WVEg

FISHERIES RESEARCH BOARD OF CANADA

Translation Series No. 2611

DeterminatiOn of the trophic limiting factor for Microcystis (Diplocystis) aeruginosa in the BMT method

by G. Bringmann, and R.',Kuhn

*Original title: Bestimmung des trophischen Begrenzungsfaktors. für Miôrocystis (DiploCystis) aeruginosa irruBMT-Verfahren -•

From: Gesundheits-Ingenieur (Health Engineer), 89:(8) : 247-252, - -196 t • ' .

:.Translated by,the Translation Bureau(SS) .Foreign Languages Division . .

Department. of .. the Secretary of State of Canada

Department of the Environment Fisheries Research Board of Canada

Great Lakes Biolimnology Laboratory Burlington, Ont.

J973

18 . pages.typescript

DETERMINATION OF THE TROPHIC LIMITING

FACTOR FOR MICROCYSTIS (DIPLOCYSTIS)

AERUGINOSA IN THE BMT METHOD

G. BRINGMANN AND R. KUHN

^,-.-...^.

DEPARTMENT OF THE SECRETARY OF STATE

TRANSLATION BUREAU

MULTILINGUAL SERVICES

DIVISION

TRANSLATED FROM - TRADUCTION DE

German

AUTHOR - AUTEUR

G. Bringnann and ^. ,dihn

TITLE IN ENGLISH - TITRE ANGLAIS

.

CANADA

SECRETARIAT D'ÉTAT

BUREAU DES TRADUCT105` C` 1 ,

DIVISION DES SERVirpl ^rr^^ A Y

MULTILINGUES

INTO - EN

English

Determination of the trophic limiting factor for fllicrocystis (Diplocystis) aeruginosa inthe BfJ nethod

TITLE IN FOREIGN LANGUAGE (TRANSLITERATE FOREIGN CHARACTERS)TITRE EN LANGUE ETRANGÉRE ( TRANSCRIRE EN CARACTÈRES ROMAINS)

f3estimmung des trophischsn Begrenzungsfaktors für fi1icrocystis (Diplocystis) aeruginosain E3MT--Uerfahren

REFERENCE IN FOREIGN LANGUAGE (NAME OF BOOK OR PUBLICATION) IN FULL. TRANSLITERATE,FOREIG:! CHARACTERS.REFERENCE EN LANGUE ETRANGERE (NOM DU LIVRE OU PUBLICATION), AU COMPLET, TRANSCRIRE EN CARACTÈRES ROMAINS.

Gesundheits-Ingenieur.

REFERENCE IN ENGLISH - REFER'cNCE EN ANGLAIS

Health Engineer

PUBLISHER - ÉDITEUR

PLACE OF PUBLICATION• LIEU DE PUBLICATION

VOLUME

DATE OF PUBLICATIONDATE DE PUBLICATION

YEAR

ANNÉE I I NUMERO

1968

L4

r:

89

REQUESTING DEPARTMENEnV1-ronI11.-'ntMINISTÉRE-CLIENT

ISSUE NO.

s

PAGE NUMBERS IN ORIGINALNUMÉROS DES PAGES DANS

L'ORIGINAL

247-252NUMBER OF TYPED PAGES

NOMBRE DE PAGESDACTYLOGRAPHIÉES

18

TRANSLATION BUREAU NO. 141281NOTRE DOSSIER NO ___ __

BRANCH OR DIVISION Inland Waters TRANSLATOR (INITIALS)DIRECTION OU DIVISION TRADUCTEUR ( INITIALES) S • S .

PERSON REQUESTINGDr. W . Glooschenko , BurlingtonDEMANDE PAR

YOUR NUMBERVOTRE DOSSIER NO

DATE OF REQUESTOct o 3 1972DATE DE LA DEMANDE ' ) /

S05-200•10-6 (REV. 2/68)

7y30-21-029-6333

,m-c, -' 6 1g 7'

UNEDITED TRANSLATIONFor information only

TRADUCTION NON REWSEEInfur(r,coi-ion Soulemenf

DEPARTMENT OF THE SECRETARY OF STATE

TRANSLATION BUREAU

MULTILINGUAL SERVICES

DIVISION

Gcsundli.us-(ngenicur Iicft 8 (89. Jahrpng 1968) - 247

^ a^^•r^

CANADA

SECRÉTARIAT D'ÉTAT

BUREAU DES TRADUCTIONS

DIVISION DES SERVICES

MULTILINGUES

Lm

CLIENT'S NO. DEPARTMENT DIVISION/BRANCH CITY

NO DU CLIENT MINISTERE DIVISION/DIRECTION VILLE

Environment Inland Waters Burlington9 0^

BUREAU NO.NO DU BUREAU

LANGUAGE

LANGUE

TRANSLATOR(INITIALS)TRADUCTEUR (INITIALES)

1412$1 German S.S.

