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Project titleBaikal endemics as extremophiles: physiological, biochemical and molecular mechanisms of resistance to hyperoxia at conditions of low temperature and mineralization

Research area 04 - BIOLOGY AND LIFE SCIENCES, 04-103 - Hydrobiology and ichthyology

KeywordsBaikal, endemics, amphipoda, antioxidants, oxidative stress, reactive oxygen species, adaptation, stress, oxygen

Annotation
The studies of extremophiles, or organisms bearing the specific adaptation to the extreme environmental factors, become especially urgent because studying such mechanisms offers interdisciplinary cooperation with other scientific fields, such as biotechnology, agriculture and pharamacology. However the number of ecosystems with an extreme environment is limited and most of those ecosystems are extensively studied. Therefore, intensive research of extremophile phyiosoligcal mechanisms is important. One unique ecosystem with such specific environmental parameters is Lake Baikal, which is the only large waterbody with complex environmental factors that occur at even at maximal depths, including high level of dissolved oxygen (especially during underice aglal bloom – underice hyperoxia), low temperatures and ultralow mineralization. This allows for the only known deep, freshwater fauna, which can survive the hydrostatic pressure of more than 160 atm under the condition of low-mineralization, low temperature and high dissolved oxygen. Due to the water's high percent transparency, light and UV-light can penetrate up to 6 and additionally enhace production of reactive oxygen species. Thus, Lake Baikal endemic species experience maximal oxygen saturation and super-saturation, and can be considered as extremophiles, making them ideal models in understanding the mechanisms of resistance to reactive oxygen species. In the frame of the proposed project, it is planning to study the mechanisms of resistance of Baikal endemic organisms to extreme conditions of hyperoxia together with the elevated level of UV-radiation, ultra-low mineralization, low temperature and the hyperbaric oxygenation due to the high level of dissolved oxygen on the big depths. On example of the broad range of Baikal endemic amphipods (Amphipoda, Custacea), which is the most diverse group within the fauna of Lake Baikal, the species-specific variability in optimal oxygen concentrations will be estimated and the peculiarities of antioxidant and pro-oxidant status of studied species will be determined. This analysis will reveal the conditions of maximal activation of target antioxidative mechanisms in Baikal endemics. Mechanisms, which will activate under this condition, will be analyzed using the most state-of-the-art methods of analysis - proteomics, transcriptomics and metabolomics. Proteomic analysis will be performed using 2D-PAGE, liquid chromatography and mass-spectrometry (LC-MS, MALDI-Tof). For the metabolomic analysis, the spectra of metabolites in aqueous and lipid phases will be obtained using liquid-, gas- and ion- chromatography; the faty-acid composition of lipids will be analyzed with chromato-mass-spectrometry. Transcriptomic analysis will be performed using the high-throughput sequencing with the sequencer HighSeq 2000 Sequencing System (Illumina). Additional target analysis of certain genes of interest will be performed by real-time qPCR. Bioinformatic analysis will be carried out using both common (e.g. NCBI Genbank, Uniprot, etc.) and custom databases (based on previously annotated transcriptomes of Baikal amphipods). The revealed mechanisms will be compared to the field observation and analyzed, taking into account evolution and ecology of studied species. The evolution of the revealed molecular mechanisms will be analyzed based on the known molecular-phylogenetic relationships of Baikal amphipods.

Expected results
Oxidative stress (induced by reactive oxygen species - ROS) is responsible for a large number of diseases and aging. Study of antioxidant system (AOS) in ROS-tolerant species can reveal new mechanisms of defense against the negative consequences of oxidative stress and will have a great impact on biomedical research, especially in areas of “oxidative diseases”, such as some cancers, neurodegenerative and cardiovascular diseases, inflammation and aging. Besides that studying of AOS can contribute the areas of biotechnology and agriculture while improving resistance of organisms. During the prosposed project a number of experimental procedures will allow to identify the conditions (oxygen concentration, UV-illumination, pressure) of the maximal activation of AOS in Baikal endemic amphipods (littoral, sublittoral and deep-water) in comparison with Palearctic fauna. The defence systems activated in these conditions will be studied by "global" methods of proteomcis, transcriptomics and metabolomics to obtain the comprehensive representation of the changes in organisms of Baikal amphipods required for maximisation of the antioxidant response. The obtained experimental data will be verified by study of amphipods in natural conditions during different seasons. The identified mechanisms will be analyzed taking into account evolution and ecology of studied species. In spite of the significant importance of such research in applied science it should be specially noted that the study of stress resistance of Baikal endemics makes a large contribution in evolutionary studies, while helping to understand the way of resistanse development. The results of such works are of the great value due to the possibility to develop new efficient methods of biomonitoring Lake Baikal ecosystems, which contains 20% liquid fresh water of the world. It is especially important during the current environmental crisis on Lake Baikal.

