EFFUSE – River Landscape, Volume 1: Scientific Education for Everybody

PAVOL JOZEF ŠAFÁRIK UNIVERSITY IN KOŠICE Faculty of Science EFFUSE - RIVER LANDSCAPE Volume 1 Scientific Education for Everybody EFFUSE team of authors Košice 2025 EFFUSE – River Landscape, Volume 1: Scientific Education for Everybody Educational text Authors: RNDr. Ivana Slepáková, PhD. Department of Microbiology, Faculty of Science, PJ Šafárik University in Košice doc. RNDr. Andrej Mock, PhD. Department of Zoology, Faculty of Science, PJ Šafárik University in Košice RNDr. Peter Ľuptáčik, PhD. Department of Zoology, Faculty of Science, PJ Šafárik University in Košice RNDr. Natália Pipová, PhD. Department of Animal Physiology, Faculty of Science, PJ Šafárik University in Košice RNDr. Monika Balogová, PhD. Department of Zoology, Faculty of Science, PJ Šafárik University in Košice RNDr. Mariana Kolesárová, PhD. Department of Microbiology, Faculty of Science, PJ Šafárik University in Košice RNDr. Lenka Maliničová, PhD. Department of Microbiology, Faculty of Science, PJ Šafárik University in Košice prof. RNDr. Martin Bačkor, DrSc. Department of Plant Biology, Faculty of Science, PJ Šafárik University in Košice This publication was produced with the financial support of the ENI CBC HUSKROUA program, through grant agreement HUSKROUA/1901/6.1/0075, with a financial contribution from the EU of 364,099.41€. doc. RNDr. Michal Goga, PhD. Department of Plant Biology, Faculty of Science, PJ Šafárik University in Košice RNDr. Dajana Kecsey PhD. Department of Plant Biology, Faculty of Science, PJ Šafárik University in Košice doc. RNDr. Yaroslava Hasynets, PhD. Department of Botany, Faculty of Biology, Uzhhorod National University doc. RNDr. Vladyslav Mirutenko. PhD Department of Entomology and Biodiversity Conservation, Faculty of Biology, Uzhhorod National University doc. RNDr. Roman Kish, PhD. Department of Botany, Faculty of Biology, Uzhhorod National University doc. RNDr. Mykhailo Vakerych, PhD. Department of Genetics, Plant Physiology and Microbiology, Faculty of Biology, Uzhhorod National University prof. RNDr. Maryna Kryvtsova, Dr.Sc. Department of Genetics, Plant Physiology and Microbiology, Faculty of Biology, Uzhhorod National University doc. RNDr. Fedir Kurtyak, PhD. Department of Zoology, Faculty of Biology, Uzhhorod National University doc. RNDr. Alexander Mateleshko, PhD. Department of Entomology and Biodiversity Conservation, Faculty of Biology, Uzhhorod National University Editor: RNDr. Beáta Valkay, PhD. This text is published under the Creative Commons 4.0 license - CC BY-NC-SA ("Attribution-NonCommercial-ShareAlike"). The authors are responsible for the professional and linguistic aspects of this publication. The manuscript has not undergone editorial or linguistic editing. Available at: www.unibook.upjs.sk Publication date: 21.10.2025 DOI: https://doi.org/10.33542/EFF-0449-1 ISBN 978-80-574-0449-1 (e-publication) CONTENTS Bioindicator species as a tool for water quality determination The importance of water for living organisms 6 Water in the region of transcarpathia 9 Water pollution 11 Bioindication as a method of environmental research 16 Assessment of the ecological status of water bodies/ watercourses and water quality by aquatic plants 25 Insects - bioindicators of water bodies 56 Fish as bioindicators of fresh waters 68 Amphibians as bioindicators of fresh waters 75 What do you need to know about water and its microbial composition to protect yourself from infections that are spread by water? 82 Significance of environmental upbringing and education 89 FOR EVERYBODY 6 THE IMPORTANCE OF WATER FOR LIVING ORGANISMS In terms of free water, the Earth is the most "water-rich" planet in the Solar System. The total volume of the hydrosphere exceeds 1454 million km3, of which almost 94% is occupied by the World Ocean, 4.12% by groundwater, 1.64% by glaciers and 0.0001% by river water. The volume of fresh water in the hydrosphere is generally 2% of its total volume, and taking into account the inaccessibility of the part of it that is preserved in glaciers, it is only 0.3% of the hydrosphere volume. For a large number of living organisms, especially in the early stages of the biosphere's development, water is the environment for the origin and development of all living things. Water in the biosphere is in constant motion, originating in the geological and biological cycles of substances. Water is life, it is the basis for the existence of life on the planet. Human civilization cannot exist without water, as people use water not only for drinking but also for sanitation and industrial and domestic needs. Water is used in many technological processes and is an integral part of many industrial and processing facilities. As for living beings, all metabolic processes in their bodies involve water. The more energetic these processes are, the greater the need for water is. A young growing organism has a higher water content than an adult. In a newborn, the amount of water reaches 75% of body weight. FOR EVERYBODY 7 Every living cell in the human body contains a healing aqueous solution of various nutrients. Water takes an active part in chemical reactions in the body, transports nutrients to each cell, removes toxins, slaggings and excess salts, and helps to lower blood pressure. Thanks to the properties of water as a solvent, blood and lymph provide an ideal medium for the most complex biochemical processes in the body. The body's ability to maintain a constant body temperature is largely due to the following three physical properties of water: • Water has a high capacity to store heat. Even cold-blooded animals are able to maintain a relatively constant body temperature in the face of short-term fluctuations in ambient temperature due to the physical properties of the fluid contained in their bodies; • Water has a high degree of thermal conductivity. This makes it easy to remove heat from deep parts of the body; • Water is constantly evaporating from the surface of the lungs and skin. This evaporation loses a considerable amount of heat, which is important for the processes of physical heat regulation. Adequate intake of water is one of the basic conditions for health. Drinking enough water is one of the best ways to prevent the formation of kidney stones. Water "lubricates" the joints, thereby acting as a cushion for the spinal cord, and also regulates body temperature and ensures skin elasticity. Water is essential for FOR EVERYBODY 8 normal digestion. Taking part in the metabolism, this unique liquid helps to reduce fat accumulation and reduce weight. Insufficient water intake disrupts the normal functioning of the body: fatigue and decreased performance, impaired digestion and assimilation of food, slowed biochemical reactions, increased blood viscosity, which creates conditions for blood clots, and disrupts the process of blood formation. Without water, it is impossible to