Effect of Cystamine on Sperm and Antioxidant Parameters of Ram Semen Stored at 4 °C for 50 Hours.

Physical and chemical changes caused by oxidative stress in the spermatozoa membrane can reduce spermatozoa function and even lead to death. Cystamine (NH2-CH2-CH2-SH, ß-mercaptoethylamine) is a natural substance that modulates the endocrine and metabolic status of animals. This substance has antioxidant and anti-apoptotic effects by inducing intracellular cysteine accumulation. Cystamine is used to treat many diseases despite its many side effects. Sheep semen is sensitive to the stressful condition of chilling storage, which restricts semen storage for artificial insemination in commercial herds. The effect of cystamine on spermatogenesis is not yet fully understood. The present study aimed to investigate the effect of cysteamine addition to the sheep sperm extender during cooling storage on semen quality parameters. Sperm samples were collected from six Edilbayevskaya rams (2 and 3 years old, 70-85 kg). The samples were diluted by extender and supplemented with different concentrations of cysteamine (0, 1, 2, 5, and 10 mM) and cooled to 4ºC for 50 h. Motility parameters, membrane integrity, viability, lipid peroxidation, and mitochondrial activity of cooled semen were evaluated at 0, 25, and 50 h of cooling storage. Although cysteamine failed to affect semen quality at start time (0 hrs), extender supplementation with cysteamine improved sperm total motility, progressive motility, and mitochondrial membrane potential during storage periods (P≤0.01). Moreover, using 1 and 2 mM cysteamine functionally and viably improved (P≤0.01) sperm membrane compared to other treatments. Antioxidant potential (AOP), lipid peroxidation (LPO), and total glutathione (tGSH) (except AOP at 50 h) were significantly different after semen storage at 4 °C. Therefore, levels of AOP and tGSH were significantly increased by using cysteamine. Cysteamine supplementation (1 and 2 mM cysteamine) leads to lower levels of LPO (p<0.01) at 0, 25, and 50 h. Therefore, finding and using the best concentrations of cysteamine in a cooling extender could be effective in saving sheep semen against damages of the cooling storage process.

Persistence of arctic-alpine flora during 24,000 years of environmental change in the Polar Urals.

Plants adapted to extreme conditions can be at high risk from climate change; arctic-alpine plants, in particular, could "run out of space" as they are out-competed by expansion of woody vegetation. Mountain regions could potentially provide safe sites for arctic-alpine plants in a warmer climate, but empirical evidence is fragmentary. Here we present a 24,000-year record of species persistence based on sedimentary ancient DNA (sedaDNA) from Lake Bolshoye Shchuchye (Polar Urals). We provide robust evidence of long-term persistence of arctic-alpine plants through large-magnitude climate changes but document a decline in their diversity during a past expansion of woody vegetation. Nevertheless, most of the plants that were present during the last glacial interval, including all of the arctic-alpines, are still found in the region today. This underlines the conservation significance of mountain landscapes via their provision of a range of habitats that confer resilience to climate change, particularly for arctic-alpine taxa.

Effect of the combined action of Quercus cortex extract and probiotic substances on the immunity and productivity of broiler chickens.

