Common borders. Common solutions.

Hydrochemical and hydrobiological

sampling guidance

CZU 543.3+574.5=135.1=111
G 49

ACADEMY OF SCIENCES OF MOLDOVA

INSTITUTE OF ZOOLOGY

The guide is a generalization of materials on the procedures and techniques of hydrochemical and
hydrobiological sampling as part of ecological monitoring of lentic and lotic water bodies, and it is
addressed to students, young researchers and all those who are interested in issues of water quality,
aquatic biodiversity and protection of aquatic ecosystems.
The work is approved and recommended for editing by the Scientific Council of the Institute of
Zoology of the Academy of Sciences of Moldova.

EDITORS:
Todera Ion, doctor habilitatus of biology, professor, academician
Zubcov Elena, doctor habilitatus of biology, professor
Bilechi Lucia, doctor of biology, associate professor
REVIEWER:
alaru Vasile, doctor habilitatus of biology, professor, corresponding member
of the Academy of Sciences of Moldova

Ghid de prelevare a problelor hidrochimice i hidrobiologice = Hydrochimical

and hydrobiological sampling guidance / Progr. Operaional Comun RomniaUcraina-Republica Moldova 2007-2013 ; ed.: Todera Ion [et al.]. Chiinu :
S. n., 2015 (Tipogr. Elan Poligraf ). 64 p. (Granie comune. Soluii comune,
ISBN 978-9975-128-28-5).
Tit., text paral.: lb. rom., engl. 200 ex.
ISBN 978-9975-66-480-6.

TABLE OF CONTENTS

I
II
III
IV
V
VI
VII
VIII
IX

Introduction
Zubcov E., Biletchi L.
General principles of sampling in aquatic ecosystems
Toderas I., Zubcov E., Biletchi L.
Sampling of water
Zubcov E., Bagrin N., Biletchi L., Borodin N., Zubcov N.
Sampling of suspensions and sediments
Jurminskaia O., Bagrin N., Zubcov N., Borodin N., Ivanov A.
Sampling of bacterioplancton
Subernetkii I., Negru M.
Sampling of phytoplankton
Ungureanu L., Tumanova D., Ungureanu G.
Sampling of zooplankton
Jurminskaia O., Subernetkii I., Lebedenco L.
Sampling of zoobenthos
Munjiu O., Toderas I., Banu V.
Sampling of ichthyologic material
Bulat Dm., Bulat Dn., Toderas I.,Usatai M.
Sources of sampling errors
Jurminskaia O.
ANNEX 1
Sampling frequencies for physico-chemical and hydrobiological parameters
ANNEX 2
General techniques for preservation of water samples
ANNEX 3
Recommended washing procedures for selected water quality elements

4
5
6
9
11
12
14
18
22
25
28
29
32

INTRODUCTION
Not for school but for life we learn (Seneca)
This work is published in the frame of the project Cross-border interdisciplinary cooperation for the
prevention of natural disasters and mitigation of environmental pollution in Lower Danube Euroregion
MIS ETC 1676, which has been carrying out during 2013-2015 by a team of researchers from Romania,
the Republic of Moldova and Ukraine.
The project targeted the enforcing of cross-border cooperation aiming to elaborate long-term solution
for pollution mitigation, sustainable development and emergency preparedness in the Lower Danube
Euroregion.
In order to fulfil the project goal, a range of objectives were set:
integration of the capacities and expertise of the partner institutions to assess the cross-border
impact of the environmental problems in the Lower Danube River basin;
creation of a mechanism capable to provide the scientific support for decision makers for the
elaboration of policy related to the prevention of the negative impact of potential natural and
anthropogenic accidents in the region;
raising the level of awareness of the population in the region regarding the environmental issues
with cross-border impact;
encouraging the involvement of young researchers, undergraduate, master and PhD students in
cross-border environmental research;
improvement of ecological situation in Lower Danube Euroregion.
The project has allowed creating by the partner institutions of an international interdisciplinary
network (INPOLDE), designed to accumulate background data, conduct complex environmental
investigations in border areas, assess the environmental impact and, finally, to offer a better scientific
support to the regional monitoring programs.
The work Hydrochemical and hydrobiological sampling guidance is part of a series of works to be
edited under the above-mentioned project, which is dedicated to the methodological aspects of research
on various environmental components, starting with the methods of filed sampling and ending with
modern analytical techniques in assessing the level of environment pollution. It is based on extensive
personal experience of the authors, and the good international practices in studying the physico-chemical
and biological components of freshwater ecosystems.
To the preparation of this work the researchers of the Institute of Zoology, the Academy of Sciences
of Moldova, have contributed, and namely: Toderas Ion, academician, Zubcov Elena, doctor habilitat
in biology, professor, Ungureanu Laurenia, doctor habilitat in biology, professor, Usatai Marin, doctor
habilitat in biology, professor, Bilechi Lucia, doctor in biology, associated professor, Jurminskaia Olga,
doctor in biology, Subernetskii Igor, doctor in biology, associated professor, Negru Maria, doctor in
biology, associated professor, Bagrin Nina, doctor in biology, associated professor, Munjiu Oxana, in
biology, associated professor, Zubcov Natalia, doctor in biology, associated professor, Bulat Dumitru,
doctor in biology, associated professor, Bulat Denis, doctor in biology, associated professor, Borodin
Natalia, Lebedenco Liubovi, Ungureanu Grigore, Tumanova Daria and Banu Vitalie.

I. GENERAL PRINCIPLES OF SAMPLING IN AQUATIC ECOSYSTEMS

Sampling is defined in ISO 5667-2 as the process of removing the portion, intended to be
representative, of a water body (or sludge, or sediment) for the purpose of examination for various defined
characteristics. Sampling is the first and very important stage in monitoring of aquatic ecosystems. The
use of sophisticated equipment and methods in the process of analysis cannot mitigate the impact of
mistakes made during the sampling.
General principles and techniques of sampling for natural aquatic ecosystems are regulated in the
set of international standards from the category Water quality Sampling (ISO 5667), approved at
the national level [1-5]. At the national level had been also approved some of international standards
(in the category of guidance) had been also approved that regulate the specific techniques of sampling
for certain groups of hydrobionts in their natural habitat [6-10]. Some important guides from this set of
European documents are not yet included in the register of national standards, making them difficult to
use in the Republic of Moldova [11 12].
Sampling conditions and requirements are quite different depending on the type of waters, program
and purpose of monitoring. It is difficult to elaborate unique recommendations, but there are always
basic principles regarding the physico-chemical and biological monitoring of aquatic ecosystems:
Sampling periodicity depends on the type of monitoring. The design of the monitoring programme
should take into account the variability of parameters, which may change both under the influence
of natural conditions and as result of human impact.
Collected material should be representative and to correspond to the purpose and programme of
monitoring.
Samples should reflect the actual state in the sampling location.
Transport, stabilization and storage of samples should have the lowest possible influence on their
physico-chemical and biological composition. It is recommended that sample containers are
tightly sealed and protected from the effects of light and excessive heat, because the characteristics
of the sample may rapidly deteriorate due to gas exchange, chemical reactions and the metabolism
of organisms which may be present.
Volume of collected samples should be sufficient to perform analysis in accordance with the used
methods and laboratory techniques.
Recipients (vessel, containers) must be made of inert materials. In most cases, collecting of samples
should be done in flasks made of polyethylene or other plastics. In the case of sampling for the
organic substances determination the glass flasks should be used.
All sampling procedures should be strictly documented, allowing an easy identification of sample
in the laboratory conditions. Each sampling point should be described. In the case of a long-term
programme, conditions which are agreed and remain unchanged need not be restarted. In this
case only results of in situ measurements and the variables such as weather conditions and unusual
observations should be recorded. When sampling is conducted for special reasons, detailed
information should be given, including the reasons for sampling and any preservation steps taken.
Samples are collected by a qualified staff and in compliance with safety standards (it is mandatory
the training of samplers in order to ensure appropriate safety conditions).
Samplers shall take into consideration that in some rivers or watercourses there are dangers of
chemical, bacteriological, viral or animal origins.
References
1. ISO 5667-1:2006 Water quality. Sampling. Part 1: Guidance on the design of sampling programmes
and sampling techniques. National version: SM SR EN ISO 5667-1:2011 Calitatea apei. Prelevare.
Partea 1: Ghid general pentru stabilirea programelor i tehnicilor de prelevare (in Romanian).
Chiinu: INSM, 2011. 35 p.
2. ISO 5667-4:1987 Water quality. Sampling. Part 4: Guidance on sampling from lakes, natural and
5

man-made. National version: SM SR ISO 5667-4:2007 Calitatea apei. Prelevare. Partea 4: Ghid
de prelevare a apelor din lacuri naturale i artificiale (in Romanian). Chiinu: MOLDOVASTANDARD, 2007. 10 p.
3. ISO 5667-6:2005 Water quality. Sampling. Part 6: Guidance on sampling of rivers and streams.
National version: SM SR ISO 5667-6:2011 Calitatea apei. Prelevare. Partea 6: Ghid pentru prelevrile
efectuate n ruri i alte cursuri de ap (in Romanian). Chiinu: INSM, 2011. 22 p.
4. ISO 5667-12:1995 Water quality. Sampling. Part 12: Guide on sampling of bottom sediments.
National version: SM SR ISO 5667-12:2007 Calitatea apei. Prelevare. Partea 12: Ghid general pentru
prelevarea sedimentelor de fund (in Romanian). Chiinu: MOLDOVA-STANDARD, 2007. 42 p.
5. ISO 19458:2006 Water quality. Sampling for microbiological analysis. National version: SMV EN
ISO 19458:2011 Calitatea apei. Prelevare pentru analiz microbiologic (in English). INSM, 2011.
30 p.
6. EN 15110:2006 Water quality. Guidance standard for the sampling of zooplankton from
standing waters. National version: SM SR EN 15110:2012 Calitatea apei. Ghid pentru prelevarea
zooplanctonului din ape stttoare (in Romanian). Chiinu: INSM, 2012. 24 p.
7. EN 15708:2009 Water quality. Guidance standard for the surveying, sampling and laboratory
analysis of phytobenthos in shallow running water. National version: SM EN 15708:2013(E)
Calitatea apei. Ghid pentru studiul, prelevarea i analiza n laborator a fitobentosului din apele
curgtoare de mic adncime (in English). INSM, 2013. 25 p.
8. EN 13946:2003 Water quality. Guidance standard for the routine sampling and pre-treatment of
benthic diatoms from rivers. National version: SM SR EN 13946:2012 Calitatea apei. Ghid pentru
prelevarea uzual i pretratarea diatomeelor bentonice din ruri (in Romanian). INSM, 2012. 22 p.
9. EN 14757:2005 Water quality. Sampling of fish with multi-mesh gillnets. National version: SM SR
EN 14757:2012 Calitatea apei. Prelevarea petilor utiliznd plase cu mrimi diferite ale ochiurilor
(in Romanian). INSM, 2012. 36 p.
10. EN 14962:2006 Water quality. Guidance on the scope and selection of fish sampling methods.
National version: SM SR EN 14962:2012 Calitatea apei. Ghid pentru domeniul de aplicare i
selecie a metodelor de prelevare peti (in Romanian). INSM, 2012. 34 p.
11. ISO 10870:2012 Water quality. Guidelines for the selection of sampling methods and devices for
benthic macroinvertebrates in fresh waters (This edition of ISO 10870 cancels and replaces ISO
7828:1985, ISO 8265:1988, and ISO 9391:1993 which have been technically revised). ISO, 2012.
26 p.
12. EN 16150:2012 Water quality. Guidance on pro-rata Multi-Habitat sampling of benthic macroinvertebrates from wadeable rivers. BSI, 2012. 18 p.

II. SAMPLING OF WATER

There are two different types of water sample that can be taken from rivers, lakes and similar surface
waters [23]: 1) instantaneous samples that are collected at a certain location, depth and time, and
2) composite or integrated samples that may be depth-integrated, area-integrated, time-integrated or
discharge-integrated.
Different types of sampler are available (dissolved oxygen sampler, depth sampler, multi-purpose
sampler), many of them designed for specific purposes.
The volume of collected water depends upon the analyses to be carried out, and ranges from 100 ml
to 5 litres (volume collected should be sufficient to insure a representative sample, allow for at least three
replicate analyses).
The temperature of the water should be measured and recorded immediately after the sample is taken.
Physical parameters (for example pH, dissolved gases, suspended solids) should be determined on site or
as soon as possible afterwards.
Containers for the transportation of samples should be used only for water samples and never for
the storage of chemicals or other liquids. Glass containers are commonly used and are appropriate for
6

samples for many analyses, but for certain chemical analyses, for biota or sediments plastic containers are
preferred. Sample containers and their recommended washing procedures for selected water ingredients
are presented in Annex 2.
A bottle/container that is to be used for transport or storage of the sample should be rinsed three
times with portions of the sample before being filled. This does not apply, however, if the transport bottle
already contains a preservative chemical. No preliminary rinsing is done before collection of suspensions
and microbiological samples. Samples should be transferred to the storage/transport bottle immediately
after collection. A small air space should be left in the storage/transport bottle to allow the sample to be
mixed before analysis. Each sample bottle must be provided with an identification label.
A range of physico-chemical parameters such as temperature, dissolved oxygen, carbon dioxide, pH,
redox potential, conductivity and turbidity are measured at the time of sampling direct in the water body,
using portable probe meters (Fig. 1 3). The instruments must be in good working condition in order to
get accurate results and have to be calibrated every time in situ. The exceptions are: temperature (check
in laboratory monthly with a certified mercury thermometer), conductivity and turbidity (calibrate at
the beginning of the sampling expedition), and redox potential (calibrate once every six months).