regulate the body's heat exchange with the environment and maintain a constant body temperature. Since the brain is 75% water, relative dehydration causes the brain cells to experience severe stress. Dehydration negatively affects the most important functions of the body, weakening it and making it vulnerable to disease. FOR EVERYBODY 15 Water quality, as in focus, reflects the complexity of aquatic ecosystems and their abiotic and biotic components. It is the result of the functioning of aquatic ecosystems, primarily biota. At the same time, the water of watercourses and reservoirs is the only possible habitat for aquatic plants and animals. In view of this, as well as the availability of the modern Methodology for Environmental Assessment of Surface Water Quality by Relevant Criteria developed in Ukraine, which contains a system of environmental classifications of surface waters of land and estuaries, it is most appropriate to determine the environmental status of surface waters using this methodology. This methodology has been adopted as a valid interagency regulatory document and is mandatory for all agencies in organizing and carrying out state monitoring. The main role in water self-purification processes belongs to aquatic life - hydrobionts. The intensity and effectiveness of these processes depend on the state of aquatic communities and the state of hydrobiocenoses. Figure 3 The Uzh River within the city of Uzhhorod (V. Pliashechnyk). FOR EVERYBODY 16 BIOINDICATION AS A METHOD OF ENVIRONMENTAL RESEARCH Due to the profound transformation of the natural environment under the influence of anthropogenic impact, which has reached the global level in terms of its scale and is outpacing the impact of natural factors in terms of strength and speed, the problems of preserving the ecosystem and the biosphere as a whole are becoming more acute and urgent. Determination of biologically significant anthropogenic loads based on the reactions of living organisms and their communities to them is associated with bioindication. The importance of vegetation cover as an indicator of ecosystem health lies in the fact that it is very sensitive to changes in environmental factors. Urban ecosystems are most affected by human activity. Therefore, it is important to monitor the state of the environment and timely analyse the pollution of the city's territory. To some extent, bioindicative assessment can solve these issues. Monitoring systems based on the study of plant and animal behaviour make it possible to assess the biological effects of air pollution, their spatial distribution, and possible accumulation over large areas. Some species of plants and animals change their developmental characteristics (growth rate, flowering process, fruit formation, colour intensity, etc.) in response to various stimuli. FOR EVERYBODY 17 These properties have long been noticed by mankind and used for practical purposes. Due to the general environmentalisation of various scientific fields and human thinking in general, bioindication methods are increasingly used by modern scientists, including in environmental quality monitoring. Bioindication (Greek: bios - life, Latin: indiso - indicate) is an operational monitoring of the environment based on observations of the state and behaviour of biological objects (plants, animals, etc.). This method is becoming more and more common because indicator organisms have such advantages as: • summarise biologically relevant environmental data; • are capable of responding to short-term and volley releases of toxicants; • respond to the speed of changes in the environment; • indicate the places of accumulation of pollutants and their migration routes; • allow us to develop early assessments of the harmful effects of toxicants on humans and wildlife and to set permissible loads on ecosystems. Bioindication is used in environmental studies as a method of detecting anthropogenic pressure on biocenosis. The bioindicator method is based on the study of the impact of changing environmental factors on various characteristics of biological objects and systems. Biological systems or organisms that are most sensitive to the factors that are studied are chosen as bioindicators. Changes in the behaviour of the test object are assessed in comparison with control situations taken as a standard. For example, when assessing FOR EVERYBODY 18 the ecological state of surface waters, observations of the behaviour of daphnia, molluscs, and some fish species are used as bioindicators. A number of indicator plants respond to increased or decreased concentrations of microand macroelements in the soil. This phenomenon is used for preliminary soil assessment and identification of possible mineral prospects. One of the specific methods of environmental pollution monitoring is bioindication, the determination of the degree of pollution of geophysical environments using living organisms which are bioindicators. Living indicators should not be too sensitive and too resistant to pollution. They should have a sufficiently long life cycle. It is important that such organisms are widespread across the planet, and each species should be specific to a particular habitat. Lichens fully meet all these requirements. They react differently to pollution than higher plants. Long-term exposure to low concentrations of pollutants causes damage to lichens that does not disappear until their slates die. This is probably due to the fact that lichens regenerate their cells very slowly, while damaged tissues in higher plants regenerate quite quickly. Due to a number of biological features, lichens are good indicators of changes in the state of the environment in terms of its pollution with sulphur dioxide, fluorides, alkaline dust, and heavy metals. FOR EVERYBODY 19 Bioindication has a number of advantages over instrumental methods. It is highly efficient, does not require large expenditures and makes it possible to characterise the state of the environment over a long period of time. Environmental factors strictly determine which organisms can live in a certain place and which cannot. It is possible to use the inverse pattern and draw conclusions about the physical environment based on the state of the organism living in it. This is how the method of bioindication of the environment emerged, which is especially widely used in forest typology, phytocoenology, and to determine the level of air pollution using lichens (lichen indication), mosses (bioindication) or fungi (mycoindication). Thus, bioindicators are a group of individuals of the same species or grouping, the presence, quantity or intensity of development of which in a particular environment is an indicator of certain natural processes or environmental conditions. Figure 4 Bioindicator lichens. FOR EVERYBODY 20 Biological indication is widely used today to assess environmental pollution, which "removes" species of lower and higher plants