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¹ 6, 2015



Authors: 1,2SukharenkoH.V. - ; 3Nedzvetsky V.S. – Doctor of Biology;1Maksimov V.I. - Doctor of Biology
1Moscow State Academy of Veterinary Medicine and Biotechnology – MVA named after Skryabin K.I., Moscow (e-mail: rector@mgavm.ru).
2Kerch State Maritime Technological University (Kerch).
3Oles Gonchar Dnepropetrovsk National University (Dnepropetrovsk).

Introduction: During the recent decades in physiological justification of xenobiotic toxicology the terrestrial animals were used as a main object in physiological justification of xenobiotic toxicology while aquatic organisms remained neglected while the mollusks which due to their attached lifestyle, filtration-based feeding type, widespread, massive scale, availability of sampling are a comfortable object for biological survey. The aim of the research was to determine formation mechanisms of response to petroleum products in hepatopancreas and gills of zebra mussel mature species and to evaluate validity of oxidative stress parameters at biological threat of industrial contaminants of aquatic environment. Methods and materials: the experiments involved adult mature Dreyssena polymorpha river bivalve mollusk living in water reservoirs challenged with xenobiotics. During 28 days the water with the experimental mollusks was contaminated with oil residuals in constant concentration of 50 mg\l. In aquariums 1\4 of water was changed twice a week. The biological material was produced by homogenization of hepatopancreas and gills. The investigation results: a reliable (P<0.05) increase of ultimate products from lipid peroxidation and enzymatic activity of antioxidant defense (catalase, enzymes superoxide dismutase (SOD), glutathione reductase, glutathione S-transferase, glutathione peroxidase) in a response to intoxication were determined. Differences oxidative stress parameters in hepatopancreas and gills of river bivalve experimental and control mollusks. A comparative analysis of antioxidant system enzymatic activity in experimental and control mollusks showed that an increase in activity of enzymes providing lower level of oxygen response compounds: catalase, enzymes superoxide dismutase, glutathione reductase in tissues of bivalve mollusks living in the environment contaminated with oil residuals are accompanied by reduced activity of glutathione S-transferase – an enzyme catalyzing utilization of inflow exogenous aromatic compounds. Conclusion: The obtained results evidence the oxidation-reduction imbalance adequately reflects the processes of biotransformation in adult mature bivalve mollusks and a higher sensitivity of hepatopacreas cells to the effect of organic xenobiotics than that of the gills. The activity of antioxidant enzymes and a level of ultimate products from lipid peroxidation - one of the oxidative stress parameters – in tissues of adult mature species of river zebra mussle indicate that the peroxide processes peculiar to oxidative stress are sharply induced in the environment polluted with organic contaminants. These parameters show a high efficacy for bioindication and biomonitoring of mollusks performance at water bioresources pollution with organic pollutants. 

Keywords: Keywords:oil residuals, petroleum refining products, pollutants, oxidative stress, bivalve mollusks.
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Authors: Makaeva A.R. – postgraduate.
Federal centre of toxicological, radiation and biological safety, Kazan (e-mail: vnivi@mail.ru).
Introduction: at constant pollution of a water body it is necessary to evaluate not only the level of toxic substances in water but also to evaluate its level in aquatic organisms. Therefore, we studied copper toxic level in water using Lymnaea stagnalis pond snail and Bebistes reticulatus guppy. These fish are useful for water toxicology because of their small size and easy maintenance. Methods and materials: for acute experiments Lymnaea stagnalis pond snails of the same size and 1-2-days-old guppies for were used. The fish survival rate and behavior was evaluated. The experiments lasted for 96 hours. Copper sulfate diluted in settled tap water was used as a source of copper ions. Results: a higher concentration of copper in water was found to have a depressing effect on aquatic organisms. The pond snails trying to avoid a toxic environment stayed at the water surface and their heads increases in size because of edema. Fish swam at the water surface staying motionless for some time and weakly responding to stimuli. Copper LC50 was 0.278 and 0.549 mg/dm3 for snails and fish accordingly. Chronic experiment was carried out on pond snails at copper concentration of 0.1 mg/dm3. Copper caused increased secretion of mucus snails, reduced food and oxygen consumption. Chronic experiments on guppies were performed at copper concentration of 0.28 mg/dm3. Weight of fish in copper group was lower than that of the controls. Conclusion: Copper reduced a backup resources of fish body. 
Keywords: Keywords: water, copper, toxicity, pond snail, fishes.
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  Infectious pathology