Gesundheits-Ingenieur 09, 8, 247-252 (1968)

Determination of the trophic limiting factor for Clicrocystis (Diplocystis)

7

aeruninosa in the 811.7 method

by Director and Prof. Dr. G. Bringmann and Dr. R. 1<6hn

from the Federal Board of Health, Institute of fjater, Soil and Air

Hygiene, Berlin-Dahlem

Introduction

The origin and distribution of substances which are biologically

active for the growth of î;licrocystis (Diplocystis) aeruginosa in West

Berlin waters were determined from preliminary studies using an in-vitro

"water bloom" test1. For a thorough quantitative analysis of the.biologically

247

active state of samples from these waters, with regard to the potential

mass development of Llicrocystis, determinations of the biomass titer (SP11T),

the trophic limiting factor, as well as the limiting effect of this factor

are necessary.

Special provisions of the method are necessary for the

1fJringmann, G.: In-vitro "water bloom" test with f'icrocystisaeruginosa. Ges. •-Ing. 88 (1967), p. 256.

UNED^T E-p YaANSLAT10tij

For in$orrnal ► an only

TRADUCTION NON REvtSEFl.,F^rmation s$ul^en^crli

f' 2

1

r1 i_

1 7 ;_,

,

:1 1:1

I 1 ' 7'

11.p.: ! ' i V'71

determination of the characteristic numbers mentioned above because

fflicrocystis test cultures in water samples unavoidably bring about an in-

stant biogenic alkalization • of' the medium; without precautionary measures

of the cultùre, an inactivation of the iron of the water samples and thus,

an inhibition of the growth of the test organisms would be the result. The

natural iron of the water samples can be converted into the chelate through

.the addition of CDTA and thus, an inactivation of the iron can be prevented

during the alkalization of the medium1

.

A . corresponding biochemical protective measure for maintaining the

phosphate of the water samples active is not given. However, it is possible

to incorporate the phosphate of the water samples into the metabolism of

the test organisms in a biochemically active state before the change of the

pH value or of the redox po .tential of the test medium brings about the in-

activation of the phosphate. Test-cultures are first held for 3 days in a

dark, thermostatically controlled room immediately after the inoculation

with storane ohophate-free cell material. In the dark, the pH value or

the redox potential of the test medium remains within the range in which

the preservation of the biochemical activity of the phosphate is guaranteed.

The phosphate is absorbed by the primarily storage phosphate-free test

organisms from the water and, at the beginning of the subsequent 12-day

test period, can be incorporated into the metabolism of fflicrocystis

(Diplocystis) aeruginosa as a now cell-specific storage phosphate which is

protected against inactivation.

hCtiUll[IIIt..Y•I11gCIIICuC IICI L n ko ^. ju.uL^ u•. h .^^^r ^*• .

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Table 1,- Determination of the biomass titer of C_ autot.r°phic nitrate

.or̂3anisms in--wâter...sam Cles. Prototype organism: C1icrocystis HDiplocHstis_.-_...____.^____.._

aeruginosa-: "

1.00 5tock-culturès

1.01 Test strain: Glicrocystis (Diplocystis) aeruginosa

1.02 Source of-the test strain: University of .'Jisconsin, Dept. of Botany

1.10 Culture vessels: Erlenmeyer flasks, 100 ml (with stopper sterilized

dry)

1.20 F.las!< stopper: cotton wool-filled metal caps

1.30 Culture medium: Nolm-Hansen, entire medium1

1.32 Volume of the culture medium: 20 ml

1.41 Pasteurization of the culture medium: fractionated.2 times 30 minswater bath at 70° C

1.50 Source of the cell suspension For inoculation of stock cultures;

10-day stock culture

1.51 filanner of inoculation: pipetted

1.52 Uolüme of the cell suspension for inoculation of stock culture: 2 ml

1.00 Culture period: 10 days

1.70 Temperature: 27° C

l^. _• Compl-eteT nu trient solution accordingtklo Flolm-Hansen (for stock

cultu'res of f:li crocystis1000 ml bi=distilled water

496 nig fJa1N03 p. a^39 mg K2FIPO4 purest7.5 mg G1g504 + 7 H20 p.a.31 mg CaCl2.40 mg Na2SiO3 + 9 !-420 pure5o mg Na2CO3 pÀëi6 nig c1tric acid p.a.6 mg iron T(Î I) citrate Erj•