AIM: This study was designed to investigate the synergistic effect of the combined action of probiotic bacterial strains (Bifidobacterium adolescentis and Lactobacillus acidophilus) and Quercus cortex extract as biologically active substances in the feed on the immunity and productivity of Gallus gallus domesticus. MATERIALS AND METHODS: For the experiment, 120 7-day-old broiler chickens were selected (4 groups, n=30, 3 replicates with 10 birds in each group). The groups were as follows: The reference group - basic diet (BD); experimental Group I - BD + Q. cortex extract (Q. cortex), 2.5 ml/kg of body weight; experimental Group II - BD + probiotic preparation based on B. adolescentis, 80.0 million colony-forming units (CFU), and L. acidophilus, 1.0 million CFU (dosage in accordance with the recommendations of the manufacturer); and experimental Group III - BD + probiotic + extract of Q. cortex. The following methods of study were used: Chemiluminescence and biochemical and hematological analysis. RESULTS: The results of the experiment showed a slight decrease in the level of leukocytes in Groups II (p≤0.05) and III, and of hemoglobin in Group III (p≤0.05), compared to the reference group. The level of alanine aminotransferase and aspartate aminotransferase in Group II was higher than both the reference group (p≤0.05) and the other groups. Introduction of Q. cortex extract into the diet increased the level of triglycerides (p≤0.05) and urea in the blood serum. The combined use of probiotic preparations and the extract resulted in an increase in the level of iron in the blood serum by 78.1% (p≤0.05) in Group III. An increase in indicators of the antioxidant system (catalase increased in Group I by 27.2% (p≤0.05) and by 3.0-12.7% in other groups; superoxide dismutase increased by 3.0-13.2%) and nonspecific immunity (ß-lysine increased by 8.8-16.0%) was noted. Introduction of the extract and probiotic preparation into the diet contributed to increasing the live weight of chickens at the age of 15 days by 5.9 and 7.4%, respectively (p≤0.05). In experimental Group II, this trend continued, and by the end of the period, the weight of animals exceeded that of their peers in other groups by 0.7-7.0%. Given the high preservation rate of poultry in the II and III Groups, and the low feed consumption per 1 kg of live weight gain (by 3.1-6.7%), the efficiency of growth was higher than in the reference group. CONCLUSION: Thus, the combined use of probiotic strains of bacteria and Q. cortex extract helped to increase the antioxidant activity of the organism and antimicrobial components of blood plasma compared with broiler chickens with similar growth rates but without the supplementation of this combination.

Characteristics of Mel Ibr Melanoma Line Subclone after Treatment with Chicken Embryo Extract.

We continue analysis of the phenotype of human melanoma cell Mel Ibr subclone obtained previously by treatment of the parental cell line by chicken embryo extract. The present study is focused on detection of markers of epithelial-mesenchymal transition that determine enhanced metastatic and invasive potential of malignant tumors of various locations. Analysis of the expression of E-cadherin and vimentin genes in the subclone and parental cells detected activation of epithelial-mesenchymal transition in the subclone. Immunological markers CD90, CD271, and CD95 were present in the parental population, but were not detected on the subclone cells. In contrast to the parental line, cells of the analyzed subclone retain viability in serum-free medium and formed vessel-like structures characteristic of vasculogenic mimicry.

[(HUMAN DIROFILARIASIS: CLINICAL DIAGNOSTIC FEATURES ASSOCIATED WITH DIFFERENT DEVELOPMENTAL STAGES OF THE PATHOGEN)].

To seek medical advice is due to the time when a person has become infected during the infection transmission season (July-August) and the duration of development of the pathogen Dirofilaria (N.) repens in his/her body Clinical manifes- tations occurred in 61% of the patients 6-10 months after infection, as confirmed by the maximum body sizes (125-160 mm) of removed females which have reached puberty. PCR-based diagnosis in conjunction with microscopic studies improves the efficiency of identifying the patients and the species of the pathogens D.repens and D.immitis. The use of these methods for the first time in 2016 could confirm D.immitis infestation in a 14-month-old infant living in the Solnechnogorsk District, Moscow Region.

[(INDUCED MALARIA)].

Induced malaria continues to be one of the most pressing public health problems in malaria-endemic countries. Ma- laria parasites remain viable in stored blood at a temperature of 2-6°C for 3 weeks. The paper presents current problems associated with transfusion-induced malaria. In the USSR and then in the Russian Federation, sporadic cases of induced malaria (Plasniodium vivax, P.malariae, rarely P.falciparum) were notified (230 cases in 1958 to 1990 and only 5 in 1991 to 2016). Current (immunological and molecular) methods for the laboratory diagnosis of malaria do not provide a 100% detection rMfteor its pathogens; therefore, it is necessary to search for highly efficient, rapid, and low-cost diagnostic methods to ensure the biological safety of donation.

[(THE IMPACT OF MIGRATION PROCESSES ON- BIOSAFETY AND HUMAN HEALTH)].

Infectious diseases are one of the serious threats to biosafety in modern society In recent years, there have been con- siderably increased international migrations: tourism and pilgrimage; labor migration flows; migration of refugees and settlers in times of armed conflicts, household migrations because of family ties and traditions. Since the number of Rus- sian tourists and migrants from endemic countries remains high, these contingents should receive special attention in the malaria epidemiological surveillance system. The paper discusses the risks of possible consequences of population migra- tions and prevention measures.

