SIST EN ISO 10710:2013

SLOVENSKI STANDARD
SIST EN ISO 10710:2013
01-julij-2013
.DNRYRVWYRGH3UHVNXV]DYLUDQMDUDVWL]PRUVNRLQEUDNLþQRPDNURDOJR
&HUDPLXPWHQXLFRUQH,62
Water quality - Growth inhibition test with the marine and brackish water macroalga
Ceramium tenuicorne (ISO 10710:2010)
Wasserbeschaffenheit - Bestimmung der Wachstumshemmung auf die marine und
ästuarine Makroalge Ceramium tenuicorne (ISO 10710:2010)
Qualité de l'eau - Essai d'inhibition de croissance sur la macro algue d'eaux marine et
saumâtre Ceramium tenuicorne (ISO 10710:2010)
Ta slovenski standard je istoveten z: EN ISO 10710:2013
ICS:
13.060.70 Preiskava bioloških lastnosti Examination of biological
vode properties of water
SIST EN ISO 10710:2013 en,fr,de
2003-01.Slovenski inštitut za standardizacijo. Razmnoževanje celote ali delov tega standarda ni dovoljeno.

---------------------- Page: 1 ----------------------

SIST EN ISO 10710:2013

---------------------- Page: 2 ----------------------

SIST EN ISO 10710:2013


EUROPEAN STANDARD
EN ISO 10710

NORME EUROPÉENNE

EUROPÄISCHE NORM
April 2013
ICS 13.060.70
English Version
Water quality - Growth inhibition test with the marine and
brackish water macroalga Ceramium tenuicorne (ISO
10710:2010)
Qualité de l'eau - Essai d'inhibition de croissance sur la Wasserbeschaffenheit - Bestimmung der
macro algue d'eaux marine et saumâtre Ceramium Wachstumshemmung auf die marine und ästuarine
tenuicorne (ISO 10710:2010) Makroalge Ceramium tenuicorne (ISO 10710:2010)
This European Standard was approved by CEN on 14 March 2013.

CEN members are bound to comply with the CEN/CENELEC Internal Regulations which stipulate the conditions for giving this European
Standard the status of a national standard without any alteration. Up-to-date lists and bibliographical references concerning such national
standards may be obtained on application to the CEN-CENELEC Management Centre or to any CEN member.

This European Standard exists in three official versions (English, French, German). A version in any other language made by translation
under the responsibility of a CEN member into its own language and notified to the CEN-CENELEC Management Centre has the same
status as the official versions.

CEN members are the national standards bodies of Austria, Belgium, Bulgaria, Croatia, Cyprus, Czech Republic, Denmark, Estonia,
Finland, Former Yugoslav Republic of Macedonia, France, Germany, Greece, Hungary, Iceland, Ireland, Italy, Latvia, Lithuania,
Luxembourg, Malta, Netherlands, Norway, Poland, Portugal, Romania, Slovakia, Slovenia, Spain, Sweden, Switzerland, Turkey and United
Kingdom.





EUROPEAN COMMITTEE FOR STANDARDIZATION
COMITÉ EUROPÉEN DE NORMALISATION

EUROPÄISCHES KOMITEE FÜR NORMUNG

Management Centre: Avenue Marnix 17, B-1000 Brussels
© 2013 CEN All rights of exploitation in any form and by any means reserved Ref. No. EN ISO 10710:2013: E
worldwide for CEN national Members.

---------------------- Page: 3 ----------------------

SIST EN ISO 10710:2013
EN ISO 10710:2013 (E)
Contents Page
Foreword . 3

2

---------------------- Page: 4 ----------------------

SIST EN ISO 10710:2013
EN ISO 10710:2013 (E)
Foreword
The text of ISO 10710:2010 has been prepared by Technical Committee ISO/TC 147 “Water quality” of the
International Organization for Standardization (ISO) and has been taken over as EN ISO 10710:2013 by
Technical Committee CEN/TC 230 “Water analysis” the secretariat of which is held by DIN.
This European Standard shall be given the status of a national standard, either by publication of an identical
text or by endorsement, at the latest by October 2013, and conflicting national standards shall be withdrawn at
the latest by October 2013.
Attention is drawn to the possibility that some of the elements of this document may be the subject of patent
rights. CEN [and/or CENELEC] shall not be held responsible for identifying any or all such patent rights.
According to the CEN-CENELEC Internal Regulations, the national standards organizations of the following
countries are bound to implement this European Standard: Austria, Belgium, Bulgaria, Croatia, Cyprus, Czech
Republic, Denmark, Estonia, Finland, Former Yugoslav Republic of Macedonia, France, Germany, Greece,
Hungary, Iceland, Ireland, Italy, Latvia, Lithuania, Luxembourg, Malta, Netherlands, Norway, Poland, Portugal,
Romania, Slovakia, Slovenia, Spain, Sweden, Switzerland, Turkey and the United Kingdom.
Endorsement notice
The text of ISO 10710:2010 has been approved by CEN as EN ISO 10710:2013 without any modification.