10 mg -t-ra0s=1;2^diamino-cycloliexane-f\i,N,f\i',f-31-tetraacetic acid (CDTA)_-_-,_1 of 1 4 24 diluted stock solution of trace elements

kr-9 Tite niè^z iii ^t Q^ T'u5 GzL^b Ye V/cYn`.s. x COC[^cl r► 0

4

«lb

- Gesutulk.e,ingenicur Hell 8 (89. Jahrgang 1968) 247

1.71 Rd. humidity: 50%

1.80 Lighting: continuous lighting

1.81 Light source: Osram L 40/32 and Osram L 40/15

1.82 Arrangement of culture flasks and lights: culture flasks arranged in a long line between 2 side fluorescent tubes in an area 60 cm wide

2.00 Preliminary cultures •

2.10 Culture vessels: Fernbach flasks, 1.8 1 (sterilized dry)

2.20 Flask stopper: cotton wool-filled plastic caps (pasteurized)

2.30 Culture medium: Holm-Hansen modified2

_

2.32 Volume of the culture medium: 1000 ml

2.41 Pasteurization of the culture medium: fractionated 2 times 45 mins water bath at 70 ° C

2.50 Source of the cell suspension for inoculation of preliminary cultures: corresp. to 1.50

2.51 Manner of inoculation of preliminary cultures: poured

2.52 Volume of the cell puspension for inoculation of preliminary cultures: entire cell suspension of a stock culture for a Fernbach flask

2.60 Culture period: corresp. to 1.60

2.70 Temperature: corresp. to 1.70

2.71 Rel. humidity: corresp. to 1.71

2Nutrient solution according to Holm-Hansen, modified (for prelim-

inary cultures of Microcysti7ST 1000 ml bi-distilled water 496 mg PaNO3 p.a. 34 mg KC1 p.a. 75 mg M9504 .1. 7 H20 p.a. 31 mg CaCl2 p.a. 40 mg Na2SiO3 .1. 9 H20 pure 58 mg Na2CO3 p.a. 6 mg citric acid p.a. 6 mg iron I (II) citrate 17-rï.

10 mg trans-1,2-diamino-cyclohexane-N,N,W,N , -tetraacetic acid (CDTA) 1 ml of 1 + 24 diluted stock solution of trace.elemants

'›e S e e -fee

5

2.80 Lighting: corresp. to 1.80

2.81 Light source: corresp. to 1.81

2.82 Arrangement of culture flasks and lights: corresp. to 1.82

2.90 Preparation of the cell suspension for inocûlation of test cultures

2.91 Filtering of the cell suspension: in sterilized filtration apparatus

over membrane filter Gr. 100

2.92 Washing of the cell material: once with sterilized NaCI solution(100 mg NaCl p.a./1 bi-distilled water)

2.94 Suspension of the cell material: in 100 mi sterilized NaC1 solution(100 mg NaCl p.a./1 bi-distilled water)

2.95 Determination of the gravimetric turbidity equivalent Oalue of the

ce11 suspension in mg/1 kieselguhr: photoelectric turbidity measure-

ment (r endorf photometer with nephelometer; filter Hg 578 nm)

3.00 Test cultures

3.10 Culture vessels: Erlenmeyer flasks, 300 ml (sterilized dry)

3.11 Number of culture flasks: 2'flasks for the determination of the basevalues; 2 flasks for the determination of the limiting effect of N;2 flasks for the determination of the limiting effect of P

3.20 Flask stopper: corresp. to 2.20

3.30 Culture medium: water-sample

3.31 Filtration: in sterilized filtration apparatus twice over membranefilter Gr. 100

3.32 Volume of the culture medium: 50 ml

3.331 Substance added for the determination of the hase value: 10 mg/l CDTA

3.332 Substance added for the determination of the limiting effect of N:10 mg/1 CDTA, 40 mg N/1 (as NaNO3 p.a.)

3.333 Substance added for the determination of the limiting effect of P:10 mg/1 CDTA, 100 T Ph (as Na2NP04 + 12 H20 p.a.)