Annotation of the results obtained in 2019
In 2019, the planned work was completed to assess the effect of pro-oxidant factors on endemic amphipods of Lake Baikal in comparison with Holarctic amphipods. The physiological reactions of the Baikal endemics and the ubiquist species Gammarus lacustris were studied under elevated levels of oxygen and ultraviolet radiation. The development of oxidative damage to amphipods under these conditions was assessed using the content of lipid peroxidation products, the level of carbonylation of hemocyte proteins, and the concentration of carotenoids. The features of the antioxidant system of the Baikal endemic amphipods in comparison with the Holarctic G. lacustris were studied in the control conditions and under the influence of prooxidant factors, partially using transcriptome data. The reaction of hemolymph parameters controlling the affinity of the respiratory pigment hemocyanin to oxygen was analyzed in the Baikal species Eulimnogammarus verrucosus under hyperoxic conditions, partially using proteomic methods. Transcripts of microsomal glutathione S-transferases were found only in Baikal endemic species of the genus Eulimnogammarus, but not in the ubiquist species G. lacustris, which may be a specialized evolutionary adaptation of Baikal endemic species. It was established that the Baikal amphipods are indeed characterized by a unique physiological response to hyperoxia, expressed in the absence of a decrease in respiratory activity, which is not characteristic of other crustaceans, including Holarctic G. lacustris. Thus, the study allowed us to confirm part of the initial hypothesis of the project, as well as raise a number of new questions regarding the mechanisms of regulation of oxygen supply to the tissues of Baikal endemic amphipods.

Publications

1. D.V. AXENOV-GRIBANOV, D.S. BEDULINA, Y.A. SHIROKOVA, V.A. EMSHANOVA, Y.A. LUBYAGA, K.P. VERESHCHAGINA, A.E. SARANCHINA, T.P. POBEZHIMOVA, M.A. TIMOFEYEV Diet influence on mechanisms of non-specific stress-response in Baikal endemic amphipod species during long-term laboratory exposure Crustaceana, DOI: 10.1163/15685403-00003951 (year - 2019) https://doi.org/10.1163/15685403-00003951

2. Ekaterina Shchapova, Anna Nazarova, Anton Gurkov, Ekaterina Borvinskaya, Yaroslav Rzhechitskiy, Ivan Dmitriev, Igor Meglinski, Maxim Timofeyev Application of PEG-Covered Non-Biodegradable Polyelectrolyte Microcapsules in the Crustacean Circulatory System on the Example of the Amphipod Eulimnogammarus verrucosus Polymers, Vol. 11, Iss. 8, P. 1246 (year - 2019) https://doi.org/10.3390/polym11081246

3. Gurkov A., Shchapova E., Nazarova A., Rzhechitskiy Y., Saranchina A., Timofeyev M. Effects of injection of non-biodegradable polyelectrolyte microcapsules into the amphipod circulatory system Book of abstracts. 18th International Colloquium on Amphipoda (ICA), P. 44 (year - 2019)

4. Lubyaga Y.A.,Yarinich L.A., Drozdova P.B., Luckenbach T., Timofeyev M.A. Изучение Р-гликопротеина байкальских эндемичных амфипод в культивируемых клетках S2 дрозофилы Тезисы докладов VII съезда ВОГиС, C. 481 (year - 2019)

5. Madyarova E.V., Shirokova Y.A, Axenov-Gribanov D.V., Bedulina D.S., Emshanova V.A., Shatilina Z.M., Timofeyev M.A. Stable indicators of energy metabolism and non-specific stress response as a strategy of biochemical adaptation in eurybathic Baikal amphipods of the genus Ommatogammarus Book of abstracts. 18th International Colloquium on Amphipoda (ICA), P. 113 (year - 2019)

