Approved by the order of Minister of Education

Science and Sport of the Republic of Lithuania

No. V-585 of 19 April 2022

RESEARCH AND EXPERIMENTAL DEVELOPMENT PROGRAMME 2022–2026

“RESEARCH ON THE STATE AND DYNAMICS OF BIODIVERSITY AND HABITATS, SCIENTIFIC BASIS FOR CONSERVATION AND RESTORATION (BIODIVERSITY)”

1. Research and experimental development programme 2022–2026 “Research on the state and dynamics of biodiversity and habitats, scientific basis for conservation and restoration (BIODIVERSITY)” (from now on – the Programme) executor – Nature Research Centre (from now on – NRC).

2. Objectives of the Programme:

2.1. Complex studies on the systematics and state of populations and their habitats of prokaryotic and eukaryotic organisms (protists, fungi, plants, animals) to gain new fundamental knowledge about biodiversity and its supporting mechanisms in the conditions of global change and anthropogenic impact.
2.2. Provision of the scientific basis for the State strategy documents and recommendations for the conservation, sustainable use and restoration of biological diversity.
2.3. Completion, systematisation and improvement of systematic national collections, metadata and other biodiversity databases of international significance, adapting them to the needs of science and society, accumulation of interactive knowledge on various components of biodiversity, promoting public involvement in these activities.

3. The tasks of the Programme:

3.1. Based on the studies of plant, fungi, prokaryotic and eukaryotic microorganism species and communitiesʼ diversity, structure and functions, to determine the signs and directions of community transformation, assess the extent of invasions of alien plants, changes related to habitat change, identify the diversity of plant, fungi, prokaryotic and eukaryotic microorganism species in natural and partially human-affected ecosystems, with an emphasis on practical conservation aspects.
Researchers and technical staff from the Laboratory of Algology and Microorganism Ecology (AMEL), the Laboratory of Economic Botany (EBL), the Laboratory of Flora and Geobotany (FGL) and the Laboratory of Mycology (ML) of the NRC Institute of Botany are involved in this task.
3.2. Based on systematic studies of insects and other arthropods, mammals and birds, characteristic and invasive species, to determine changes in species distribution, population abundance, trends and regularities caused by climate change and anthropogenic stress, determine the impact of these factors on ecosystems and their functions, assess prospects for rational management of populations, the use and conservation of species resources.
Researchers and technical staff from the Laboratory of Entomology (EL), the Laboratory of Avian Ecology (AEL) and the Laboratory of Mammal Ecology (MEL) of the NRC Institute of Ecology are involved in this task.
3.3. To gain new knowledge about the diversity, life cycles and phylogeographic relationships of eukaryotic parasitic organisms (protists and helminths) based on integrated modern experimental, morphological, ecological, cariological, molecular and statistical research. Based on this knowledge, to assess the state of biodiversity of the studied pathogens, the dynamics of global change and the virulence and epidemiology of diseases in the wild, determine the patterns of species introduction and justify preventive measures for the spread of new infections in natural ecosystems of the Baltic Region.
Researchers and technical staff from the P. B. Šivickis Laboratory of Parasitology (PL) and the Laboratory of Molecular Ecology (MEL) participate in implementing this task.