and fauna that are unstable to pollution factors from their natural ecological niches. Bioindication has certain advantages as a method of obtaining direct information about changes in the state of biota under specific pollution conditions, but it should be combined with chemical and geophysical experiments to obtain not only qualitative but also quantitative data. Thus, due to the need for global monitoring, the use of indicator capabilities of biological objects is becoming increasingly important. Indicator plants are used both to detect individual pollutants and to monitor the general state of the environment. Basic principles of bioindication All biological systems - organisms, populations and biocenoses - have adapted to the complex of factors in a particular area in the course of their development. They have occupied a certain area within the biosphere, an ecological niche, in which they find optimal living conditions and can feed and reproduce normally. Each organism has a genetically determined, phylogenetically acquired, unique physiological tolerance range for each factor that affect it, within which this factor is suitable. If the factor is too low or too high, but not yet lethal, the organism is in physiological pessimism. In the examined area of the factor's intensity, which is particularly favourable for a given person, the body exists in physiological optimum conditions. FOR EVERYBODY 21 There are different forms of bioindication. If two identical reactions are caused by different anthropogenic factors, then we speak of nonspecific bioindication. If certain changes can be attributed to only one factor, we are talking about specific bioindication. If a bioindicator reacts with a significant deviation of vital index from the norm, it is a sensitive bioindicator. Accumulative bioindicators accumulate anthropogenic impact without rapid manifestations of deviation. Such significant accumulation of pollution, is gradually exceeding the normal level, and most often occurs at the level of ecophysiological or biocenotic processes. In nature, all types of bioindication are included in a chain of sequential reactions and processes. If an anthropogenic factor acts directly on a biological element, then we are talking about direct bioindication. But often bioindication becomes possible only after a change in state under the influence of other directly involved elements. In this case, we are dealing with indirect bioindication and a bioindicator. It is often preferably to detect the biological effect of an anthropogenic factor in advance in order to be able to influence this effect under certain conditions The presence of very bioindicators leads to early indication, when the reaction occurs at minimal doses in a short period of time and finishes in a short period of time and takes place at the site of the factors effect on elementary molecular and biochemical processes. FOR EVERYBODY 22 Six different types of sensitivity can be distinguished depending on development time of bioindication reactions: Type I: the bioindicator gives a single strong reaction after a certain time, during which it did not respond to the action (no effective level) and loses sensitivity (above the upper effective level). Type II: as in the first case, the reaction is immediate and strong, but lasts for some time after which it abruptly disappears. Type III: the bioindicator reacts from the moment of detection of the deviated action with the same intensity over a long period of time. Type IV: after an instant strong reaction, its cessation is observed, first rapidly, then more slowly. Type V: when a disturbed action occurs, a reaction begins, which is becoming more and more intense until it reaches a maximum, and then gradually stops. Type VI: the type V reaction is repeated many times; oscillation of bioindicator parameters occurs. Bioindication can be used at different levels of living organisation (macromolecule, cell, organ, organism, population, biocenosis). As the level of organisation of biological systems increases, so does their complexity, as their interrelationships with location factors become increasingly complex. FOR EVERYBODY 23 At the same time, bioindication at lower levels is dialectically included in bioindication at higher levels, acting in a new way. While direct and more often specific types of bioindication prevail at the lower levels of organisation of biological systems, indirect bioindication dominates at the higher levels. Due to the complexity of biological systems, only nonspecific bioindication is often possible. However, this is where the pathways to detecting complex stressors and thus to assessing the permissible loads on a complex ecosystem open up. Sometimes bioindication methods that are easily used at lower organisational levels become so complex in more complex systems that it becomes impossible to distinguish the impact of a factor. On the other hand, bioindicators that are detected at a higher organisational level are linked to corresponding changes at the previous levels. This pattern should be taken into account when looking for opportunities of early bioindication. Compared to individual organisms, ecosystems often respond to stressful impacts with a delay and in a highly altered form. According to the organisational levels of biological systems, different levels of bioindication can be identified: Level 1 biochemical and physiological reactions; Level 2 anatomical, morphological, biorhythmic and behavioural abnormalities; Level 3 floristic and faunal changes; Level 4 cenotic changes; Level 5 biogeocenotic changes; Level 6 changing landscapes. FOR EVERYBODY 24 There are two proper methods of bioindication: passive and active monitoring. In the first case, visible or invisible damage or abnormalities are studied in free-living organisms, which are signs of stress. In active monitoring, the same impacts are detected on test organisms that are under standard conditions in the researched area. There are four main requirements to consider when conducting bioindication: 1 the relative speed of the transaction. 2 obtaining sufficiently accurate and reproducible results. 3 the presence of objects used in bioindication, if possible in large quantities and with homogeneous properties. 