3

---------------------- Page: 5 ----------------------

SIST EN ISO 10710:2013

---------------------- Page: 6 ----------------------

SIST EN ISO 10710:2013

INTERNATIONAL ISO
STANDARD 10710
First edition
2010-06-15

Water quality — Growth inhibition test
with the marine and brackish water
macroalga Ceramium tenuicorne
Qualité de l'eau — Essai d'inhibition de croissance sur la macro algue
d'eaux marine et saumâtre Ceramium tenuicorne




Reference number
ISO 10710:2010(E)
©
ISO 2010

---------------------- Page: 7 ----------------------

SIST EN ISO 10710:2013
ISO 10710:2010(E)
PDF disclaimer
This PDF file may contain embedded typefaces. In accordance with Adobe's licensing policy, this file may be printed or viewed but
shall not be edited unless the typefaces which are embedded are licensed to and installed on the computer performing the editing. In
downloading this file, parties accept therein the responsibility of not infringing Adobe's licensing policy. The ISO Central Secretariat
accepts no liability in this area.
Adobe is a trademark of Adobe Systems Incorporated.
Details of the software products used to create this PDF file can be found in the General Info relative to the file; the PDF-creation
parameters were optimized for printing. Every care has been taken to ensure that the file is suitable for use by ISO member bodies. In
the unlikely event that a problem relating to it is found, please inform the Central Secretariat at the address given below.


COPYRIGHT PROTECTED DOCUMENT


©  ISO 2010
All rights reserved. Unless otherwise specified, no part of this publication may be reproduced or utilized in any form or by any means,
electronic or mechanical, including photocopying and microfilm, without permission in writing from either ISO at the address below or
ISO's member body in the country of the requester.
ISO copyright office
Case postale 56 • CH-1211 Geneva 20
Tel. + 41 22 749 01 11
Fax + 41 22 749 09 47
E-mail copyright@iso.org
Web www.iso.org
Published in Switzerland

ii © ISO 2010 – All rights reserved

---------------------- Page: 8 ----------------------

SIST EN ISO 10710:2013
ISO 10710:2010(E)
Contents Page
Foreword .iv
Introduction.v
1 Scope.1
2 Terms and definitions .1
3 Principle.2
4 Test organisms, nutrients, media, and materials.2
5 Cultivation.6
6 Apparatus.6
7 Procedure.7
7.1 Algae material.7
7.2 Preparation of dilution water.8
7.3 Choice of test concentrations.8
7.4 Start of test.9
7.5 Incubation.9
7.6 Measurement.9
7.7 Test with reference substances.9
8 Validity criteria.9
9 Evaluation of results .10
9.1 Calculations.10
9.2 Determination of E C .10
r x
9.3 Expression of results.10
9.4 Interpretation of results .11
10 Reproducibility .11
11 Test report.11
Annex A (informative) Growth pattern of Ceramium tenuicorne in 7S and 20S.13
Annex B (informative) Results of ring test with Ceramium tenuicorne at 20 S.16
Bibliography.17

© ISO 2010 – All rights reserved iii

---------------------- Page: 9 ----------------------

SIST EN ISO 10710:2013
ISO 10710:2010(E)
Foreword
ISO (the International Organization for Standardization) is a worldwide federation of national standards bodies
(ISO member bodies). The work of preparing International Standards is normally carried out through ISO
technical committees. Each member body interested in a subject for which a technical committee has been
established has the right to be represented on that committee. International organizations, governmental and
non-governmental, in liaison with ISO, also take part in the work. ISO collaborates closely with the
International Electrotechnical Commission (IEC) on all matters of electrotechnical standardization.
International Standards are drafted in accordance with the rules given in the ISO/IEC Directives, Part 2.
The main task of technical committees is to prepare International Standards. Draft International Standards
adopted by the technical committees are circulated to the member bodies for voting. Publication as an
International Standard requires approval by at least 75 % of the member bodies casting a vote.
Attention is drawn to the possibility that some of the elements of this document may be the subject of patent
rights. ISO shall not be held responsible for identifying any or all such patent rights.
ISO 10710 was prepared by Technical Committee ISO/TC 147, Water quality, Subcommittee SC 5, Biological
methods.
iv © ISO 2010 – All rights reserved