3.50 Source of the cell suspension for inoculation of,test cultures:prepared cell suspension of the 10-day preliminary culture (see 2.90to 2.95)

1 Î]

]

1-Tr,

...1.1.:

I: -T ,

];e:14

GCS1111(111,-.1S-111gC111Cilr II el t 5 W. I

6

3.51 Manner of inoculation of test cultures: corresp. to 1.51

3.53 Gravimetric turbidity equivalent value of test cultures after inoc-ulation: 20 mg/1 kieselguhr

3.60 Culture period: 3 days + 12 days

3.70 Temperature: corresp. to 1.70

3.71 Rel , humidity: correap. to 1.71

3.80 Lighting: 3 days incubation in a dark, thermostatically-controlled room following 12 days lighting corresp. to 1.80

3.81 Light source: corresp. to 1.81

3.82 Arrangement of the culture flasks and lights: corresp. to 1.82

4.00 Assay ...cul,t.ures of the reaction norm of the cell material of the preliminary culture

4.10 Culture vessels: corresp. to 3.10

4.11 Number of culture flasks: . 5 for the assay of the cell material of the . preliminary culture in the absence of biologically active storage P; 5 for the assay of the constancy of the physiological reaction norm of the cell material of the preliminary culture

4.20 Flask stopper: corresp. to 2.20

4.301 Culture medium for the assay of the cell material of the preliminary culture in the absence of biologically active storage P: Holm-Hansen modified 3

3 Nutrient solution according to Holm-Hansen, modified (for the assay of the cell material of the preliminary culture of Microcystis in the ab-sence of biolonically active storage P 1000 ml bi-distilled water 496 mg NaNO3 p.a. 34 mg KC1 p.a. 75 mg 89504 + 7 H20 p.a.. 31 mg Ca012 p.a. 40 mg Na25iO3 + 9 H20 pure 58 mg Na2CO3 p.a.

•

6 mg citric acid p.a. 6 mg iron I (II) citrate

10 mg . trans-1, 2-diamino 7cyclohexane-N, N , N' ,N -tetraacetic acid (CDTA) 1 ml of 1 + 24 diluted stock solution of trace elements

->e 7L.c, k- 01,1- p ,

4.302 Culture medium for the assay of the constancy of the physiological reaction norm of the cell material of the preliminary culture: Holn-Hansen modified 4

4.32 Volume of the culture medium: corresp. to 3.32

4.41 Pasteurization of the culture medium: 30 mins water bath at 70 0 C

4.50 Source of the cell suspension for inoculation of assay cultures: corresp. to 3.50

4.51 Manner of inoculation of assay c ultures: corresp. to 1.51

4.53 Cravimetric turbidity equivalent value of assay cultures after ineculation: corresp. to 3.53

4.60 Culture period: corresp. to 3.60

4.70 Temperature: corresp. to 1.70

4.71 Rel. humidity: corresp. to 1.71

• 4.60 Lighting: corresp. to 3.80

4.81 Light source: corresp. to 1.81

4.82 Arrangement of culture flasks and lights: corresp. to 1.82

6.00 Treatment of test and assay cultures during the 12-day test period: shaken once daily

4Nutrient solution according to Holm-Hansen, modified (for the

assay of the constancy of the physiological reaction norm of the cell mate-rial of the preliminary culture of Microcystis)

1000 ml hi-distilled water 49.6 mg NaM03 p.a. 3.9 mg K2HPO4 purest

75 mg 1119604 4. 7 H20 p.a.

31 mg CaC12 p.a.

40 mg Na2SiO3 4. 9 H20 piire

58 mg Na2CO3 p.a.

6 mg citric acid p.a.

6 mg iron I (II) citrate Er.

10 mg trans-1,2-diamino-cyc1ohdxane-N,N,W,N'-tetraacetic acid (COTA)

1 ml of 1 4, 24 diluted stock solution of trace elements

7

Stock solution of trace elements 1000 n1 bi-distilled water

. 2.66 g H3603 p.a. 0.08 g CuSO4 4- 5 H20 p.a. 1.61 g MnC12 4- 4 H20 p.a. 0.024 g Na2Mo04 2 H20 p.a. 0.22 g ZnSO4 4. 7 H20 p.a. 0.04 g CoC12 4. 6 H20 p.a. .

.3,2z iee -hte-te r,

1

7

]

8

7.00 Culture room: conditioned room, daylight excluded.

7.10 Background: white Resopal plate

8.00 C1easurement of test and assay cultures

8.20 fi,'lethod of measurement: photoelectric turbidity measurement( pPendort' photometer with nephelometer added; filter I-1g 578 nm)

8.30 Values: gravinetric equivalent value of the biomass of the testorganism in mg/1 kiesel.guhr ( biomass titer) as measure of thebiological activity of the nutrients for f:licroc^^siis ^!^i?lec}^stisaerEqinosa contained in the water sanples,T

The biological_ activity of the natural nutrients contained in thewater samples is expressed i n the base value of the bio.mass titer.The selective increase of the biomass titer (â.n ng/l !<ieselgl.lhr)obtained over the base value of the I^est cultures of the homologouswater sar„p7.es enriched either with F,'03- or P04`'° indicates :uhetherin the water sample nitrate or phosphate is active as limitingfactor for the growth of the prototype organis;a, ft1icrocystis.