Authors: Pokrovskaya E.S. - Candidate of Biological Sciences; Shuralev E.A. - Candidate of Veterinary Sciences; Mukminov M.N. - Doctor of Biology, professor; Elizarova I.A. - research assistant; Faizov T.Kh. – Doctor of Veterinary Medicine, professor.
Federal Center for Toxicological, Radiation and Biological Safety, Kazan (e-mail: vnivi@mail.ru).

Introduction: caprine arthritis encephalitis is a highly contagious slowly flowing viral infection of goats which spread on all continents. It is not sufficient to make a decision on basis of virus isolation on cell culture, and the diagnosis through the detection of specific antibodies may become more effective. Currently available with open access bioinformatic databases allow analysing protein amino acid sequences that would greatly reduce the costs of labor-intensive experimental surveys. The aim of this work is to study Caprine arthritis encephalitis virus antigens with immunogenic epitopes that have the greatest similarity among different strains of the pathogen. Methods and materials: using the methods of bioinformatics, database of proteins (NCBI PubMed) and immunogenic epitopes (IEDB), the antigenic determinants of Caprine arthritis encephalitis virus proteins were investigated. Results: two specific groups of the virus which are 1) protein epitopes encoded by env gen, and 2) protein epitopes encoded by gag gen were discovered. Using BLAST analysis of amino acid sequences of the antigens it was found that the majority of epitopes of studied proteins have significant overlaps with Caprine arthritis encephalitis virus and group specific small ruminant lentiviruses. In terms of a score and amount of alignments the epitopes were divided into three conventional groups with high, medium and low degree of matching. Conclusion: It has been established that the greatest diagnostic value is provided by two epitopes of transmembrane protein TM gp38 and one epitopes of capsid protein p25. That is expressed in high probability, sufficient degree and maximum number of matches of the amino acid sequences with proteins of both most strains of Caprine arthritis encephalitis virus, and the group specific small ruminant lentiviruses.

Keywords: Keywords: small ruminant lentiviruses, CAEV, glycoproteins, immunogenic epitope, BLAST analysis.
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Authors: À.G.Spiridonov – Candidate of Biological sciences, senior staff scientist; D.D. Nasertdinov – research assistant; H.N. Makaev – Doctor of Veterinary Medicine, professor; G.N. Spiridonov – Doctor of Biology, head of laboratory.
Address: 2Federal Centre for Toxicological, Radiation and Biological Safety, Kazan (e-mail: vnivi@mail.ru).
Cl.perfringens and their associations with other enterobacteria particularly E.coli are of growing significance in etiology of gastrointestinal diseases in calves. The aim of the investigation was to develop an associated vaccine from hyperimmune serum to prevent and treat anaerobic enterotoxemia and Escherichiosis diarrhea and to evaluate its efficacy in laboratory and fields studies. Methods and materials: for vaccine development and hyperimmune serum inactivated antigens to Cl. perfringens bacteria A, C, D serotypes and E.coli synthesizing adhesive K99, A20 antigens were used. In the infected farms the biopreparations were evaluated on spot. Vaccine was twice subcutaneously injected into down-calving cows in dosage of 10cm2 50-60 days before calving with interval of 14 days. The calves born to vaccinated cows were vaccinated at the age of 18-20 days old in dosage of 3cm3 twice with interval of 14 days. The hyperimmune serum was intramuscularly introduced into newborn calves in 2 cm3/kg dose for prevention and 2.5 cm 3/kg dose for treatment. The investigation results: an associated vaccine was determined to have high antigenic and immunogenic activity. Using the vaccine for prevention purpose reduces sickness rate for 4.6 times and increases survival rate for 16.6% in permanently infected farms. Hyperimmune serum has an expressed treatment and prevention properties. Using the serum in treatment allowed to cure 93.0% sick calves that if higher for 13.6% than in the control group.  Conclusion: an associated vaccine and hyperimmune treatment and prevention serum against anaerobic enterotoxemia and E.coli-borne bovine diarrhea were developed and evaluated. Positive results of evaluation allow their recommendation for veterinary use in the Russian Federation. 