---------------------- Page: 10 ----------------------

SIST EN ISO 10710:2013
ISO 10710:2010(E)
Introduction
The red macroalga Ceramium tenuicorne belongs to Ceramiaceae, Rhodophyta. The species can be used as
a model organism for the near coastal ecosystem. This species is found in temperate marine waters in both
the northern and southern hemispheres and is thus relevant for large areas. As primary producers, they are a
food source for many invertebrates and serve as living habitat for bacteria, invertebrates, and juvenile fish.
They also serve as substrate for many oviparous fish species.

© ISO 2010 – All rights reserved v

---------------------- Page: 11 ----------------------

SIST EN ISO 10710:2013

---------------------- Page: 12 ----------------------

SIST EN ISO 10710:2013
INTERNATIONAL STANDARD ISO 10710:2010(E)

Water quality — Growth inhibition test with the marine and
brackish water macroalga Ceramium tenuicorne
WARNING — Persons using this International Standard should be familiar with normal laboratory
practice. This International Standard does not purport to address all of the safety problems, if any,
associated with its use. It is the responsibility of the user to establish appropriate safety and health
practices and to ensure compliance with any national regulatory conditions.
IMPORTANT — It is absolutely essential that tests conducted according to this International Standard
be carried out by suitably trained staff.
1 Scope
This International Standard specifies a method for the determination of the inhibition of growth of the
macroalga Ceramium tenuicorne by substances and mixtures contained in seawater or by waste water with
salinities between 4S and 32S. This method is applicable to substances that are easily soluble in water.
[4] [2]
NOTE With modifications as described in ISO 14442 and ISO 5667-16 , the inhibitory effects of poorly soluble
organic and inorganic materials, volatile compounds, metals, waste water, marine water samples, and elutriates of
sediments can be tested.
2 Terms and definitions
For the purposes of this document, the following terms and definitions apply.
2.1
algal length
length from the first division to the most distant tip of the plant
NOTE The algal length is expressed in millimetres.
See Figure 1.
2.2
control medium
combination of dilution water and/or nutrient medium used in the test
[5]
[ISO 20079:2005 , 3.6]
2.3
control batch
control medium including organisms used for testing
[5]
[ISO 20079:2005 , 3.5]
2.4
effective concentration
E C
r x
concentration of test sample at which an effect of x % is measured, if compared to the control
[5]
[ISO 20079:2005 , 3.9]
© ISO 2010 – All rights reserved 1

---------------------- Page: 13 ----------------------

SIST EN ISO 10710:2013
ISO 10710:2010(E)
2.5
growth medium
mixture of natural seawater and nutrients in which algal plants are cultivated, which is used for pre-cultures
[3]
NOTE Adapted from ISO 8692:2004 , 3.3.
2.6
growth rate
µ
〈water quality〉 proportional rate of increase in algal length per day
NOTE See Clause 9.
2.7
salinity
practical salinity
S
〈seawater〉 ratio K of the electrical conductivity of the seawater sample, at the temperature of 15 °C and a
15
pressure of one standard atmosphere, to that of a potassium chloride solutions in which the mass fraction of
−3
KCl is 32,435 6 × 10 , at the same temperature and pressure
NOTE Adapted from Reference [14], p. 12.
2.8
test medium
mixture of seawater, nutrients and test sample
[3]
NOTE Adapted from ISO 8692:2004 , 3.5.
2.9
test sample
〈water quality〉 aqueous sample, e.g. chemical substance, mixture of chemicals or waste water, for which the
inhibitory effects on the growth of algae are determined
[3]
NOTE Adapted from ISO 8692:2004 , 3.4.
3 Principle
Algal tips from monocultures of ceramium female gametophytes are grown in defined test conditions and in a
defined medium containing a range of concentrations of the test sample. The test solutions are incubated for a
period of 7 d after which the increase in length is measured and the growth rate is calculated. The growth
inhibition is determined as a reduction in growth rate, relative to control cultures grown under identical
conditions.
When toxicity of samples is to be compared to the toxicity of other chemicals or waste waters, tests can be
performed in artificial seawater. If the purpose of the testing is to assess and to predict effects in a specific
receiving water body, the tests can be conducted with algae adapted to the salinity in the receiving water body.
In this case, natural seawater from an uncontaminated site of the same properties is used.
4 Test organisms, nutrients, media, and materials
Unless otherwise stated, use only reagents of recognized analytical grade and distilled or demineralized water
or water of equivalent purity.
2 © ISO 2010 – All rights reserved