4. Methodological substantiation of research

Wildlife biodiversity is a living network made up of the many life forms that exist on the planet to provide food, shelter and protection for humanity.
The only way to preserve the quality and continuity of human life on Earth is to preserve and restore biodiversity, as emphasised in the EUʼs 2030 Biodiversity Strategy. Biota is a critical factor in ensuring a healthy life for the well-being of society. However, despite the exceptional importance of wildlife for human existence, efforts to create well-being have contributed to the rapid and reckless degradation of biodiversity – 60 per cent of the worldʼs wildlife population has declined over the past 40 years, and nearly a million species are on the verge of extinction. Furthermore, we lose trillions of euros a year due to lost access to ecosystem services. Biodiversity loss, leading to declining ecosystem services and the imminent economic crisis, has been identified as humanity’s most significant challenge in the coming decades. Considerable investment in natural capital is urgently needed to halt biodiversity loss and the crisis affecting most natural populations. This requires a pool of experts from countries, their citizens and biodiversity-based research, fundamentals and expertsʼ efforts to develop research in areas relevant to society. The integrated biodiversity assessment programme will bring together the best experts in protistology, mycology, botany and zoology and gather a wealth of new fundamental data on biodiversity and its state in Lithuania and the world, which will provide a better understanding of the patterns of the existence of organisms in ecosystems and will provide a basis for assessing the stability of communities and the likelihood of change in the context of intensifying climate change and growing anthropogenic impacts. Furthermore, by applying the most modern research methods, the systematics of organisms will be specified, without which it is impossible to identify living organisms and make predictions of the state and development of the environment. Experience, data from long-term research and synergies between specialists of various organisms in the NRC are the basis for the effective development of the objectives of this Programme and to make a significant contribution to the implementation of Lithuaniaʼs strategic goals related to biodiversity. Long-term studies by the NRC researchers are designed to determine the patterns of species structure, population structure, the genetic structure of populations, animal and plant ecology of various systematic groups of organisms, including prokaryotes, fungi, plants, invertebrates and vertebrates. The biodiversity research methods developed by the NRC researchers are extensive, encompassing many modern fields of research that are constantly evolving, including morphometric, population, community composition, species phylogeny, microevolution, and large-scale data collection and analysis. The NRC has modernly equipped laboratories and field research bases, i.e. infrastructure adapted to the study of wild fauna and flora, and systematic large-scale and complex research allows for the accumulation of new scientific knowledge based on which the growth of scientific production is observed – the number of scientific papers published in high-level international journals is steadily increasing. To achieve the Programmeʼs objectives, the NRC laboratories will carry out molecular and traditional studies on various systematic groups, using a solid experimental base combining renovated stationary stations and laboratories, a modern experimental aquarium, two marine research vessels, GIS infrastructure and two EU-licensed vivariums. As a result, during the benchmarking exercise (comparative assessment) in 2018, the NRC zoological science infrastructure received a good rating. Specialists working at the NRC, combining many years of experience in the collection of research material under expedition, using specific research methods requiring high technical qualifications, combining light and electron microscopy with DNA fragment amplification, sequencing, genomic and proteomic studies, using unique databases and moderated data analysis methods will receive new knowledge for science during the Programme period and, based on their analysis, will provide scientifically based conclusions on the current real state of biodiversity, species populations and habitats, and will prepare scientific forecasts and recommendations for nature protection. By making available and systematically replenishing the accumulated collections of organisms of national importance (some of which are the largest or the only in the country and some have no analogues in Europe) and large data sets that are not only a national asset but also a prerequisite for current and future research, preconditions for mutually beneficial, interactive cooperation between scientists representing the NRC and interested groups of public representatives, universities, state institutions, which must ensure the countryʼs competencies and obligations to preserve and improve the state of biodiversity, ensure cooperation between scientific institutions, domestic and foreign specialists. The complexity of research and the diversity of methods are essential for modern biodiversity research. To reveal the peculiarities of the functioning of aquatic ecosystems under changing environmental conditions and develop recommendations for a strategy for the conservation and restoration of sustainable biodiversity, research carried out under natural conditions will be combined with experimental work in the laboratory with pure cultures of microorganisms. The diversity of planktonic blue-green algae and algae species, morphological and biological features, interactions with other organisms in the aquatic ecosystem will be studied by traditional hydrobiological (assessment of physicochemical conditions, light microscopy) and molecular biology (PCR (polymerase chain reaction), sequence of different genes analysis, population studies). Cyanotoxins are analysed by chemical (LC-MS/MS), enzyme-linked immunosorbent assay (ELISA) and molecular (specific sxtA, cyrA, cyrJ genes responsible for toxin synthesis) methods. Modern DNA and RNA sequencing and bioinformatics analysis, classical methods of molecular biology (quantitative PCR) and microbiological (isolation, purification and cultivation of microorganisms), and chemical analysis (LC-MS/MS) will be used for the analysis of freshwater planktonic bacteriophages. For research and experimental work, the equipment available at the NRC as part of the open access infrastructure will be used in cooperation with researchers from other Lithuanian institutions or foreign countries. The research objects for the experiments will be selected in the Lithuanian freshwater blue-green algae and algae collection maintained by the AMEL. Plant community studies will be carried out by using morphometric plant measurements, extraction of essential oils by hydrodistillation, gas chromatography, spectrophotometry, high-performance liquid chromatography (HPLC), mass spectrometry (MS), in vitro model systems for free radicals, anatomical microscopy, analysis of scientific publications, modern methods of mathematical statistical analysis. Detailed phytosociological descriptions of vegetation will be performed by recording both the species of plants and their cover and the values of some of their functional properties (description study fields or linear transect method); population surveys will be conducted on the basis of representative samples, disposable and stationary test fields, transects, field experiments and other field research methods; dendrochronological samples will be used for woody plant research; the assessment of grassland productivity will be carried out using unmanned aerial vehicles both for the geomorphological environment and for the determination of vegetation diversity; the assessment of the microclimate is carried out using passive data loggers on the temperature of the air, the soil surface and the topsoil; The GEST method (Greenhouse Gas Emission Sites Type) is based on the identification (mapping) of vegetation unit (area) types, the assessment of the ecological conditions (humidity, trophicity) and land use of the habitats. The laboratory will perform a morphometric and microscopic examination of collected plant samples, taxonomic identification of samples using morphological and genetic methods; an analysis of the basic physical and chemical properties of the soil samples is performed. Multidimensional analysis, various methods of digital classification of communities, species distribution models, and data mining in international databases will be used for statistical analysis and data management.
Standard field (sampling, habitat assessment) and laboratory (macro- and micromorphometric measurements) investigations will be used to study the diversity of fungi and fungal-like organisms. In addition, isolation of pure cultures of fungi and molecular genetic research (DNA Sequencing, Next Generation Sequencing) will also be performed, and data analysis from online citizen science platforms will be used.
Morphological microscopy (light microscopy, electron microscopy), comparative morphology, morphometric, histological as well as molecular (PCR, sequencing) methods will be used to study insects and other arthropods and their trophic relationships. Phylogenetic, statistical and trophic analysis of the spectrum, determination of the composition of communities, collection and analysis of large data arrays will be performed. Monitoring of dispersal and alien species, including invasive, arthropod species, and the compilation of a targeted database will be carried out with the participation of public members.
Mammalian abundance and species diversity research will use traditional methods, including night accounting of dormouse, a quantitative analysis of the study of killed on the roads mammals and data obtained using surveillance cameras and drones combining with the results from registries, GIS and neural (self-organisig) network, GLMM models and the inclusion of the society representatives.
Point surveys, sector accounts, route accounts, transectic accounts will be used for bird watching in nature; search and description of nests; registration of breeding parameters; telemetry (remote) surveys of birds: surveys of bird movement and behaviour using data loggers and remote bird tracking transmitters (GPS-GSM, satellite), light geolocators, etc.; monitoring of bird nests will be carried out using remote video cameras, automatic recording and identification of bird voices will be carried out, and research on the genetic structure of bird populations will be carried out using molecular research methods. In addition, spatial data analysis, statistical modelling and forecasting are performed.
Traditionally, an essential part of biodiversity research at the Nature Research Centre is associated with cosmopolitan parasitic organisms, with a high diversity of species, complex life cycles and morphological plasticity. They affect many ecological and evolutionary processes. Humans, vertebrates and invertebrates are still dying en masse and weakening from parasitic diseases. Outbreaks of new parasitic zoonotic infections have become more frequent in recent decades. Research into the biodiversity of wildlife pathogens is significant in nature, agriculture, medicine, veterinary, epizootiology and other areas. Despite a long history of research in Europe, the diversity, virulence, host cycle, life cycles, and molecular biology of most parasitic protists and helminths, including pathogens, are unknown, or knowledge is inaccurate and sometimes even erroneous. Basic knowledge about the diversity of these pathogens is still based on traditional morphological analysis, which is often insufficient to differentiate species and determine their phylogenetic relationships. Molecular methods for identifying parasites show that the diversity of protists and helminths is significantly greater than previously thought.
Analysis of parasite DNA sequences, integrated with morphological studies, provides an opportunity to reliably link the different stages of their life cycle development and elucidate the characteristics of parasite circulation in ecosystems. The planned research will provide fundamental knowledge about parasitic protists and helminths, their systematics, host specificity, life cycles, developmental features, phylogenetic and phylogeographic relationships, intraspecific and interspecific diversity, its status, population formation patterns and potential changes in ecosystems. The planned work deals with fundamental issues of parasitology, ecology and evolution, the solution of which will provide new information on the peculiarities of the formation of these species and populations and possible changes in ecosystems in the context of global climate change and anthropogenic activities.
The research methodology for parasitic protists and helminths has been developed, tested, and described in the NRC research publications in recent years. The research will comprehensively apply classical methods (light microscopy, histology, cariology, experiments, analysis of digital images of parasites, statistical analysis of data using the R-package) and modern molecular technologies (chromogenic in situ hybridisation, real-time PCR, innovative second-generation sequencing, etc.), which will allow the linking of individual databases and allow for a reliable assessment of biodiversity and the refinement of existing knowledge. Experimental studies will elucidate the peculiarities of the spread of parasites, the mechanisms of pathogenicity and interactions.
Studies on food-borne pathogens that may infect humans and are essential for animal health will focus on the morphological, genetic and ecological characteristics of the parasitic organisms Sarcocystis spp., Toxoplasma gondii (protists), Trichinella spp., Alaria alata (helminths). Pathogens will be identified using light and electron microscopy techniques, preparing various intermediate and final host tissues, and analysing avian and mammalian muscle, blood, liver, spleen, lung, CNS, and intestine. Predator excrement and environmental samples, including water, soil and feed samples, will also be examined. PCR-RFLP, and microsatellite marker assays will be used to identify genotypes of important pathogens. Genetic testing will use nuclear, mitochondrial, and apicoplast DNA markers. Population genetic studies will be based on DNA sequencing of neutral loci and genotyping of microsatellites. Loci responsible for adaptive polymorphism will also be analysed in intraspecific genetic diversity studies. Data will be processed using statistical and specific genetic variability assessment programmes and databases.
Through the Programme, and based on the latest primary research findings, the NRC experts will focus on biodiversity conservation, population and habitat assessment and restoration based on the latest essential research findings. The results will be significant in identifying mechanisms for the stability of ecosystems. In addition, the research will reveal new interspecific relationships and explain the impact and importance of biodiversity on ecosystem sustainability in the context of increasing pollution, climate change and the spread of parasitic pathogens and pests.