Keywords: Keywords: calves, anaerobic enterotoxemia, Escherichiosis diarrhea, vaccine, hyperimmune serum treatment, prevention

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Authors: 1Baratov M.O. - Candidate of Veterinary Sciences; 2Ahmedov M.M. - Doctor of Veterinary Medicine, professor; 2Sakidibirov O.P. - Candidate of Veterinary Sciences.
1Caspian Zonal Research veterinary institute (e-mail: alama500@rambler.ru).
2M.M.Dzhambulatov Dagestan state agrarian university  

Introduction: A mixed antigen from M.scrofulaceum #12-C and M.intracellulare #13-H are widely used in veterinary practice to discriminate cattle non-specific responses to tuberculine. To improve its discriminatory functions the antigen content was enlarged with sensitines from N.asteroides VKM Ac1077 and R.bronchialis IMV Ac737 strains. The aim of the work was to improve specificity and efficiency of mycobacteria complex antigen by including corynebacterium sensitin. Materials and methods: To produce tuberculosis allergen a culture of Corinebacterium xerosis N1911 was used.  The threshold sensitivity for allergen was determined using guinea pigs inoculated with corynebacteria (the protein was used in 6 concentrations). The determined threshold level of 0.0003mg protein was further used to develop an allergen , i.e. 1350 units of activity of each component (atypical mycobacteria, nocardia, rodoccoci) was tested in 0.2 mg and 0.0003 mg corynebacteria proteins. Results: The discriminatory ability of allergen was determined using guinea pigs at 27 days after inoculation with mycobacteria, corynebacteria, nocardia, rodoccoci. The animals from the experimentally challenged group (C.xerosis, M.scrofulaceum, Nasteroides and R.Bronchialis) responded to the allergen under the trial in more expressed way than to the complex allergen. In guinea pigs inoculated with M.BCG response to complex allergen was slightly higher. A significant discriminatory ability of the tested allergen was also detected during examination of 136 cattle herds from tuberculosis-free farm where previously nonspecific responses were regularly registered. Conclusion: sensitin from corynebacteria as component of complex allergen was experimentally proved to enhance efficiency of simultaneous testing to discriminate nonspecific responses caused by corynebacteria.  

Keywords: keywords: PPD-tuberculin for mammals, associated allergen for mycobacteria, sensitization, Corynebacterium, Rodococcus, monoallergen, atypical.
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Authors: 1Gulyukin A.M. – Candidate of Biology Sciences; 1Shabejkin A.A.- Candidate of Veterinary Sciences; 1Zaykova O.N. – researcher; 1Polyakova I.V. - Candidate of Biology Sciences; 1Zaberezhny A.D. - Doctor of Biology, professor;2Khismatullina N.A. - Dîctor of Biology, professor; 2 Samerkhanov I.I. - research assistant.
1Y.R.Kovalenko All-Russian Institute of Experimental Veterinary Medicine (VIEV), Moscow (e-mail: plych@mail.ru).
2Federal Centre for Toxicological, Radiation and Biological Safety, Kazan (e-mail: vnivi@mail.ru).
Introduction: the article describes the epizootic situation on rabies in the Republic of Tatarstan. Methods and materials: the methods of molecular epizootology the species and number content of the animals with the laboratory-based confirmed infection and phylogenetic analyses of the rabies agents isolated in various regions of the republic of Tatarstan were used. As the results of rabies epizootical monitoring were analyzed the sickness rate, coefficient of focality, and infection rate were calculated. The parameters were rather high, thus confirming the endemic situation in Tatarstan and allowing the local veterinary service performing complex analysis and risk forecast. The G gene fragment of rabies field isolates from Tatarstan was sequenced and compared with similar fragment from reference strains. The investigation results: a comparative analysis showed no genomic differences in nucleotide and amino acid sequences between the field isolates. The difference in nucleotide sequence between field and reference strains was as high as 8%, while in their amino acid sequences no difference was found. This suggests high selective pressure on the amino acid structure of the analyzed fragments. Since live vaccines are widely used to control rabies in wild animals, it is necessary to check potential reversion of vaccine strains to the virulent one. Conclusion: in the enzootic area of Tatarstan rabies was determined to be eventual and natural focal infection. Nozoologic maps were developed in a computed geo-informational system and the vector of epidemic process in the Republic of Tatarstan within 2013-2014 was determined.

