Systems and technologies of the early biological notification of the general toxicity of water

One of actual problems of ensuring ecological safety of constructions of the waterworks (W) and water supply of the population is creation, development, improvement of methods and means of ensuring of sufficient completeness and reliability of information on a time history of a condition of water of the reservoirs and water currents which are sources of the centralized water supply which experience a continuous anthropogenic load, and in which danger of emergence of technogenic accidents and the emergency situations (ES) is big. Questions of development of new methods and warning facilities (prevention), identification and mitigation of consequences of an emergency not only a technogenic, but also natural origin belong to priority. It is especially important for estimates in real time of level of toxicity of water of sources of the centralized drinking water supply.

Characteristic features of an emergency are external unexpectedness of their emergence with the subsequent fast development of the negative processes conducting to significant economic and/or ecological damage. At the same time for decrease in level of the subsequent material inputs on compensation of damage there is a basic need of the fastest adoption of organizational and managerial decisions in the conditions of uncertainty of a situation, complexity of forecasting of the further course of events. 

 For this reason for the benefit of the solution of practical problems of ensuring ecological safety of water intakes of AF from impact of technogenic accidents, an emergency and their effects the important place is taken development and deployment of methods and hardware components on their early identification.

In particular, in most cases early, early detection of an emergency is not only necessary, but also sufficient for a complete elimination or minimizing of influence of effects of an emergency on life or human health by temporary switching of life support systems of the population on reserve resources.

 Practice shows that environmental monitoring on the basis of automatic stations of continuous action is the safest method of receiving objective and sufficient information about a time history of a condition of quality of water of a source for preparation and adoption of reasonable managerial decisions. One of necessary conditions of success of practical implementation of a control system of water treatment by criteria of ecological safety is the organization on constructions of the first raising of W (before ingress of water on treatment facilities of W) continuous analytical and bioanalytical control of natural and drain waters in real time by means of specialized automatic stations. It allows to provide information support of adoption of the managerial decisions directed to minimization of environmental risks of operation of constructions of AF and water supply of the population including in case of natural and technogenic emergencies.

The number and the list of the measured characteristics usually are defined by purpose, specifics of quality of controlled water, and also economic opportunities. In most cases the most effective it is also information sufficient at economic availability restriction of the monitoring system with continuous measurements of very small number of non-selective, integral quality characteristics of water. As shows domestic and foreign experience, knowledge time history of such integral characteristics of water as the maintenance of the dissolved organic substance (DOS) characterizing its "chromaticity", a turbidity (content of suspended matters), the chemical oxygen uptake (COU), pH, Eh redox potential, concentration of oxygen, a direct-current conductivity, temperature, as a rule, is sufficient for preparation and justification of adoption of managerial decisions on ensuring ecological safety not only the population, but also aquatic ecosystems.

Implementation at the water user enterprises of rather inexpensive means of automatic production monitoring and self-checking not only is economic (sharply reduces labor costs), but also sharply increases ecological safety of the population, for example, at the expense of continuous, in real time, preparation of information on a time history of quality of the water of a source necessary for adoption of managerial decisions in contingencies. As practice shows, the range of changes of quality of surface waters caused only by natural factors can be rather wide in size and stochastic on time. The existing engineering devices and systems on the basis of sensors of measurement of physical and chemical characteristics of surface waters are provided only for monitoring of specific characteristics of water, but do not allow to monitor other harmful effects (HE) on water which can unexpectedly appear at the dangerous level.

Besides, they do not give the chance to objectively define degree of danger of these changes to live organisms, including the person. For this reason observed datas only of physical and chemical characteristics of natural waters are, as a rule, insufficient for an assessment of level of danger of these influences to safety of water supply of the population. The objective assessment of level of danger of toxicological water pollution is impossible without use on water intakes of AF of these or those biological methods of production monitoring of a source capable in the integrated type, taking into account a synergism of the operating factors, to reveal and predict any negative changes of quality of water. And for each specific source it is essentially important to use as bioindicators of native representatives of fauna which are part of its ecosystem. It is essential difference of methods of bioindication from biotesting methods in which by determination (GOST 27065-86) the test organisms prepared in laboratory, as a rule, adapted for dwelling in the supported specific aqueous medium with a small range of change of its physical and chemical characteristics are used. For this reason only native organisms chosen as bioindicators can carry out function of ecological "target" for the most objective integral criterion of danger of changes of quality of surface waters as a result of their pollution. However in most cases they can be used only as operational signaling devices of emergence of ecologically dangerous water pollution level. By preparation of decision making about a type of HE of alternative to use of these purposes of methods and hardware components of the physical and chemical analysis of characteristics of water does not exist now.

The direction based on use for measurement in real time of reactions of a biota of physiological and behavioural ecotoxicological biomarkers is of essential interest to development of biological methods of quality control of surface waters. The ecotoxicological biomarker is a biochemical, cellular, physiological or behavioural change which can be measured in fabrics or in samples of liquids of an organism, or at the level of the whole organism which provides the proof of exposure to influence and/or action of one or bigger number of chemical contaminants and/or physical factors.