[(EXPERIENCE WITH GEOGRAPHIC INFORRMATION SYSTEMS USED IN EPIDEMIOLOGICAL STUDIES (AS AN EXAMPLE OF MALARIA AND DIROFILARIASIS)].

The effective application of GIS against parasitic diseases requires the patterns of spread of parasitic diseases in certain natural, climatic, and socioeconomic conditions of different regions where there are epidemically effective carriers. The epidemiological parameters defining a high risk of an epidemiological process after P.vivax importation into Russia were calculated using the developed HealthMapper controlled module with a database. Analysis of the average long-term air temperatures in the administrative territories of Russia over 78 years, namely July, the warmest month of the year when there are the largest numbers of all types of mosquito vectors and high levels of microfilaria in the peripheral blood of in- fected dogs (definitive hosts), has shown that the northern border of the maximum possible area of dirofilariasis in Russia is most fully described by the +14C July isotherm.

[THE CLINICAL AND EPIDEMIOLOGICAL CHARACTERISTICS OF MALARIA CONCURRENT WITH OTHER INFECTIONS AND INVASIONS].

The present review considers malaria infection concurrent with different species of helminths, bacterial and viral infections, as well as mixed malaria pathogens in the subtropical and tropical countries of the world, causing the clinical picture and epidemiological situation to be different. Malaria co-infections with different pathogenic micro-organisms, such as HIV, tuberculosis, viral hepatitides, and others, affect almost one third of the planet's population. It is known that people who are at risk for malaria may be also at risk for other parasitic and infectious diseases, most commonly helminthisms.

[MALARIA IMPORTATION BY RUSSIA'S CITIZENS AND FOREIGNERS, INTO THE CITIES AND TOWNS OF THE RUSSIAN FEDERATION].

A total of 436 malaria cases, including 12 from the CIS countries and 424 from far foreign countries (of Africa and Central and South-East Asia), were imported into the Russian Federation in 2010-2014. Most (96.6%) cases were notified in the urban areas of 52 administrative subjects of Russia. The largest number of the imported cases were seasonal workers (39.2%), tourists (31.3%), students and foreign postgraduate students (19.5%), and ship or aircraft crews (10%). During a short malaria transmission season (June to August), there were 150 cases of different types, out of them there were only 63 cases of tertian malaria (its pathogen is Plasmodium vivax, to which malaria mosquitoes of Russia's fauna are susceptible). The relatively small number of infection sources in the short transmission (June to August) season of malaria, its importation into low-susceptibility large towns, and a small proportion of imported vivax malaria cases substantially reduce the risk of malaria in the highrisk areas of the country.

MOSQUITOES (DIPTERA, PSYCHODIDAE, PHLEBOTOMINAE) IN THE CRIMEA: SPECIES COMPOSITION, SPREAD, ENVIRONMENTAL FEATURES.

Crimea was known to be a mosquito-borne fever (pappataci fever) endemic area in the past. There is information on sporadic cases of visceral leishmaniasis (VL), the vectors for which are also mosquitoes. This disease was registered in Crimea in both the 20h century and the early 21st century. The data available in the literature on the mosquitoes in Crimea, which were published mainly in the first half of the 20th century, have been analyzed. Based on the classification of these insects, which has been proposed by M.M. Artemyev, the authors have established that there are 8 mosquito species: P.papatasi, P.alexandri, P.similis, P.neglectus, P.perfiliewi, P.balcanicus, P.longiductus, and S.dentata in Crimea. There is evidence on the landscape and biotopic association of individual species and on their environmental features.

[The improvement of methodical approaches of hygienic regulation of pesticides in water bodies].

The article highlights the methodical features of hygienic regulation of pesticide preparation in water bodies, concretizes and supplements (specified) number of positions of the acting current methodical document on hygienic evaluation of new pesticides. From the position of the comprehensive hygienic standardization there are marked principles of the study of MPC for pesticides in water bodies, there is substantiated the expediency of expanding the arsenal of methods for determination of threshold concentrations of pesticides on the impact on organoleptic properties of water and sanitary regime of reservoirs, shows the role and importance of hydrolysis processes of active ingredients ofpesticides in water, which should be considered in the assessment of the impact of drugs on the processes of self-purification of water bodies.

Changes in the Morphological and Immunological Characteristics of Mel Ibr Melanoma Cells in Response to Chicken Embryo Extract.

