Act& hq-drorhim. hydrobiol. 15 (1989) 1 , 81-85

A411-Union Centre of Space and Viicuum Research, USSR State Committee for Standards, Moscow

Water pollutant classification 011 the basis of biological object rcsponse

Sumrucrryr For. a speedy solut~ion to the problem of water resources preservation it is necessary ;a t present to enliance the level of research aimed at eatablislling a system of classification of water pollutants. Control of the state of the environment, must be based on wide use of achievements by different branches of scientific research. Watching the change of t,he biological component of t,he biosphere (biomonitoring) is to ascert.ain t,hc response by susceptiblc biotic forms to enviroiiinental pollut~nnt influence. Microorganisms are a,mong the organismus which itre most susceptible to clivironmcnt influerice. Test cult,ure responses to influences from different pollutants were evaluat- rd by building mathematical models. The principles worked out in drawing up response models could he used to develop clavsificixtion syst'cnis.

Introduction

The inat,erial ill biotesting hased on t'he L I S ~ of experimental inaterial is qiiite enough to attempt solving t,he problem of classifying water pollut,ant.s. Both in this country and elsewhere a lot of factual inaterial has beeii accumulated covering differelit as- pects of t~iotesting. 1 1 1 this country niiich was done t'o estahlish the applicability of t,he methods suggested to assess toxicky levels in water ecosystems, for inst,ance, by the Provisional Scientific and Technological ('ommission, State Committee on Science and Technology, that preseiit.ed its findings a t the Pirst All-Union Conference on the. hio- indicating and biotesting of iiat)ural waters (Kostov-on-the-l)on, September 30-Octo- her 4, 1986). The approval of methods by the cornmission was coiiducted nsing such parameters as sensitivit'g, reproducibility, speediness, lo\\- cut, level of equipment, automation. A s a result of such approval work itiformation was gathered on t,he hio- assays and t.oxicants which are cominon to inany researchers.

Most of the bioassaya proposed were said t'o be efficieiit etrorigh. Aniutrg t,hese were hiniiiiesc.ence, photjosynthrsis, behavioral and eiizyine responsesj et>c. 111 most, works on bioassajririg test objects of different systemat'ic groups mere used, such as microorga- nisms, multicellular algae, protozoa, lower crustaceans, moll uses, fish, and others.

Unfort~nnately, we think that among the general shortcomings is the fact, that it is difficult' because of different conditions under which the assays were held, t'he lack of a unified approach. I n the contrary case we would have more uniformity in pro- cessing observation data. This latter could permit the data processing programme to be duplicated and the pollutant classification to he facilitated.

6 Acta hydrochim., Bd. 17, H. 1

GEXJATULIN, K. V. : Pollutant Clsssificntiori

Objec ts and me thod

JIicroorpanisms oii 11 hich our own research is based a re promiilcnt among t h e various test objects. Interest iii microorganisms is st imulated hy t h e evaliiatioii of t h e degree of reservoir pollution which should be given in dependence on t h e attit,iide of hydrobio- logists aiid hygienists. \Vater pollutioii hy cliemicals hririgs a certain influence t o bear on t h e life of niicroflora. Stiidyiiip the chaiige occiirring in microorganismsand brought about hx chemicals, t h e infliieiice of these cheiiiicals could he tested. These were t h e probleilis facing 11s. Their soliitioii \\-as based i i p o i i a matheniatic~al description of t h e response 1)y different toxic rrsistant mic.roorganisnis.

Work i i i this direct ioii was arraiiged a i d carried out rising as tes t ohjects cultures of niicroorpnisms aiid iiitestiiial virnws (EschrrichI'tr co l i : phages T, E. Coli and polyo virus). Siirvival \\as r ep i s t e rd as a test iwictioii.