4 error range compared to other testing methods is no more than 20%. FOR EVERYBODY 31 Determining water quality by aquatic plants According to the developed methods (Karpova et al., 2011; Maltsev, Karpova, and Zub, 2011), bioindication of the ecological status of a water body or watercourse by aquatic plants is carried out by assessment of: • species composition of aquatic plants in the reservoir/watercourse; • the number (abundance) of individuals of certain species; • the degree of development of individual species or aquatic plant communities/habitats (projected coverage - PC*); • the presence of individual indicator species and indicator groups; • the spatial distribution of the thickets on the water body. When surveying a water body, it is advisable to pay special attention to the most abundant (dominant) plant species and their communities, as they reflect the overall picture of the ecological status of the water body/watercourse. However, it is also necessary to take into account species that are not numerous or even rare, which, in the case of periodic observations, i.e. monitoring, may indicate the direction of processes occurring in the water body. An indicator of the ecological status of a water body can be not only the species composition of aquatic plants, but also the abundance of species and the specifics of the spatial distribution of aquatic vegetation, etc. The easiest way to do this is to study the species composition of aquatic plant communities in a particular section of a watercourse or reservoir. Determining the species composition involves compiling a complete FOR EVERYBODY 32 list of plant species, which includes all species that occur in the area under study. Several recommendations are offered for the practical implementation of the description of aquatic vegetation and the compilation of a general list of aquatic plant species. To determine the water quality of a water body/watercourse in general, it is necessary to select the most typical sites. For a comprehensive assessment of water quality, one should try to cover a variety of habitats in the water body, e.g., for a river - riffles, rifts, bays. If the goal is to study the impact of a single source of pollution on water quality, then sites should be selected upstream and downstream of the source of pollution. The length of the surveyed areas depends on the size of the water body. For example, for a small river or pond, it is necessary to survey 50 m of the coastline in 2-3 sections and make 3-4 descriptions. For a mediumsized river and a small pond (lake), 100 m of coastline (5-8 descriptions). The survey should be carried out both from the shore and from the pond (knee-deep or from a boat), always looking inside the thickets for interesting findings. Be sure to examine all possible belts and layers of aquatic vegetation: the upper surface, the water surface itself, and its thickness. It is important to note that during the surveys and collection of material, it is not allowed to collect or disturb communities of protected species - those included in the Red Data Book of Ukraine, international and regional red lists. After compiling a general list of plants, indicator species and indicator groups are identified among them, which will be used in further work. FOR EVERYBODY 33 Assessment of the ecological status of a water body/watercourse and determination of water quality by indicator species of higher aquatic plants (Macrophyte Index (MI)) Based on the patterns of change in the species composition of aquatic plants in a water body/watercourse, their abundance and spatial distribution, which occurs due to changes in the hydrological regime, trophic state, increased pollution and deterioration of water quality (mainly due to an increase in the concentration of nutrients and trophic level), the state of the water body and water quality can be determined. The hydrological regime and trophic state of a water body/watercourse is determined by the specific species composition, primarily by sensitive indicator species, taking into account the phytoindicative ecological scales established for most macrophyte species (Didukh, 2011). A preliminary assessment of the ecological status of a water body/watercourse or a separate site/area and determination of water quality can be made by the presence of indicator species through the establishment of an aquatic plant index or Macrophytic (biotic) index (MI), the methodology for determining which was developed by domestic hydrobiologists and botanists (Karpova, Zub, Melnychuk, and Protsiv, 2011; Maltsev, Karpova, and Zub, 2011). The methodology is based on the natural change in indicator groups of aquatic plant species that occurs in a water body in response to increasing pollution and deteriorating water quality. FOR EVERYBODY 34 By determining the presence of species of a certain indicator group in a water body/watercourse and calculating the total number of aquatic plant species growing in it, it is possible to obtain the Macrophyte Index (MI), which will be an indicator of the ecological status of the water body/watercourse and water quality. The Macrophyte Index is determined using a special table (Table 1). After examining the water body/watercourse section, we identify aquatic plants and compile a general list of them. Then, to assess the condition of the water body, the table should be used to calculate the approximate number of all aquatic plants found in the water body under assessment, taking into account all species found in the water body, not just those listed in the table. We use the table to determine which indicator species occur in the study area. First, we look for species sensitive to pollution in our list, which are listed in the first (second) rows of the table. If there are such species from the first (second) indicator group, the other rows below are not considered, and the assessment is carried out within this group. If these species are not in our description, we move on to the next line and look for species from the next indicator group, ignoring the rest, etc. Next, within this line, we set the total number of species of this group in the study water body and at the intersection with the column with the corresponding number of species, we find the value of the Macrophyte Index in the table. FOR EVERYBODY 35 Table 1 Determination of water quality by aquatic plants through the establishment of the Macrophyte Index Indicator species Total number of presented species <5 6-10 >11 Isoëtes lacustris, water moss (Fontinalis) (Fig. 5.1), algae Chara sp. (Fig. 5.2), Myriophyllum alterniflorum 10 9 - Potamogeton complex (except Stuckenia pectinata), Persicaria amphibia (Fig. 5.3), Ranunculus aquatilis 9 8 7 Potamogeton complex (Potamogeton perfoliatus (Fig. 5.4), P. lucens, P. crispus and Potamogeton with floating leaves (Fig. 6.3), Nuphar lutea (Fig. 7.6), Elodea canadensis, Sagittaria sagittifolia (Fig. 7.5), Myriophyllum verticillatum (Fig. 61, 6.2), Ceratophyllum submersum - 7 8 Nymphaea alba (Fig. 7.4), Myriophyllum spicatum, Batrachium circinatum (Fig. 7.3), Stuckenia pectinata 4 5 6 Stratiotes aloides, Utricularia vulgaris (Fig. 8.1), Hydrocharis morsus-rапае (Fig. 8.2). 