Fig. 1 Portable pH/EC/ TDC/

Temperature Meter

Fig. 2 Portable DO/Barometric

pressure/Temperature Meter

Fig. 3 Portable CO2 Meter

The dissolved oxygen (DO) can be determined with the traditional iodometric Winkler-method. This
method is used for calibration of different portable DO Meters, and as a reliable laboratory method, often
used in the aquatic ecology. For this method the most important step is the collection of the sample into
the narrow-mouthed glass flasks with stoppers (Winkler flasks, or any other suitable flasks) of capacity
between 130 and 350 ml, calibrated to the nearest 1 ml (Fig. 4).
Each flask and its stopper shall bear the same identification number. During sampling
precautions must be taken to ensure that no bubble is left under the stopper. Flasks
should be filled up to overflowing, taking care to avoid any change in the concentration
of dissolved oxygen. After filling, the dissolved oxygen should be immediately fixed
with manganese (II) sulphate solution and alkaline reagent as stipulated this method
[13]. The flask is inverted several times to mix the contents thoroughly. Allow the
precipitate which is formed to settle for at least 5 min and then mix by inversion again
Fig. 4 Winkler
to ensure that the mixture is homogeneous. In this state the flasks may be transported
oxygen flask
to the laboratory (Fig. 5).
If protected from light, the samples may be stored for up to 24 h. If in water is present the suspended
matter capable of fixing or consuming iodine, it is preferable to use the electrochemical probe method
[14]. All measurements taken in the field and all supporting information (the weather, growth of algae,
the presence of dead fish floating in the water or oil slicks, etc.) must be recorded in the field notebook
before leaving the sampling station.
The separate portions of the water sample should be set aside for pH and conductivity determinations.
7

The same portion must not be used for both determinations because of the possibility of potassium
chloride diffusing from the pH probe.
For the determination of proportion between the soluble and insoluble forms of metals the water
samples are filtered immediately after collection through filters with pore size of 0.45 m (Fig. 6). The
filtered water is preserved with nitric acid. Filtering does not apply if the membrane filter may keep one or
more constituents analyzed. It is also essential that the filtration system can be easily washed prior to filling.

Fig. 5 Oxygen box and flasks with fixed dissolved

oxygen ready for transport

Fig. 6 Filtering water for metal analysis

in field conditions

The samples collected for determination of pesticides

are preserved according to standard methods by using
different solvents.
The basic requirements for sample storage without
deteriorating are the follows: sample bottles should be
sealed and stored in a clean, cool, dark environment and
protected from recontamination. Additional methods of
preservation include freezing, solvent extraction and the
addition of chemical preservatives. However, in each case
the preservation and storage method shall be compatible
with the used analytical techniques. Suggested chemical
preservatives and maximum permissible storage times are
Fig. 7 Portative spectrophotometer
for chemical analysis
regulated by the international standard ISO 5667-3 [15].
The ideal devises that can be used for water sampling are the
Van Dorn Sampler and bathometers (Fig. 8). Van Dorn sample bottles are well suited for general purpose sampling
at any depth. They are made both in horizontal and vertical design. The horizontal bottles are descending parallel
to the bottom. They are ideal for sampling various stratification levels of water, the thermocline or just above the
bottom sediments. The vertical bottle design is typically used for stratification studies with multiple or single
samplers suspended by cable. The Alpha Van Dorn bottle is used for general physical, chemical and biological
sampling. It has a wide open mouth that does not restrict water flow and is ideal for collecting microplankton.
The Beta Van Dorn Sampler can be used for trace metals and organic pollutants.
Sometimes, when the sampling point is situated on the bank of a river or lake, the sample is collected
directly in vessel for transportation by using any vessel (as example, a bucket) made from a material,
which is not able to contaminate the sample (Fig. 9).

Fig. 8. Van Dorn sampler [16]

Fig. 9. Prelevation of water in a shallow lake

References
13. EN 25813:1992 Water quality. Determination of dissolved oxygen: iodometric method. (ISO
5813:1983) National version: SM SR EN 25813:2011 Calitatea apei. Determinarea coninutului de
oxigen dizolvat. Metoda iodometric (in Romanian). Chiinu: INSM, 2012. 12 p.
14. ISO 5814:1990Water quality. Determination of dissolved oxygen: electrochemical probe method
National version: SM SR EN ISO 5814:2014 Calitatea apei. Determinarea coninutului de oxigen
dizolvat. Metoda electrochimic cu sonda (in Romanian). Chiinu: INSM, 2014. 25 p.
15. EN ISO 5667-3:2003 Water quality. Sampling. Part 3: Guidance on the preservation and handling of
water samples. National version: SM SR EN ISO 5667-3:2011 Calitatea apei. Prelevare. Partea 3: Ghid
pentru conservarea i manipularea probelor de ap (in Romanian). Chiinu: INSM, 2012. 31 p.
16. http://shop.sciencefirst.com/wildco/567-van-dorn-water-samplers.

III. SAMPLING OF SUSPENSIONS AND SEDIMENTS

Collection of suspensions and sediments
The collection of suspensions is of high importance in the research of migration of chemical substances
in the water suspensions - bottom sediments system. As rule, the filtration methods are used for their
collection. The filters are chosen in dependence of pore size. Mainly the paper, membrane or glass filters
are used, in accordance with the methods of determination of chemical substances.
In order to analyse the content of metals in suspensions, a determined volume of water is filtered (200-1000
ml). The filter with suspensions in placed in a special vessel and further analysed in laboratory conditions.
Sediments are a recommended matrix for the assessment of chemical status in water ecosystems,
especially lakes and pond [17].
Bottom sediments can be used as a matrix for three types of investigation: chemical, physical and
biological. A biological survey generally involves classification and number of bacterio-, phyto- and
zoobenthos groups in sediment.
The main principles of the sediment sampling procedure (including choice of sampling site and
sampling points, choice of sampling method, etc.) are described in the international guideline document
ISO 5667-12 [18]. As a general principle, the sampling procedure should not alter the properties of the
sediment (e.g. by contamination or disturbing the sample). Analysis of sediments can be a cost-effective
approach for initial screening of areas for contamination, to compare contaminant concentrations in
different areas and to identify possible sources of contaminants.
The minimum frequency required for water monitoring of priority substances is once per month
(once every 3 months for river-basin-specific pollutants), but for sediment the monitoring frequency can
be once per year, unless technical knowledge and expert judgement justify another interval.
Sediments can be collected using either a grab or core sampler. A wide range of sampling devices is available
(Fig. 10). The choice of equipment should be made depending on the local conditions at the site of sampling,
e.g. water depth and type of sediment. Core samplers are tube-like devices that penetrate the sediment by
gravity (free-fall), vibration, hydraulic pressure or by hand. Either a grab sampler or a corer may be used to
sample the top layer of the sediment, while in smaller and shallow rivers, scoops can also be employed.

Fig. 10: a grab sampler [19], b grab sampler [20], c hand corer [21], d gravity corer [22], e box corer [21]

When the migration of metals is investigated, the upper 5 cm layer of sediments is collected. The
content of metals is determined per total sediments or separately in different sediment granulometric
fractions. A part of sample is centrifuged for 30-45 min with a speed of 2500-3000 rotation/min in order
to obtain the water extract, where the most mobile forms of metal migration in the water bottom
sediments system occur.
Sediment samples sieving and transport
Sediment samples must be sieved over 2 mm mesh to remove large detritus and benthic organisms.
Otherwise, during subsequent sample handling and processing, such as storage, freezing or ultrasonic
treatment, biotic material will deteriorate and become part of the sediment sample. In order to minimise
the disturbance of the sediment/water equilibrium wet sieving is best performed at the sampling point
with ambient water. The same water should be reused to prevent changing the equilibrium. In the case of
saline samples it is particularly important to sieve with water with approximately the same salinity as at
the sampling location. In general, sieving should be performed during the first 24 hours after collection
of sediments, but it is preferably to do it in field conditions. If field sieving is not possible, sieving should
take place in the laboratory under controlled conditions.
Sieves are traditionally made of corrosion-resistant brass (rim and mesh). Today, stainless steel is
preferred for organics analyses (Fig. 11). However, these must not be used for the analysis of trace metals.
In this case, polymer sieves are recommended: PVC or acrylic rim, with nylon or polyester mesh.
Samples (sieved or not) are transferred preferably into wide-mouth bottles of amber glass (or
aluminium, or other non-contaminating material) for organic analysis or into plastic bags or bottles
for trace element analysis. Alternatively, amber glass jars can be used for all kinds of contaminants (Fig.
12). Sampling containers should be filled to the top (minimal headspace) to reduce the likelihood of
oxidation and loss of acid volatile sulphide during transport. It is preferable to transport samples as
soon as possible to the laboratory under refrigerated conditions (about 4C). Refrigeration is easily
accomplished with cooling boxes (Fig. 13).

Fig. 11. Stainless sieves [23]

Fig. 12. Amber glass jar [24]

Fig. 13. Cooling box [25]

Sediment sample storage

Storage begins when the samples are taken. All storage methods will affect the sample to some extent,
and the choice of preservation technique depends mainly on the objective of the sample collection.
Because the first few hours after sampling are the most critical for changes to occur in the sample,
preservation steps should be taken, where possible, immediately upon sample collection. Particularly,
this is very important in the case of investigation of pesticides and volatile compounds in sediments.
Containers for storing lyophilised or dried sediment samples are preferably wide-mouth bottles with
a screw cap. Samples taken for the analysis of organic contaminants must be stored in amber glass, PTFE
(polytetrafluoroethylene) containers or stainless steel and aluminium containers. Sediments collected
10

for analysis of metals can be stored in closed plastic or glass containers. It is possible to use a amber glass
jar for all determinations in order to simplify sediment characterisation.
For mercury, samples must be stored in acid-washed borosilicate glass or quartz containers. For
organic compounds, storage of samples is preferably done in amber glass bottles, but containers of other
materials such as polycarbonate or aluminium are also suitable. Maximum suggested time of storage of
freeze dried sediment before analysis is about 180 days (30 days for Hg) if stored in a freezer.
Sediment samples can be archived. If metals are investigated, then a simple archiving procedure
may be applied, which consists of drying sample at 30C or 105C and storing of dried sediments in
polyethylene containers.
References
17. Guidance on chemical monitoring of sediment and biota under the Water Framework Directive.
Luxembourg: Office for Official Publications of the European Communities, 2010. 82 p.
18. ISO 5667-12:1995 Water quality. Sampling. Part 12: Guidance on sampling of bottom sediments.
National version: SM SR ISO 5667-12:2007 Calitatea apei. Prelevare. Partea 12: Ghid general
pentru prelevarea sedimentelor de fund (in Romanian). Chiinu: MOLDOVA-STANDARD,
2007. 41 p
19. http://www.envirotools.hu/english/benthic_grabs.html.
20. http://www.gserentals.co.uk/details.aspx?product=93
21. http://clu-in.org/programs/21m2/sediment
22. http://www.osil.co.uk/Products/RentalEquipment/tabid/59/agentType/View/PropertyID/314/
Default.aspx
23. http://www.gemplers.com/product/134572/Stainless-Steel-Sieve-Set
24. http://www.inglass.com/amberjars.html
25. http://www.waecofridges.co.uk/coolbox/mobicoolu26.htm

IV. SAMPLING OF BACTERIOPLANKTON

For microbiological analysis the thick-walled glass bottles with a minimum capacity of 300 ml should
be used. Before autoclaving, the bottle caps should be covered by Kraft paper to protect them from
contamination during handling. To prepare sample bottles, they should be washed with a non-ionic
detergent and rinsed at least three times with distilled or deionised water before autoclaving. New bottles
require the same preparation.
Selection of the mode and place of water sampling depends on the research purposes and local
conditions. The main requirements are [5, 26-27]:
Bottles for water samples are sterilized in laboratory dryer (Fig. 14) for 1 hour at 180C or
autoclaved for 20 minutes at the 1 atm pressure that corresponds to 121C (Fig. 15).
Samples from the surface layers of water cannot be collected in the vicinity of a pollution source
(except of cases foreseen in research programme).
Necessary quantity of water varies from 500 ml to 1000 ml, in dependence of the type of
performed analysis.
Bottles for sample collection should be open only prior to sampling.
Surface samples are collected at a depth of 15 - 30 cm.
In the case of sampling from small stagnant water bodies it is necessary to prevent disturbing the
bottom sediments.
Bottles are filled with collected water so that the distance between the water level and the bottle
cap is at least 2 cm.
During the sampling any possibility of accidental contamination of water should be avoided.
After sampling the bottles are labelled by indicating the place, date, time and eventual
characteristics of sampling points. All samples are listed in the register.
11

Fig. 14. Laboratory dryer

and sterilizer [28]

Fig. 15. Vertical-loading

laboratory autoclave [29]

Fig. 16 Transport box for

bacterioplankton samples [30]

If the delay between sample collection and bacteriological analysis will be less than 2 hours, samples
should simply be kept in a cool, dark place. When more than 2 hours will elapse, samples should be chilled
rapidly to about 4C by placing them in a cooling box, where they should remain during transportation
(Fig.16). If the time between collection and analysis exceeds 6 hours, the report of the analysis should
include information on the conditions and duration of sample transport.
The collection of samples in the running waters is made in the same way as in stagnant surface waters.
In some cases, particular at the collection of water samples at the water treatment stations, a special
sampler is necessary to be used.
Analysis of the bacterioplankton samples should be carried out as fast as possible (within 6 hours)
after their collection. If it is not possible to return samples to the laboratory within this period, they may
be stored in the fridge, but never exceeding 24 hours [26].
References
26. Toderas I., Negru M., Ionica D., Nicolescu D., Simon-Gruita A. Ecology of aquatic microorganisms.
Original title: Todera I., Negru M., Ionic D., Nicolescu D., Simon-Grui A. Ecologia
microorganismelor acvatice (in Romanian). Chiinu: tiina, 1999. 281 p.
27. Kopilov A., Kosolapov D. Bacterioplankton of reservoirs of Upper and Middle Volga. Moscow:
Original title: A., .
(in Russian). : , 2008. 377 .
28. http://eprus.pl/farma-play-suszarki-sterylizatory/.
29. http://labimpex.com.ua/p42078351-sterilizator-parovoj-avtoklav.html.
30. http://www.who.int/water_sanitation_health/resourcesquality/wqmchap5.pdf.