There is a sufficient number of qualified researchers to carry out the planned research and obtain the intended results – 15 chief researchers, 25 senior researchers, 16 researchers, 4 junior researchers, 20 specialists. It is also expected that 23 doctoral students will participate in the implementation of the Programme.

5. Research stages and their characteristics

Five stages of the research planned for the Programme in 2022–2026 are envisaged:

Stage I (2022)

• Studies on the diversity, distribution and change of cyanobacterial species and bacteriophages in Lithuanian freshwater aquatic ecosystems affected by eutrophication.
• Estimation of the rate of spread of alien species of the genus Solidago. Development of a structural model for the national database of wild relatives of cultivated plants in accordance with the principles approved by the ECPGR programme and the data exchange format.
• Studies on the diversity of natural habitats of anthropogenic origin and the reproductive properties of their constituent components.
• Preliminary studies on mycobiota in coniferous forests and hornbeam-woods and freshwaters, in particular on macromycetes of the family Tricholomataceae, micromycetes of the family Mycosphaerellaceae and on fungal-like organisms (stramenophyllums or oomycetes) found in freshwater fish.
• Inventory of biological diversity of arthropod organisms and search for new organisms for science, identification of new trophic relationships, monitoring of propagating and alien arthropod species, studies on soil community structure, identification of changes in the development and significance of blood-sucking bivalve insects in the distribution of pathogens, development of endobionic diagnostic tools for Nepticulidae, completion and digitisation of the scientific insect collection.
• Study on long-term dynamics of demographic characteristics of populations of model species of birds of prey. Summary of ecological research on breeding and migrating duck species in Lithuania and Eastern Europe. Studies on the specifics of food habitats for foraging birds in the agro-landscape and identification of threats in the changing anthropogenic landscape. Studies on the migration phenology of Great Reed Warbler (Acrocephalus arundinaceus), assessing the influence of age and sex.
• Influence of road deaths of ungulates (elk, red deer and roe deer) on population abundance.
• Studies on the seasonal dynamics of small mammalian parasite communities in the context of climate change. Determination of the role of dormice in the diet of different systematic and ecological groups of predators (mammals, birds, reptiles). Evaluation of metal accumulation in small mammals in commercial gardens. Summary of small mammal ecology studies in large cormorant colonies.
• Collection of parasites (haemosporidian protists and helminths) in natural habitats and experimental studies of model eukaryotic parasite organisms. Introduction and application of new molecular methods for the analysis of genetic diversity, diagnostics, phylogeny and phylogeography of parasites of different taxonomic groups. Experimental study of sporogony in blood-sucking insects. New carriers of haemosporidian parasites and ornithophilic species of blood-sucking bivalves will be identified. Investigation of avian blood samples and internal organs to identify new to science species of blood parasites and the pathologies caused by these parasites. Experimental work to determine the peculiarities of parasite development, microbial composition and changes in healthy and malaria-infected hosts.
• Helminth specificity of hosts and life cycle studies based on molecular phylogenetic analysis. Studies on intermediate hosts in helminth communities assess biodiversity status and dynamics of selected freshwater ecosystems.
• Collection of intermediate and final host tissues and environmental samples in the wild and experimental studies on model organisms to determine the prevalence of parasitic protozoa of the genus Sarcocystis in natural populations. Selection of diagnostic markers for research on animals and their parasites, development of species-specific PCR primers. Development of test methods including preparation of biological samples, isolation of parasitic microscopic cysts, extraction of DNA from sarcocysts, suspension of sporocysts from water and soil samples, optimisation of PCR conditions, sequencing, and data analysis.