Keywords: Keywords: rabies, Republic Tatarstan, epizootic monitoring, epizootic process, sequencing, geo-information system, phylogenetic analysis, immune pressure, molecular epizootology.
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  Non-transmittable diseases


Authors: Buchenova A.V. – postgraduate; Usha B.V. – Doctor of Veterinary Sciences, professor, RAS academician.
Moscow State University of Food Production, Moscow (e-mail: anuta-bychenova@mail.ru).

The aim of the research was theoretical and experimental validation of new methods for the differential diagnosis of nephrotoxic ARF (acute renal failure) in dogs, and using a number of techniques for effective treatment. Methods and materials: using the traditional diagnostic methods (clinical examination, biopsy, blood and urine tests, ultrasound) 46 dogs of various breeds having acute renal failure were initially selected for further study. Blood for laboratory analyses (number of leucocytes, erythrocytes, hemoglobin) was taken from forearm subcutaneous vein. Abdominal organs and kidneys were examined using ultrasound scanner. Results: in some dogs etiology of nephrotoxic ARF was confirmed by differential diagnosis based on direct determination of nephrotoxicants in blood and urine using the "Abraxis OP / C"™ test system for in vitro inhibition of acetylcholinesterase. This method allow detecting OPS (organophosphorus compounds) and carbamates. Endogenous acetylcholinesterase was inactivated during the implementation of this technique. Immune-microarray technology showed that antigens specific to nephrotoxicants responded to monoclonal antibodies immobilized on the biochips, and the final results were registered using automatic chemiluminometer Randox™. This method allowed both qualitative and quantitative detection of nephrotoxicants having medical origin (antibiotics, sulfonamides, antiparasitic drugs). During the study of dogs with ARF, nanogram amounts of neomycin, cefoperazone (medotsef) and diazinon were found out, and intoxication by these substances could occur either due to treatment of different kinds of diseases or to fodders containing residual amounts of these substances. Conclusion: Basing on the obtained data and confirmation of the etiology, the dogs affected with ARF were treated using a comprehensive system including a consistent application of hemodialysis and drugs recovering nitrogen and amino-acid balance. The treatment efficiency was confirmed by the normalization of biochemical and physiological parameters.

Keywords: Keywords: dogs, nephrotoxic, acute renal failure, inhibition, acetylcholinesterase, differential diagnosis, treatment, hemodialysis, nitrogen and amino acid balance.
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Authors: Zhitlova E.A.- postgraduate; Shakirova F.V.- Doctor of Veterinary Sciences; Achtyamov I.F.- Doctor of Medical Sciences, professor.
Kazan State Academy of Veterinary Medicine, Kazan(E-mail: largo-kazan@mail.ru).Kazan State Medical University, Kazan(E-mail: yalto60@mail.ru).

Introduction: the problem of compensation of bone defects is one of the important problems in veterinary orthopedics. The issue can be solved by using various implantable materials compatible with the composition and properties of native bone tissues. The aim of the research was to study the effect of “Inrok” drug based on lanthanide and calcium ions after it is injected into a bone with experimentally induced defect. Methods and materials: using computed tomography, density parameters of the bone regenerate in the area of the experimentally made local defect of the tibial cortical layer were evaluated. The experimental researches were carried at the beardless rabbits. Animals were divided on two groups: the comparison and experimental. First animal group were double inoculate with the “Inrok” preparation in distress of the cortical plate. For the local change control on the injury zone researches were conducted on the computer tomography Brilliance 64(Philips). A cut processing of reparative zone was conducted in the multiplanaris reconstruction mode in sagittalis plate. Results: the average value of the density in the first 10 days after surgery in rabbits of the test group (where the drug "Inrok" was injected into the experimentally made bone defect) is 1.5-2 times higher than that of the rabbits from the compared group. Conclusion: further terms of the experiment were characterized by a continuing trend towards increasing the density of regenerate in both groups. 