6. Pekkoeva S.N., Voronin V.P., Shatilina Zh.M., Madyarova E.V., Aksenov-Gribanov D.V., Shirokova Yu.A., Nemova N.N., Timofeyev M.A., Murzina S.A. Липидный и жирнокислотный спектр двух видов амфипод-некрофагов Ommatogammarus albinus и Ommatogammarus flavus, обитающих в градиенте глубин оз. Байкал XII Съезд Гидробиологического общества при РАН: тезисы докладов, С. 379-380 (year - 2019)

7. Rzhechitskiy Y.A., Gurkov A.N., Shatilina Zh.M., Emshanova V.A., Bobkova V.A., Larina O.A., Timofeyev M.A. Воздействие гипероксии на байкальских эндемичных и голарктических амфипод XII Съезд Гидробиологического общества при РАН: тезисы докладов, - (year - 2019)

8. Rzhechitskiy Y.A., Gurkov A.N., Shatilina Zh.M., Emshanova V.A., Bobkova V.A., Larina O.A., Timofeyev M.A. Воздействие гипероксии на частоту дыхательных движений и показатели окислительного стресса у байкальских эндемичных амфипод Eulimnogammarus cyaneus (Dybowsky, 1974) Биология – наука ХХI века: 23-я Международная Пущинская школа-конференция молодых ученых, С. 307 (year - 2019)

9. Shirokova Y.A., Madyarova E.V., Rzhechitskiy Y.A. Влияние градиентного изменения температуры среды на активность антиоксидантных ферментов у байкальских амфипод Ommatogammarus flavus Материалы 57-й Международной научной студенческой конференции, С. 38 (year - 2019)

10. - Как развивать Байкал по-новому Ведомости, - (year - )

11. - Больше, чем море N+1, - (year - )

12. - Ученые выяснили, как повышение температуры воды в Байкале повлияет на обитателей озера ТАСС, - (year - )

Annotation of the results obtained in 2017
During the first year of the project, the monitoring evaluation of Lake Baikal in the studied species habitats was carried out. Mechanisms of resistance of Baikal endemic amphipods to hyperoxia at the conditions of high hydrostatic pressure were investigated on physiological, biochemical and molecular levels. More than 200 field and laboratory experiments were carried out. More than 22 000 crustaceans were fixed and not less than 15 000 biochemical measurements were performed. The influence of normoxia (10-12 mg O2 / l) and hyperoxia (30-45 mg O2 / l) on Baikal endemic and Holarctic amphipods was estimated. The investigations of 2017 year included several directions. Temperature and oxygen regimes was monitored during the year, using the autonomous programmable dataloggers iButton (Maxim Integrated, USA), portable oximeter ExtecHDO and underwater video equipment. The hypothesis of the presence of subglacial hyperoxia in Lake Baikal, with oxygen exceeding 100% saturation in water of the littoral zone, has been confirmed. Using classical methods of hydrobiology, sexual structure of amphipods population at different seasons was studied. For the first time, the limits of metabolic tolerance of amphipods to high oxygen content were investigated. The level of oxidative damage of macromolecules, the activity of the antioxidant enzymes in amphipods under hyperoxia conditions were evaluated. To estimate the limits of metabolic tolerance, Baikal and Holarctic amphipods were exposed at different levels of oxygen concentration. Limit of metabolic tolerance was investigated by motor activity and pleopod beating frequency. It has been shown that Baikal amphipods have the increased tolerance to acute hyperoxia (up to 30-45 mg O2 / L) and are able to survive in such conditions for more than ten days; Moreover, hypoxia caused the significant changes in physiological parameters (motor and respiratory activities) in Baikal species G. fasciatus which considered to be a tertiary relict within Baikal fauna. At the same time, hyperoxia but does not have a significant effect on representatives of the younger fauna of amphipods from genus Eulimnogammarus. For the first time, it has been proved that G.lacustris is more sensitive to hyperoxia in contrast to Baikal endemics. The fluctuations in levels of macromolecules oxidative damage and nonspecific stress response in littoral amphipods, associated with periods of maximum subglacial hyperoxia, were found. The significantly higher level of lipid peroxidation in littoral amphipods, compared to deep-water species was found. The influence of season of the year on stress proteins content and stress-induced changes in their content in amphipods, exposed at a gradual temperature increase were assessed. Additionally, the PCR primers for the most important metabolic genes expression in deepwater amphipods were developed and their efficiency was evaluated. During the present study, for the first time the effect of season on amphipods proteomes was investigated and the specific stress-induced compounds in Baikal amphipods were evaluated. The possibility of using two-dimensional gel-electrophoresis and mass-spectrometry for seasonal changes in amphipods proteomes and metabolomes was confirmed. During the mass-spectrometric analyses the new unknown compounds were found in the exposed at the hyperoxia amphipods. These compounds may serve as potential candidates in specific stress-response to hyperoxic conditions. Particularly, mass-spectra of compounds in amphipods from Eulimnogammarus genus were similar and the hyperoxia did not cause the qualitative changes in natural compound compositions. However, experimental hyperoxia caused a two-fold increase of the compound with the mass 763,53344 Da, ([M+H] 764,54124 Da; [M+Na] – 786.50427) in tissues of amphipod species E.verrucosus after 48 hours of hyperoxia exposure (30-45 mg/L dissolved oxygen). The changes in this metabolite amount was not found in the congener species E.cyaneus. It is worth to mention, that this mass is not registered in the largest database of natural compounds - Dictionary of Natural products (СRС press, 2017). Thus, the found compound may serve as a new biomarker of stress conditions as well as a new antioxidant, which is specifically synthetized in Baikal amphipods. Thus, during the first year of the project, first data about resistance mechanisms of Baikal amphipods to hyperoxia and a high pressure on physiological, biochemical and molecular levels were obtained. These results require the further research and development.