-4 qiiaiititative matheinstical statistical int.thnd \\as iistd for describing micro- orgariisnis esposcd t o t h e actioii of chemical silt-)stance (copper sa l t : petroleum; siirfac.e-activc sultstnilces (S-4 S), a i d other, a ~r-icle range of conceiitratioiis). T h e quantitati i- t . deter iiiiiiatioii of t he response of microorganisiiis was based in our case 011 drawing t h e yc>po~ise fuiictioii a s a polynoiiiial:

h K

ri ?/ = bo + 2bil.i - zbij.~i~j ( 1 )

where y is the \.slue of t h e outpiit paraiiieter: x-t.he factor iiivestigatcd (chemical products): b-the repressioii coefficieiits. T h e values of t h e yield paraiiiet'er (y) were ohtained \isiiig t he resiilts of t he f i i l l factor experiment, 2 k . The significance of t h e effects was tested proceeding from dispersioti analj.sis and special graphical methods (GFNJATULIS 1984).

Results

-Ahout 300 repressioii equations were o1)taiiied reflectiiig t h e res])oiise function of selected test cd t i i res i i i clifferetit concentrat'ion ranges of cheinicd water pollatants. Every regression coefficieiit expressed t h e depeiiderice of the yield parameter on only a significant impact of the factor under iii\mtigatioti. For esample, t'he iiifliieiice of copper salt (C'uSOG 5H,O) in t h e conceiitratioii of 0.5 LC14 (LCA =0.1 mg/l according t o Cuz+ : LC'&-\: limit concentration allo~ved) on t h e pliage was expressed in t h e follow- ing dependence :

( 2 ) !/ = 50.6 - 14.92, - 3 . 4 ~ 3 + 5 . 9 ~ 3 + 4.4~2~:: where x i : ( a p p e r salt concentrat ioii ; x 2 : water mediuiii fac tor (river/sea water), .rj: phage niimerical factor (10" ... 106 cells per litre).

Other test ciiltnres (bacteria and viruses) were subject'ed to t)he same pol lu tan t concentration (concentration range 0 ... 0.5 LCA). Copper had a significant effect iipon the bacteria ( the equation reflected th i s in t'he fac tor x1 effect), while viruses did not respond t o t h a t coiiceritratioir, exhibiting resistence to toxic agent,s (as expect- ed from reference da ta ) .

By comhiriing t h e responses from all tes t cultures to t h e action hy one and t h e same

Acta hyclrochim. hydrobiol. 17 (1989) 1 8 3

range of pollutant concentration, we obtain a more definite pollutant representation : let i t be the folloiving in matrix notation :

12.2 - 3.7 0 3.7 0 0 0 [:+[50.6 -14.9 -3.4 0 5.9 0

?j,! 38.7 0 -1.5 0 0 7.1 0

X'X3

This system (upper line, matrix one: regression coefficients determining the effect of the factors given in the matrix t w o on the bacteria: the second line: regression coefficients for the phages: the lover line: for the viruses) is a model of the response by the three ciiltiire systems to the impact of the heav:, inrtal salt.

In nccordance with this effect, the model could be used to classify the pollutants. The saint' model IS used in other response models. I)etailed ~nformation on them

is given (GENJATULIN 1986). The niforn~ation obtained should be verified (on recommendation from IbIoscow

State University) to use the regularities obtained in natural water monitoring (and for sev age, identifying priority pollutants).

Discussion

We hare clet,erniinecl the response by three kinds of niicroorganisms to the action by the most frequent water pollutants in mathematical notation. The list of micro- organism test cultures used in different researches and registration of the dat'a heconies still more variegated over the years. A case in point may be provided by the observa- tions of organic si11)stance assimilation by bacterial plankton under the influence of heavy metal salts (ziiik, copper, cadmium) added t,o river wat'er (TUMANOV et al.). The salts caused a slowing down of the process of orgaiiic substance assimilat'ion by bacterial plankton.

High sensitivity to phenol compounds was established on the basis of one-cell algae movement at, the spore stage (ATAVINA and STOM). Microbiological inethods of water quality aiialysis coiild be recommended for use everywhere. For instance, it could be heterotrophic bacteria in the same of organic pollution; spore bacteria in the case of the hardly disintegrahle organic conipounds; intestinal microorganisms i n the case of the heavy iiietal salts, S A S and a number of others (both isolated arid in mixture).

