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Medicinal leeches (Hirudo verbana Carena, 1820) used in veterinary medicine, human medicine and pharmacology, are currently bred artificially at bio-factories. The technology of leech culture as a component of aquaculture biotechnics is based on the method of accelerated growth of leeches due to intensive feeding, which allows you to get high-quality products in a fairly short time (8-10 months). Nevertheless, the problem of assessing the physiological status of raised individuals upon artificial reproduction remains not fully addressed. The findings we present herein for the first time allow us to suggest that the age-specific amino acid composition of tissues can be used to assess the physiological state and well-being of medicinal leeches in nature and in aquaculture. The aim of the work is to study the age-dependent content of the free amino acids in the tissues of medicinal leeches artificially bred in aquaculture. Studies were conducted in 2012 on the leech (Hirudo verbana Carena, 1820) individuals without signs of diseases which were purchased at the biofactory International Center for Medicinal Leeches (Moscow Province, p. Udelnaya). Leeches were kept in the laboratory in glass vessels with dechlorinated water at 20-22 oC and fed once a month with fresh bovine blood from healthy animals. The experimental specimens of H. verbana was: 5 days (newborn filaments, 0+), 1, 3, 5, 7 and 9 months old, with an average body weight of 0.029, 0.09, 0.25, 0.61, 1.33 and 1.81 g, respectively. Newborn individuals served as a control. The concentration of free amino acids (AA) in the skin and muscle tissue was determined on the analyzer AAA-339M (Mikrotechna, Czech Republic). A total of 105 leech specimens were used, 30 bioassays were prepared, and 660 element determinations were performed. The amino acid composition of the tissues of medicinal leeches H. verbana was represented by 23 AA and their derivatives. The dominant AA with an antioxidant orientation, regardless of age, were glutamine and glutamic acid, alanine, valine, leucine, and glycine, the total content of which was 65 % in filaments, and 58 % in 9-month — old individuals of the total fund AA. In the tissues of medicinal leeches, as well as in warm-blooded animals, the full composition of essential amino acids (EAA) was revealed: threonine, valine, lysine, leucine, isoleucine, histidine, arginine, methionine, phenylalanine, tryptophan. With age, the total concentrations of free amino acids decreased (r = -0.98 at p = 0.000), due to a sharp decrease in the content of arginine (24.0-fold), proline (13.4- fold), isoleucine (12.2- fold), glycine (8.5- fold), lysine (6.8- fold), histidine (6.6-fold), leucine (5.2-fold), ornithine (4.0- fold), glutamic acid and glutamine (3.8-fold), and alanine (2.9-fold). At the age of 5, 7 and 9 months, leeches had a trace content of secondary metabolites: taurine, citrulline and tryptophan (p < 0.001). However, the amino acid balance of nitrogen and protein metabolism was not disturbed, since the ratio of essential / non-essential AА did not change significantly and was 0.60 in newborn filaments, 0.73 — in 9-month-old leeches. It was found that the growth and development of H. verbana was accompanied by a significant decrease in the indicator of maturity (glycine/alanine) from 0.75 (newborn filaments) to 0.25 (9 months). The information available in the literature and the data of our research allow us to put forward the concept of using functional amino acids as biomarkers in the development of the scientific bases of technologies for the industrial breeding of these amphibionts.

Objective.To describe the difficulties of diagnosis of Waldenström macroglobulinemia in a patient with atypical clinical picture.Materials and methods.Patient K., 57 years old, came in outpatient department with complaints of unilateral increase in submandibular lymph nodes on the right, discomfort in the nasopharynx, cough without sputum, increased fatigue. During the examination she was consulted by: an infectious disease specialist, otolaryngologist, surgeon, dentist, phthisiologist, hematologist and oncologist consulted her. To confirm the diagnosis conducted diagnostic activities: dynamic assessment of laboratory parameters, examination program cancer search (including multislice computed tomography and magnetic resonance imaging of various areas), with the exception of tuberculosis, monogenically study proteins in the blood and urine tests, biopsy of the ileum, immunohistochemistry trepanobiopsy.Results.During the 4‑year examination in the patient’s blood, an M-gradient in the gamma zone was detected, monoclonal immunoglobulin М-κ 19.3 g / l, Bens-Jones-κ protein in urine (daily proteinuria 0.45 g) was detected during immunofixation. In the myelogram at the light-optical level, the number of cell elements was significantly reduced with a pronounced lymphoid proliferation of 40 %, the granulocytic series and erythropoiesis were relatively narrowed, and plasma cells 6 %. Immunomorphological picture trepanobiopsy our patient based on the data of laboratory methods of research corresponds to the defeat of the bone marrow in Waldenström’s disease-κ, secretion of M-paraprotein. An important feature that allowed to go on the right path of diagnosis was almost pathognomonic for tumor lymphoproliferation, detection of Bens-Jones protein in the urine, which was absent in the onset of the disease.Conclusion.This clinical case is interesting not only by the complexity of the diagnosis of macroglobulinemia of Waldenström in General, but also by the atypical, erased clinical disease in our patient.