Stage II (2023)

• Investigations of the biology, morphology and ecology of blue-green algae and microalgae genotypes and species in natural and experimental conditions. Studies on the seasonal and spatial distribution of planktonic bacteriophages in Lithuanian lakes and the Curonian Lagoon.
• Evaluation of antioxidant activity and its marker potential in Solidago raw material. Evaluation of the influence of growth regulators (according to the revised scheme) on the morphometric, anatomical, physiological and biochemical parameters of T. × citriodorus plants. The organisation of a network of providers of data on the prevalence of wild relatives of cultivated plants, data collection and processing according to ECPGR (European Cooperative Programme for Plant Genetic Resources) approved principles and data exchange format, testing of national database model, submission of data to the European catalogue EURISCO (European Search Catalogue for Plant Genetic Resources).
• Analysis of the study of relevant target species of medicinal plants.
• Identification of the interaction of native and alien plants in the habitats of natural and anthropogenic origin.
• Surveys of terrestrial and underground fungal communities in coniferous forests and hornbeam woods according to the age and structure of the forest. Determining the prevalence and state of rare Europe fungi in Lithuania. Detection of stramenopilic fungi (oomycetes) – fish saprolegnia pathogens – found in fresh Lithuanian waters.
• Identification of interactions between biological diversity of arthropods, trophic analysis of endobionic organisms and elucidation of new nutritional relationships, monitoring of alien and protected arthropod species, comprehensive studies on pedobiont communities, documentation and description of new Diptera and Lepidoptera species, development and demonstration of innovative endobionic diagnostic tools for Nepticulidae for the consumer, testing of blood-sucking bivalve insects as pathogen vectors, adaptation of the database of arthropod organisms to public needs, further completion and digitisation of the scientific collection of insects.
• Investigations of the dependence of the distribution of model species of birds of prey in ecosystems on the structure of the environment, biotic and anthropogenic factors. Studies and analysis of species diversity and dynamics of indicator forest and agro-landscape bird species (Columbidae, Turdidae). Evaluation and analysis of seasonal migration characteristics of birds under the influence of biotic and abiotic factors and changes in migration characteristics in the age gradient of birds. Analysis of the prevalence of blood parasites of bearded reedling (Panurus biarmicus) in different European populations.
• Studies of model species – the common duckweed, fish – the common perch, roach, eel and re-introduced species – the Atlantic sturgeon and capercaillies using neutral and adaptive polymorphic markers to determine the population genetic structure of the studied species, to identify evolutionarily significant and environmentally important management units.
• Use of neural network models to predict road deaths of ungulates. Anthropogenic studies on small mammalian parasite communities. Peculiarities of habitat selection of the European edible/fat dormouse in Lithuania, outside the northern range of the European beech, and assessment of the anthropogenic impact on habitat status. Influence of microhabitats on the spatial distribution of species in the small mammal community. Analysis of the results of the use of surveillance cameras to assess the prevalence of invasive mammals.
• Collection of research material in nature and experimental research of model eukaryotic parasite organisms to achieve the programme’s objectives. Identifying previously undetected organisms and species new to science in Lithuania and the Palearctic by determining their systematic position and molecular diagnostic markers. It is planned to gain new knowledge about the development patterns of the stages of haemosporidian parasites in birds (sparrows and predators) and to elucidate the mechanisms of pathogenesis of infected internal organs.
• Identification of new haemosporidian parasites and helminthic vectors. Investigating the effect of temperature on haemosporidian sporogony using blood-sucking insects and experimental birds. Detection of links between malaria agents and microbiota, studies on changes in bacterial populations in vectors and effects on the development of malaria parasites.
• Based on morphological, karyological and DNA sequence studies and comparative analysis, it is planned to determine the systematic position and phylogenetic relationships of helminth larvae of unknown taxonomic position (with only provisional names) and to link them with adult forms parasitising the final hosts. It is planned to gain new knowledge about the specificity of helminths for hosts and life cycles. Assessment of the state of biodiversity and its dynamics in freshwater ecosystems based on studies of intermediate hosts in helminth communities.
• Carrying out morphometric, histopathological and molecular studies of parasitic protozoa found in intermediate and final hosts it is planned to gain new knowledge about distribution, intraspecific and interspecific genetic variability and life cycles of representatives of Sarcocystidae family.