Publications

1. Axenov-Gribanov D.V., Bedulina D.S., Shirokova Y.A., Emshanova V.A., Voytsekhovskaya I.V., Lubyaga Y.A., Gurkov A.N., Vereshchagina K.P., Shatilina Zh.M., Timofeyev M.A. Diet influence on mechanisms of non-specific stress-response in Baikal endemic amphipod species during long-term laboratory exposure Limnologica, - (year - 2017)

2. Axenov-Gribanov D.V., Protasov E.S., Voytsekhovskaya I.V., Shatilina Z.M., Rzhechitskiy Y.A., Dimova M.D., Kostka D.V. Lubyaga Y.A., Timofeyev M.A. Hemolymph of cold adapted Baikal endemic amphipods as a promising source for screening of novel natural products Journal of Stress Physiology & Biochemistry, Vol. 13 No. 4, pp. 74-80 (year - 2017)

3. Jakob L., Bedulina D.S., Axenov-Gribanov D.V., Ginzburg M., Shatilina Zh.M., Lubyaga Y.A., Madyarova E.V., Gurkov A., Timofeyev M.A., Pörtner H.-O., Sartoris F.J., Altenburger R., Luckenbach T. Uptake kinetics and subcellular compartmentalization explain lethal but not sublethal effects of cadmium in two closely related amphipod species Environmental Science & Technology, 51 (12), pp 7208–7218 (year - 2017) https://doi.org/10.1021/acs.est.6b06613

4. Protasov E.S., Axenov-Gribanov D.V., Voytsekhovskaya I.V., Shatilina Z.M., Vereshchagina K.P., Lubyaga Y.A., Rzhechitskiy Y.A., Shirokova Y.A., Timofeyev M.A. Secondary metabolites detected in deep-water endemic amphipods of Lake Baikal: bacterial or crustacean origin? Journal of Stress Physiology & Biochemistry, Vol. 13 No. 4 2017, pp. 104-109 (year - 2017)

5. Shchapova E.P., Axenov-Gribanov D.V., Lubyaga Y.A., Shatilina Z.M., Vereshchagina K.P., Protasov E.S., Madyarova E.V., Timofeyev M.A. Crude oil at concentrations considered safe promotes rapid stress-response in Lake Baikal endemic amphipods Hydrobiologia, № 805, Is. 1, P.189-201 (year - 2018) https://doi.org/10.1007/s10750-017-3303-3

6. Axenov-Gribanov D., Protasov E., Rebets Y., Voytsekhovskaya I., Tokovenko B., Bedulina D., Luzhetskyy A., Timofeyev M. The diversity and antibiotic properties of actinobacteria associated with endemic deepwater amphipods of lake Baikal Proceedings of the 17th International Colloquium on Amphipoda in Biodiversity Journal, 8 (2) pp. 675–676 (year - 2017)

7. Bedulina D., Meyer M., Gurkov A., Kondratjeva E., Baduev B., Timofeyev M. Intersexual differences of thermal sensitivity and heat shock response proteins in Baikal endemic amphipods Proceedings of the 17th International Colloquium on Amphipoda in Biodiversity Journal, 8 (2) pp. 641–642 (year - 2017)