V. SAMPLING OF PHYTOPLANKTON
Phytoplankton is represented by microscopic organisms, free leaving in the water, capable to
photosynthesis. Researches of phytoplankton, determined by its main role as producer of organic substance
and as a factor of formation of water quality, have a great importance in the elaboration of scientific bases
of the management of aquatic ecosystems, building of high productive hydrobiocenoses, relatively stable
despite of the influence of anthropogenic factors. Structure and functional characteristics of phytoplankton
to the highest extent determine the character of biological productivity of aquatic ecosystems.
The productivity of phytoplankton, in contrast with hydrobionts, which refer to higher trophic levels,
can be determined without analysing the phytoplankton.
The method of collecting samples of phytoplankton is relatively simple, but their processing requires
high qualifications of engaged personnel.
Selection of sampling points
The selection of horizontal sampling points and the frequency of their collection in time are determined
in accordance with the goal and objectives of hydrobiological investigations. In all cases the collection
12

of samples and the determination of phytoplankton primary production are performed synchronically
in the same points.
The choice of vertical sampling points of phytoplankton will be made in accordance with the
characteristics of the physical factors, which influence the development of algae (light, temperature).
In most of water bodies (except of those very deep) the number of points of collecting samples at
different depths (h), which are necessary to record the phytoplankton at different levels, may be limited
to six [31]:
1) surface area of the highest illumination and heating, concomitantly withstanding the influence
of the wind;
2) horizon located at the half of the value of transparency;
3) horizon located at the depth of transparency basically coincides with the middle of the trophogenic
layer;
4) horizon corresponding to the double depth of transparency; as a rule, it coincides with the limits
of trophogenic area (the point of compensation lies here, and the thermocline at the setting of
summer stratification);
5) horizon corresponding to the half of tropholitic zone; it is determined as the half way from the
bottom of the water body up to the double value of transparency;
6) horizon located at the 0.3 0.5 m from the bottom.
In the case of the vertical sampling it is necessary to take into consideration that in the relatively small
water bodies vertical migrations of phytoplankton occur, influenced by changes of aquatic ecosystems
within 24 hours and weather conditions. In rivers sampling shall be carried out at 0.2 - 1 m, due to the
relatively homogenous vertical distribution of phytoplankton.
Methods of sampling and equipment
Methods of sampling and investigation of algae are very
diverse, being determined as by the ecological and morphological
peculiarities of representatives of different phyla and ecological
groups, as the diversity of goals and modes to approach the
investigations. There will not be described methods that are
designed to achieve specific goals, but only the sampling
and investigation methods of algae from continental aquatic
ecosystems, designed for floristic - systematic and hydrobiological
researches.
In order to study the specific composition of the phytoplankton
during
its intense development,
Fig. 17. Plankton net [32]
it is sufficient to sample water
in a bottle and then to analyse the sample under a microscope. But
in most cases different methods of prior concentration are applied.
One of these methods consists of filtering water through the plankton
nets of different construction (Fig. 17).
Water is taken from surface layers (up to 15 20 cm of deepness)
with a sample bottle and drawn through a net, and thus filtering out
50 100 litre of water. In large rivers and lakes the plankton samples
shall be taken from a boat. It is recommended to drag the plankton
net with a thin rope behind the moving boat for 5 10 min. Devices
Fig. 18. Ruttner sampler for vertical
with the special construction are used for the vertical sampling of
plankton sampling [32]
phytoplankton (Fig. 18).
In the small water bodies plankton samples can be collected from
the bank by advancing gradually into the water and filtering water from the front side through the net,
or the net is threw into the water and carefully pulled by string.
13

After sampling was done, the plankton net shall be rinsed well by sinking it few times in the water up
to circle in order to remove the remaining algae from the internal surface of the net.
Concentrated in such a way, the plankton sample is poured from the cup of the net, through the
evacuation tube, into a bottles prepared in advance. Before and after sampling the net must be well
rinsed, and after completion of samplings dried and stored in a special pouch. Plankton samples can
be analyzed alive or fixed. The fixation of samples should be done as soon as possible after the sampling,
but should always be done on dry land with a water supply on hand in case of splash/spillage.
In order to estimate the quantitative indices of phytoplankton, samples are collected with a specified
volume. In this case also the net can be used, providing that the quantity of water filtered through the net
and the volume of collected water is taken in evidence. But the safest way of collection of phytoplankton
samples is the method, which consists of application of the sampler. In this last case the phytoplankton
samples can be used for assessment of both taxonomic structure of algae communities and quantitative
indices of phytoplankton.
The volume of sampled water for phytoplankton investigation shall be not less than 1 L in water
bodies with poor phytoplankton and 0.5 0.25 L in those rich in phytoplankton.
Labeling of samples
Each sample shall be labelled. The label shall indicate the date of collection,
the place, the name or number of the station (Fig. 19). Label is fixed on the
bottle. In the register of collected samples, in addition to the mentioned above
data, some other characteristics are recorded: air and water temperature, water
transparency, pH, speed, visual estimation of water body state (e.g. level of
development of macrophytes, etc.).
Fig. 19. Labelled
Nalgene bottle

References

31. Algae. Handbook / Vasser S., Condratieva N., Masiuc N. et al. Kiev: Naukova Dumka, 1989 (in
Russian). Original title: . / .., ..,
.. . : , 1989. 608 .
32. http://www.duncanandassociates.co.uk/equip.htm

VI. SAMPLING OF ZOOPLANKTON

Zooplankton is represented by heterotrophic plankton organisms drifting in oceans, seas and
terrestrial fresh water bodies. Along with phytoplankton, zooplankton is key component of marine and
ocean ecosystems. The Republic of Moldova territory does include neither sea, nor ocean. Therefore,
the national monitoring programs and scientific investigations are concentrated on the zooplankton
communities of lentic and lotic terrestrial ecosystems. Sampling of zooplankton in rivers and standing
water bodies (lakes, ponds and reservoirs) is the target of this part of the Guidance.
Location of sampling
Sampling site: general area within the water body, where samples are collected.
Sampling point: the exact position within the sampling location where samples are collected.
Littoral zone: the area near shore where sunlight penetrates to the bottom and allows macrophytes
to grow (Fig. 20).
Pelagic zone: free water body beyond the littoral zone.
Limnetic zone: the well-lighted near-surface open water surrounded by the littoral zone where
plankton is dominated (both phytoplankton and zooplankton).
Stratified water: the thermal layering of water in the fresh and salt water bodies. It is characterized
14

by the vertical density gradient. Cold water is denser than warm water and the epilimnion generally
consists of water that is not as dense as the water in the hypolimnion.
Thermocline (metalimnion): layer of a thermally stratified water body in which the temperature
gradient has the maximum value.
Epilimnion: water layer positioned over thermocline in a stratified water body.
Hypolimnion: layer positioned under water thermocline in a stratified water body.
Identification of sampling points within
the stationary hydrological network allows
collecting comparative samples for creation
of multiannual databases. In most cases,
the sampling points can be easily found by
making reference to the hydrological and
morphological features of the water body.
Usually, in order to water quality control,
zooplankton samples should be collected at
the same place where other hydrobiological
(as well as hydrochemical) samples should
be sampled.
Selection of sampling points within
the Operational Monitoring Programmes
Fig. 20. Zonal distribution of habitats in the lake [33]
primarily depends on the location of
pollution source (household and industrial wastewaters, livestock farms, agricultural lands, etc.). For
the assessment of the reference conditions in the sampling location, the sampling points shall be set
upstream the pollution source. Other points should be selected in the zone of contamination: in the case
of small rivers, the distance from the source of pollution shall not be closer than 50 m, for large rivers
than 200 m [34].
Sampling points within the Scientific Investigation Programmes are determined in accordance with
the purposes of research and morphological characteristics of the water body. In general, the selected
points must be representative for the studied area. Zooplankton is unevenly distributed in streams and
more uneven in standing water bodies, due to the diurnal migration of hydrobionts both horizontally
and vertically.
In littoral zone samples of zooplankton are collected, as rule, from a depth of not more than 1 m. It
is recommended to collect separate samples: from zones with aquatic vegetation (protected shore) and
from zones with scant vegetation (unprotected shore).
Pelagic samples are taken at selected depths or from entire column of water, depending on the objectives
of the study. If the depth of the water body is not known, it should be estimated using a portable sounder or
another tool for measuring depth. In general, in the thermally stratified water bodies, samples of zooplankton
from pelagic zones have to be collected at the depth of thermocline, since most of plankton organisms is
found in epilimnion. For studying the distribution of zooplankton in epilimnion and metalimnion, the
samples shall be collected at depth intervals not larger than 2 m. Depth intervals in hypolimnion may be
more than 2 m. In order to get reliable results, in each zone of the water body the minimum number of
sampling points should be at least three, including littoral area with macrophytes [35].
Frequency and time of sampling
Annual frequency of sampling depends on the type of monitoring. Within the National Surveillance
Monitoring Programme, aiming to assess the state of aquatic ecosystems, the frequency of collection for
planktonic samples (bacterioplankton, phytoplankton and zooplankton) is set as of 3 4 times during
the vegetation season (April-September) [36]. Within one day, sampling period is determined by the
vertical daily migration of most species of zooplankton. Normally, zooplankton should be sampled
between 10 am and 4 pm.
15

Scientific investigations aimed to study the processes of aquatic ecosystems functioning (provides
information on species composition, density of hydrobionts and individual productivity), should
comprise of at least monthly samples collected during the vegetation season. An additional important
consideration is the number of samples required to characterize the zooplankton in a manner that
accounts for its variation in vertical space, horizontal space, and time.
The general procedures of zooplankton sampling in standing waters for the purposes of water
quality assessment are described in the European guidance EN 15110:2006 [37]. These procedures are
recommended for the collection of zooplankton that inhabits the pelagic and littoral zones of lakes,
reservoirs and ponds. They are inappropriate for the collection of species that primarily live on the
surface of sediments and on the surface of aquatic plants.
Sampling techniques
Depending on the size of planktonic organisms, two types of sampling techniques are used: filtration
and sedimentation. Principle of filtration (directly or indirectly) is used in various types of equipment:
plankton nets, plankton traps, tube samplers, and pump samplers. Each of these devices has pros and
cons and the choice depends on characteristics of the water body (depth, density of algae, etc.) and the
zooplankton itself.
The hydrobionts of mesoplankton and microplankton can be collected by filtration of water through
various models of plankton nets (Fig. 21). Plankton net is a conical device made of fine nylon mesh that
is pulled through the water either vertically or horizontally at a known distance. Animals are captured by
the nylon walls of the net and collected in the bottom bucket. There are versions of the plankton net with
non-closing and closing system. The latter is used to sample the specific stratum in the water column.
The main advantage to using a net is that large volume of water can be filtered quickly. Nets are used
most often, yet they have serious limitations in regard to obtaining good quantitative data, especially in
nutrient and algae-rich waters. For qualitative sampling it is recommended to use planktonic nets with
different mesh size: 45 m for rotifers, 90 m for most of crustaceans, and 150 m for predator species
of copepods.

a
b
c
d
Fig. 21: a Prelevation of zooplankton, b Wisconsin plankton net [38], c closing plankton net [39],
d Bongo double net combined as one sampler [40]

A tube sampler is made of common PVC or Tygon. A tube is lowered

into the water column, and when the bottom reaches the desired depth, a
line is pulled to close the bottom with a rubber stopper or other device. The
tube is raised into the boat and the collected water poured through a net
to collect the zooplankton. This device provides a high degree of certainty
about volume of water sampled, but it may not be an effective way to sample
large animals that occur at a low density, or animals that can detect and
escape from a narrow sampling device.
16

Fig. 22. Trap sampler [41]