Stage III (2024)

• Investigations of the peculiarities of the change of blue-green algae and microalgae communities, their functional potential and significance for the stability of freshwater communities under climate change and increasing anthropogenic impact. Determining changes in bacteriophage activity using virus-host systems and assessing the dependence of infections on environmental conditions in natural aquatic ecosystems.
• Collection of plant raw material samples of Achillea species and determination of species and intraspecific differences in the composition of phenolic compounds. Study on the impact of alien species of the genus Solidago on the species diversity of plant communities (2024–2025). Evaluation of the influence of growth regulators (according to the revised scheme) on the morphometric, anatomical, physiological and biochemical parameters of T. × citriodorus plants. Analysis of the distribution of specific bioactive substances in the selected taxonomic group of plants.
• Evaluation of the importance of natural and anthropogenic factors for expressing functional properties of vegetation components.
• Research on the diversity of fungal species in rare forest habitats. Initial analysis of citizen science data related to fungal diversity. Studies on the diversity of species of fungi-like organisms (oomycetes) and the composition of communities in some natural aquatic ecosystems.
• Integrated biotaxonomy studies (including new species for scientific purposes) to provide qualitatively new knowledge on biodiversity and interactions, continuous monitoring of alien arthropod species, new data on the structure of pedobiont communities, completion and presentation of endobionic diagnostic tools for the user, complex blood vessels research on insects as pathogen vectors, filling in the database of arthropod organisms, filling in and digitisation of the scientific collection of insects.

• Disclosure of the impact of legislative and institutional changes in Lithuania on managing red deer and roe deer populations. Studies on the parasitic infections of predatory mammals and their transmission to humans and domestic animals. Analysis of changes in mortality/survival of the hazel dormouse population and their determinants. Investigations of infection of small mammals with Sarcocystis and other pathogens parasitising them.
• Investigations of the identification, genetic diversity and distribution of parasitic protozoa, studies of the population genetic structure of naturally occurring species typical of the Baltic Region and introduced species, analysis of data, and publication of results.
• Complex study of space use in various stages of the annual cycle of model bird species (predators, storks, cranes) and identification of threats in the conditions of global change and anthropogenic impact. Investing Sarcocystis and other pathogenic organisms in birds and analysing the genetic diversity of populations of indicator bird species. Determining the influence of the width of the habitats of model sparrow bird species on their energy needs and the change of isotopic niches.
• It is planned to summarise the life cycle studies of parasites (protists and helminths) using experiments and molecular markers. Comparative and phylogenetic analysis of molecular data. Identification and more detailed studies of problematic parasite species and species complexes. Experiments are planned to determine whether different host habitats are an ecological barrier to transmitting haemosporidian parasites. Detection and experimental investigation of blood parasite vectors (haemosporidian parasites and trypanosomes) will be performed.
• Systematic revisions of model species of parasites using morphological, ecological and molecular data interfaces will be developed. We will seek to research and analyse the diversity of species of hidden (secondary) model organisms. We will summarise the ongoing and ongoing new virulence studies of the parasites to gain new knowledge about the effects on the host and the interactions between these parasites. It is planned to study and re-evaluate the diversity of suckers in selected species of freshwater molluscs and helminth communities in fish populations to identify changes related to climate change and anthropogenic activities.
Stage IV (2025)
• Identification of the importance of interactions between cyanobacteria and microalgae species with other aquatic organisms for the functioning and sustainability of the ecosystem in the context of increasing pollution and climate change. Investigations of the dependence of biogeochemical processes on bacteriophage infection and lysis under experimental conditions.
• Evaluation of differences in the quantitative and qualitative composition of plant raw materials of Achillea species under changing environmental conditions, evaluating raw materials collected in Lithuania and other geographical regions. Determining the Indicative Value of Correlations between Terpene Compounds and Changes in Environmental Conditions. Installation and implementation of T. puleigioides fertilisation experiment in the field collection, evaluating the influence of Na and S fertilisation on aboveground biomass, quantitative and qualitative composition of essential oils and anatomical structures (size and density of germinal and important oil glands). Overview of relevant methods used in research on medicinal plants.
• Determining the consequences of anthropogenic activities on vegetation functioning and highlighting the directions of changes in its main components.
• Identification of the diversity of fungal species in rare forest habitats. Analysis of citizen science data related to fungal diversity Assessment of species diversity and community composition of fungal-like organisms (oomycetes) in some natural freshwater ecosystems.
• Comprehensive morphological, phylogenetic and ecological studies of insects, including dicotyledons and specialised phytophagous insects, continuous monitoring of alien arthropod species, collection of additional data on the structure of pedobiont communities and collection of blood-sucking bivalve insects, digitalisation of the database of arthropods.

• Analysis of long-term structural changes in the populations of indicator bird species (Anatidae, Columbidae, Turdidae). Investigation of the dependence of the complex use of space on the characteristics of individual model species of birds of prey. Studies and identification of threats to the use of bird habitats during the winter in the context of global change and anthropogenic impacts. Analysis of the factors influencing the success of the great reed warbler breeding, the influence of the course of breeding season on migration.
• Quantitative assessment of road deaths of semi-aquatic mammals (otter, mink, beaver, muskrat). Studies on parasitic infections in ungulates in the context of anthropogenic effects. Determination of changes in the population structure of the hazel dormouse in the conditions of long-term reduced population abundance. Studies of differences in the nutritional niche of small mammals depending on habitat transformation. Comparison of the results of the surveillance chamber and ungulate accounting transects method.
• Diagnosis of parasites from environmental samples. Investigations of secondary genetic variability in animals using adaptive loci affected by natural selection. Peculiarities of the formation of the population genetic structure of the model and re-introduced species and their importance in the context of biodiversity conservation.
• Identification of vector species involved in sporogony associated with the transmission of haemosporidia in birds of prey and sparrows. Investigations into the exo-erythrocyte development and pathology of avian malaria parasites and related haemosporidians in European birds of prey and sparrows. Identifying and associating helminthic larval forms with adults and determining life cycle specificity and host range in the studied ecosystems based on comparative analysis of newly acquired DNA sequences deposited with the Gene Bank (USA).