8. Bedulina D.S., Kalkof S., Gurkov A., Drozdova P.B., Timofeyev M.A., Luckenbach T. Идентификация белков стресс-ответа у байкальской амфиподы Eulimnogammarus cyaneus Dyb. методами протеогеномики тезисы докладов Всероссийской научной конференции "Фундаментальные проблемы экологии России", С. 24. (year - 2017)

9. Dimova M.D., Madyarova E.V., Gurkov A.N., Lubyaga Y.A., Kondrateva E., Adelshin R.V., Timofeyev M.A. Genetic diversity of microsporidia in some endemic amphipods from lake Baikal Abstract book of Young biologists science week-2017, С. 125-131 (year - 2017)

10. Gurkov A., Rivarola-Duarte L., Casas I.F., Michael H., Bedulina D., Stadler P., Timofeyev M., Luckenbach T. Population structure of amphipods Eulimnogammarus verrucosus (Gerstfeldt, 1858) and E. cyaneus (Dybowsky, 1874) endemic to Lake Baikal Proceedings of the 17th International Colloquium on Amphipoda in Biodiversity Journal, 8 (2), pp. 625–626 (year - 2017)

11. Lubyaga Y.A., Trifonova M.C., Vereshchagina K.P., Kondrateva E.C., Shatilina Z.M., Axenov-Gribanov D.V., Zadereev E.S., Gurkov A.N., Timofeyev M.A. Сравнительное исследование влияния изменения температуры среды на выживаемость и энергетический метаболизм пресноводных амфипод Gammarus lacustris и Gmelinoides fasciatus из отдаленных популяций сборник материалов VI Всероссийской конференции по водной экотоксикологии, посвященной 80-летию со дня рождения д.б.н., проф. Б.А. Флерова, с приглашением специалистов из стран ближнего зарубежья, С. 62-63. (year - 2017)

12. Lubyaga Y.A., Trifonova M.S., Emshanova V.A., Gurkov A.N., Axenov-Gribanov D.V., Timofeyev M.A. Comparative study of nonspecific mechanisms of stress-adaptations in representatives of remote Gmelinoides fasciatus populations under hyperthermic conditions Abstract book of Young biologists science week-2017, С. 299-305 (year - 2017)

13. Lubyaga Y.A., Trifonova M.S., Emshanova V.A., Madyarova E.V., Axenov-Gribanov D.V., Shatilina Zh.M., Timofeyev M.A. Влияние изменения температуры на биохимические механизмы резистентности у представителей различных популяций Gmelinoides fasciatus (Stebb., 1899) тезисы III Международной конференции «Биоиндикация в мониторинге пресноводных экосистем», С. 204-206. (year - 2017)

14. Madyarova E.V., Bedulina D.S., Baduev B.K., Axenov-Gribanov D.V., Dimova M.D., Emshanova V.A., Shirokova Y.A., Shatilina Z.M., Timofeyev M.A. Роль белков теплового шока в адаптации глубоководных амфипод озера Байкал к большим глубинам тезисы докладов Всероссийской научной конференции "Фундаментальные проблемы экологии России", С. 117. (year - 2017)

15. Vereshchagina K., Jakob L., Gurkov A., Bedulina D., Sartoris F.J,. Pörtner H.O, Timofeyev M., Lucassen M. Effects of rising temperatures on expression patterns of genes related to universal stress-response and energy metabolism in Baikal endemic and Holarctic amphipods Proceedings of the 17th International Colloquium on Amphipoda in Biodiversity Journal, 8 (2), pp. 629–630 (year - 2017)

16. Vereshchagina K., Lubyaga Y., Shatilina Z., Zadereev E, Timofeyev M. Can adaptation to habitats with different salinity regimes modulate the energy metabolism, cellular stress responses and thermal tolerance in amphipod Gammarus lacustris Sars? Proceedings of the 17th International Colloquium on Amphipoda in Biodiversity Journal, 8 (2), pp. 627–628 (year - 2017)

17. Vereshchagina K.P., Bedulina D.S., Gurkov A.N., Axenov-Gribanov D.V., Timofeyev M.A. Gene expression profile in Baikal and Holarctic species amphipod in response to raising temperatures материалы X Всероссийского с международным участием Конгресса молодых ученых-биологов «Симбиоз-Россия 2017», pp. 135 – 136. (year - 2017)