The most common trap sampler is the Schindler-Patalas trap (Fig. 22),
named after the two scientists who invented the device. This is a clear plastic box
that is lowered to a desired depth in the water column and then quickly closed
(upper and lower doors) by pulling upward in the water. It traps zooplankton
inside the box. When device is lifted, the water is allowed to exit a small mesh
net that is attached to the wall of the box, and zooplankton is collected inside a
sampling bucket at the end of that net. This device also provides a high degree
of certainty regarding the actual volume of water sampled.
The pump sampler consists of a readily interchangeable nylon-mesh sleeve
mounted on the submersible suction pump (Fig. 23). The unit is compact and
robust, with the mesh protected from external damage. Built-in flow meters
Fig. 23 Pump sampler [42] are essential for accurate volume estimation. Although designed primarily for
use in the intakes and outfalls of power station, this sampler could find more
general application. The incorporation of a submersible pump circumvents the problems associated
with plankton travel through long hose. Less than 3% of the zooplankton members so collected showed
damage or had died within 48 hours of capture. Attaching the suction pump to the end of a rigid tow
net is suggested as a method of surmounting the potential errors when density of sampling organisms is
below 1 - 5 ind./m3.
The technique of sedimentation is used for collection of nanoplankton. For this purpose the sample
is collected from a certain horizon of the water body in a bottle with a determined volume, is fixed and
stored for 7 10 days. After mentioned period of time the water above the sediment is removed using a
siphon (rubber tube, which inferior part is closed by the net with mesh size of 45 50 m). The remained
sediment is processed according to the established procedure [34].
In dependence of the objective of monitoring, sampling may be qualitative or quantitative. For
quantitative sampling can be used volumetric samplers such as Schindler-Patalas trap, pump samplers with
built-in flow meters, tube samplers, and bottle samplers. The Student plankton net (Fig. 17) can also be
used for quantitative sampling in the following way: 50 100 litres of water are filtered through fine mesh
net. The material collected in the bottom bucket is quantitatively transferred to sample storage bottles.
Sample preservation and storage
When sampling equipment is removed from the water, all collected material is transferred into the
sample recipient (plastic container or flasks with stoppers). It is important to fill almost completely the
sample recipients. A sample volume smaller than the recipient volume can cause the loss of some species:
the organisms can just remain trapped on the walls of the recipient and can get dried during the storage.
Preservative liquid may be added previously into the sample containers. If this was not fulfilled, it
should be added immediately after sampling. Non-preserved zooplankton samples should be stored in
the dark at the temperature 2 5C and no more than 48 h. Samples for quantitative analysis should be
preserved immediately. As a rule, the samples are fixed with solution of formalin, or ethyl alcohol, or
Lugols solution [37].
Lugols solution is added in the amount of 0.5 1.0 ml at 100 ml sample. If the samples contain large
amounts of plankton organisms and other organic materials (especially in the case of sampling from
the coastal zones), the Lugols solution volume should be in the amount of 3 to 5 ml per 100 ml sample.
Samples should have the color of cognac. Samples preserved with Lugols solution should always be
stored in the dark and preferably chilled below 5C place, except those to be analyzed within the week
(in this case they can be stored in the dark at the room temperature). Storage in the dark is required in
order to prevent photo-oxidation. It should regularly check the sample to determine if it is need to add
preservative solution.
If ethanol is used for samples preservation, the concentration should be no less than 70%. If samples
will be used for genetic analysis (based on DNA extraction) or for measure of reproduction parameters,
it is recommended ethanol concentration 90%. A high concentration of ethanol helps to maintain the
17

shape of the body (cladocerans) as well as prevents the release of eggs. If samples have been stored
properly, they can retain its capacity for long time. Samples for genetic analysis should be stored in a
refrigerator or biologic freezer.
If formalin is used, its concentration in the preserved sample should be of about 4 %. For example,
20 ml 20% formalin solution is added per 100 ml sample. Preservation in the formalin-sucrose solution
that is chilled to about 6 C, prevents the release of eggs. Formalin preserves the animals by preventing
bacterial decomposition. For best results the preserved samples should be stored in air-tight containers
(small plastic Nalgene bottles, Fig. 19) under refrigerated conditions. If properly stored in suitable
containers, the samples will remain in good state for several years.
Formalin (formaldehyde 37%) is a carcinogen and should be handled with the care. Proper safety
equipment includes gloves and plastic goggles.
References
33. http://www.lakeaccess.org/ecology/lakeecologyprim9.html.
34. Guidelines on hydrobiological monitoring of freshwater ecosystems / ed. by Abakumov V.
St. Petersburg: Gidrometeoizdat, 1992 (in Russian). Original title:
/ . . : , 1992. 318 .
35. Zooplankton and its production. Methodical recommendations on collecting and processing of
materials in hydrobiological investigations on freshwater bodies. St. Petersburg: GosNIORH, 1984
(in Russian). Original title: .

. : , 1984.
36. Regulation on monitoring of surface and underground water state. Decision of the Government of
the Republic of Moldova no. 932 on 20.11.2013. Published on 29.11.2013 in the Official Monitor
of the Republic of Moldova no. 276-280, art. no.1038 (in Romanian). Original title: Regulamentul
privind monitorizarea strii apelor de suprafa i apelor subterane. Hotrrea Guvernului
Republicii Moldova nr. 932 din 20.11.2013 n: Monitorul Oficial al Republicii Moldova Nr. 276 280, art. Nr. 1038.
37. EN 15110:2006 Water quality. Guidance standard for the sampling of zooplankton from standing
waters. Chiinu: INSM, 2012 (in Romanian). National version: SM SR EN 15110:2012 Calitatea
apei. Ghid pentru prelevarea zooplanctonului din ape stttoare. Chiinu: INSM, 2012. 24 p.
38. http://shop.sciencefirst.com/wildco/575-wisconsin-sampler.
39. http://shop.sciencefirst.com/wildco/466-birge-closing-nets.
40. http://www.kc-denmark.dk/products/plankton-nets/bongo-net-2-x-oe60-cm.aspx.
41. http://shop.sciencefirst.com/wildco/plankton-samplers-and-processing/6326-replacement-netfor-schindler-patalas-nitex-153m.html.
42. http://www.kc-denmark.dk/products/plankton-pumps.

VII. SAMPLING OF ZOOBENTHOS

Benthic invertebrates are animals living at the bottom of water bodies, in the bottom water layer
and on the various types of substrates and, mainly, are represented by the following taxonomic groups:
oligochaetes, chironomids, molluscs, crustaceans, caddisflies (Trichoptera), mayflies (Ephemeroptera),
stoneflies (Plecoptera), etc. Depending on the body size of animals, the benthos is classified into
microbenthos (under 0.5 mm), meiobenthos (0.5 - 2.0 mm) and macrobenthos (more than 2 mm).
Collection of samples of benthic invertebrates is performed accordingly to international and national
standards ISO 5667-6:2005, ISO 5667-12:1995, ISO 7828:1985, ISO 8265:1988, ISO 9391:1993,SM SR
EN ISO 8689-1:2011, SM SR EN ISO 8689-2:2011 [3-4, 11, 43 - 44].Also, the methods unanimously
accepted in hydrobiology are used [45 46].
18

Benthic invertebrates are met in different types of water bodies (running and stagnant, deep and
shallow) and populate both the various types of substrates (stones, gravel, sand, silt, stands of macrophytes,
periphyton, etc.) and bottom water layer; therefore, there are no universal tools for collecting of benthic
invertebrate samples.
In function of investigated type of substrate and collected group of invertebrates, the following
sampling devices and methods are applied:
1. Grabs: as example, Petersen and Ekman grabs, with a capture area of 0.025 m2 (Fig. 24). At a
sampling station at least 2 samples are taken from an area of 0.025 m2, and respectively 4 5 samples
from the area of 0.01 m2. Grabs are used both for quantitative (density, ind./m2, biomass, g/m2)
and qualitative (taxonomic diversity) assessment
of macrozoobenthos.
2. Trawls, dredges, as example the rectangular
dredge with a capture area of 8 m2, which is used
by the researchers of the Institute of Zoology
of the Academy of Sciences of Moldova for
quantitative (Mysidae) and qualitative assessment
of zoobenthos (Fig. 25, 1 2). The usage of dredge
on stone, gravel and xylal (dead wood, branches,
roots) substrates should be avoided.
a
b
3. Frames are used for quantitative and qualitative
Fig. 24. Grab samplers: a Petersen, b Ekman [47]
analysis of molluscs (Fig. 25, 3).
4. Nets (usually made by silk or nylon cloth with mesh size of 333 m) are suitable for thickets of
submerge macrophytes (Fig. 25, 4). If the method of mowing is used, the net is dragged for three
minutes opposite the water stream or on a one meter distance. The procedure is repeated three
times. In such case and if a net with square opening of 2525 cm is used, than the total picked up
area is equal to 0.75 m2.
5. Bottom scrapers (mesh cloth of 500 1000 m) are used for sampling on hard substrates, including
hydrotechnical constructions (Fig. 25, 5).
6. Artificial substrates (Fig. 25, 6 7).
7. Air-lift sampling devices are used in big rivers, with fast running and deep, exceeding 7 meters,
waters (Fig. 25, 8 9).

e
f
g
h
Fig. 25 Devices for sampling of benthic invertebrates: a triangle bottom dredge, b rectangle bottom dredge, c frame,
d hydrobiological scraper, e hand net [48], f air-lift sampler [49], g and h different types of artificial substrates [50]

19

8. Hand sampling of benthic invertebrates is used for different types of substrates for both qualitative
and quantitative analyses.
If the sampling is performed with net or scraper, operator should move opposite the water current,
dislodging by heel of boot the substratum and the fauna and holding the tool downstream from his feet. If
benthic invertebrates are sampled in thickets of macrophytes or among tufts of filamentous algae, the pieces
of plants that penetrated in the net should be rinsed thoroughly before being thrown out of the net.
In the case of stone substrate, the clingers and sessile organisms are captured by using a brush or
pincers. In order to calculate the area of stone first its projection is made on a paper and after the whole
projection and 1 square centimetre are weighted. By dividing the weight of whole projection to those of
1 square centimetre the approximate stone area is obtained.
The selection of sampling stations depends on investigation objectives. The biological monitoring
requires the selection of substrates with the most various benthic fauna, taking in account the mosaic
distribution of organisms and their affiliation to different substrates [45 46, 51-61]. It should be
reminded that sampling is performed from the downstream end of the stream reach to upstream one.
In rivers the benthic assemblages are faunisticaly more reach at the border between riffles and pools
[45], in natural and man-made lakes in littoral zones [2], and also on artificial substrates [11]. Usually,
benthic invertebrates are sampled at depths up to 1.5 m. In rivers sampling is performed in the littoral
zone, in riverbed, in thickets of macrophytes and in the system of riffle and pool areas. In small rivers
collection is performed on a distance at least 50 m and in big once at least 200 m along the river. In
the case of big rivers it is recommended to collect at least 3 4 replicates (subsamples), from which the
sample is composed, per sampling site proportionally to the presence of different microhabitats (lithal,
psammal, pelal, phytal, etc.).
In lentic ecosystems the samples of benthic fauna are collected in littoral zone, in deep sectors of the
water body and among macrophytes [2].
In the case of faunisticaly poor habitats, it is recommended to increase the number of collected
replicates up to 20 in order to obtain the highest possible number of species [11]. A sample allows
obtaining of a statistically significant hydrobiological material, if it consists of at least 700 individuals
[52]. A special attention should be paid to areas where the alien species are met. Rare species and also
relict species require a separate treatment, because their withdrawal from the water body may affect their
populations.
The frequency of sampling depends on investigation objectives and may be annual (in the case
macrobenthos it is preferred in September-October), seasonal, monthly, decadal, but in addition the
exceptional situations should be taken in account [62].

a
b
c
Fig. 26: a - transferring the benthic sample into a bucket with water [63], b - container for transportation and storage,
c - transferring the benthic sample into a container

20

In general, no benthic samples should be taken during or shortly after floods, droughts (only if this
is not foreseen by investigation objectives), or in the case of a maninduced disturbance, for example,
during the dredging works in the riverbed at the given sampling site [52]. In order to establish the
dynamics of density and natural changes of population taxonomic structure, it is recommended to take
samples repeatedly during the year and during few consecutive years.
After collection the benthic sample is transferred into a container with water (Fig. 26) by using a net
or a sieve the large fraction is removed from the sample. The sample is transferred from the net into a
glass or plastic container with a volume of 200 1000 ml and preserved. For preservation (fixation) the
96 % ethanol or 37 % formalin is used, which is added to the sample till a 70% and, correspondingly,
3.7% final concentration is obtained [15, 51 52]. If the sample is not preserved, then it should be stored
cool, at a temperature of 1 5 and processed during the first 24 hours after collection [15].
Each container with sample is labelled by indicating the number of sample, name of water body, site
and data of collection, deepness and type of substratum, number of replicates.
References
43. SM SR EN ISO 8689-1:2011 Water quality. Biological classification of rivers. Part 1: Guidance
on the interpretation of biological quality data from surveys of benthic macroinvertebrates (in
Romanian). National version:: SM SR EN ISO 8689-1:2011 Calitatea apei. Clasificarea biologica
a rurilor. Partea 1: Ghid pentru interpretarea datelor biologice de calitate obinute din studierea
macronevertebratelor bentonice.
44. SM SR EN ISO 8689-2:2011 Water quality. Biological classification of rivers. Part 2: Guidance
on the presentation of biological quality data from surveys of benthic macroinvertebrates (in
Romanian). National version: SM SR EN ISO 8689-2:2011 Calitatea apei. Clasificarea biologica
a rurilor. Partea 2: Ghid pentru prezentarea datelor biologice de calitate obinute din studierea
macronevertebratelor bentonice.
45. Guidelines on the methods of hydrobiological analysis of surface watersand bottom sediments
/ Abakumov V.A. (editor). Leningrad: Gidrometeoizdat, 1983. 240 p. (in Russian). Original title:

/. .. .: , 1983. 240 .
46. Jadin, V.I. Methods of hydrobiological investigations. Moscow: Vysshaya shkola, 1960. 190 p.
(in Russian). Original title: . .: ,
1960. 190 . . .: , 1960. 190 .
47. http://www.anhydre.eu/wildlife.html.
48. http://www.nhbs.com/browse/subject/516/aquatic-samplingnets.
49. http://www.uwitec.at/html/river_benthos.html
50. http://www.iad.gs/index.php?item=zooplanktonzoobenthos&PHPSESSID=bfd9bce62c0a
64891c9902f73d9f61b9
51. Unified methods of water quality investigation. Part III: Methods of biological analysis of waters.
M.: SEV, 1983. 227 p. (in Russian). Original title:
. III: . .: , 1983. 227 .
52. Manual for the application of the AQEM system. A comprehensive method to assess European
streams using benthic macroinvertebrates, developed for the purpose of the Water Framework
Directive. Version 1.0,February 2002.
53. Jadin, V.I. Molluscs of fresh and brackish waters of the USSR. .-L. AN SSSR, 1952. 376 p. (in
Russian). Original title: .. . .; .:
- , 1952. 376 .
54. Unified methods of water quality investigation. Part III: Methods of biological analysis of waters.
M.: SEV, 1983. 227 p. (in Russian). Original title:
. III: . .: , 1983. 227 .
55. Identification guide into the freshwater invertebrates from the European parts of the USSR
21