3 4 5 Ceratophyllum demersum, Lemna sp. ПП*<50% 2 3 4 ПП*>50% 1 2 - Filamentous algae 1 2 - *Projected plant coverage on the water surface in percentage FOR EVERYBODY 36 After surveying the area of the water body/stream section, we identify macrophyte species and compile their general list. To assess the condition of the water body using the table, it is necessary to estimate the approximate number of all aquatic plants encountered in the water body being evaluated. All species found in the water body are considered, not just those listed in the table. According to the table, we determine which indicator species are present in the surveyed area. Initially, in our list, we look for species sensitive to pollution, indicated in the first (second) rows of the table. If these species are present in our description, the subsequent rows, including indicator groups, are not considered, and the assessment is carried out within this group. If these specified species are not present in our description, we move on to the next row and look for species from the next indicator group, disregarding the rest, and so on. Then, within this row, we determine the total number of species in this group in the surveyed water body. At the intersection with the column containing the corresponding number of species, we find the value of the Macrophyte (Biotic) Index (MI) in the table. The Macrophyte (Biotic) Index (MI) has values that coincide with the water quality classes commonly used in Ukraine: 9-10 points (blue color) - Class I water quality, very clean; 7-8 points (green color) - Class II, clean; 56 (yellow color) - Class III, polluted; 3-4 (orange) - Class IV, dirty; 1-2 (red color) - Class V, very dirty. The higher the Macrophyte Index value, the better the water quality and ecological conditions in the water body/stream. FOR EVERYBODY 37 1 Water moss Fontinalis 2 Chara sp. 3 Persicaria amphibia 4 Potamogeton perfoliatus Figure 5 Species of aquatic plants: 1 2 3 4 FOR EVERYBODY 38 1 Myriophyllum verticillatum 2 Myriophyllum verticillatum 3 Potamogeton natans 4 Potamogeton natans Figure 6 Species of aquatic plants: 1 2 3 4 FOR EVERYBODY 39 1 Sagittaria sagittifolia 2 Nuphar lutea 3 Batrachium circinatum 4 Nymphaea alba Figure 7 Species of aquatic plants: 1 2 3 4 FOR EVERYBODY 40 Figure 8 Species of aquatic plants: 1 Utricularia vulgaris 2 Hydrocharis morsus-rапае 1 2 FOR EVERYBODY 47 1 2 Figure 11 Species of infusoria: 1 Frontonia angusta 2 Colpidium colpoda FOR EVERYBODY 48 Figure 12 Species of infusoria: 1 Halteria chorelligera 2 Histobalantium natans 1 2 FOR EVERYBODY 49 1 2 Figure 13 Species of infusoria: 1 Ophryoglena flava 2 Trithigmostoma cucullulus FOR EVERYBODY 50 Figure 14 Species of infusoria: 1 Chilodonella uncinate 2 Epistylis coronata 1 2 FOR EVERYBODY 51 Figure 15 Species of infusoria: 1 Paramecium caudatum, Carchesium polypinum 2 Tokophrya quadripartite 1 2 FOR EVERYBODY 52 Figure 16 Species of infusoria: 1 Urostyla grandis 2 Vorticella aquadulcis 1 2 FOR EVERYBODY 53 Figure 17 Species of infusoria: 1 Paramecium bursaria 2 Metopus es 1 2 FOR EVERYBODY 54 Figure 18 Species of infusoria: 1 Tokophrya lemnarum 2 Opercularia articulate 1 2 FOR EVERYBODY 55 Figure 19 Species of infusoria: 1 Oxytricha chloreligera 2 Paramecium caudatum 1 2 FOR EVERYBODY 56 INSECTS AS BIOINDICATORS OF WATER BODIES Many species of insects are associated with fresh water. Some of them (water beetles, bedbugs) live in water bodies permanently, while others, amphibians (mayflies, dragonflies, springtails, damselflies, damselflies, damselflies) live in water bodies only in the pre-immature stages of life. Insects play an important role in aquatic biocenoses and human activity. They are part of various trophic links and play a sanitary role in water bodies. Some species kill mosquito larvae and pupae, while others are pests of fisheries. Insects also provide abundant food for fish and other aquatic organisms. Inhabiting almost all types of water bodies, insects are able to live in wet places outside of them and fly from one body of water to another, which indicates their high ecological plasticity and wide distribution. At the same time, they often react sharply to changes in environmental factors, which is why the entomofauna of each particular water body accurately reflects its type and the processes taking place in this water body. Hence, insects play an important role as environmental indicators, which is essential for a comprehensive assessment of continental water bodies. Insects are a very promising group of animals for bioindication research. They are distinguished by a large species and ecological diversity, different resistance to anthropogenic impacts and different characteristic reactions to them. FOR EVERYBODY 63 Figure 21 Calopteryx dragonfly larva (source) Figure 22 Nemoura springtails larva (source) FOR EVERYBODY 64 Figure 23 Nepa water scorpion (source) Figure 24 Twilight spinneret Orectochilus (source) FOR EVERYBODY 65 Figure 25 The water beetle Hydrobius (source) Figure 26 The Platambus floating beetle (source) FOR EVERYBODY 66 Figure 27 Dryops diving beetle (source) Figure 28 The larva of the Rhyacophila caddisfly (source) FOR EVERYBODY 67 When selecting insects as model bioindication objects, we are guided by the generally accepted requirements for bioindicator organisms such as sufficiently well-studied species and intraspecific taxa, wide range, low migration activity, high indicative plasticity of the species, ease of collection in nature, and sufficient numbers for sampling. Figure 29 Chironomus mosquito larva (source) FOR EVERYBODY 68 FISH AS BIOINDICATORS OF FRESH WATERS Many The transformation of aquatic ecosystems caused by anthropogenic changes at the basin level has led to a significant deterioration of fish populations in most reservoirs of Ukraine. The goal of any reservoir reconstruction is to increase productivity or expand the spectrum of its use, but the structural complexity of ecosystems does not allow predicting all the negative changes and processes that will develop as a result of human activity. In this context, the problem of identifying environmental risks and indicators at the biocoenotic and population levels that will allow for negative changes in ecosystems is acute. At the moment, the level of ichthyological research does not always allow us to clearly determine what processes in reservoirs are associated with changes in the species composition or structure of fish populations. That is why there is a need to develop theoretical approaches to the use of fish as indicators of the state of hydroecosystems. The use of structural features of fish populations and communities as bioindicators has both advantages and disadvantages compared to aquatic invertebrates, algae, and higher aquatic plants. The advantages of this technique include the relatively large size of objects, the relative simplicity of determining the species belonging to fish, the possibility of conducting research with minimal use of laboratory equipment, as well as the fairly simple determination of the structural characteristics of fish populations. The most significant defects are: the difficulty of determining reliable indicators of the number of populations of various fish species; mobility of representatives of the ichthyofauna, which allows them to avoid