Glethod (see 'fable 1)

Stock cultures of the test strain of f!1icrocystis (Diplocystis)

aeruginosa are held in 100-m1 'Erlenneyer flasl<s (plugged with cotton wool-

filled raetal caps) in pasteurized nutrient solution according to Ho?m-

Hansen (20 ml) (nutrient solution 1 of the iable,) under exclusion of day-

light with continuous lighting between a pair of fluorescent tubes (Osram

l_ 40/32 and Osram L 40/15), The room temperature is adjusted at 27° C with

a thermostat and the humidity of the culture room at 50;:) relative hunidity

with a hygrostat. After a,10-day culture period, stock cultures are always 250

inoculated in fresh nutrient solution (2 ml cell su-sponsion/20 ml nutrient

solution).

The cell material of a 10-day old stock culture is always used for

the inoculation of a preliminary çulture in 1000 ml phosphate-free

9

pasteurized (2 times 45 mine water bath 70 0 C) Holm-Hansen nutrient.solu-

tion 2 of the table) in Fernbach flasks (provided with cotton wool-filled

plastic caps). Through transfer over this phosphate-free nutrient solution,

algal material which is free of physiologically active storage phosphate

is obtained. Under exclusion of daylight, the preliminary cultures remain

10 days under continuous lighting between a pair of fluorescent tubes

(Osram L 40/32 and Osram 40/15) at 270 C and 50% rel. humidity (see above).

The cell material of the preliminary cultures is separated from the culture

solution.by membrane filter Group 100 (in a sterilized filtration apparatus),

washed once with sterilized 0.01% NaC1 solution and taken up in 100 ml

sterilized 0.01% NaC1 solution. In a suitable dilution of the sample, the

turbidity equivalent value of the cell suspension of the preliminary cul-

ture is determined by the photoelectric measurement of the extinction of

the monochromatic radiation, Hg 578 nm, with a turbidity measuring apparatus

(Eppendorf) calibrated in mg/1 kieselguhr.

For the analysis of best cultures, water samples are filtered

twice over membrane filter Group 100 in a sterilized filtration apparatus.

rri each case, 50 mi each of the water samples is poured into six 300-ml

Erlenmeyer flasks (provided with cotton wool-filled plastic caps). For

the determination of the base value of the biomass titer of Microcystis,

10 mg/1 CDTA is added to 2 test flasks of each water sample; for the

determination of the trophic limiting factor, 10 mg/1 CDTA and 40 mg/1 N

(as Nee'3.

pea') are added ta two test fiasks of each water sample, and

La mg/1- COTA and 100Y/1 P (as Na2HP0 4 + 12 H 20 p.a.) to two additional

test flasks of each water sample. The prepared water samples are inoc-

ulated with the prepared Cell suspension of the 10-day preliminary culture

i 10

••

" —71

.taat

of known turbidity equivalent value so that their gravimetric turbidity

equivalent value corresponds to 20 mg/1 kieselguhr; the volume of the cell

suspension used for the inoculation should not exceed 0.6 ml. Immediately

after the inoculation,the test cultures are placed for 3 days in a dark,

thermostatically controlled room (27° C) and for the following 12 days,

under exclusion of daylight, held at 27 ° C and 50% relative humidity with

continuous lighting between a pair of fluorescent tubes (Osram L 40/32 and

Osram L 40/15). Each flask is shaken vigorously once daily during the

12-day culture period.

At the end of the test period , the turbidity of the cell suspensfon

of each test culture is determined by photoelectric measurement of the ex-

tinction of the monochromatic radiation, Hg 578 nm. In accordance with the

calibration of the measuring - apparatus with monochromatic radiation, Hg 578

nm, With a kieselguhr suspension of known content, the qravimetric turbidity

equivalent value of the. cell suspension is given in mg/1 kieselguhr as

.equivalent value of the biomass of the test organisms.

The gravimetric equivalent value of the biomass'of the test organisms

in mg/1 kieselguhr (biomass titer) is considered a measure of the biological

'activity of the nutrients in the water samples for rlicrocystis (Diplocystis)

aeruginosa.