---------------------- Page: 14 ----------------------

SIST EN ISO 10710:2013
ISO 10710:2010(E)
4.1 Test organisms
Use either of the following marine algal clones:
a) Ceramium tenuicorne Kützing Waern (7S clone originating from the Baltic Sea);
b) C. tenuicorne Kützing Waern (20S and 30S clone originating from the Oslo fjord).
This alga is a widely distributed macroalgae species (phylum Rhodophyta) in estuarine and coastal areas. The
1)
strains recommended are available in unialgal, non-axenic cultures .
NOTE 1 This growth inhibition test is based on two clones, which were formerly regarded as two different species.
These species were the marine Ceramium strictum Harvey sensu Kylin and the brackish water species C. tenuicorne
Kützing Waern. Complete interfertility (References [11][12]) and DNA data (Reference [10]) have shown that the two
entities belong to the same species, with C. tenuicorne as the valid name. The marine clone (former C. strictum) used in
this test was isolated in 1973 and originates from the Oslo fjord (20S to 25S). It has been maintained as a laboratory
culture for over 30 years. The brackish water clone was isolated in 1995 and originates from the Baltic Sea, 20 km south
of the Askö laboratory in northern Baltic proper (6S to 7S). Cultures can be maintained in the medium specified in Clause 5.
Regular subculturing is necessary.
NOTE 2 Among the red algae, changes occur between haploid and diploid generations. In the growth inhibition test,
the female gametophytic generation is used since it has an even dichotomous growth pattern and the fastest growth rate.
In nature it is difficult to distinguish between male and female plants. This can be done in the laboratory where
spermatangia are found on the branches of the male plants and trichogynes can be seen on the tips of the claws on the
female plants.
NOTE 3 The Baltic Sea clone can be adapted and used in tests in salinities between 4S and 12S. The marine clone can
be used as test organism in salinities between 12S and 32S.
4.2 Natural and artificial seawater
4.2.1 General
Natural seawater is used for the cultivation of the algae and either natural or artificial seawater should be used
for testing. The type of seawater to be used depends on the objective of the test. When natural seawater is
used, care shall be taken to ensure that it is not polluted. Take special care to avoid contamination of the
water by inorganic or organic substances during preparation and storage. Equipment made of copper shall not
be used.
4.2.2 Artificial seawater
Prepare the stock solutions for artificial seawater according to Table 1.
Start with about one third of the desired volume of water, add the weighed quantities of chemicals in
accordance with Table 1 and make up to volume with water.
This stock solution with a salinity of 100S (equivalent to 10 % mass per volume) has a durability of at least six
months, if stored in darkness at room temperature. Before use, the stock solution should be diluted with water
to the desired salinity. Adjust the pH to 8,0 ± 0,2 with 1 mol/l HCl or 1 mol/l NaOH.
The artificial seawater shall be sterilized by autoclaving or sterile filtration (pore size, 0,2 µm) before use.
Re-check the pH after sterilization, and adjust if necessary to 8,0 ± 0,2 with 1 mol/l HCl or 1 mol/l NaOH,
before use.

1) Suitable suppliers are: a) ITM, Department of Applied Environmental Research of Science, Stockholm University,
S-106 91 Stockholm, Sweden; b) University of Oslo, Department of Biology, P.O. Box 1066 Blindern, N-0316 Oslo,
Norway. This information is given for the convenience of users of this document and does not constitute an endorsement
by ISO of these suppliers.
© ISO 2010 – All rights reserved 3