Stage V (2026)

• Evaluation of the potential of functional adaptations of blue-green algae and microalgae in the conditions of change of aquatic ecosystems and their application to the conservation or restoration of biological diversity. Assessing the effects of viruses on the diversity of functional food webs.
• Determination of biological activity and antioxidant efficiency of Achillea species in model free radical systems and evaluating their potential significance for the use of plant raw materials. Execution of T. puleigioides fertilisation experiment in a field collection, assessing the influence of Na and S fertilisation on aboveground biomass, quantitative and qualitative composition of essential oils and anatomical structures (size and density of glandular epithelium and important oil glands). Statistical analysis and processing of collected data. Analysis of in situ achievements in research and storage of medicinal plants.
• Evaluation of the effectiveness of active nature conservation and simulation measures for vegetation components’ diversity and functional properties.
• A final assessment of the diversity of fungi in some forest ecosystems, focusing on the status of rare and protected species and providing recommendations for their protection. Diversity assessments of freshwater stramenopilic fungi (oomycetes). Evaluating citizensʼ scientific data on fungal diversity by identifying gaps and emerging issues.
• Summary of the results of the inventory of biological diversity of arthropods, phylogeny and diagnosis of new organisms, interpretation of trophic analysis, a summary of multiannual monitoring of alien, invasive, protected arthropod species, interpretation of studies on the structure and condition of pedobiont communities in a changing environment, a summary of new data on changes in the development and role of blood-sucking insects in pathogen distribution, final digitisation of the scientific insect collection.
• Analysis and forecast of the impact of climate change and anthropogenic factors on the populations of indicator bird species (Anatidae, Columbidae, Turdidae). Investigation of the substantiation of the management of threats relevant to the viable functioning of populations of model species of birds of prey. Establishment of a model for the use of bird habitats in the annual cycle, identification of threats, analysis and forecast of their impact on bird populations, and scientific substantiation of threat management. Evaluation of phenological parameters of birds and their long-term changes.
• Study on the dynamics of perennial road deaths of small and medium-sized predators (fox, mangrove, marten, ferret, etc.). Evaluation of parasitic infections in predatory mammals and their transmission to humans and domestic animals. Analysis of the influence of climate change on the population of the hazel dormouse. Evaluation of boar abundance in Lithuanian regions after measures on applying the OMC (Olen Method of Coordination).
• Summary of the obtained results on the prevalence of pathogenic parasites, emphasising food safety and prevention of infection, preparing recommendations to the institutions of the Republic of Lithuania. The systematisation of molecular data generated by species identification of parasitic protists and genetic structure studies of model and introduced species populations and creation of a molecular database. Identification of changes in the genetic structure of vulnerable and model species affected by intensive anthropogenic activities and global climate change, a summary of research results, publication, and preparation of recommendations for preventive measures for the conservation of biological diversity.
• Preparation of data obtained during the implementation of the Programme for publication and preparation of manuscripts for publication. Summary of the work performed and presentation of the scientific basis for the state strategic documents and recommendations on the conservation of biological diversity. It is planned to provide information about reference collections of parasitic organisms and molecular diagnostic markers on the NRC website and create open-access databases for these data.
Detailed implementation plan and indicative financial allocation

Stage I (2022)
Preparation of new projects to ensure the implementation of the Programme objectives, joining international projects. Updating and developing new databases, reviewing the latest research, implementing and adapting new methodologies, setting up tests, sampling and analysis, and collecting and analysing data. Participation in conferences and seminars; reading reports and lectures. Preparation of publications.
The planned need for the stage is 20% of the funds allocated for the Programme.

Stage II (2023)
Conducting tests and observations, sampling and analysis, data collection and analysis, modelling; preparing strategies, programmes, methodologies, methods, publications, and publication of results. Participation in conferences and seminars; reading reports and lectures. Preparation of publications.
The planned need for the stage is 20% of the funds allocated for the Programme.

Stage III (2024)
Summary of interim results, preparation of forecasts, updated strategies, programmes, recommendations, methods and methodologies, submission of the interim report, publication of results, adjustment of planned research. Participation in conferences and seminars; reading reports and lectures. Preparation of publications.
The planned need for the stage is 20% of the funds allocated for the Programme.

Stage IV (2025)
Conducting tests and observations, sampling and analysis, data collection and analysis, modelling; preparing strategies, programmes, methodologies, methods, publications, and publication of results. Participation in conferences and seminars; reading reports and lectures. Preparation of publications.
The planned need for the stage is 20% of the funds allocated for the Programme.

Stage V (2026)
Summary of the results generated during the implementation of the Programme, preparation of forecasts, updated strategies, programmes, recommendations, methods and methodologies, publication of results, and adjustment of planned research. Participation in conferences and seminars; reading reports and lectures. Preparation of publications. Preparation and submission of the final report.
The planned need for the stage is 20% of the funds allocated for the Programme.