The biological activity of the natural nutrients in the water samples

is expressed in the base value of the biomass titer. The slective increase

of the biomass titer (in mg/1 kieselguhr) obtained . against the base value

of the test cultures of the homologous water samples enriched with either

NO 3 or PO 43-

indicates'whether nitrate or phosphate is active as the limit-

ing factor for the growth of the prototype organism,Sicrocystis, in the

gi■

water sample.

The physiological control of the cell material of the preliminary

culture is carried out by means of assay cultures- For testing the absence

of biologically active storage phosphate in the cell material of the pre:-

liminary culture, 50 ml each of modified phosphate-free Holm-Hansen nutrient

solution (nutrient solution 3 of the table) is poured into five 300-ml

Erlenmeyer flasks (provided with cobton wool-filled plastic caps). For

testing the constancy of the physiological reaction norm of the cell mate-

rial of the preliminary culture, 50 ml each of modified Rob-Hansen nutrient

solution containing standardized amounts of mineral nutrients (nutrient .

solution 4 of the table) is poured into 5 additional Erlenmeyer flasks.

A 11 flasks are pasteurized 30 mins in the water bath at 70 0 C. Inoculation

and further treatment for assay cultures are similar to that for test

cultures.

Results

For 287 water samples from 41 sampling sites of the West Berlin

water system, the BMT of Microcystis (Diplocystis) aeruginosa, the trophic

limiting factor, as well as the limiting effect of the trophic limiting

factor were determined in 1722 test cultures.

Mean values of BMT of Microcystis (0iplocystis) aeruginosa as well

as mean values of the limiting effect of nitrate from seven sampling series .

in the period from June to October 1967 were taken as a basis of the eval-

uation of the findings.

In the flow region of the Spree (Fie. 1), water samples from the

upper and lower courses of the Panke were found to be extremely heavy in

11

12

1800

1600

1400

1 200

- 600

400

200 h

^ -2i

m ^ ô o x

3 Ÿ -o F ^é ^ W ^a N u

¢ N ^ ^ ^ ÿ s ^ ^-5 ûs ÿ E â _ _ - ô

_

^ û n s` i â s> c v U ^ U Û.` c.

as - - - ô - , V z m o d m in 3^ ^

^ G1ean values of E3f.1T of 51licr.ocystis June/October 1967Ej Glean values d' the limiting effect of nitrate for

lKIT of (11icrocystis June/October 1967

Fig. 1. f;lean values of the biomass titer of -Microcystis and mean valuesof the limiting effect of nitrate for. the bioi;iass titer of P,1i crocystis. inwater samples from the flow region of the Soree.

Spree, unterh. ;bztileigung, C1n.-Spand.Schiff.-Kanal = Spree, below diffluence,Oln.-Spand. navigation canal

Spree, oberh. f:lündung, Landwehrkanal = Spree, above estuary, Landwehr canalLandwehrkanal, Lohnühlenbrucke = Landwahr canal, Lohmuhle bridge

Neukoilner Schiff.-•I;anal, oberh, Lohmühlenbr, = fleukolln navigation canal,above Lohmuhle bridge

Landwehrkanal, f0l6ckernbrucke = Landwehr canal, Mocker bridgeLandwehrkanal, ;.Iiüncung = Landwehr canal, estuary

Chlbg. Uerbindungskanal, Mundüng - Chlbg. connecting canal, estuary

aln.-Spandauer Schiff.-Kanal = Dln.-Spandau navigation canalPanke, Oberlauf _ Panke, upper coursePanke, Unterlauf = par,ke, lower course

Bln.-Spandauer. Schiff.-Kanal, Seestrassenbrücke = B1n.-Spandau navigationcanal, Seestrasse bridge

IJesthafenkanal -. 'Jesthafen canalSpree, Schleuse Ch1bg. = Spree, Chlbg. lockSpreen 'ûndung = Spree estuary

] 1- -''''

'i .

is '

1 •s .,7, 1,..,....

251

••-/••••.1, • {.• ■ •• • tr,•• C•••

13

sewage-bearing substances; their biological activities are expressed in

correspondingly high SMT values of Microcystis (1210 and 1219 mg/1 kiesel-

guhr, respectively). In water samples from the Berlin-Spandau Schiff-

fahrtskanal [navigation canal] at Seestrassenbrücke [Seestrasse bridge]

(260 mg/l.kieselguhr) - main outfall of the Panke, as well as in water

samples from the Westhafenkanal [Westhafen canal] (230 mg/1 kieselguhr),

and from the Spree estuary (215 mg/1 kiesélguhr), relatively high SMT values

of Microcystis were measured in comparison to the measured values of OMT

of Picrecystis in water samples from other sampling sites in the flow

region of the Spree. From this the presence of sewage-bearing substances

in water which promote the growth of Microcystis may be inferred at these

sampling sites of the flow region of the Spree.