adverse FOR EVERYBODY 69 conditions; the factor of removing fish for fishery purposes, which disrupts the structure of populations and groups. Most Western European countries use biotic indices for standard water quality control for bioindication. In the last decade of the 20th century in Europe and the USA, there was a tendency to develop biological assessment methods within the ecosystem integrated approach. In Ukraine, researchers are currently showing interest in studying different approaches to using fish as indicators of the state of hydroecosystems. At the same time, it should be noted that there is a certain difficulty in using fish as indicators, which is primarily connected with the following disadvantages: 1 empirical data have some ambiguity; 2 lack of reliable criteria for choosing absolutely adequate biological indicators for the purpose of assessing impact on ecosystems; 3 the problem of choosing a "benchmark" for comparing assessment results; 4 about 2/3 of biotic indices are based on benthic macroinvertebrates; 5 fish are considered very rarely as bioindicators; 6 the possibility of bioindication based on the structural features of fish populations in the reservoirs of Ukraine has not been sufficiently investigated; 7 the vast majority of research on the problem of bioindication in Ukraine is carried out on large rivers and reservoirs, small rivers have not been studied in this aspect; 8 problems of assessing the quality of the environment from anthropocentric and ecosystemic positions, and problems of determining the optimal level of anthropogenic transformation of hydroecosystems; 9 new threats to the stability of ecosystems are constantly arising - this requires expanding the possibilities of bioindication. FOR EVERYBODY 70 The detection of ichthyological indicators at the biocenotic and population levels, that characterize the state of hydroecosystems, may in the future be the basis for research aimed at predicting changes and preventing ecological risks in the reservoirs of Ukraine. The detailing of the dependencies between quantitative indicators characterizing the structure and dynamics of ichthyocenoses and fish populations and the influence of the main factors on ichthyofauna, on the other hand, allows us to reveal the peculiarities of the structure of ichthyocenoses and the quantitative characteristics of the size-mass, sex structure of fish populations, which correspond to a certain level of negative changes in hydroecosystems. Analyzing the existing approaches and methods, in our opinion, there are 5 indicators of population and coenotic levels that will allow us to judge transformations in the reservoir: Diversity of the population. As you know, each age group is represented by individuals of different sizes, which depends on the quality of the environment, so their size distribution will differ. To assess the current state of environmental quality, analyzes performed on juveniles and short-lived fish will be most informative, as the size diversity of older age-groups of long-lived fish may be the result of actions that took place in past periods. The coefficient of variation can also be an indicator. Thus, if the ecological conditions of the environment are favorable for the development of fish, then individuals of the same species with a wide range of biological characteristics survive and coexist, for example, fish with different body lengths and weights. If the ecological conditions of the environment are unfavorable and negative factors influence them, then the action of stabilizing selection is observed, which cuts off extreme variants and supports a certain phenotype with a narrow range of indicator variations. Accordingly, the coefficient of variation of each indicator will be low in its value. FOR EVERYBODY 71 Size and mass structure of the population. Indicators of the population structure of a species can be an indirect reflection of the influence of negative factors. Thus, significant dynamics of indicators in the size-mass structure of individuals makes it possible to talk about the facts of overfishing and undermining the population size. The use of this indicator is possible for species with sexual dimorphism in size . Ratio of sexes is a particularly important indicator for fish with sexual dimorphism. Under certain conditions, there may be sharp deviations from the "normal" sex ratio as a result of various natural and/or anthropogenic factors. The sex structure of individual fish species varies considerably, but mostly the ratio is about 1:1. Taking this dependence into account, it can be stated that the preference of females over males can be an indicator of the level of extraction and the state of the fish population in the studied reservoirs. Individual morphological variability of individuals and the presence of phenodeviations. To determine the level of variability when studying a natural population as a complete genetic-evolutionary system, accounting is perspective for the stability of individual development by such features as the level of fluctuating asymmetry and the number of phenodeviations. The latter, as a peculiar group of changes that occupy an intermediate position between qualitative and quantitative signs and indicate hereditary deviations from the norm, are very changable and occur with different frequency. As a rule, in natural populations there are different levels of deviations, the frequency of which is small, but in some cases their frequency increases significantly. In addition, there is another approach to assessing the stability of individual fish development in the conditions of anthropogenic pressure on ecosystems - the analysis of morphological bilateral signs, in which the variability of these signs on the left and right sides of the body is clarified. FOR EVERYBODY 72 Among the clearly expressed signs, that is, those that do not require a very close inspection of fish, there are various violations in the structure and topography of the organs of the lateral line. The main types of fish anomalies in the studied region were: curvature of the spine, underdevelopment of one gill cover, bent fins. Their share was 1%, and the main reason for such changes is inbreeding. Summing up, it should be noted that the frequency of appearance of any phenodeviant depends significantly on the living conditions of the fish. The most important environmental factors that affect the frequency and degree of manifestation of these anomalies are temperature, excessive or deficient supply of food for fish, the gas regime of the reservoir, water pH, and the level of pollution. The presence of phenodeviants in the population can be considered as an indirect indicator of a decrease in genetic diversity and developmental homeostasis. Genes or a combination of genes that are not detected in a well-balanced genotype and optimal living conditions are determined when the genetic imbalance occurs and when