allows carrying out the rapid control of water and aqueous extracts toxicity in laboratory and field conditions.

EFFECT: improved method for biotesting.

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