. In water samples from all sampling sites of the flow region of the

Spree, a considerable increase of the BMT of Microcystis over the base

addition of phosphate. Correspondingly, in water samples from all sampling

sites, phosphate was present in physiological excess and nitrate was a

trophic 1imiting factor for the growth of Microcystis. The relative limiting

effect of nitrate was especially marked in water samples from the [following]

sampling sites: Spree below the diffluence of the Serlin-Spandau Schiff-

fahrtskanal (101:346 mg/1 kieselguhr), Berlin-Spandau Schiffahrtskanal at

Seestraesanbriicks. (260:570 mg/1 kieselguhr), Westhafenkanal (230:495 mg/1

kieseiger) and Spree estuary (215:532 me/1 kieselguhr).

In the flow region of the Havel (Fie. 2), the biological state of

the, water: samples from the upper Havel is characterized by extremely low

val_ue of" Microcystis (31 mg/1 kieselguhr). in striking contrast to

• 1 value was obtained through the addition of nitrate, however, not through

.w 7-'I711'g'.TIVg- Lii7

1800

1600

600

400

200

0

1400

1 200

9

' Mean values of BMT of f;licrocystis June/October 1967Glean values of the limiting effect of nitrate forNIT of filicrocystis lune/October 1967

14

Fig. 2. Gïean values of the biomass titer of Cîicroc;:s^'.:is and mean valuesof the limiting effect of nitrate for the biomass titer of G1icrocystis inwater samples from the flow region of the 111avel,

Oberhavel = upper HavelTegeler Fliess, Oberlauf = Tegeler flow, upper course

Tegeler Fliess, Unterlauf = Tegeler flow, lower course

Nordqraben, Oberlauf = Nordgraben, upper courseNIordgraben, UnL-erlauf = Nordgraben, lower course

Tegeler See, Lindwerder - Tegeler lake, LindwerderHohenzollernkanal, G,ündung = Hohenzollern canal, estuaryHavel, OU Schleuse Spandau Havel. OW lock SpandauSüdhafen = 3iidhafen

Havel-GemUnd = Havel opening

Havel, Grunewa].dturm = Havel, GrunewaldturmHavel, Kladow = Havel, Kladow

Havel, I:ëlberwarder = Havel, KalberwerderHavel, Krughorn = Havel, Krughorn

,

Iiet 15

• t2.3

I

" -efr".7,,,,,,?:. • . •` , . ..,

this, in water samples from the Nordgraben and t.he Togeler Fliess [flow],

extremely high OMT values of Microcystis (1257/1124 and 941/498 mg/1

kieselguhr, resp.) were measured. The bioloeical activity of the sewage-

bearing substances in water of the above-mentioned outfalls brings about,

in addition, the high 0MT values of Microcystis in mater samples from the

Tegeler See [kake] (401 mg/1 kieselguhr).

Alse, the substantially higher - compared with SMT values in water

samples . from the upper Havel - DMT values of Microcystis in water samples

from Südhafen and Havel estuary (192/164 mg/1 kieselguhr) as well as From

the lower Havel (205/183/153/ 177 mg/1 kieselguhr) could be due to the Inflow

of sewage-bearing substances in water.

In water samples from all sampling sites of the flow region of the

Havel, a considerable increase of the OMT of Micrecystis over the base value

was obtained through addition of nitrate, however, not through addition of

phosphate. It follows from this that in water semples from all sampling

sites, phosphate was present in physiological excess and nitrate is re- .

garded as a limiting trophic factor for the growth of Microcystis. This

applies especially to water samples from the [following] sampling sites:

lower course of the Tegsler Fliess (498:1218 mg/1 kieselguhr), Tegeler See

(401:906 mg/1 kieselguhr), Südhafen (192:575 mg/1 kieseiguhr), Havel estuary

(164:456 mo/1 kieselguhr) and Havel-Grunewaldturm (205:501 mg/1 kieselguhr).