environment is unfavorable. A large number of asymmetric manifestations in fish indicates a decrease in the viability of their natural populations being influenced by powerful anthropogenic pressure, pollution in particular, and can be used as indicators of the state of the environment. Anomalies of the skeleton. The connection between the frequency of skeletal anomalies of aquatic vertebrates and pollution was confirmed experimentally. The organochlorine pesticide Kepon, for example, caused scoliosis in lampreys, and when exposed to heavy metals, spinal curvatures and fractures were observed in fish. Therefore, monitoring involves a careful examination of the fish for obvious abnormalities, with possible follow-up fluoroscopy to detect hidden deformities, such as vertebral adhesions. It is difficult to examine the gill stamens and dorsal fins. Plankton can be of great benefit to detect pathologies in larvae and anomalies in young fish. FOR EVERYBODY 79 The striata morph is characterized by a higher level of oxidation-reduction processes, hemoglobin content, reduced sodium permeability and the content of a number of metals at a higher body weight. A change in the phenetic structure of amphibian populations in a polluted water body is associated with different adaptive value of phenotypes, which is manifested in their selective mortality. Thus, the ratio of striata and maculata phenotypes in amphibian populations can be a convenient feature for bioindication of pollution. Morphological indicators of amphibians against the background of anthropopressing The sizes of morphological features are formed largely under the influence of the environment, and the average values of many features can be reliable markers of negative changes occurring in the habitat of amphibians. Living in polluted water bodies is usually associated with changes in the exterior parameters of amphibians (body length and weight). It is known that amphibians are smaller in size in a polluted water body. This may be due to the accumulation of toxic substances in their body, metabolic disorders. For bioindication, it is advisable to use both the entire complex of signs (males and females) and individual most informative ones, since with the increase in the concentration of heavy metals in water, a reliable change of all morphological signs occurs. Standard morphological parameters must be measured using a caliper with an accuracy of 0.1 mm. FOR EVERYBODY 80 Teratogenesis of amphibian populations. Under conditions of environmental stress, the variety of types of anomalies and the general frequency of aberrations changes, so the variety and frequency of anomalies can be an indicator of the degree of transformation of the natural environment. The high frequency and variety of types of anomalies , in our opinion, may be an indicator of chronic stress due to the increased content of heavy metals in the water. According to the literature, anomalies are the result of a critical disturbance in the stability of development (Kurtyak F.F., 2010). The frequency and diversity of congenital morphological abnormalities can be highly effectively used in environmental monitoring systems. The stability of development is evaluated using the indicator of fluctuating asymmetry. The degree of pollution of the water environment relative to the norm is determined by a violation of the stability of development based on fluctuating asymmetry. The indicator of fluctuating asymmetry reflects morphogenetic homeostasis. For operational primary bioindication of the ecological state of reservoirs and watercourses, it is advisable to use the population characteristics of tailless amphibians, the ratio of males to females, the proportion of the striata morph, and the assessment of fluctuating asymmetry. For an extended, in-depth assessment of the ecological state of reservoirs and watercourses, the entire complex of tested population, organismal and cytological characteristics sensitive to environmental pollutants can be recommended. FOR EVERYBODY 81 This complex also includes (in addition to the characteristics for primary bioindication) variations of morphometric parameters, indexes of internal organs and fatness, detection of the spectrum of phenodeviants, assessment of ontogenetic homeostasis by the level of stability of nuclear structures . A high birth rate and a wide variety of phenodeviants can be recommended as indicators of heavy metal pollution of water bodies and watercourses. FOR EVERYBODY 82 WHAT DO YOU NEED TO KNOW ABOUT WATER AND ITS MICROBIAL COMPOSITION TO PROTECT YOURSELF FROM INFECTIONS THAT ARE SPREAD BY WATER? At different times of the development of human civilization, water was one of the crucial conditions for human existence and, at the same time, a factor in the spread of infections. With the invention of sewerage and treatment facilities, the situation has changed drastically, at the same time due to the intense deterioration of the ecological situation, the water of open reservoirs and other objects of the environment can be a factor in the transmission of pathogenic microorganisms. Microorganisms are mandatory inhabitants of water bodies, which play a key role in their vital activity, providing closed cycles of the main biogenic elements. Water is a natural habitat for various microorganisms. The totality of all aquatic microorganisms is designated by the term microbial plankton. Its qualitative composition depends on the season, meteorological factors, the degree of remoteness of the reservoir from populated areas, the chemical composition of the source, the nature of the soil of the banks, the presence and composition of hydrobionts, sources of pollution. The quantitative composition of microorganisms in water is affected by its bioand physico-chemical composition (content of organic and inorganic substances, temperature, pH value, irradiation, saturation of CO 2 and O 2 , etc.), depth of reservoirs, flow rate, flora and fauna, inflow of sewage water and other factors. On the surface of the water, microorganisms are adversely affected by the rays of the sun, and at depth - high pressure, low temperature, lack of oxygen. FOR EVERYBODY 83 Water has its permanent inhabitants, at the same time, many microorganisms fall from the soil, especially after rains. So, if 1 ml of lake water contains about 10 microbial cells, then after rain their number increases to 1,200. These are representatives of microorganisms that participate in the cycle of nitrogen, sulfur, and fiber decomposition. Photosynthesizing bacteria develop in water bodies rich in hydrogen sulfide. When a large amount of organic substances enters, bacteria appear that can be the causative agents of intestinal infections and other diseases. The number of microorganisms in water is also affected by the proximity of settlements - towns and villages. There are significantly fewer microorganisms in the water upstream of settlements