---------------------- Page: 15 ----------------------

SIST EN ISO 10710:2013
ISO 10710:2010(E)
Table 1 — Artificial seawater with a salinity of 10 % mass per volume or 100S
(adapted from Reference [13])
Quantity per 1 l medium Quantity per 5 l medium Quantity per 10 l medium
Substance
g/l g/5 l g/10 l
NaCl 70,1 351 702
Na SO 11,7 58,7 117
2 4
KCl 2,03 10,2 20,3
KBr 0,293 1,47 2,93
Na B O ⋅10H O 0,113 0,567 1,13
2 4 7 2
MgCl ⋅6H O 31,7 158 317
2 2
6,6 33 66
CaCl ⋅6H O
2 2
0,066 0,334 0,668
SrCl ⋅6H O
2 2
4.2.3 Natural seawater
Natural seawater shall be collected from an uncontaminated site. Filter to remove larger particles. Dilute as
necessary with water. Salinity should be increased by addition of natural seawater of a higher salinity or with
artificial seawater (see Table 1). Check the pH and adjust if necessary to 8,0 ± 0,2 with 1 mol/l HCl or
1 mol/l NaOH.
The natural seawater shall be sterilized by autoclaving or sterile filtration (pore size, 0,2 µm) before use.
Re-check the pH after sterilization, and adjust if necessary to 8,0 ± 0,2 with 1 mol/l HCl or 1 mol/l NaOH,
before use.
NOTE 1 A paper filter of around 30 µm mesh size is sufficient.
NOTE 2 Natural seawater can be stored frozen at temperatures below −18 °C for several years before use.
4.3 Nutrients
Prepare the six nutrient solutions in water, with the compositions given in Table 2.
Solutions 1, 2, 3, 4 and 6 in Table 2 are prepared in 100 ml one-mark volumetric flasks (6.14). A 1 l one-mark
volumetric flask (6.14) is recommended for the preparation of stock solution 5, due to the low masses of the
trace element reagents. Precipitation in the trace element solution is avoided by adjustment with NaOH to
pH 8. The trace element stock solution (solution 5) shall be diluted 10 times before use in the cultivation media
(see Table 3). After this dilution, a freshly prepared iron solution 3 may be added to trace element solution 5 to
increase the durability of the iron.
The iron solution shall not be older than one month.
These stock solutions will eventually be diluted according to Table 3 to obtain the final nutrient concentrations
in the growth and test media. The final concentrations in the media are given in the two rightmost columns of
Table 2.
4.4 Media
Additions of stock solutions to salt water for preparation of media are shown in Table 3. For cultivation, testing
in natural seawater, and testing in artificial seawater, additions shall be made in accordance with columns A, B,
and C, respectively.
4 © ISO 2010 – All rights reserved

---------------------- Page: 16 ----------------------

SIST EN ISO 10710:2013
ISO 10710:2010(E)
Table 2 — Nutrient stock solutions (adapted from Reference [9])
Levels in the medium after additions
according to Table 3 as
Compound mass
Reagent
elemental mass elemental amount of
concentration
concentration substance concentration
µg/l µmol/l
1 — Nitrogen solution
KNO 5 000 mg/100 ml 3 462 (N) 247 (N)
3
2 — Phosphorus solution
KH PO 680 mg/100 ml 775 (P) 25 (P)
2 4
3 — Iron solution
FeCl ·6H O
100 mg/100 ml 103 (Fe) 1,9 (Fe)
3 2
4 — Carbon solution
NaHCO 5 760 mg/100 ml 16 480 (C) 1 370 (C)
3
5 — Trace element solution
Na EDTA 6 000 mg/l
2
MnSO ⋅H O 620 mg/l 10 (Mn) 0,18 (Mn)
4 2
250 mg/l 2,84 (Zn) 0,043 (Zn)
ZnSO ⋅7H O
4 2
Na MoO ⋅2H O 130 mg/l 2,58 (Mo) 0,027 (Mo)
2 4 2
CoSO ⋅7H O 4 mg/l 0,042 (Co) 0,000 7 (Co)
4 2
4 mg/l 0,05 (Cu) 0,000 8 (Cu)
CuSO ⋅5H O
4 2
6 — Vitamins
Thiamine (B ) 10 mg/100 ml 50
1
Cyanocobalamin (B ) 0,1 mg/100 ml 0,5
12
Biotin 0,1 mg/100 ml 0,5
Table 3 — Additions of stock nutrient solutions to seawater
for preparation of growth and test medium
A B C
Growth medium Test medium Test medium
Stock solutions
in natural seawater in natural seawater in artificial seawater
for cultivation
ml/l ml/l ml/l
1 — Nitrogen solution 0,5 0,5 0,5
2 — Phosphorus solution 0,5 0,5 0,5
3 — Iron solution 0,5 0,5 0,5
4 — Carbon solution — — 2
5 — Trace element
0,5 — —
solution diluted 10 times
6 — Vitamin solution 0,5 — —
© ISO 2010 – All rights reserved 5