The rapidity and exact.ness of the bioassaying could be increased by using inatru- mental hioassaying methods. The physiological state of bacteria and one-cell algae could be characterized by a wide range of indices many of which could be easily con- trolled by instrumental methods.

The response registration system has also been improved. One of such highly- sensit~ive systems with high velocity is provided by the luminescence inet'hods tha t find still wider use in practice (BARENBOIM et a].). Some of the most sensitive charac- t.eristics reflecting change in theobject are theluminescence spectrum and the qimntum o*

84 GESJATI-LIT, K. V. : Pollutitiit Classification

yield. Curves of liiminescence decrease could be charact,erized hy great setisitivity and rapidity. This ctharacteristic is used i n hioassayiiig both tiat iiral and waste waters.

This assay could he also used together u-it'll others, as for example when the Dcrpl~n.itr, perish atid the determitiatioii of growth rate of h'/odrir. sprouts. The evaluat,ion of the effects of sewage from a proteiii aiid vitamiti coticetitrates (l'\-C')fact.ory was performed using hioassays 1)ased on tlie estitiguisliiiig of the light of liiminescerit bacteria, as well as the stopping of protoplasma movement i t i algae, cell inimobilizatioii arid change of forin ( K a s r r r s a and SAMUSESOK). Toxicological control of the 1'F.C industry sewage with the iise of these assays could identify tlie qitality of t h e sewage dropped into the reservoir aiid t.lie tlecessilry diliitioii rate i n order to prevent its iicgative influence 011

hydrohiot its. -4s we see, matiy microorganism species and many parameters of their life activity

on t,he hasis of registeriiig t h i s or that test react ion wit'li the iise of some equipment could he implemented i n the system of hioassayiiig chemical pollutants in water.

Otie of the pro1)lems here is the deterniiriatioti of tlie iiifluence of t'he chemical pollu- tarit (,ail isolated siilistaucr or their niixtwc) accordiiig to the respoiisc by the t,est ohject iised 1)). ivhich the classification (or diagnosis) of the pollutant could he per- f oriii etl .

In order to iiiipro\.e the hiottlsting qiiality the use of modern teclitiicd aiid computer aids is required as well as the wide applicatioii of matheinatical met'hods in describing hiotesting processes atid resiilts.

It is irnportant that teclinical illeatis slioiild he used to measure such physical characteristics that norild full>- describe the biological ohject stat'e.

&As a rule, measuring a separate parameter in test ohject life activit'y to determine t,he response to polliitant does not fully describe the polliit~aiit specificit'y. To achieve a descriptioii of polliitaiit itifluence that \voiild he inore reliable and stable in its reprodiicibi1it.j. it. is n.orth\vhilc~ having iiiformatioii on t'he response h s e d on the change iri several paranicters. For esaiiiple, a test sj-stein coiild be cited hnsed on one t.est object, a i d iticliiditiy tlie follo\viiig hiotcsts: niorpliologicd eliariges in cells, elianges iti tlie character of cell nio\-eiiietit, cliarige i n I)elliclc solidit).. kweakdowi oii contractile vacuole activitJ-, atid others (SIKANOROV et al.).

Pollritioii cuiiltl 1)e classified i l l a tliffr!rc.lit' \\.a)- 1,). lookiitg r r t i-espoiiscs hy several test ohjects, too. All test ot)jects c:oiisirlcreci corild presetit chaiiges in otie and t,he same parameter (as, for iiistance, srirr-iral or Iiiniinescence or others) n-hicli could make t'liings easier for a graphical (easilj, riiscernihle) representation of hiotesting results (the graiih 1)reseIits resimtises by set~eral test ohjects t o the same coiicent#ration of a. chemical pollutant) (GENJITULIS, 1986). The suni total of responses from all the test objects u s e d n.ould yield H "portrait" (image) o f tlie pollutant.