We present new data of the comparison of the geographical variability of heavy metal concentrations (Cu, Zn, Mn, Fe, Cd, Pb, Ni) in tissues of medical leeches (Hirudo medicinalis, Hirudo verbana) and in bottom deposit from places of their inhabitation in various regions of Russia and Ukraine. The geographical factor was found to have a significant impact on the elemental status of medical leeches and bottom sediments. It was shown that, irrespective of geographic and biotopic timing, both types of medicinal leeches are macroconcentrator Zn and deconcentrator Mn and Fe. A high cumulative activity of H. medicinalis and H. verbana in relation to ecotoxicants Cd, Pb and Ni in aquatic ecosystems of industrial regions was revealed. A decrease in bioaccumulation activity towards heavy metals in specimens of medical leeches living in adverse climatic conditions on the northern and eastern borders of the range is shown. The ability of leeches H. medicinalis and H. verbana to regulate the level of essential metals Mn and Zn in the body is established, while the content of Cu, Fe, Cd, Pb, Ni is determined by the concentration of these elements in the habitat (bottom sediments). A high variability of the studied parameters and the general laws of bioaccumulation activity of leeches in relation to heavy metals are caused of significant correlation with climatic and geographical features of the studied regions, and a degree of anthropogenically modified territories. It is shown that the microelemental status of leeches as a whole objectively reflects regional specificity of the ecological conditions of hydrobiocoenosises. Thus, the performed investigation can be used in monitoring actions for preservation of water resources and protection of natural biological resources of the hirudofauna.

We analyze here for the first time the plasma free amino acid profile in pond bats (Myotis dasycneme Boie, 1825) living in the boreal Ural region and exposed experimentally to low positive and near-zero temperatures during their preparation for hibernation. Pond bats were caught in their mass habitation territory in the Middle Ural near the Smolinsky cave (N 56°28’, E 61°37’) in the third decade of September 2015. Qualitatively, blood plasma in pond bats contains 21 amino acids. In a model experiment carried out on pre-hibernating animals at a regular hibernation temperature (0–2°C), the total plasma pool of free amino acids increased significantly by 42% (irrespective of sex) and reached 1561.4 ± 112.6 μmol/L (p = 0.01). Under these experimental conditions, the fraction of glucogenic amino acids rose by 34% (p = 0.01) and that of essential ones by 80% (p = 0.001). Both in control and experimentally cooled pre-hibernating animals, the plasma was found to lack tryptophan, suggesting its utilization as a substrate in the synthesis of serotonin, a biogenic amine directly involved in the maintenance of hypothermia and hypometabolism in these chiropterans.

The amino acid spectrum of blood plasma in three bat species (Myotis dasycneme, Pipistrellus nathusii, Vespertilio murinus) inhabiting the Urals has been studied for the first time. The bats were trapped in the zone of their high abundance in Chelyabinsk oblast (2013–2014). Free amino acids were determined by liquid ion exchange chromatography (a total of 384 determinations). It has been shown that the plasma amino acid spectrum consists of 22 amino acids in subadult bats of all three species, but there are species-specific differences in their concentrations. The total amino-acid pool concentration in migratory P. nathusii and V. murinus exceeds that in resident M. dasycneme by factors of 2.9 and 1.8, respectively. Migratory species are characterized by a high concentration of plasma arginine: it is six times higher in V. murinus than in M. dasycneme, and in P. nathusii arginine accounts for 25.4% of the amino acid pool. The group of glucogenic amino acids is prevalent in the blood plasma of migratory species (75% in V. murinus and 79% in P. nathusii), while in M. dasycneme the total proportion of lysine, glycine, and glutamic acid is 2.3 times lower than in P. nathusii and 1.7 lower than in V. murinus (p < 0.05). These results provide evidence for significant differences in the contents of free blood plasma amino acids between migratory and resident bat species.

The article presents the results of a new comparative analysis of free amino acids in the blood plasma of representatives of insectivorous Chiroptera (Mammalia: Vespertilionidae) in the fauna of the Ural Mountains: the pond bat (Myotis dasycneme Boie, 1825) and the parti-colored bat (Vespertilio murinus Linnaeus, 1758). This is the first study to show the species variability of free amino acids in resident and migratory species of bats from different ecosystems of the Ural Region.

Ecological features and species composition of leeches were studied in 25 water bodies located in the city of Yekaterinburg and its vicinity. Specific features of their occurrence and distribution in water bodies of various types were analyzed. Dominant, mass, common, and rare species were distinguished.