18. Vereshchagina K.P., Lubyaga Y.A., Shatiina Z.M., Zadereev E.S., Gurkov A.N., Timofeyev M.A. The influence of mineralization in environment on thermotolerance and energy metabolism in amphipods Gammarus lacustris Sars Abstract book Young biologists science week-2017, pp. 540-545 (year - 2017)

19. Vereshchagina K.P., Lubyaga Y.A., Shatilina Z.M., Zadereev E.S., Gurkov A.N., Timofeyev M.A. Salinity modulates thermotolerance, energy metabolism and stress response in amphipods Gammarus lacustris Sars Book of abstracts of 13TH INTERNATIONAL CONFERENCE ON SALT LAKE RESEARCH (ICSLR 2017), P. 25-26 (year - 2017)

20. - СЛУЖИЛИ ДВА ЭНДЕМИКА. БАЙКАЛЬСКИЕ БИОЛОГИ НАШЛИ ПРИМЕНЕНИЕ УНИКАЛЬНОЙ ЖИВНОСТИ Газета Поиск, - (year - )

21. - СЕКВЕНИРОВАНИЕ ЭКЗОМА, ПЕРЕПРОГРАММИРОВАНИЕ КЛЕТОК И ТРАНСФЕР ТЕХНОЛОГИЙ Наука и технологии России, - (year - )

Annotation of the results obtained in 2018
During the second year of the project, new data characterizing long-term dynamics of the influence of high oxygen concentration and UVA irradiation on physiological parameters in Lake Baikal endemic amphipods have been obtained. In addition, markers of cellular oxidative damage have been shown to be highly stable in Baikal amphipods under the mentioned environmental influences. Multilevel comparison of differences in the antioxidant defence system between different species of Baikal and Holarctic amphipods has been performed with global approaches such as proteomics, metabolomics, transcriptomics and lipidomics. Differences in expression of antioxidant enzymes, metabolic and lipid composition have been found. Proteins that participate in the antioxidant response the of Baikal endemic amphipod species E. cyaneus have been deeply characterized. Metabolic mechanisms that underline the adaptation of Baikal deep-water amphipods to a higher pressure than the littoral species have been analyzed. The obtained data allow us to propose the accumulation of small antioxidant molecules as the most probable major mechanism that ensures the tolerance of Baikal amphipods to hyperoxia.

Publications

1. Axenov-Gribanov D.V., Shatilina Z.M., Lubyaga Y.A., Emashanova V.A., Vereshchagina K.P., Lozovoy D.V., Kondratieva E.S., Timofeyev M.A. Estimation of experimental cohabitation between Golarctic and Baikal endemic amphipods species: G. lacustris against G. fasciatus Journal of Stress Physiology & Biochemistry, Vol. 14, No. 1. – Р. 5-11. (year - 2018)

2. Bedulina D.S., Shatilina Z.M., Axenov-Gribanov D.V., Lubyaga Y.A., Timofeyev M.A. Estimation of basal hsp70 levels in two thermotolerant baikal amphipod species from different thermal habitats Journal of Stress Physiology & Biochemistry, Vol. 14, №. 2. – Р. 26-35 (year - 2018)

3. Bowman L. L., Kondrateva E. S., Timofeyev M. A., Yampolsky L. Y. Temperature gradient affects differentiation of gene expression and SNP allele frequencies in the dominant Lake Baikal zooplankton species Molecular ecology, 27 (11), pp. 2544-2559 (year - 2018) https://doi.org/10.1111/mec.14704

4. Dimova M. , Madyarova E., Gurkov A., Drozdova P., Lubyaga Y., Kondrateva E., Adelshin R., Timofeyev M. Genetic diversity of Microsporidia in the circulatory system of endemic amphipods from different locations and depths of ancient Lake Baikal PeerJ., 6:e5329 (year - 2018) https://doi.org/10.7717/peerj.5329

5. Gurkov A., Borvinskaya E., Shchapova E., Timofeyev M. Restraining small decapods and amphipods for in vivo laboratory studies Crustaceana, 91 (5), P. 517-525 (year - 2018) https://doi.org/10.1163/15685403-00003778