7. Expected results

Research on algae, blue-green algae and viruses will assess the diversity of native and alien species, interspecific competition and biotic relationships, patterns of adaptability to different environmental conditions, importance for ecosystem functioning and stability, and conservation and restoration of biodiversity. The research results will be used to comprehensively assess biodiversity, the state and health of aquatic ecosystems, and predict their changes.
Experimental research of pure algae and blue-green algae cultures carried out by NRC AMEL will provide new knowledge about the plasticity and biological features of the populations of species common in Lithuanian freshwaters, competitiveness in communities and relevance in food webs, i.e. characteristics that determine the stable functioning of the ecosystem. Modelling tools will be used to assess trends in the natural environment, emerging threats, and the potential to adapt aquatic ecosystems to global climate change. The AMEL collection of pure cultures of blue-green algae and microalgae is maintained and expanded. The database of the diversity of these groups of microorganisms in Lithuania is updated. Scientific information will be adapted to the needs of society to change consumer attitudes and increase their involvement in the conservation of natural ecosystems.
Researching the diversity and functional potential of prokaryotic and eukaryotic aquatic microorganisms in the context of water habitat change will provide guidance material based on new fundamental scientific knowledge relevant to the state, preparing relevant state strategic documents on biodiversity conservation, sustainable use and restoration, eutrophication aquatic ecosystems, measures for their ecological restoration and the development of “blooms” management strategy. Specialists of the Center for Health Education and Diseases Prevention of the Republic of Lithuania will be consulted on assessing the impact of toxic water “blooms” on human health in the implementation of Directive 2006/7 / EC of the European Parliament and of the Council concerning bathing water quality.
Researchers from the Laboratory of Algology and Microorganism Ecology (AMEL) of the NRC Institute of Botany, together with co-authors, plan to publish 16 CA WoS papers (in publications referenced and indexed in the “Clarivate Analytics Web of Science” database) and 6 popular science publications.
It is planned to prepare and defend 3 doctoral dissertations at the AMEL.
After the implementation of the Programme, a functional structure of the national database of wild relatives of cultivated plants in accordance with European standards will be created, and recommendations for the conservation and rational use of medicinal plant populations will be prepared.
Researchers from the Laboratory of Economic Botany (EBL) of the NRC Institute of Botany plan to publish 12 CA WoS papers on the topic of the Programme.
It is planned to prepare and defend 1 doctoral dissertation at the EBL.
It is planned to supplement the database of Lithuanian vegetation (EU-LT-001) with at least 500 phytosociological descriptions; the Lithuanian database of alien plants (INVA) – not less than 5 thousand records.
Revision of the NRC Institute of Botany Herbarium (BILAS) collections – not less than 10 thousand herbarium specimens.
Researchers from the Laboratory of Flora and Geobotany (FGL) of the NRC Institute of Botany, together with co-authors, plan to publish 14 CA WoS papers, 13 other scientific publications, prepare 21 reports at scientific conferences, disseminate scientific results in 11 popular articles, formalised interviews, and more.
It is planned to prepare and defend 3 doctoral dissertations at the FGL.
New knowledge on the diversity of fungal and fungal-like species, the specificity of their communities, and the number of different species of fungi will be clarified; new species will be registered for individual sites, reducing the currently declared severe lack of data on biodiversity, which is an obstacle to practical environmental issues. In addition, new knowledge of the links between fungal communities and individual habitats, their condition and changes will ensure better protection and contribute to the better designation of new protected areas and the further monitoring and assessment of their condition.
The analysis of the impact of CivicScience on the accumulation of knowledge about fungal diversity and the identification of existing gaps, and the development of measures to address them will at least partially alleviate the lack of data due to the critically declining number of biodiversity professionals in the country.
Researchers from the Laboratory of Mycology (ML) of the NRC Institute of Botany, together with co-authors, on the topic of the Programme, plan to publish 8 CA WoS papers, about 20 popular articles, and present reports at four international conferences. The data will also be used in COST activity CA17122 (ALIEN CSI). It is planned to prepare and defend 1 doctoral dissertation at the ML.
The database of arthropod organisms and the scientific collection of insects will be supplemented. Innovative diagnostic tools for Microlepidoptera mining in plant assimilation tissues will be developed and presented to the modern consumer in various activities. New knowledge will be gained about the diversity, taxonomy, phylogeny, trophic relationships of insects and other arthropods, data on the peculiarities of their development and their role in the spread of pathogens in a changing environment.
Researchers from the Laboratory of Entomology (EL) of the NRC Institute of Ecology, together with co-authors, plan to publish 30 CA WoS papers on the topic, 1 monograph will be prepared.
It is planned to prepare and defend 4 doctoral dissertations at the EL.
New research on the abundance and distribution of bird populations in different habitats, their changes, genetic diversity and their role in diverse terrestrial and aquatic ecosystems will allow for more effective protection and management and forecasting of bird populations. The results of the genetic testing of birds will be deposited in the International Gene Bank (USA) for open access. Data from remote bird tracking will be published on the international open-access animal telemetry research portal Movebank. The general public will be informed about research results through publications promoting science and disseminating information on social networks and in the media. The new knowledge will be used to teach university students and doctoral students.
Researchers from the Laboratory of Avian Ecology (AEL) of the NRC Institute of Ecology, together with co-authors on the topic of the Programme, plan to publish 15 CA WoS papers and 10 popular science articles.
It is expected to prepare and defend 2 doctoral dissertations at the AEL.
Mammalian research will assess changes in their populations over time and in the geographical gradient, the significance of these changes for the protection and sustainable use of mammals, and the impact of small mammals on the transmission of human pathogens to natural ecosystems. Recommendations will be provided to reduce road deaths of mammals, and hunting limits for ungulates and predatory mammals will be clarified.
Researchers of the Laboratory of Mammalian Ecology (MEL) of the NRC Institute of Ecology, and co-authors, on the topic of the Programme, plan to publish 28 CA WoS papers and make 25 reports at conferences.