Keywords: Keywords: rabbit, reparative bone regeneration, X-ray computed tomography.
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Authors: Mezhenskiy A.A. – Candidate of Veterinary Sciences.
State Scientific and Research Institute of Laboratory Diagnostics, Veterinary and Sanitary Expertise (e-mail: mezhaavet@gmail.com).

Introduction: the article provides the investigation results of etiological structure of equine uveitis in Ukraine. Methods and materials: at ophthalmologic examination of 615 horses uveitis of various course was diagnosed in 87 animals. Blood serum taken from the sick animals was tested for leptospirosis, brucellosis, viral arteritis, influenza and eqiune herpes virus of I and IV type, also, orthopedic and dental examinations were carried out. Investigation results: equine uveitis was determined to be a polyetiological disease having the same type of clinical signs and changes in the eye. In Ukraine leptospira-borne uveitis (50.6%), idiopathic uveitis (29.9%) and uveitis of non-contagious etiology, emerging after physical activity (5.7%) and closed mechanical trauma of eyeball (4.6%) were the most spread. Leptospira Grippotyphosa group prevail in uveitis cases of this etiological structure – its serological prevalence was 79.5%. In positively responding horses with acute Leptospira uveitis single responses for microagglutination test prevailed (75%), whereas at subacute and chronic uveitis polyreactions were frequently recorded – 83.3 and 59.1%. Conclusion: etiological detection of equine uveitis should be comprehensive and include a general clinical, ophthalmic, orthopedic and dental examination as well as laboratory studies of biological material from sick horses for the most common infectious diseases, primarily for leptospirosis. The results of Leptospira etiological range should be considered at developing prevention measures against uveitis in horse breeding.

Keywords: KEYWORDS: horse, eye disease, uveitis, etiology of uveitis, leptospirosis, leptospiral uveitis.
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Authors: Mosseva A.I. – postgraduate.
Nizhny Novgorod State Agricultural Academy, Nizhny Novgorod, Russia, Gagarin Avenue 97, 603137, e-mail: anatomifarmitox@mail.ru.

Introduction: Two experiments were carried out on 20-30-days old calves in a vivarium All-Russian Research Institute of Physiology, Biochemistry and Nutrition of Animals, Borovsk, Kaluga region, and a farm "Tsentralnoe" in Nizhny Novgorod region. The aim of the research was to study the effect of mixture of nucleates and interleukin-2 (Roncoleukin) drugs on calve leukopoiesis during the lactation period and on the formation of calves non-specific resistance comparing to that of the thimogen drug. Methods and materials: the level of blood immunoglobulins and morphological parameters in calves parenterally introduced nucleic acids and interleukin-2, as well as Òhymogen drugs was studied. Blood samples were taken from the jugular vein at 10 days after the drugs were administerd and in the second experiment - at 30 days. Results: using nucleic acid-base drugs increased the level of white blood cells (+ 7.7%), protein (+ 23.6%) and immunoglobulins (+ 24.1%) in the calve blood and was similar to the effect of thymogen. Using IL-2 resulted in a significant increase of leucocytes level in calve blood (+ 31.2%), and gamma-globulins (+ 57.6%) comparing to thimogen (21.5% and 33.8% correspondingly) but both drugs promoted the formation of non-specific resistance, that was demostrated by a higher blood phagocytic and bactericidal activity parameters. An increased non-specific resistance parameters were accompanied by a higher live weight during the milk-feeding period in newly born calves. Conclusion: The results of the study allow clarifying some aspects of the formation of calves immune system and non-specific resistance during the milk-feeding period.