(plankton and benthos) / Kutikova L.A., Starobogatov E.I. (editors). Leningrad: Gidrometeoizdat,
1977. 510 p. (in Russian). Original title:
( ) / . .., ..
: , 1977. 510 .
56. Identification guide into the freshwater invertebrates of Russia and adjacent countries, Vol. 1.
Lower invertebrates / Tsalolikhin S.J. (ed.). Saint-Petersburg: Nauca, 1994. 394 p. (in Russian).
Original title:
/ . .. 1. . .: , 1994. 394 c.
57. Identification guide into the freshwater invertebrates of Russia and adjacent countries, Vol. 2.
Crustaceans / Tsalolikhin S.J. (ed.). Saint-Petersburg: Nauca, 1995. 627 p. (in Russian). Original
title: /.
.. 2. . .: , 1995. 627 c.
58. Identification guide into the freshwater invertebrates of Russia and adjacent countries, Vol. 3.
Arachnids / Tsalolikhin S.J. (ed.). Saint-Petersburg: Nauca, 1997. 439 p. (in Russian). Original
title: /.
.. 3. . .: , 1997. 439 c.
59. Identification guide into the freshwater invertebrates of Russia and adjacent countries, Vol.
4. Diptera / Tsalolikhin S.J. (ed.). Saint-Petersburg: Nauca, 2000. 997 p. (in Russian). Original
title: /.
.. 4. . .: , 2000. 997 c.
60. Identification guide into the freshwater invertebrates of Russia and adjacent countries, Vol. 5.
Higher insects / Tsalolikhin S.J. (ed.). Saint-Petersburg: Nauca, 2001. 836 p. (in Russian). Original
title: / .
.. 5. . .: , 2001. 836 c.
61. Identification guide into the freshwater invertebrates of Russia and adjacent countries, Vol.
6. Shellfish. Polychaete. Nemerteans. / Tsalolikhin S.J. (ed.). Saint-Petersburg: Nauca, 2004.
528 p. (in Russian). Original title:
/ . .. 6: . . . .:
,2004. 528 c.
62. ISO 5667-14:1998 Water quality. Sampling. Part 14: Guidance on quality assurance of
environmental water sampling and handling. National version: SMV ISO 5667-14:2011 Calitatea
apei. Prelevare. Partea 14: Ghid pentru asigurarea calitii la prelevarea i manipularea probelor
de ap natural (in English). Chiinu: INSM, 2011. 18 p.
63. http://shop.sciencefirst.com/wildco/551-standard-ekman-grab

VIII. SAMPLING OF ICHTHYOLOGICAL MATERIAL

Today in Europe are mainly used the following methods of fish sampling:
reversible electronarcosis, especially useful in small, medium-sized rivers and some lakes (this
method is prohibited by law in the Republic of Moldova);
classic mesh tools, used mostly in lakes (drag nets, gillnets installed stationary or left free to drift,
etc.);
strings of hooks with bait, used in addition to other research tools.
To be sure that the data on abundance and age structure are valid, a sufficient number of samples shall
be collected. The selected stations for scientific fisheries shall be as representative as possible, in order to
reflect the status of the whole ecosystem. In lakes with large depths the existence of vertical stratification
(hypo-, meta- and epilimnion) is taken into account, thus the surface nets, nets for water layer and
bottom nets are used [64, 68, 70, 74-75].
Number of samplings should be large enough to ensure statistically the obtained results - the error
should not exceed 20%. The total length of the used nets is determined by the species abundance and
diversity of size groups intended to be captured. The sampling should last at least 12 hours or 24 hours.
22

The efficiency of collection is appreciated by determining the fishing effort (catch per unit effort,
CPUE), which expresses the number of individuals or biomass of fish in the capture (total value or
separately for each species), which was collected on a certain net area (e.g. 100 m2) in 12 h or 24 h.
In the case of fishing by electronarcosis the determination of numeric (individuals/100 m2 or
individuals/ha) and gravimetric stocks is relatively simple, as it is possible to determine easily the total
sampled surface of water body, and then to express the sampled quantity or number of individuals to
conventional units of surface (100 m2, 1 ha, 100 linear meters of river, etc.)
Examination of fishis performed usually by their counting, measuring and weighing. Indentification
of species is made on the base of morphological caracters, by using identification keys for each sistematic
unit [67, 69, 71-72]. In order to determine fish age the sampling of scales, or other hard tissues (operculum,
otolithes, sections of bony fin rays) is made. Also, the identification of sex and stage of development of
gonads is performed.
The sex of fish is determined by studying the gonads. The determination of sex is easily to be done
at mature stages of development: at adult females the ovules are easily distinguished in the ovaries, but
males have smooth, whitish, apparently non-granulated testes. In order to facilitate the identification of
various stages of the sexual cycle, the different scales of the degree of gonad maturation were drawn up.
All of them are based on highlighting the gonad maturation at different ontogenetic stages [73].
In order to put in evidence the fish growth rate a series of biometric measurements are done [70, 73- 75].
The following characteristics are determined via biometric studies:
metric characteristics: absolute or standard body length, head length, thickness, the highest and
the lowest body height, the distance between fins, eye diameter, etc.;
gravimetric characteristics: body weight or the weight of some organs, which is determined by
weighing;
meristic characteristics: number of scales in the linea lateralis, number of transversal rows of scales
in the highest part of the fish body, number of fin rays, number of gill rakers, formula of pharyngeal
teeth, etc.
Afterwards, all these data may be statistically processed in order to calculate average values, the limits
of variation, the existence or non-existence of certain correlations, etc.
A set of math methods, which are called generically synecological analysis, are used for deciphering the
relationship between different species in the ecosystem, and hierarchies established in the ichthyocenosis.
This type of analysis allows identifying precisely the species, which have the highest share in the given
ecosystem in view of the exchange of energy with environment, which species are characteristic for given
biotope, or which species penetrated accidentally in studied area. Also, it is possible to establish with
sufficient accuracy interrelations between species, which composed the biocenosis.
Depending on how they are calculated, there are two distinct categories of indices: 1. analytical
ecological indices (operations are made with raw data collected in the field); 2. synthetic ecological
indices (operations are made on the ground of analytical indices; used for highlighting the interrelations
between species, communities, or cenoses).
The most frequently there are used the following ecological indices: dominance (D), constancy (C),
ecological significance index (W), index of species similarity (S), diversity index or Shannon - Wiener index,
equitability (e), Simpson index (Is), etc. [64-66].
When evaluating the age structure of populations, it is enough to undertake ichthyological
measurements of a samples composed by at least 30 individuals/species, however the accuracy of data
representation increases with the increase of the number of measurements [64].
Determination of numeric and gravimetric stocks offers accurate and comparable information on
the number and biomass of each population across the species and the whole ichthyocenosis at given
sampling point. Moreover, these indices provide information of utmost importance in the case of the
ecological reconstruction.
With the exception of individuals for museum collections and those kept for identification and
processing in the laboratory (confuse characters, genetics and molecular analysis, histological analysis,
identification of trophic spectrum, etc.), all fish will be released once processing has been terminated.
23

Processing and release will be done after each collected fish tone or covered distance.
Details about the physical characteristics of hydrobiotop, fishing team, fishing effort, fishing objectives,
equipment, catches, etc., shall be recorded in a typical factsheet. It will be prepared and distributed by the
organizer of the fishing, following a unique template.

References
64. Davideanu G. (2013) Methodological guide for monitoring the ichthyocenoses structure. Ed.
Performantica, Iasi, 57 p.
65. Dediu I. (2007) Treaty of theoretical ecology: monographic study of synthesis. Chisinau: Ed.
Academia Nationala de Stiinte Ecologice, 557 p. (in Romanian). Original title: Dediu I. (2007)
Tratat de ecologie teoretic: Studiu monografic de sintez. Chiinu: Ed. Academia Naional de
tiineEcologice, 557 p.
66. Gomoiu M.T., Skolka M. (2001). Ecology. Methodologies for ecological studies. Constanta: Ed.
Ovidius University Press, 173 p. (in Romanian). Original title: Gomoiu M.T., Skolka M. (2001)
Ecologie. Metodologii pentru studii ecologice. Ed. Ovidius University Press, Constana, 173 p.
67. Koblitskaia A.F. (1981) Identification guide of freshwater fish fingerlings. Moscow: Legkaia i
piscevaia promishlennosti, 209 p. (in Russian). Original title: .. (1981)
. : . , 209 c.
68. Kotlear O.A. (2004) Methods of fish farming researches. Tutorial. Ribnoe, 180 p. (in Russian).
Original title: O.A. . .
, 2004, 180 .
69. Kottelat M., Freyhof J. (2007) Handbook of European Freshwater Fishes, Ed. Delemont,
Switzerland, 646 p.
70. Navodaru I. et al. (2008) Assessment of fish stocks and fisheries. Methods of assessment and
prognosis of fishery resources. Ed. Dobrogea, 298 p. (in Romanian). Original title: Nvodaru I.
.a. (2008) Estimarea stocurilor de peti i pescriilor. Metode de evaluare i prognoz a resurselor
pescreti. Editura Dobrogea, 295 p.
71. Otsel V. (2007) Atlas of fish from the Danube Delta Biosphere Reserve. Tulcea: Ed. Centrul de
informare tehnologic Delta Dunarii, 481 p. (in Romanian). Original title: Oel V. (2007) Atlasul
petilor din Rezervaia Biosferei Delta Dunrii. Tulcea: Ed. Centrul de informare tehnologic
Delta Dunrii, 481 p.
72. Popa L. L. (1977) Fish of Moldova. Reference book identification guide. Chisinau: Cartea
Moldoveneasca, 200 p. (in Russian). Original title: . . (1977) .
. : , 200 c.
73. Pravdin I.F. (1966) Guidelines on fish study. Moscow: Piscevaia promishlennosti, 376 p. (in
Russian). Original title: .. (1966) . :
, 1966, 376 .
24

74. Pricope F., Stoica I., Battes K. (2013) Secondary production of aquatic ecosystems. Bacau: Ed.
Alma Mater, 151 p. (in Romanian). Original title: Pricope F., Stoica I., Battes K. (2013) Producia
secundar a ecosistemelor acvatice. Bacu: Ed. Alma Mater, 151 p.
75. Shibaev S.V. (2007) Industrial ichthyology. Saint Petersburg: Prospect nauki, 2007, 399 p. (in
Russian). Original title: . . (2007) . -:
, 399 .

IX. SOURCES OF SAMPLING ERRORS

Sources of errors associated with sampling
The errors associated with sampling can be caused by various factors, the main of which are the
following [62]:
Contamination. Contamination can be caused by the inappropriate sampling equipment materials,
transport arrangements or storage conditions.
Sample instability. The type of sampling vessels and containers can affect the stability of the
determinants in the period between sampling and analysis due to the inherent instability of the
sample itself and the conditions in which samples are stored and transported.
Incorrect preservation. The choice of options for preservation, as detailed in ISO 5667-3:2003 [15],
can affect the concentration of ingredients in the sample.
Incorrect sampling. Deviation from the sampling procedure or sampling from non homogenized
water bodies may be a source of error.
Quality control techniques
Quality control measures in sampling have three main objectives: 1) to provide a way of monitoring
and detecting sampling errors with aim at ejecting the invalid or misleading data; 2) to demonstrate that
sampling errors have been controlled adequately; 3) to indicate the variability of sampling and thereby to
control this important aspect of error. The main quality control techniques are the following:
a) Collection of replicate samples as a check on the precision of sampling. This technique can be used
to caver a range of approaches to quality control which aim to assess the random error associated
with different levels of the sampling process. The relationship between the different sampling variances is illustrated in the Fig. 27. The difference between A1 and A2 gives an estimate of total sampling variance (sampling,
containers, storage and
analysis). The difference
between B1 and B2 gives an
estimate of analytical plus
sampling variance (including storage, excluding
sampling container).
b) Rinsing of equipment. This
technique can be used to
identify any errors relating to contamination of
sampling devices and the
sampling process caused
by incomplete cleaning of
the sampling vessels. At
the laboratory a sample of
Fig. 27. Quality control on the precision of sampling
deionized water is divid25

ed into two parts: Part A is

retained in the laboratory.
Part B is transported into
the field and subdivided into
portions B1 and B2. Portion
B1 should be returned to
the laboratory without any
processing in the field. Portion B2 should be processed
as real sample (Fig. 28). The
difference in results between
Part A and portion B1 identifies errors due to sample
transportation. The difference in results between Part
A and portion B2 identifies
Fig. 28. Quality control of sampling equipment with deionized water blank
errors due to sampling, processing and transportation.
c) Filtration recovery. This technique can be used to identify any errors relating to contamination of
sampling devices and sampling process associated with filtration of samples. At the laboratory a
sample of deionized water is divided into two parts: Part A is retained in the laboratory. Part B is
transported into the field and subdivided into portions B1 and B2. Portion B1 should be returned to
the laboratory without any processing in the field. Portion B2 should be processed using the sampling
devices and filtration equipment as real sample (Fig. 29). The difference in results between Part A
and portion B2 identifies errors due to sampling, filtration, processing and transportation. The comparison of results of portions B1 and B2 identifies
errors due to contamination of sampling containers
and sampling process associated with filtration.
d) Spiked samples. For estimating the systematic error
of the sampling processes
which is due to the sample
instability (volatilization of
organic compounds, adsorption of trace metals or
consumptions of nutrients
by bacterio- and phytoplankton) the spiked samples can be used on similar
Fig. 29. Quality control of sampling and filtration equipment
technique.
with deionized water blank
Analysis and interpretation of quality control data
The most widely form used for interpretation of quality control datais the Shewhart control chart
[76]. In this case the form of a chart is used on which the difference between duplicate determination
(d = R1 R2)1 is plotted sequentially. The measured values are compared with the limit values (warning
1

R1 is the result of the first sample analysed and R2 is the result of the second sample analysed

26

upper/lower limit and action upper/lower limit) and result is analysed by a visual examination of the
control chart (Fig. 30).