The methods based on assignment of cardioactivity of benthic invertebrates with a tough outside cover, for example, of crayfish, crabs, shell mollusks are the most developed from such methods so far. The slow-moving way of life of these benthic organisms does them by convenient object for tool systems of bioindication. It is necessary to emphasize that in each specific water area, different representatives of benthic communities can act as a type - "target".

Taking into account the above in laboratory of experimental ecology of water systems of the SRCES RAS  the fiber-optical method of assignment of cardioactivity of the benthic invertebrates (BI) having a tough armor was developed: Crustacea (Decapoda) and Mollusca which allows continuously, in real time to carry out remote (to hundreds of meters) noninvasive control of a functional condition of BI. At the same time diagnostics of a functional condition of animals - "targets" is carried out with use of the method of the variation pulsometriya (VP) adapted for backboneless animals. The flowchart of installation for registration of cardioactivity of BI, and also the main stages of mathematical processing of selection of cardiorhythms (normal not less than 100) for obtaining characteristics of VP are provided on Fig. 1.

The essence of the VP method consists in studying of the distribution law of cardiointervals (KI - time frames between two next cycles of a cardiorhythm) as random values in the studied number of their values. At the same time is under construction variation curve (histogram), reflecting probabilistic distribution of KI in the analyzed selection of cardiorhythms. According to the histogram by mathematical methods calculate numerical characteristics and the VP indexes. The mathematical sense of the calculated characteristics and indexes is that they reflect features of a form of the histogram. Physiological interpretation of the above-stated characteristics and the VP indexes is still known only in relation to a cordial rhythm of the person.

The VP method was developed and is quite successfully applied in space medicine to estimates of adaptive opportunities of an organism, "health level" of conditionally healthy people - astronauts, and a time history of their functional state when testing during the periods of selection, preflight preparation and work in an orbit. In medical practice this method well proved as a prenosological method (i.e. "to an illness") diagnostics of conditionally healthy people, that is the method allowing to reveal levels of external influence, harmful to an organism, at the earliest stages, stages of emergence of risk of a disease. It allowed to develop the "Traffic light system" (see fig. 2) which well proved at its use for diagnostics of a condition of conditionally healthy people. At the same time, as medical practice showed, for people with reduced adaptive opportunities of an organism (as a result of this or that illness) the VP method not always allows to receive reliable results of diagnostics of reactions of an organism to stress influencing factors. This lack of a method is connected, apparently, that availability at the person of these or those diseases is the constant spring of own, internal stressful action on the general condition of its organism against which to reveal objective level of significance of the interesting source of stressful impact on a functional condition of an organism it is already difficult.

The analysis of the data which are available in open literature and results of ecophysiological experiments with BB allowed to establish that use of the VP method in the analysis of cardioactivity of native BI is rather effective remedy for the solution of the important production tasks connected with need of detection for real time for water of a source dangerous to live organisms, including the person, levels of its chemical pollution on quantitative assessments of a functional condition of native animals.

According to the original fiber-optical method of studying of cardioactivity of BI developed in SRCES RAS, information on a condition of an organism is output to the registering system (fig. 3) located out of water by means of fine optical fiber which practically does not stir life activity of an observed animal bioindicator - a native crawfish (fig. 4). This physiological method was the basis for the bioanalytical block of the new information measuring system intended for continuous production biomonitoring of quality of water of a source in real time on the basis of the analysis of cardioactivity of a crawfish. The bioanalytical block includes also the system based on the analysis in real time of behavioural reactions of native fishes.

   
Fig. 1. Flowchart of assignment and registration of cardioactivity of cancer.
On the scheme the main stages of mathematical processing of selection of cardiorhythms (normal not less than 100) for obtaining characteristics of VP are provided: average size of cardiointervals Ki_a, average square deviation of SD, variation scope VS and stress index of SI.

Fig. 3. Diagrammatic representation of one of options of system of monitoring of quality of water of the water area containing the  biosensing module on the basis of assignment and the analysis of cardioactivity of a native animal where 1 - a site for placement of the container 2 with recording systems, the analysis and data transmission, 3 - a mesh cell for an animal, 4 - a slot for optical fibers, 5 - the transferring optical fiber, 6 - receiving optical fiber, 7 - the tested animal.

Fig. 4. A crawfish with the fiber optic sensor strengthened on external part of a carapace. 

Assignment and structure of the bioanalytical block of System of production biological monitoring of quality of water Bioargus-W

 The developed aquarian biotouch information measuring system is intended for ensuring technical capability of detection in real time in the water of a source arriving on constructions of the first growths of waterworks, water pollution levels high toxic substances which day regulation of consumption is life-threatening the person. As a key indicator variability of a cordial rhythm of crawfishes is used: the heart rate (HR), square deviation (SD) of distribution of cardiointervals on duration and the stress index (SI).