than near the settlement and downstream. The number of microorganisms also changes depending on the depth. Thus, 70-100 microbial cells are determined on the surface of the reservoir in 1 ml of water; at a depth of 5 m - 140-150; 10 m ‒~200; 20 m ‒~150; 40 m ‒~50; 50 m - only 5-10 cells. Microorganisms that enter the water from the soil can survive there for different periods of time. It depends on the type of microorganism and the nature of the water. Cholera vibrios, typhoid and dysentery bacteria, causative agents of tularemia, anthrax bacilli (anthrax) and many other microorganisms dangerous for humans and animals are detected in the water. It is believed that about 25% of infectious diseases are transmitted through water. For example, in clean well water, salmonella can be stored from 2 days to 3 months, shigella - 5-9 days, leptospira - 7-150 days. The quality of water is determined, first of all, by the total number of microorganisms found in it. FOR EVERYBODY 84 Since water is used during the production of any type of product, as well as directly for food, the compliance of its quality with sanitary and microbiological indicators is extremely important. Intestinal infections such as cholera, typhoid and paratyphoid, salmonellosis, dysentery, hepatitis A, poliomyelitis, as well as leptospirosis, anthrax, tularemia, tuberculosis, thrush, Ku fever, and various fungal diseases can be transmitted by water. In order to prevent the spread of infections by water, there is a system of sanitary and microbiological research . The purpose of sanitary and microbiological research is: • detection of pathogenic microorganisms and products of their metabolism (toxins) in environmental objects; • assessment of the sanitary and hygienic quality of these objects; • creating recommendations for the "remediation" of environmental objects by influencing the pathogenic microbiota. Figure 32 Determination of ZMCH of a water sample from a natural reservoir. FOR EVERYBODY 85 All and sanitary indicator microorganisms (SPM) are considered as indicators of biological water pollution . Sanitary indicator microorganisms: are constantly and in large quantities contained in human and animal secretions; do not reproduce intensively in the environment; are easily detected by modern research methods. Sanitary indicators of the state of the environment. Total microbial count (TQM) is the presence of mesophilic aerobic and facultatively anaerobic microorganisms in environmental objects, regardless of the species composition in a certain volume or mass of the product. The species composition of the main sanitary-indicative microorganisms: • Escherichia coli bacteria • enterococci, • proteins, • clostridium botulism and tetanus, • thermophilic bacteria, • salmonella, • bacteriophages of enterobacteria (coliphages) FOR EVERYBODY 86 Indicators of fecal pollution, which means, possibly contamination of water with pathogenic microorganisms, are bacteria of the Escherichia coli group (BGKP). Bacteria of the genera Echerihia, Enterobacter, Klebsiella, Citrobacter and other representatives of the Enerobacterium family, which are Gram-negative rods that do not form spores and capsules, belong to BGKP. They ferment glucose and lactose with the formation of acid and gas at a temperature of 37 °C for 24–48 hours. Water quality is also assessed by the presence of E. coli in it. Determine coli-titer and coli-index. The coli index is the number of individuals of Escherichia coli found in 1 liter of water. Coli titer is the smallest amount of water in which at least one E. coli is found. The presence of the latter in the water indicates its contamination with the contents of the gastrointestinal tract. Tap water is considered to be of good quality if the coli index is 2-3 and the coli titer is 300. Well water should have a coli index of no more than 10 and a coli titer of at least 100. Figure 33 Exceeding the indicators of sanitary norms of water quality according to the microbiological indicator: ZMCH and the number of bacteria of the Enterobacteriaceae family. FOR EVERYBODY 87 The total microbial count indicates contamination of the object with substances of organic origin. The greater the value of ZMCH, the greater the probability of the presence of pathogenic microorganisms in the object. But this number is not always a correct indicator of object pollution, since its value may be high due to saprophytic microorganisms, while pathogenic ones may be absent. In this regard, ZMCH is used as an indicator of the intensity of environmental pollution by organic substances. General rules for water sampling Water samples for analysis are taken from open water bodies with the help of special devices - bathometers, which are lowered to a certain depth, where the device opens and water is collected in it, which is then brought to the surface. Certain rules must be followed when taking samples. 1 If there is a source of pollution, then three water samples are taken from such a reservoir : above the source of pollution, opposite it and downstream. 2 Water samples from wells are taken 2 times: in the morning before water analysis and in the evening after analysis. 3 From ponds, lakes and rivers, water samples are taken from a depth of 0.5 - 1 metres and at a distance of 1 - 2 metres from the shore. FOR EVERYBODY 88 Water samples must be taken in sterile containers . For microbiological analysis 2 litres of water are needed. An accompanying note is attached to each sample, in which the following is noted: the name of the source and its location; year, month and date of sampling; the exact place of sampling; air temperature, data on precipitation on the day of sampling and for the last 10 days; water temperature; a brief description of the reservoir; for which purpose it is directed. Samples are delivered to the laboratory immediately.The content of mesophilic aerobes and facultative anaerobes is determined in 1 liter of water . Two volumes are inoculated from each sample, so that the number of colonies that will grow varies between 30 and 300. When analyzing tap water, 1 ml of contaminated water - 0.01 and 0.001 ml is added to each of the two cups . When determining the microbial count of heavily polluted and wastewater, the studied samples are diluted with sterile water, making serial dilutions. Sanitary and microbiological examination of water is carried out during: • selection of the source of centralized economic and drinking water supply and periodic control of this source; • control of the effectiveness of disinfection of drinking water of centralized water supply; • during monitoring of underground sources of centralized water supply (artesian wells, groundwater, etc.); • determination of the state and degree of suitability of water sources of individual water use (wells, springs, etc.); • monitoring the sanitary-epidemiological state of water in open bodies of water: reservoirs, ponds, lakes, rivers.