---------------------- Page: 17 ----------------------

SIST EN ISO 10710:2013
ISO 10710:2010(E)
Preparation of 1 l of growth medium or test medium is shown in Table 3. For the cultivation of stock cultures,
sterile natural seawater shall be used. For the growth inhibition test, either sterile artificial or natural seawater
may be used depending on the objective (see Clause 4).
5 Cultivation
Stock cultures of female gametophytes of C. tenuicorne are cultivated in natural seawater (see 4.2.3) where
nutrients have been added according to Table 3, column A. The brackish water clone is cultivated at a salinity
of 7S and the marine clone (former C. strictum) at salinities of 20S and 30S.
The maintenance of the cultures is facilitated by working under aseptical conditions. C. tenuicorne should be
grown in sterile plastic or glass Petri dishes (e.g. 90 mm diameter, 15 mm height). The algae are cultured at
−2 −1 −2 −1
22 °C ± 2 °C, at a light intensity of 35 µmol m s ± 7 µmol m s and a light regime of 14 h light and 10 h
darkness. These conditions can be achieved on a laboratory bench with an ordinary time-regulated lamp at
about 350 mm distance.
To maintain actively growing female gametophytes, plants have to be transferred to fresh medium each week.
The procedure is to fill sterile Petri dishes with approximately 25 ml sterile culture medium according to
Table 3 column A. Approximately 20 to 30 tips of length 10 mm to 20 mm from female plants are transferred to
each dish.
If the test is intended to be performed at a salinity other than 7S, 20S or 30S, the algae need to be adapted to
the new salinity prior to the start of the test. Seawater of other salinities is prepared according to 4.2.2 and
4.2.3. The algae should be adapted successively by transferring the algae every other day into fresh media
with an increase or decrease in salinity of 3S. The algae shall be cultivated for at least two weeks in the final
test salinity prior to the start of the test.
NOTE 1 To increase the stability of iron, iron solution 3 can be added to the diluted trace element solution.
NOTE 2 Back-up cultures can be held for up to two months without refreshing if cultivated under lower light intensities
at room temperature. If the temperature is at approximately 10 °C, the algae can be kept for up to three months.
6 Apparatus
All equipment that has contact with the test medium shall be made of glass or other chemically inert material.
Usual laboratory equipment and in particular the following.
6.1 Temperature-controlled cabinet or room, with a white fluorescent light providing even illumination,
suitable for the lighting requirements specified for the test in 7.5.
6.2 Fluorescent tubes of the daylight or “warm white” type, capable of providing a light intensity of
−2 −1 −2 −1
(70 ± 7) µmol m s for the exposure period, and around (35 ± 7) µmol m s for cultivation.
6.3 Timer, to control a light regime of 14 h light and 10 h darkness.
6.4 Light meter, which measures photons or energy (expressed in micromoles per square metre per
second), within the photosynthetic range 400 nm to 700 nm, is preferred.
6.5 pH meter, for the measurement and adjustment of pH during the preparation of culture and test
solutions and to measure pH at the termination of a test.
6.6 Salinity meter or conductivity meter, to measure and adjust the salt content of the culture and dilution
water.
6 © ISO 2010 – All rights reserved

---------------------- Page: 18 ----------------------

SIST EN ISO 10710:2013
ISO 10710:2010(E)
6.7 Stereomicroscope, with a magnification of 6 times to 10 times, for cutting female plants prior to a test
and for measuring the length of the plants.
6.8 Sterilization equipment. All equipment and solutions should be sterile to prevent contamination. This
can be done in an autoclave. Solutions can also be sterilized by filtration (pore size, 0,2 µm). Glassware can
be sterilized in a muffle furnace at 150 °C for 3 h.
6.9 Gas burner or spirit lamp. To maintain the plants as a monoculture, sterile microbiological techniques
should be employed for all work in connection with the algae. Bottles and instruments should be flamed when
working with the cultures and preparing solutions.
6.10 Scalpel, scissors, and a pair of pincers are useful tools for c
...