Fig. 30. Shewhart control chart for duplicate control samples

In general, the quality assurance/quality control procedures (QA/QC) are necessary to ensure
the quality of the sampling activities of a monitoring programme, including care to preserve sample
integrity. Quality assurance of sampling including selection of sample, pre-treatment, subsampling,
preservation, storage and transport is essential for the quality of final results of the chemical analyses.
The effort expended on sampling quality control is dependent on the objectives of the programme, but it
is recommended that at least 2 % of analytical efforts should be devoted to quality control for sampling.
References
76. ISO 8258:1991 Shewhart control charts. This standard has been revised by: ISO 7870-2:2013
Control charts. Part 2: Shewhart control charts. National version: SM SR ISO 8258+C1:2012 Fie
de control Shewhart (in Romanian). Chiinu: INSM, 2012. 44 p.

27

ANNEX 1. Sampling frequencies for physico-chemical and hydrobiological parameters [36]

Quality
Elements

Rivers (once during the

mentioned interval)

Lakes/reservoirs (once during the

mentioned interval)

Physico-chemical Quality Elements

Conditions thermal

1 month

1 month

Conditions gaseous

1 month

1 month

pH and redox potenialul

1 month

1 month

Conductivity

1 month

1 month

Main ions and mineralization

1 month

1 month

Turbidity and total suspended solids

1 month

1 month

Waters smell and color

1 month

1 month

Condition of eutrophication:
nutrients

1 month

1 month

Chemical and biochemical oxygen

demand (CODCr and BOD5)

1 month

1 month

Acidification conditions: alkalinity

1 month

1 month

Priority persistent organic substances

1 month

1 month

Heavy metals

1 month

1 month

Other pollutants

1 month

1 month

Biological Quality Elements

Bacterioplankton

3 times per year during the growing

season (April to September)

3 times per year during the growing

season (April to September)

Zooplankton

3 times per year during the growing

season (April to September)

3 times per year during the growing

season (April to September)

Phytoplankton

3 times per year during the growing

season (April to September)

3 times per year during the growing

season (April to September)

Chlorophyll A

3 times per year during the growing

season (April to September)

3 times per year during the growing

season (April to September)

Benthic invertebrates

2 times per year during the growing

season (April to September)

2 times per year during the growing

season (April to September)

Macrophytes

2 times per year during the growing

season (April to September)

2 times per year during the growing

season (April to September)

2 times per year

(spring and autumn)

2 times per year

(spring and autumn)

Fishes

28

ANNEX 2. Suggested methods of preservation and maximum permissible storage times [15]
Quality
Elements
Acidity and Alkalinity
Aluminium
Ammonia free and
ionized

Recommended
container material2

Conservation
techniques

Permissible storage time

prior to analysis

P or G

Cooling to 1 5C

24 h

P or BG

Acidification to pH 1 2
with HNO3

1 month

Cooling to 1 5C
Acidification to pH 1 2
with H2SO4

24 h
21 days

P or G

Filtered before preservation

Arsenic

Acidification to pH 1 2
with HNO3

1 month

Cooling to 1 5C

24 h

Biochemical Oxygen
Demand (BOD)

P or G

Bromide

P or G

Cooling to 1 5C

1 month

Calcium

P or G

Cooling to 1 5C
Acidification to pH 1 2
with HNO3

7 days
1 month

Carbon:
carbon dioxide

P or G

Cooling to 1 5C

24 h

Carbon:
Total Organic Carbon
(TOC)

P or G

Chemical Oxygen
Demand (COD)

P or G

Chloride

P or G

Chlorophyll
Chromium total
Chromium (VI)
Color of water

P or G
washed with acid

P or G
(brown containers)
P or G
washed with acid
P or G
washed with acid

Samples are stored in the absence of light

The preferred determination is made in situ

Cooling to 1 5C
Acidification to pH 1 2
with H2SO4

24 h
7 days

If the presence of volatile organic compounds is

suspected, acidification is not used;
the analysis should be carried out in 8 hours
Acidification to pH 1 2
1 month
with H2SO4
Cooling to 1 5C
24 h

1 month

Cooling to 1 5C

24 h

Acidification to pH 1 2
with HNO3

1 6 months

Cooling to 1 5C

24 h

Cooling to 1 5C

5 days

P or G

Samples are stored in the absence of light

P polyethylene (PTFE - polytetrafluoroethylene, PVC - polyvinyl chloride, PET - polyethylene terephthalate),

G - glass, BG - borosilicate glass

29

Compounds of metals
(Cu, Pb, Zn, Ni, Cd, Co,
V, Mn, Se)
Conductivity

P or BG
washed with acid

Acidification to pH 1 2
with HNO3

1 6 months

Cooling to 1 5C

24 h

P or BG

The preferred determination is made in situ

Cooling to 1 5C
24 h
Alkalization to pH > 12
7 days
with NaOH
Samples are stored in the absence of light

Cyanide:
total cyanide

Cyan chloride

Cooling to 1 5C

24 h

G washed with
methanol

Acidification to pH 1 2
with H2SO4,
cooling to 1 5C

2 days

G washed with
methanol

Cooling to 1 5C

2 days

P but not PTFE

Cooling to 1 5C

7 days

Detergents:
anionic surfactants
Detergents:
cationic surfactants
Fluoride
Hydrocarbons
Iron (II)
Iron total
Magnesium

Mercury
Nitrogen:
nitrate nitrogen
Nitrogen:
nitrite nitrogen
Nitrogen:
nitrogen total
(Kjeldahl)
Oil and Grease
Oxygen (Winkler)

G washed with the

solvent used for
extraction
P or BG
washed with acid
P or BG
washed with acid
P
BG
washed with acid
BG
washed with acid
P or G

1 month (if possible, the

extraction is carried out
in situ)
Acidification to pH 1 2 7 days (isolating from air
with HCl
oxygen)
Acidification to pH 1 2
1 month
with HNO3
Cooling to 1 5C
7 days
Acidification to pH 1 2
1 month
with HNO3
Acidification to pH 1 2
with HNO3 and adding
1 month
K2Cr2O7 to the final
concentration 0,05 %
Cooling to 1 5C
24 h
Acidification to pH 1 2
7 days
with HCl
Acidification to pH 1 2
with H2SO4 or HCl

P or G

Cooling to 1 5C

24 h

P or G

Cooling to 1 5C
Acidification to pH 1 2
with H2SO4

24 h
1 month

G
washed with solvent

Acidification to pH 1 2
with H2SO4 or HCl

1 month

Oxygen is fixed in situ

4 days

P or G

Samples are stored in the absence of light

30

Pesticides:
carbamate pesticides

G
P
washed with solvent

Pesticides:
organophosphorus,
organochlorine,
organoazotate

G with PTFE cap

Phenols

BG brown
with PTFE cap
washed with solvent

pH

P or G: completely
fill the container to
remove air

Phosphorus dissolved

Phosphorus total

Potassium
Silicates:
dissolved silicates
Silicates:
total silicates

G or BG

G or BG

P
P

Cooling to 1 5C
7 days
Freezing at 20 C
1 month
If the sample is chlorinated, add 80 mg Na2S2O3
5H2O per 1000 ml sample container prior to
analysis
Extraction
5 days
(in 24 h after collection)
(storage time for extract)
Cooling to 1 5C
If the sample is chlorinated, add 80 mg Na2S2O3
5H2O per 1000 ml sample container
prior to analysis
Acidification to pH < 4
with H3PO4or H2SO4

3 weeks

If the sample is chlorinated, add 80 mg

Na2S2O3 5H2O per 1000 ml sample container
prior to analysis
Cooling to 1 - 5C
6h
The measurement is preferably performed in situ
immediately after sampling
Filtration in situ
24 h
(0,45 m filter)
Cooling to 1 - 5C
Filtration in situ
(0,45 m filter)
Acidification to pH 1 - 2
with H2SO4
Cooling to 1 - 5C
Acidification to pH 1 - 2
with HNO3
Filtration in situ
Cooling to 1 - 5C

1 month

7 days
1 month
7 days

Cooling to 1 - 5C

1 month

Sodium

Cooling to 1 - 5C

7 days

Sulphides

Sulphate

P or G

Cooling to 1 - 5C

1 month

Suspended solids

P or G

Cooling to 1 - 5C

2 days

Turbidity

P or G

Cooling to 1 - 5C
24 h
Samples are stored in the absence of light

Cooling to 1 - 5C
7 days
The samples are fixed immediately after sampling
with zinc acetate 10%

31

ANNEX 3. Recommended washing procedures for selected water quality elements [30]
Quality elements

Washing procedure

Colour, Turbidity, pH, Acidity, Alkalinity,

Calcium, Potassium, Sodium, Magnesium,
Carbon organic total, Nitrogen (ammonia,
nitrite, nitrate, total), Phosphorus, Chloride,
Fluoride, Sulphate

Rinse three times with tap water once with

chromic acid3 three times with tap water once
with HNO3 (1:1) three times with distilled water

Aluminium, Cadmium, Cobalt, Copper, Iron,

Zinc, Nickel, Lead, Molybdenum, Manganese,
Mercury, Selenium, Strontium, Vanadium

Rinse three times with tap water once with

chromic acid three times with tap water once
with HNO3 (1:1) three times with ultrapure
distilled water4

Organophosphorus, Organochlorinated
pesticides, Polychlorinated Biphenyls (PCBs),
Pentachlorophenol, Phenolics, Phenoxy acid
herbicides

Rinse three times with tap water once with

chromic acid three times with organic-free water
once with special grade5 acetone twice with
pesticide grade hexane dry (un-capped) in a hot
air oven at 360C for at least 1 h

Fig. 31. a Glass Distillation Unit [78], b Millipore Super Q Water System [79]

References
77. GOST 4517-87 Interstate standard. Reagents. Methods for preparation of accessory reagents and
solutions used for analysis (in Russian) Original title: 4517-87
. . ,
. M.,1988
78. http://www.indiamart.com/avm-scientific/scientific-laboratory-equipments.html
79. http://www.medicalexpo.com/prod/merck-millipore/product-70876-658188.html

Chromic acid: solution of potassium dichromate with a mass concentration 5% in sulphuric acid [77].
Ultrapure distilled water is obtained by passing distilled water through an all-glass distillation unit (Fig. 31 a) and then
through a Millipore Super Q Water System (Fig. 31 b) containing a pre-filter cartridge, an activated carbon cartridge and
a mixed bed deionisation cartridge.
5
Special grade acetone: pesticide grade when GC analysis to be performed, UV grade for LC analysis
3
4

32

Programul Operational Comun

Romnia-Ucraina-Republica Moldova 2007-2013

Granie comune. Soluii comune

Ghid de prelevare a probelor

hidrochimice i hidrobiologice

Proiect finanat de
UNIUNEA EUROPEAN

Proiect MIS ETC 1676

implementat de Universitatea Dunrea de Jos din
Galai, Romnia, Institutul de Zoologie, Academia de
iine a Moldovei, Institutul Geologie i Seismologie,
Academia de iine a Moldovei,Centrul tiinific
.Ucr[nean pentru Ecologia Mrii, Odesa, Ucraina

33

CUPRINS

I
II
III
IV
V
VI
VII
VIII
IX

Introducere
Zubcov E., Bilechi L.
Principii generale de prelevare n ecosisteme acvatice
Todera I., Zubcov E., Bilechi L.
Prelevarea apei
Zubcov E., Bagrin N., Bilechi L., Zubcov N., Borodin N.
Prelevarea suspensiilor i sedimentelor
Jurminskaia O., Bagrin N., Zubcov N., Borodin N., Ivanov A.
Prelevarea bacterioplanctonului
ubernekii I., Negru M.
Prelevarea fitoplanctonului
Ungureanu L., Tumanova D.,Ungureanu G.
Prelevarea zooplanctonului
Jurminskaia O., ubernekii I., Lebedenco L.
Prelevarea zoobentosului
Munjiu O., Todera I., Banu V.
Prelevarea materialului ihtiologic
Bulat Dm., Bulat Dn., Todera Ion, Usati M.
Surse de erori n prelevare
Jurminskaia O.
ANEXA 1
Frecvenele prelevrii de probe pentru parametrii fizico-chimici i hidrobiologici
ANEXA 2
Metodele recomandate de conservare i durata de pstrare maximum admisibil
ANEXA 3
Proceduri recomandate de splare pentru elemente selectate ale calitii apei