Keywords: Keywords: calves, immunoglobulins, nonspecific resistance.
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Authors: Shukanov R.A., applicant
K.I.Scriabin Moscow State Academy of Veterinary Medicine and Biotechnology 
(e-mail: shukanovr@rambler.ru).
The aim of the work was to study a correlation between a condition of in boar metabolic, natural resistance, performance parameters and the level of their adaptation in the Volga biogeochemical area of Chuvashia using kizelgur, «Suvar», «Polistem». Materials and methods: II series of researches using 60 weaned boars-analogs with 15 animals in each group were performed. In two groups the boars at the age from 60 days to 300-days old (duration of experiments) were fed the basic diet (BD) with the main parameters balanced according to the norms for diets and feeding recommended by the Russian Academy of Agrarian Sciences. During the research the animals from the experiment groups were fed both BD kizelgur natural zeolite at the rate of 1.25 g/kg of body weight (b.w.) and «Suvar» biogenous substance in a dose of 25-50 mg/kg of b.w. for 20 days with 10-day intervals until the animals were 240-days old (I series) or intramuscularly were injected «Polistem» biological product twice at the age of 60-and 240-daysold at the rate of 0.1 and 0.03 mg/kg of b.w. accordingly (II series). The correlation of metabolic, immune and growth parameters in experimental pigs was analyzed according to the standard techniques used in Biology.The investigation results showed that under the experimental conditions the number of significant relations (r=0.70) among the studied metabolic, natural immunity and growth parameters of the experimental boars varied, and in the second group animals the range of fluctuations was narrower. Thus, inthe control animals the correlation relations changed from 3±0.12 to 7±0.40 (I series) and from 4±0.17 to 9±0.41 (the II series), and in their experimental counterparts – from 5±0.09 to 6±0.24 and from 4±0.11 to 7±0.20 accordingly. Moreover, the metabolic, immunological and growth processes showed better physiological parameters in pigs fed diet enriched with both zeolite kizelgur and «Suvar» bioactive additive, while the associated using of both kizelgur and «Polistem» biological product had less effect on optimization of physiological parameters.Conclusion: The different levels of adaption revealed in the control and experimental animals areaffected by the tested biogenous compounds. Therefore, using kizelgur natural zeolite and «Suvar» bioactive agent in the main diet promotes a more expressed eurybiontic features of organism than an associated using zeolite kizelgur and «Poliståm» biological product regarding the biogeochemical specificity of Volga region of Chuvashia.

Keywords: KEYWORDS: boar, kizelgur, «Suvar», «Polistem», blood, correlation.
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Authors: Vasilev R.O. – postgraduate.
Address: Izhevsk State Agricultural Academy, Izhevsk (e-mail: veterenar4ik@mail.ru).
The aim of the research was to study comparative efficiency of radioprotective features of "Monklavit–1" and potassium iodide iodine-containing drugs at a single external gamma irradiation of guinea pigs at half-lethal dose. Methods and materials: "Monklavit–1" and potassium iodide containing 20g of iodine elements was introduced twice introgastrically  through a probe. The experiments were carried out using male guinea pigs grown in biogeochemical province lacking iodine in soil, water, and fodders. During the experiment clinical, haematological, pathological and immunological methods were used. Results: during the experiment an acute irradiation sickness of moderate severity was experimentally restored. A positive radioprotective effect of iodine-containing drugs at intragastric administration to guinea pigs a day before the exposure and three days after: animal sickness rate reduced and velocity of peripheral blood parameters improved, thyroid gland functions stabilized at first days after exposure to irradiation with consequent increase of its activity by the 26th day. At 26 days a number of leukocytes, erythrocytes and platelets was lower in pigs receiving "Monklavit–1» and potassium iodide than that of the intact by 25%, 12%, 33% and 31% 17%, 49% accordingly. Conclusion: the experiments showed that radioprotective properties of "Monklavit–1" were more that of potassium iodide, but these changes are not reliable. By the moment of exposure to irradiation to some extent the state of thyroid gland as a leading organ of endocrine system can determine the level of nonspecific resistance.

Keywords: Keywords: radiation sickness, gamma irradiation, Monklavit¬1, potassium iodide, thyroid hormones, guinea pigs.
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