Coriclusiuii

-Th~is, iiiformat.iou obtained oirly from studying responses of t'he three microorganism test cultures, enlarged hy data from other researchers on the same types of biological objects, allied t o the capabilities of tecliiiological eqiiipment could sett,le the question on water po1lut.a nt classificat ion.

Using itiforination gathered iti hioassa>, studies together with other of biological

Acta hydrochini. hydrobiol. 17 (1989) 1 85

objects arid using the iiiveiitory of means indicated, incltidiiig computers, chemical pollutants could he classified (ident>ified) successfully.

Due to t.he achievemeiits of geiietic engineering great possibilities i t i creating standard samples might he opened 1111. .4 dat'a hank based on st'aridard samples could become the kernel for a complex syst'eni of obtaining solutions t'o problems of water pollritant classificat~ion. Such a classificatiori syst'em for the main hulk of water chemi- cal pollutants could be founded on the using of aut'omatic recognition systems.

I n part'icular, into the dat'a bank there coiild be included equation systems which reflect the quantitative influerice of a chemical upon test ol-jjects. The inathematical modelling of the influence hy a certain pollutant (single chemicals or t,heir mixture) on a given test object (or a system of test objects, as noted above) produces a classi- ficat'ion of the pollutant effects on the basis of response.

A s noted above, work has already started In this direction. Microorganisms in test cultures could he ilsed for c.ontrolling chemical as well as

biological polliition of wa.t.er (GENJATULIN 1982).

References

ATAVINA. T. G., and D. 1. STOM: Interruption of zoospore movement as method for the operative indication of toxicants of water medium. -Theses of the All-Union Conference on Bioindicatiori and Biotesting of Natural Water, Rostov on t,he Don, September 30-October 4, 1986 (1988), 119.

BARENBOJM, G. M . , A. N . DOMANSKIT and K. K. TUROVEROV: Luniinescence of biopolymers imd cells. Moscow, Science (1966).

GENJATVLIN, K. V. : Problem of biological pollntion and its control. Hygiene and Sanitation (1982), 1, .5842.

-: Using graphic mtithemat'ical statistic methods t'o anelyse the impact by chemical environnienti~l polllitants on microorganisms. Hygiene and Sanitation (1984), 3, 66-70.

-: Diiignosis of chemical pollutants on the basis of biotesting results. Thesis of the All-Union Conf~rence on Bioindication and Biotesting of Natural Water, Rostov 011 the Don, September 30 ... October 4, 1986 (1986), 119.

KASHINA, X. F., and L. V. SAMUSXNOK: Estimation of toxic effects of sewage RS product of albnmi- nous-vitamin concentrat,e. Thesis of tlic: 811-Union Conferenre on Bioindicatiori iind Bio- testing of Natural LVwter, Rostov on the Don, September 30 ... Ortoher 4, 1986 (l986), 170.

KIKAKOROV, A. &I., L. V. BRAZNIKOVA, N. M. TRUNOV e t 81.: System of biological t'oxic t'ests oi l Pnrnrnmium caudatum. Thesis of the S11-Union Conference on Bioindication and Biot,esting of Nat,iiral Wat'er, Rostov on t,he Don, Sept'ember 30 ... October 4, 1986 (1986), 120.

TUNIANOV, A. A, M. S. GLUHOVA, S. &I. FROLOVA :%,id D. V. MUSLIN: Spore-niicroorgenisii~s as malyticn,l indicators on silicon organic phenols. Thesis of the All-Union Conferencc on Bio- indicidon and Biot.esting of Natural Watcr, Rostov on t.he Don, September 30 ... October 4. 198G (1986), 182.

~~ariuaX-ri~teiu~lrcny: 23. 11. 1957.

Anschrrft des V~rfnssers.

Dr. B. V. GEXJATULIN, All-'l'nion Centre of Spitcc and Vncuuni Research, USSR State Committee for Standards; 117049 Moscow, Leninskiy prospect 9, USSR.