34

35
36
37
40
42
43
45
50
54
56
59
60
63

INTRODUCERE
nvm pentru via, nu pentru coal (Seneca)
Lucrarea de fa este publicat n cadrul proiectului Cooperare interdisciplinar transfrontalier
pentru prevenirea dezastrelor naturale i reducerea polurii mediului n Euroregiunea Dunrea de Jos
MIS ETC 1676, realizat n perioada anilor 2013-2015 de un colectiv de cercettori din Romnia, Republica Moldova i Ucraina.
Proiectul a vizat ntrirea cooperrii transfrontaliere n scopul elaborrii soluiilor pe termen lung
pentru reducerea polurii, dezvoltarea durabil i pregtirea pentru situaii de urgen n Euroregiunea
Dunrea de Jos. Pentru atingerea scopului, au fost stabilite un ir de obiective:
integrarea capacitilor i experienei instituiilor partenere pentru aprecierea impactului transfrontalier al problemelor de mediu n bazinul Dunrii de Jos;
studiul de fezabilitate pentru elaborarea planurilor de dezvoltare a localitilor - pilot i a prevenirii
dezastrelor;
crearea unui mecanism capabil de a oferi suport tiinific pentru factorii de decizie pentru elaborarea politicilor legate de prevenirea impactului negativ al eventualelor accidente antropice i
naturale n regiune;
sporirea nivelului de contientizare a populaiei din regiune privind problemele de mediu cu impact transfrontalier;
ncurajarea implicrii tinerilor cercettori, studenilor, masteranzilor i doctoranzilor n cercetrile de mediu transfrontaliere;
ameliorarea situaiei ecologice n Euroregiunea Dunrea de Jos.
Proiectul a permis crearea de ctre instituiile-partenere a unei reele internaionale interdisciplinare
(INPOLDE), menit de a acumula date de fond, efectua cercetri de mediu complexe n regiunile de
frontier, evalua impactul de mediu i, ntr-un final, de a oferi un suport tiinific mai bun pentru programele de monitoring regionale.
Lucrarea Ghid de prelevare a probelor hidrochimice i hidrobiologice este parte component a unei
serii de lucrri, ce urmeaz a fi editate n cadrul proiectului mai sus-menionat, care este dedicat aspectelor metodologice ale cercetrii diferitor componente de mediu, ncepnd cu metodele de prelevare
a probelor n cmp i sfrind cu aplicarea tehnicilor moderne analitice n evaluarea nivelului de poluare
a mediului nconjurtor. Ea are la baz experiena personal bogat a autorilor, dar i practicile bune internaionale n domeniul studierii componentelor fizico-chimice i biologice ale ecosistemelor acvatice
dulcicole.
La alctuirea acestei lucrri i-au adus contribuia cercettorii Institutului de Zoologie, Academia
de tiine a Moldovei i anume: Todera Ion, academician, Zubcov Elena, doctor habilitat n biologie,
profesor cercettor, Ungureanu Laurenia, doctor habilitat n biologie, profesor cercettor, Usati Marin,
doctor habilitat n biologie, profesor cercettor, Bilechi Lucia, doctor n biologie, confereniar cercettor, Jurminskaia Olga, doctor n biologie, ubernekii Igor, doctor n biologie, confereniar cercettor,
Negru Maria, doctor n biologie, confereniar universitar, Bagrin Nina, doctor n biologie, confereniar
cercettor, Munjiu Oxana, doctor n biologie, confereniar cercettor, Zubcov Natalia, doctor n biologie,
confereniar universitar, Bulat Dumitru, doctor n biologie, confereniar cercettor, Bulat Denis, doctor
n biologie, confereniar cercettor, Borodin Natalia, Lebedenco Liubovi, Ungureanu Grigore, Tumanova
Daria i Banu Vitalie.

35

I. PRINCIPII GENERALE DE PRELEVARE N ECOSISTEME ACVATICE

Prelevarea este definit n ISO 5667-2 drept un proces de nlturare a unei pri, care se presupune a
fi reprezentativ, dintr-un bazin acvatic (sau nmol, sau sediment) n scopul examinrii diferitor caracteristici strict delimitate. Colectarea probelor este prima etap n monitoringul ecosistemelor acvatice,
avnd o importan deosebit. Utilizarea echipamentului i a metodelor sofisticate n procesul de analiz
nu sunt n stare s diminueze impactul erorilor fcute n procesul colectrii probelor.
n cazul ecosistemelor acvatice naturale, principiile generale i tehnicile de prelevare sunt reglementate
printr-un set de standarde internaionale din categoria Calitatea apei. Prelevare (ISO 5667), aprobat la
nivel naional [1-5]. De asemenea, la nivel naional au fost aprobate unele standarde internaionale (de
tipul ghidurilor), care reglementeaz tehnicile specifice de prelevare a anumitor grupuri de hidrobioni n
habitatul lor natural [6-10]. n registrul standardelor naionale nc nu sunt incluse unele ghiduri importante din acest set de documente europene, ceea ce ngreuiaz utilizarea lor n Republica Moldova [11-12].
Condiiile i cerinele naintate fa de prelevare difer ntr-o anumit msur n dependen de tipul
apelor, programul i scopul monitoringului. Este greu de elaborat un ghid sau recomandri unice, dar
ntotdeauna sunt cerine sau principii de baz privind recoltarea probelor pentru monitorizarea fizico-chimic i biologic a ecosistemelor acvatice:
Peridiocitatea prelevrii depinde de tipul monitoringului. Coninutul programului de monitorizare trebuie s ia n consideraie variabilitatea parametrilor, care se pot modifica att sub influena
condiiilor naturale, ct i ca rezultat al impactului uman.
Materialul colectat trebuie sa fie reprezentativ i s corespund scopului i programului de monitorizare.
Probele trebuie s reflecte situaia real n locul de colectare.
Metodele de conservare, transportare i pstrare trebuie s se reflecte ct mai puin asupra componenei fizico-chimice i biologice a probelor recoltate. Se recomand nchiderea ermetic a recipientelor i protejarea de aciunea luminii i cldurii excesive, deoarece caracteristicile probei se
pot deteriora rapid datorit schimbului de gaze, reaciilor chimice i metabolismului organismelor
prezente n prob.
Volumul materialului colectat trebuie s fie suficient pentru a efectua analiza n conformitate cu
metodele i tehnicile de laborator utilizate;
Recipientul (vesela, containerul) trebuie s fie confecionat din materiale inerte. De cele mai multe
ori recoltarea se face n flacoane de polietilen sau alt material plastic. n cazul determinrii substanelor organice, probele prelevate sunt depozitate n flacoane de sticl.
Toate procedurile de prelevare a probelor trebuie s fie strict documentate, fapt care permite identificarea fr dificultate a probei n laborator. Fiecare loc de colectare trebuie descris. Dac programul de monitorizare este de lung durat, atunci nu este necesar de a descrie repetat elementele
stabile ale mediului. n asemenea caz doar rezultatele msurrilor in situ i aa condiii variabile
precum condiiile meteorologice, precum i fenomenele neobinuite observate trebuie nregistrate. Dac prelevarea vizeaz careva obiective speciale, atunci informaia se prezint detaliat, inclusiv
cauzele recoltrii probelor i fiecare etap de conservare a acestora.
Probele sunt recoltate de un personal calificat i n corespundere cu standardele de siguran (este
obligatorie instruirea personalului n scopul asigurrii unor condiii corespunztoare de siguran).
Persoanele care preleveaz probe trebuie s ia n consideraie c n unele ruri i cursuri de ap
exist pericole de origine chimic, bacteriologic, viral i animal.
Bibliografie
1. ISO 5667-1:2006 Water quality. Sampling. Part 1: Guidance on the design of sampling programmes
and sampling techniques. National version: SM SR EN ISO 5667-1:2011 Calitatea apei. Prelevare.
Partea 1: Ghid general pentru stabilirea programelor i tehnicilor de prelevare (in Romanian).
Chiinu: INSM, 2011. 35 p.
36

2. ISO 5667-4:1987 Water quality. Sampling. Part 4: Guidance on sampling from lakes, natural and
man-made. National version: SM SR ISO 5667-4:2007 Calitatea apei. Prelevare. Partea 4: Ghid
de prelevare a apelor din lacuri naturale i artificiale (in Romanian). Chiinu: MOLDOVASTANDARD, 2007. 10 p.
3. ISO 5667-6:2005 Water quality. Sampling. Part 6: Guidance on sampling of rivers and streams.
National version: SM SR ISO 5667-6:2011 Calitatea apei. Prelevare. Partea 6: Ghid pentru prelevrile
efectuate n ruri i alte cursuri de ap (in Romanian). Chiinu: INSM, 2011. 22 p.
4. ISO 5667-12:1995 Water quality. Sampling. Part 12: Guide on sampling of bottom sediments.
National version: SM SR ISO 5667-12:2007 Calitatea apei. Prelevare. Partea 12: Ghid general pentru
prelevarea sedimentelor de fund (in Romanian). Chiinu: MOLDOVA-STANDARD, 2007. 42 p.
5. ISO 19458:2006 Water quality. Sampling for microbiological analysis. National version: SMV EN ISO
19458:2011 Calitatea apei. Prelevare pentru analiz microbiologic (in English). INSM, 2011. 30 p.
6. EN 15110:2006 Water quality. Guidance standard for the sampling of zooplankton from
standing waters. National version: SM SR EN 15110:2012 Calitatea apei. Ghid pentru prelevarea
zooplanctonului din ape stttoare (in Romanian). Chiinu: INSM, 2012. 24 p.
7. EN 15708:2009 Water quality. Guidance standard for the surveying, sampling and laboratory
analysis of phytobenthos in shallow running water. National version: SM EN 15708:2013(E)
Calitatea apei. Ghid pentru studiul, prelevarea i analiza n laborator a fitobentosului din apele
curgtoare de mic adncime (in English). INSM, 2013. 25 p.
8. EN 13946:2003 Water quality. Guidance standard for the routine sampling and pre-treatment of
benthic diatoms from rivers. National version: SM SR EN 13946:2012 Calitatea apei. Ghid pentru
prelevarea uzual i pretratarea diatomeelor bentonice din ruri (in Romanian). INSM, 2012. 22 p.
9. EN 14757:2005 Water quality. Sampling of fish with multi-mesh gillnets. National version: SM SR
EN 14757:2012 Calitatea apei. Prelevarea petilor utiliznd plase cu mrimi diferite ale ochiurilor
(in Romanian). INSM, 2012. 36 p.
10. EN 14962:2006 Water quality. Guidance on the scope and selection of fish sampling methods.
National version: SM SR EN 14962:2012 Calitatea apei. Ghid pentru domeniul de aplicare i
selecie a metodelor de prelevare peti (in Romanian). INSM, 2012. 34 p.
11. ISO 10870:2012 Water quality. Guidelines for the selection of sampling methods and devices
for benthic macroinvertebrates in fresh waters (This edition of ISO 10870 cancels and replaces
ISO 7828:1985, ISO 8265:1988, and ISO 9391:1993 which have been technically revised). ISO,
2012. 26 p.
12. EN 16150:2012 Water quality. Guidance on pro-rata Multi-Habitat sampling of benthic macroinvertebrates from wadeable rivers. BSI, 2012. 18 p.

II. PRELEVAREA APEI

Exist dou tipuri de probe de ap, care pot fi prelevate din ruri, lacuri i ape de suprafa similare
[23]: 1) probe instantanee, care sunt colectate ntr-un anumit loc, la o anumit adncime i or i 2)
probe compuse sau integrate, care pot fi integrate n dependen de adncime, suprafa, timp i scurgere.
Exist diferite tipuri de prelevatoare (prelevator de oxigen dizolvat, prelevator de adncime, prelevator
multifuncional), cele mai multe fiind concepute pentru atingerea unor obiective specifice.
Cantitatea de ap recoltat depinde de analizele care trebuie efectuate, aceasta variind de la 100 ml
pn la 5 litri (volumul colectat trebuie s permit efectuarea analizei n cel puin 3 repetri).
Temperatura apei se msoar i se nregistreaz imediat dup ce proba a fost colectat. Parametrii
fizici ai apei (de exemplu, pH, gaze dizolvate, suspensii) se determin n punctul de colectare ct mai
curnd posibil dup recoltarea probei de ap.
Recipientele pentru transportarea probelor se folosesc numai pentru probele de ap i nici ntr-un
caz pentru depozitarea substanelor chimice sau a altor lichide. De regul, sunt utilizate recipiente de sticl, care sunt potrivite pentru transportarea probelor n cazul mai multor analize, ns, pentru anumite
analize chimice, hidrobioni sau sedimente este preferabil de a utiliza recipiente de plastic. Recipientele
37

i procedurile recomandate de splare a acestora n dependen de compusul chimic analizat sunt prezentate n anexa 2.
Un vas/recipient, care urmeaz a fi utilizat pentru transportarea sau pstrarea probei, se cltete de
trei ori cu poriuni de prob nainte de a fi umplut. Totui, aceast procedur nu se aplic dac recipientul deja conine un conservant chimic. Nu se efectueaz cltiri preliminare i n cazul colectrii suspensiilor i probelor microbiologice. Probele sunt transferate n recipientul destinat pstrrii/transportului
imediat dup colectare. n recipient se las un mic spaiu cu aer, pentru a permite amestecarea probei
nainte de analiz. Fiecare vas trebuie s aib o etichet de identificare.
O serie de parametri fizico-chimici, precum temperatura, oxigenul dizolvat, dioxidul de carbon, pH,
potenialul redox, conductivitatea i turbiditatea sunt msurate n momentul prelevrii probei direct n
bazinul acvatic, folosind instrumente de msurare portabile cu senzori (Fig. 1 3). Instrumentele trebuie s fie n stare bun de lucru, pentru a obine rezultate precise, fiind supuse calibrrii in situ de fiecare
dat. Exist urmtoarele excepii: temperatura (verificat lunar n laborator cu ajutorul unui termometru
cu mercur certificat), conductivitatea i turbiditatea (calibrate la nceputul expediiei de colectare a probelor) i potenialul redox (calibrat o dat la fiecare ase luni).