In the flow region of the Teltowkanal (Fin. 3), the relatively high

081 values of Microcystis in water samples from sampling sites at Kreuzung

(177 mg/1 kieselguhr) as well as from other sampling sites in the flow

region of the Teltowkanal (156/196/179/179 mg/1 kieselguhr), as reported

elsewhere, are especially due to the influence of biologically active

16

800

600 t_T;

- 400

-, 200

0

- g

- -5

• T N -§

à

T1- ; , - .1,

1

71 • • z I ul

O 'Mean values of BMT of Microcystis June/October 1967 • Mean values of the limiting effect of nitrate for

onr of Microcystis June/October 1967

Fig. 3. Mean values of the biomass titer of Microcystis and mean values of the limiting effect of nitrate for the biomass titer of Microcystis in water samples from the flow region of the Teltowkanal,

Hafen Rudow-Ost = Hafen Rudow east Kreuzung = Kreuzung Oritzer Zweigkanal = Oritz branch canal Neuk.cillner Schiffahrtskanal = Neuk'Clln navigation canal Hafen Mariendorf = Hafen Mariendorf Schilfluchgraben = Schilfluchgraben Teltowkanal Mündung = Teltow canal estuary Prinz-Friedrich-Leopold-Kanal = Prinz Friedrich Leopold canal

substances in water of the Rudower Fliess.

In water samples from all sampling sites of the flow region of the

Teltowkanal, a considerable increase of the BMT of Microcystis over the

base value was obtained through addition of nitrate, however, not through

addition of phosphate- Therefore, in water samples from all sampling

sites, phosphate was present in physiological excess and nitrate was a

limiting trophic factor for' the growth of Microcystis. This applies

Gesundh^,.,s-lnnenieur Heft 3(39. Jahrgang 1968) •?47

17

particularly to water samples from the sampling sites: Kreuzung (177:523

mg/l kieselguhr), fJeuk^lln Schiffahrtskanal (156:3[36 ng/1 kieselguhr),

Hafen Glariendorf (193:418 mg/1 kieselguhr), Schilfluchgraben (179:300 mg/1

kieselguhr), estuary of the Teltowkanal (179:410 mg/1 kieselguhr) and

prinz-Friedrich-Leopold-=Kanal (197:457 mg/1 kie.selguhr).

.A

a

Summary

For the quantitative determination of the biologically active con-

dition of water samples from West Berlin waters with regard to the potential

mass development of blicrocystis, the biomass titer (BC?T), the trophic limit-

ing factor, as well as the trophic limiting effect of this factor were

determined for the prototype organism, Microcystis (Diplocystis) aeruginosa.

Test cultures of Microcystis (Diplocystis) aeruginosa for determinations

of the biomass titer and the trophic limiting factor in water samples from

waters alkalize the culture medium instantly; inactivation of iron could be

associated with alkalization. The natural iron in water samples can be con-

verted into the chelate through addition of CDTA and thus can be protected

against inactivation during the biogenic alkalization.

The natural phosphate of water samples can be guarded against in-

activation if it is possible for the primarily stora2e phosohate-frse test

organisms to absorb and to store the phosphate. The absorption of the

phosphate takes place in a period of 3 days in the dark preceding the

12-day test time in which a change of the pli value or the redox potential

of the water samples detrimental for the preservation of the biochemical

activityof the phosphate is not expected. The phosphate-storage capacity

of Clicrocystis (Diplocystis) aeruginosa is adequate for purposes of the

18

Gestindlt,ts-Ingenieur Heft 8 (89. Jahrgang 1968) • 247

SOT procedure in all cases. Storage phosphate-free Cell suspensions

(initial turbidity equivalent value 20 mg/1 kieselguhr) in neutral water

samples in a dark, thermostatically centrolled room are capable of storing

so much phosphate in 3 days that in the following 12-day test pnriod, with

a correspondingly high natural nitrate content of the water, BUIT values of

> 1300 mg/1 kieselguhr, or with a further nitrate addition, OMT values of

> 2500 mg/1 kieselguhr are reached.

Results of BMT of Microcystis in 287 water samples from 41 sampling

sites of the Uest Berlin water system in the months from June to October

1967 confirm that the biologically active substances in water which promote

the growth of Microcystis originate essentially from the main outfalls

carrying communal sewage, Nordgraben, Tegeler Fliess, lower Spree, Panke,

as well as Rudower Fliess; in water samples from the slowly flowing waters

Tegeler See, lower Havel, Berlin-Spandau Schiffahrtskanal as well as

Teltowkanal, influenced by the above outfalls, they are detectable in in-

creased concentration. In water samples from ail sampling sites of the

West Berlin water system, nitrate was found to ,be a limiting factor for

Microcystis (Diplocystis) aeruginosa, while phosphate was pressnt in phys-

iological excess.

Address of author: 1 Berlin 33, Corrensplatz 1.