For carrying out these measurements the fiber-optical method of registration of cardioactivity of benthic invertebrates developed in laboratory of experimental ecology water systems of the SRCES RAS is used. The continuity and uninterrupted operation of measurements of these physiological biomarkers is provided by means of the special flowing aquarian systems supporting three couples of crayfish. Time of complete removability of water in each aquarium has to be comparable or slightly less similar time of complete removability of water in a construction of the first growth of the relevant waterworks. The scheme of installation for complex biomonitoring of toxicity of water of a water supply source is provided in fig. 5.

Fig. 5. Scheme "Bioargus-W".

Installation includes: aquariums 1 and 2 - for keeping of crayfish whose cardiorhythm is analyzed in real time; aquariums 3, 4, 5 and 6 - for keeping of the crayfish used as "doublers" of crayfish in aquariums 1 and 2; an aquarium 7- for keeping of native fishes (in this option it is not used). 

Animals in aquariums 3 - 7 carry out function of additional test objects behind which visual examination needs to be made at tool detection of toxicity of water by means of the animals who are in aquariums 1 and 2. 

Fig. 6. Diagrammatic representation of a rack with three shelves on which are located: 1-receiving rapredelyayushchaya capacity with the source water proceeding through it; 2 - computer; 3 aquarium with small fishes; 4 - 6 aquariums with crayfish (in this option it is not used); 5 - ATsP; 6 - fiber-optical photoplethysmograph.

 For any system of biomonitoring based on use of physiological biomarkers, not only changes of toxicity of water to levels, life-threatening aquatic organisms, but also all rather fast and considerable changings of any physical and chemical characteristics of water essential to ensuring normal functioning of their separate bodies and an organism in general will be significant factors from the point of view of the reaction chosen for indication of change of quality of water. For the chosen biosensors (fishes and crawfishes) first of all it is necessary to carry to such factors: water temperature (defines the general intensity of all processes), the general content of salts (defines a condition of all cellular membranes and processes of ion exchange through them), activity of hydrogen ions (rn), the content of separate anions and cations. Some of them influence exchange processes at the used aquatic organisms directly (type chloride ions), and others as, for example, an ammonium ion, can bear "information" character, indirectly confirming change of a biological effeciency of water, and can be used by aquatic organisms, for example, crawfishes when questing.

 For taking note of possible abrupt changes of such "natural" factors, as the reason of manifestation of alarm reaction (an initial stage of a stress) the biosensors used as a part of the bioanalytical block have significantly important and necessary a continuous registration parallel (in real time) at least of the main physical and chemical characteristics of that water which comes to aquariums of the bioanalytical block. Data processing of measurements of these integral characteristics (water temperature, rn, conductivity, quantity of suspended particles, chromaticity, contents chloride - nitrate - and ammonium ions) has to be processed in real time and together with data on cardioactivity of crayfish. Only in this case it is essentially possible to separate the "false" operations of system of bioanalytical control connected with reaction of the crawfish to abrupt changes of the integral characteristics of water stated above, but not on manifestations new who suddenly arose in the analyzed water of the chemical making strong impact on these or those vital systems of a test organism (availability signs in water of considerable level of content of high toxic substance).

  The above described systems are installed and work, since 2005 at all 11 intake structures of waterworks of St. Petersburg, and since 2010 also at intake structures of waterworks of Vodokanal Municipal Unitary Enterprise of Khabarovsk.

Two-fold sea Mollusks as biosensors of an ecological condition of water areas

 Besides, for a reliable assessment of an ecological condition of water water areas widely use the automated bioelectronic systems of early diagnostics and the prevention of threats of ecological safety where as biosensors native shell mollusks on which shutters in addition to tiny sensors of cardioactivity also motion sensors of shutters are installed are used.

 Fig. 7. The submersible module with sensors of measurement of cardioactivity (1) and the movement of shutters (2) mussels.

  Now does not raise doubts that two-fold sea mollusks are convenient object for researches of influences of different abiotic factors of a circumambient (in particular, changes of temperature and pollution of an aqueous medium different pollyutant, etc.) on live organisms, their systems, bodies and fabrics. Besides, thanks to the high density of settlements and rather large sizes, clams are the leading component of the natural biofilter and, therefore, can be considered as a key type of an ecosystem in which they live.

 The developed systems of simultaneous registration of functional characteristics of clams (heart rate - HR) and their physical activity (the size of disclosure of shutters - SDS) allow to obtain in nature in situ data on the general metabolism of an organism and features of respiratory activity in the conditions of availability in water of contaminants. Besides there is a possibility of an assessment of integral answers of a complete organism to a stress factors, owing to change of quality of a circumambient. The combined use of the characteristics of a condition of aquatic organisms and their biochemical and cytologic biomarkers received thus is able to afford to move to essentially new level of an assessment of a state of health of populations of key types of ecosystems, and as a result to integral criterion of a state of health of the studied ecosystems in general.