6. Khomich A. S., Golubev A. P., Axenov-Gribanov D. V., Bodilovskaya O. A, Shirokova Y. A., Lubyaga Y. A., Shatilina Z. M. The dynamics of contents of the total protein, haemocyanin and activity of the antioxidant enzymes under condition of acute hyperthermia in pulmonate mollusk Lymnaea stagnalis Journal of stress physiology and biochemistry, Vol. 14, No 1. - P. 19-26. (year - 2018)

7. Khomich A.S., Golubev A.P., Axenov-Gribanov D.V., Bodilovskaya O.A., Shirokova Y.A., Loshakova Y.V., Lubyaga Y.A., Shatilina Z.M. The Effect of Acute Hiperthermia on Levels of Hsp70 and Lipid Peroxidation in Laboratory-Cultured Pulmonate Mollusk Stagnicola corvus Сontemporary problems of ecology, Vol. 11, No 1. - P. 64-71. (year - 2018) https://doi.org/10.1134/S1995425518010067

8. Vereshchagina K.P., Kondrateva E.S., Axenov-Gribanov D.V., Shatilina Zh.M., Khomich A.S., Bedulina D.S., Zadereev E. S., Timofeyev M.A. Nonspecific stress response to temperature increase in Gammarus lacustris Sars with respect to oxygen-limited thermal tolerance concept PeerJ., 6:e5571 (year - 2018) https://doi.org/10.7717/peerj.5571

9. Woolway R. I., ... Timofeyev M. A., ... Weyhenmeyer G. A. Lake surface temperature [in "State of the Climate in 2017", Blunden J., D. S. Arndt, G. Hartfield, Eds.]. Bull. Amer. Meteor. Soc., Vol. 99, No. 8, P. 13-15 (year - 2018) https://doi.org/10.1175/2018BAMSStateoftheClimate.1

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12. Drozdova P. B., Bedulina D. S., Saranchina A. E., Timofeev M. A. Поиск молекулярных механизмов формирования окраски тела Eulimnogammarus cyaneus (Dybowsky, 1874) Международная конференция «Пресноводные экосистемы - современные вызовы», 10-14 сентября, 2018. Тезисы докладов и стендовых сообщений, С. 134. (year - 2018)

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16. Lubyaga Y.A., Trifonova M.S., Vereshhagina K.P., Madyarova E.V., Shatilina Z.M., Gurkov A.N., Kondrateva E.S. Сравнительное исследование неспецифических механизмов стресс-адаптации у представителей отдаленных популяций Gmelinoides fasciatus и Gammarus lacustris в условиях гипертермии. Международная конференция «Пресноводные экосистемы - современные вызовы», 10-14 сентября, 2018. Тезисы докладов и стендовых сообщений, С. 220-221. (year - 2018)

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18. Nazarova A.A., Gurkov A.N., Shapova E.P., Dmitriev I.A., Timofeev M.A. Изучение иммунного ответа амфипод на имплантируемые оптические микросенсоры с помощью первичной культуры гемоцитов Международная конференция «Пресноводные экосистемы - современные вызовы», 10-14 сентября, 2018. Тезисы докладов и стендовых сообщений., С. 257. (year - 2018)

19. Nazarova A.A., Gurkov A.N., Shhapova E.P., Dmitriev I.A. Первичная культура гемоцитов, как инструмент для изучения клеточного иммунного ответа амфипод Eulimnogammarus verrucosus Материалы Международного молодежного научного форума «ЛОМОНОСОВ-2018», - (year - 2018)

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24. Vereshhagina K. P., Kondrateva E. S. Холодовая адаптация как ключевой механизм уникальности байкальских эндемичных амфипод Материалы Международного молодежного научного форума «ЛОМОНОСОВ-2018», - (year - 2018)

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26. Vereshhagina K. P., Kondrateva E. S., Aksenov-Gribanov D. V., Timofeev M. A. Устойчивость к холоду как ключевой механизм уникальности байкальских эндемичных амфипод Международная конференция «Пресноводные экосистемы - современные вызовы», 10-14 сентября, 2018. Тезисы докладов и стендовых сообщений., С. 355. (year - 2018)

27. - Бокоплав-кузнечик выживает в «горячей» воде за счет неверного жиросжигания Чердак, https://chrdk.ru/news/bokoplav-kuznechik-vyzhivaet-v-goryachei-vode-za-schet-nevernogo-zhiroszhiganiya?fbclid=IwAR0C_f1z9JTyGSOSFMf8h_yHNNf14wkPq9id5u4S4nikP8pLI4OZtIgEVwM (year - )