It is planned to prepare and defend 3 doctoral dissertations at the MEL.
After the implementation of the Programme, the prevalence, abundance, morphological characteristics, species composition, host specificity, and pathogenicity of the studied parasitic protozoa of birds and mammals will be determined, and phylogenetic relationships revealed. The genetic diversity, genetic structure, related population size, effective population size of the studied model and introduced species will be assessed, and phylogeographic characteristics and historical demographic dynamics of the populations will be revealed.
Diagnostic techniques for the tested parasites will be refined and developed in various samples: muscle, brain and blood samples from intermediate hosts, small intestine, and environmental samples. The study of the different stages of the life cycle of parasites in hosts and the environment will provide new fundamental knowledge on biodiversity, quantitative parameters of infections, the phenomenon of host-co-evolution of parasites and practical knowledge on the possible spread and prevention of infectious diseases. New species of Sarcocystis parasites are expected to be described. Units essential for the environmental management of the studied animal species will be identified, and genetic monitoring and anthropogenic impact assessment systems will be proposed.
Researchers from the Laboratory of Molecular Ecology (MEL) of the NRC Institute of Ecology, together with co-authors, on the topic of the Programme, plan to publish 25 CA WoS papers and make 15 reports at scientific conferences.
It is planned to prepare and defend 3 doctoral dissertations; it is also scheduled to train 10 students for the first and 5 for the second cycle at the MEL.
New knowledge will be gained on the diversity and life cycles of eukaryotic parasitic organisms (protists and helminths) and the virulence and epidemiology of the resulting diseases in the wild, as foreseen in the research stages. New DNA sequencing data will be deposited at the International Gene Bank (USA). The research will contribute to a better understanding of the spread of parasitic diseases and the search for preventive measures. The acquired new knowledge and collected material will serve for long-term research on Nordic biodiversity in the context of climate change. The obtained data will significantly supplement the knowledge about the diversity of parasite species in Lithuania and the world, life cycles, dispersal pathways, specificity for hosts, virulence, phylogeographic relationships and circulation in ecosystems.
The new knowledge will deepen the understanding of ecosystem biodiversity, help implement targeted conservation programmes and anticipate potential ecosystem changes. In addition, the results of the experiments will provide new knowledge about the developmental characteristics of parasitic organisms. They may improve the diagnosis of parasitic diseases and develop prevention measures in a changing environment.
The systematics of parasitic organisms (protists and helminths) of individual taxa will be reviewed and improved, and species new to science will be described and registered.
It is planned to develop NRC open-access databases to characterise helminth and haemosporidian parasite species. Deposit DNA sequencing information in open access (International Gene Bank, USA).
Researchers from the P. B. Šivickis Laboratory of Parasitology (PL) of the NRC Institute of Ecology, together with co-authors, plan to publish about 20 CA WoS papers on the topic of the Programme.
It is planned to prepare and defend 3 doctoral dissertations at the PL by participating in the educational process, to teach students, postgraduate students and postdoctoral fellows to apply modern and traditional research methods and to recognise parasites, understand their biology and impact on ecosystems. It is planned to train 10 students for the first and 5 for the second cycle.
During the implementation of the Programme it is planned:
• To make recommendations for selecting gene conservation areas for the conservation of wild relatives of cultivated plants and integration into the European network.
• To provide subject matter information to the Ministry of Environment of the Republic of Lithuania on fungi conservation issues.
• To suggest updating the Global Crop Diversity Trust portal Crop Wild Relatives.
• To maintain databases on the abundance, use and road deaths of wild mammal populations.
• To recommend wild mammal limits to the Ministry of Environment of the Republic of Lithuania.
• To provide recommendations to the Ministry of Environment and the Lithuanian Road Administration under the Ministry of Transport and Communications on protecting mammals and improving traffic safety.
• To prepare recommendations for the protection of birds in Lithuania and the Eastern Baltic Region and to form the basis for the protection of birds in the changing climate in Lithuania (threat analysis, forecasting, management, preventive measures and methods, new protection policy and strategy).
• To prepare scientific recommendations for the development of a strategy for the management of problematic phytopathogenic microorganisms.
• To supplement and revise the collections of the Herbarium of the Institute of Botany (BILAS) by including the Suvalkija Herbarium (HSUD) transferred to the NRC by the decision of the Government of Lithuania.
• To supplement and maintain living microorganisms collection and update the electronic database.
• To develop a scale for assessing aquatic ecosystems to assess the developmental processes of blue-green species that cause aquatic “blooms” and synthesise toxins.
• To make recommendations to the Center for Health Education and Diseases Prevention on assessing the effects of toxic aquatic “blooms” on human health in the implementation of Directive 2006/7/EC of the European Parliament and of the Council concerning the quality of bathing water.
• To prepare and update methodological tools for I, II, III cycle study programmes.
• To provide factual information to the Ministry of Environment on the development and improvement of biota resources legislation.
• To prepare a new edition of the Lithuanian Red Data Book.
The main results of the research are expected to be published in more than 150 international journals with a citation rate listed in the WoS database.
After implementing the Programme, at least 23 doctoral dissertations will be prepared and defended.
8. Means of dissemination of results
• Depositing new information in the open access International Gene Bank (USA), Zoo Bank and other databases (Queensland Museum, Australia; US National Museum; Geneva Museum of Natural History, Switzerland, etc.).
• The research results on the topic of the Programme will be presented to the scientific community in Lithuania and at international conferences and world congresses.
• Lectures for students and schoolchildren, presenting relevant information to the Lithuanian society using mass media.
• Publications promoting science are being prepared.
• The results will be presented at Lithuanian and international scientific events.
• The acquired new knowledge will be used in the teaching of higher education subjects “Protistology”, “Biodiversity”, “Ecology and Environmental Science”, “Invertebrate Zoology”, “Parasitology”.

9. Funds allocated for the implementation of the Programme 9.553.000.00 EUR (nine million five hundred and fifty-three thousand euros):

10. The programme manager is posted on the Centreʼs website www.gamtostyrimai.lt