Exposure of Mel Ibr human melanoma cells to chicken embryo extract resulted in the appearance of a subclone with morphological and growth characteristics similar to those of embryonic stem cells. The subclone differed from the parental line cells by a sharply reduced percentage of HLA-DR(+) and CD54(+) cells, a significantly elevated percentage of CD63(+) cells, and appearance of CD133(+) and Oct-4A(+) cells. Hence, the subclone cells were characterized by the same features as stem tumor cells and could be responsible for further progress of tumor growth.

[Human dirofilariasis: clinical and diagnostic signs and diagnostic methods].

The clinical and diagnostic signs caused by the tissue location and migration of adult Dirofilaria in the human body determine the use of different methods for the diagnosis of dirofilariasis. During their investigations, the authors modified polymerase chain reaction (PCR): they chose and synthesized primers and selected amplification regimens for them and obtained agarose gel bands that corresponded to PCR fragment length nucleotide sequences that were equal to 245 bp for D. (N.) repens and 656 bp for D. immitis. There was 100% agreement in the results of PCR and microscopic examination of sera from 32 dogs and 1 female patient with low parasitemia and in the blood nucleotide sequence characteristic of D. repens.

[Results of studies of human dirofilariasis in Russia].

Human dirofilariasis is a pressing health problem in Russia. By 2014, there have been as many as 850 Dirofilaria repens-infested people living in 42 subjects of the Russian Federation. One of the favorable factors for circulation and spread of invasion is a temperature of above +20-24 degrees C; when the latter is maintained during at least 20 days there may be 1.-1.5 circulations of invasion in the carrier and a 2.8-fold increase in transmission intensity. The border ofa dirofilariasis area with a temperate climate is southward to 58 degrees N in European Russia and West Siberia and southward to 50 degrees N in the Far East. The conditions in the human body have been found to be more favorable for the development of Dirofilaria than considered before and allow the helminth to achieve sexual maturity and to propagate. If man has microfilaremia, he may be a source of invasion. It is necessary to examine venous blood by the enrichment method and, if possible, polymerase chain reaction and enzyme immunoassay, which make it possible to establish a diagnosis in occult invasion and to identify a pathogen species.

[Use of geographic information systems in public health].

To enhance the efficiency of epidemiological surveillance by the countrywide use of current information telecommunication technologies, diagnostic systems based on monitoring is one of the tasks of the Russian Federal Service for Consumer Rights Protection and Human Health Control in the control and prevention of parasitic diseases. The epidemiological surveillance system for parasitosis encompasses not only the monitoring and assessment of the situation, but also necessary measures if epidemic complications occur. Geographic information systems (GIS) may be successfully used for this purpose. GIS-based interactive health atlases have been created and put on the Internet and researches made.

[The current status of the resistance of malaria pathogens to antimalarials].

The review presents the results of trials of the clinical efficacy of a test antimalarial drug for each malarial parasite species, which were published in 2000-2013 and supplemented by the data of in vitro studies or investigations using the molecular markers of resistance. There are data on the resistance of each medicament since many of the drugs are used in combination with artermisinin derivatives.

[Assessment of the risk of expansion of the habitats of the mosquitoes AEDES aegypti L. and Aedes albopictus Skus. in Russia].

Based on climatic characteristics, the authors show that an invasive Aedes albopictus species is likely to emerge in new areas of the Russian Federation.

[Epidemiological alertness as a component of the malaria surveillance system].

Malaria vigilance is a continuation of the functions of the malaria surveillance system in the countries that have achieved malaria elimination on their territories. The main function of the malaria vigilance during the post-elimination period is a continuous monitoring and evaluation and implementation of necessary prevention measures against the possible re-establishment of local malaria transmission due to the importation of malaria from its endemic countries by residents and/or migrants. Various epidemiological aspects of malaria vigilance, such as standard identification of a malaria case during the post-elimination period; notification procedures and reporting systems; training of personnel; free access to malaria diagnosis and treatment; mandatory laboratory confirmation of malaria diagnosis; and prompt efficacious treatment, are discussed. Other aspects include epidemiological classification of cases, seasonal pattern of imported cases, its territorial confinement, time-lag between the detection of a case, its diagnosis and treatment initiation, evaluation of the efficacy of individual chemoprophylaxis and treatment.