Fig. 1. Instrument portabil de

msurare a pH-ului, conductivitii
electrice, cantitii totale de
suspensii i a temperaturii

Fig. 2. Instrument portabil de msurare

a oxigenului dizolvat, presiunii
barometrice i a temperaturii

Fig. 3. Instrument portabil de

msurare a dioxidului de carbon

Oxigenul dizolvat poate fi determinat prin metoda tradiional metoda iodometric Winkler. Aceast
metod este folosit pentru calibrarea diferitor instrumente portabile de msurare a oxigenului dizolvat i,
fiind o metod de ncredere de laborator, adesea este utilizat n ecologia acvatic. Cea mai important etap n aplicarea acestei metode const n prelevarea probei n vase de sticl cu deschiztura ngust, cu dop
(vase Winkler sau oriice alte vase potrivite) i cu un volum de 130-350 ml, gradate la fiecare 1ml (Fig. 4).
Att fiecare vas, ct i dopul su trebuie s poarte acelai numr de identificare. n
timpul colectrii probei trebuie luate msuri de precauie pentru a nu permite formarea bulelor de aer sub dop. Vasele se umplu pn la refuz, avnd grij de a evita orice
modificare a concentraiei oxigenului dizolvat. Dup umplerea vasului, oxigenul dizolvat este fixat imediat cu soluie de sulfat de mangan i reactiv alcalin [13]. Vasul este
inversat de mai multe ori, pentru a amesteca bine coninutul. Se ateapt cel puin 5
min formarea precipitatului i sedimentarea lui, apoi vasul este din nou inversat, penFig. 4. Vas de
oxigen Winkler tru a asigura omogenitatea amestecului. n aceast stare vasele pot fi transportate la
laborator (Fig. 5).
Dac sunt protejate de lumin, atunci probele pot fi pstrate pn la 24 de ore. Dac n ap se conine
materie n suspensie capabil de a fixa sau de a consuma iod, atunci se recomand utilizarea metodei
electrochimice [14].
Toate msurrile efectuate n cmp i ntreaga informaie de suport (condiii meteo, nivelul de dezvoltare a algelor, prezena petilor mori sau a petelor de ulei, etc.) trebuie notate n carnetul de cmp nainte
de a prsi staiunea de prelevare.
38

Fig. 5 Fixarea oxigenului dizolvat

Fig. 6. Filtrarea probelor de ap

n condiii de cmp

Fig. 7. Spectrofotometru portativ pentru

analize chimice

Anumite pri ale probei de ap trebuie prevzute pentru

determinarea pH-ului i conductivitii. Nu se permite utilizarea aceleeai porii de ap pentru ambele determinri, datorit posibilitii de difuzie a clorurii de potasiu din membrana
electrodului pH-metrului.
Pentru determinarea coraportului dintre formele solubile
i insolubile ale metalelor, probele de ap sunt filtrate imediat
dup recoltare prin filtre cu mrimea porilor de 0,45 m (Fig.
6). Apa filtrat obinut se conserveaza cu acid azotic. Filtrarea
nu se aplic dac filtrul membranar poate reine unul sau mai
muli analii. De asemenea, este important ca sistemul de filtrare s poat fi splat uor nainte de umplere. Probele colectate
n scopul determinrii pesticidelor sunt conservate conform
metodelor standard, cu utilizarea diferitor solveni.
n ceea ce privete pstrarea probelor, exist anumite cerine
de baz, care previn deteriorarea acestora: vasele cu probe trebuie bine nchise i depozitate ntr-un loc curat, rcoros, ntunecat
i protejat de contaminare. Metodele suplimentare de conservare includ congelarea, extracia cu solveni i adugarea conservanilor chimici. Totui, n fiecare caz metoda de conservare i
pstrare trebuie s fie compatibil cu tehnicile analitice utilizate.
Standardul internaional ISO 5667-3 sugereaz utilizarea anumitor conservani chimici i perioada maxim admis de pstrare
a probelor [15].
Prelevatorul Van Dorn i batometrele sunt instrumentele ideale pentru colectarea apei (Fig. 8). Vasele pentru probe
Van Dorn sunt bine adaptate pentru prelevarea probelor la
orice adncime, atunci cnd se urmrete un scop general.
Ele pot avea att poziie orizontal, ct i vertical. Vasele
orizontale sunt coborte paralel cu fundul bazinului acvatic.
Ele sunt ideale pentru diferite niveluri de stratificare a apei,
regiunea termoclinului sau stratul de ap situat nemijlocit
de asupra sedimentelor de la fund. Modelul cu vase verticale
este, de regul, folosit pentru cercetarea stratificrii; n acest
caz unul sau mai multe prelevatoare sunt suspendate de un
cablu. Vasul Alpha Van Dorn este utilizat pentru probe fizice, chimice i biologice. El are o deschiztur larg, care
nu restricioneaz fluxul de ap i este ideal pentru colectarea microplanctonului. Prelevatorul Beta Van Dorn poate
fi utilizat pentru studierea microelementelor i poluanilor
organici.

Fig. 8. Prelevatorul Van Dorn [16]

Fig. 9. Colectarea probelor de ap la mal

39

Uneori, cnd locul de prelevare este situat pe malul unui ru sau lac (Fig. 9), proba este recoltat direct n vasul pentru transportare cu ajutorul oricrui vas (de exemplu, o cldare) confecionat dintr-un
material, care nu are proprietatea de a contamina proba de ap.
Bibliografie
13. EN 25813:1992 Water quality. Determination of dissolved oxygen: iodometric method. (ISO
5813:1983) National version: SM SR EN 25813:2011 Calitatea apei. Determinarea coninutului de
oxigen dizolvat. Metoda iodometric (in Romanian). Chiinu: INSM, 2012. 12 p.
14. ISO 5814:1990Water quality. Determination of dissolved oxygen: electrochemical probe method
National version: SM SR EN ISO 5814:2014 Calitatea apei. Determinarea coninutului de oxigen
dizolvat. Metoda electrochimic cu sonda (in Romanian). Chiinu: INSM, 2014. 25 p.
15. EN ISO 5667-3:2003 Water quality. Sampling. Part 3: Guidance on the preservation and handling of
water samples. National version: SM SR EN ISO 5667-3:2011 Calitatea apei. Prelevare. Partea 3: Ghid
pentru conservarea i manipularea probelor de ap (in Romanian). Chiinu: INSM, 2012. 31 p.
16. http://shop.sciencefirst.com/wildco/567-van-dorn-water-samplers.

III. PRELEVAREA SUSPENSIILOR I SEDIMENTELOR

Prelevarea suspensiilor i sedimentelor
Prelevarea suspensiilor are o mare importan n cercetarea migraiei substanelor chimice n sistemul
ap suspensiii depuneri subacvatice. De regul, pentru colectarea lor se utilizeaz metoda filtrrii. Filtrele sult selectate n dependen de mrimea porilor. n majoritatea cazurilor, sunt utilizate filtre de hrtie,
filtre membranare sau din sticl, n dependen de metodele de determinare a substanelor chimice.
Pentru analiza coninutului de metale n suspensii, se filtreaz un volum determinat de ap (200-1000
ml). Filtrul cu suspensii este transferat ntr-un vas special i ulterior analizat n condiii de laborator.
Studierea sedimentelor este recomandat n cazul aprecierii strii chimice a ecosistemelor acvatice, n
special, lacuri i heleteie [17].
Sedimentele pot fi utilizate drept matrice n cazul a trei tipuri de investigaii: chimice, fizice i biologice. Natura i cantitatea substanelor din sedimente se determin prin investigaii chimice. Un studiu
biologic include, n general, grupele i efectivul numeric al bacterio-, fito- i zoobentosului n sedimente.
Principiile de baz ale procedurii de colectare a sedimentelor (inclusiv selectarea staiunii de colectare,
punctelor de colectare, a metodei de colectare, etc.) sunt descrise n standardul internaional ISO 5667-12
[18]. Drept principiu general poate fi considerat evitarea deteriorrii proprietilor sedimentelor n timpul prelevrii (de exemplu, prin contaminare sau tulburare a substratului). Cercetarea sedimentelor poate
reprezenta o abordare cost-eficient pentru screening-ul iniial al nivelului de poluare a bazinelor acvatice,
compararea concentraiilor poluanilor ntre diferite staiuni i identificarea surselor de poluare poteniale.
Frecvena minim de prelevare a apei pentru monitoringul substanelor prioritare este de o dat pe
lun (o dat la fiecare 3 luni n cazul poluanilor, care sunt specifici pentru bazinul rului dat), ns
frecvena monitoringului sedimentelor poate fi de o dat pe an, cu excepia cazurilor cnd informaiile
tehnice i opiniile experilor justific un alt interval.

a
b
c
d
e
Fig. 10: a prelevator de tip ben [19], b prelevator de tip ben [20], c prelevator manual de tip Corer [21],
d prelevator de tip Corer cu gravitaie [22], e prelevator de tip Corer cu cutie [21]

40

Sedimentele pot fi colectate fie prin utilizarea unui prelevator de tip ben, fie de tip Corer (Fig. 10).
Exist o gam larg de dispozitive de prelevare a sedimentelor. Alegerea echipamentului trebuie fcut
n funcie de condiiile locale la staiunea de prelevare, de exemplu, adncimea apei i tipul de sedimente.
Prelevatoarele de tip Corer sunt dispozitive sub form de tub care ptrund n sedimente datorit gravitaiei (cdere liber), vibraiei, presiunii hidraulice sau puterii manuale. Att prelevatoarele de tip ben, ct
i cele de tip Corer pot fi folosite pentru recoltarea stratului superior al sedimentelor, pe cnd n rurile
mai mici i puin adnci pot fi folosite cue, lopele.
Pentru studierea migraiei metalelor, se colecteaz stratul superior de 5 cm al sedimentelor. Coninutul metalelor poate fi determinat per total pe sedimente, sau n diferite fracii granulometrice ale acestora. O parte din proba de sedimente este centrifugat pentru 30-45 min, cu o vitez de 2500-3000 rotaii/
min, pentru obinerea extractului apos, n care se afl cele mai mobile forme de migraie a metalelor n
sistemul ap depuneri subacvatice.
Cernerea i transportarea probelor de sedimente
Probele de sedimente trebuie cernute prin site cu mrimea ochiului de peste 2 mm, pentru a nltura detritul i organismele bentonice de dimensiuni mari. Altfel, pe parcursul etapelor ulterioare de manipulare i
procesare a probei, precum depozitarea, nghearea sau tratarea cu ultrasunete, va avea loc descompunerea
materialului biologic, substanele rezultate devenind pri componente ale sedimentelor. Pentru a minimiza
perturbarea echilibrului sediment/ap, se recomand cernerea umed a sedimentelor n punctul de prelevare
a probei, folosind apa din bazinul de ap respectiv. Se utilizeaz aceeai ap, pentru a preveni schimbarea
echilibrului. n cazul probelor cu o salinitate nalt, este important, n special, de a folosi n timpul cernerii
sedimentelor o ap cu aproximativ aceeai salinitate ca i n locul de colectare a probei. n general, cernerea
se efectueaz pe parcursul primelor 24 de ore dup colectarea sedimentelor, de preferin n condiii de cmp.
Dac cernerea n condiii de cmp nu este posibil, atunci ea se face n laborator, n condiii strict stabilite.

Fig. 11. Site din inox [23]

Fig. 12. Borcan de sticl

ntunecat [24]

Fig. 13. Cutie de rcire [25]

De obicei, sitele sunt confecionate din alam rezistent la coroziune (rama i plasa). Actualmente, n
cazul analizei compuilor organici, sunt preferate sitele confecionate din oel inoxidabil (Fig. 11).Totui,
acestea nu trebuie utilizate dac sunt analizate microelementele. n cazul dat se recomand sitele din
polimeri, cu ram din policlorur de vinil (PVC) sau acril i plas din nailon sau poliester.
Pentru analizele organice, probele (trecute sau nu prin sit) sunt, de preferin, transferate n vase din
sticl ntunecat (sau din aluminiu, sau din alt material necontaminant) cu deschiztur larg, iar pentru
analiza microelementelor n sculee sau butelii de plastic. n mod alternativ, borcanele de sticl ntunecat pot fi utilizate pentru toate tipurile de contaminani (Fig. 12). Recipientele pentru probe trebuie
umplute cu vrf (cu un spaiu liber minim), pentru a diminua probabilitatea oxidrii i pierderii n timpul transportrii a sulfurilor, care pot degaja hidrogen sulfuros la interaciunea cu acizii. Se recomand
transportarea probelor ct mai curnd posibil ctre laborator; pe durata transportrii se asigur o temperatur de aproximativ 4 C. Rcirea probelor se efectueaz uor cu ajutorul cutiei de rcire (Fig. 13).
41

Depozitarea probelor de sedimente

Depozitarea ncepe imediat dup prelevarea probelor. Toate metodele de pstrare vor afecta proba
ntr-o oarecare msur, de aceea, alegerea tehnicii de conservare depinde, n special, de obiectivul colectrii probei. n legtur cu faptul c primele ore dup prelevare sunt cele mai critice n vederea modificrilor, care pot avea loc n componena probelor, conservarea trebuie efectuat, atunci cnd este posibil,
imediat dup colectarea probelor. ndeosebi, acest lucru este important n cazul analizei n sedimente a
pesticidelor sau compuilor volatili.
Recipientele pentru depozitarea probelor de sedimente liofilizate sau uscate, de preferin, sunt vase
cu deschiztur mare i capac filetat. Probele prelevate pentru analiza poluanilor de natur organic se
depoziteaz n recipiente de sticl ntunecat, PTFE (politetrafluoretilen), oel inoxidabil sau aluminiu.
Sedimentele colectate pentru analiza metalelor pot fi pstrate n recipiente nchise de plastic sau sticl.
Indiferent de analizele chimice efectuate ulterior, se admite utilizarea borcanelor de sticl ntunecat, n
scopul simplificrii caracterizrii sedimentelor.
Pentru analiza mercurului, probele trebuie depozitate n recipiente de sticl borosilicat sau cuar,
splate preventiv cu acid. Pentru analiza compuilor chimici, este de dorit de a pstra probele n vase de
sticl ntunecat, ns, de asemenea, este binevenit utilizarea recipientelor din alte materiale, de exemplu, policarbonat sau aluminiu.
Perioada maxim de pstrare a sedimentelor liofilizate pn la momentul analizei este de cca 180 de
zile (30 de zile pentru mercur), cu condiia c probele sunt depozitatea ntr-un congelator.
Probele de sedimente pot fi arhivate. n cazul analizei metalelor se aplic o procedur simpl de arhivare,
care const n uscarea probei la 30C sau 105C i depozitarea sedimentelor uscate n recipiente de polietilen.
Bibliografie

17. Guidance on chemical monitoring of sediment and biota under the Water Framework Directive.
Luxembourg: Office for Official Publications of the European Communities, 2010. 82 p.
18. ISO 5667-12:1995 Water quality. Sampling. Part 12: Guidance on sampling of bottom sediments.
National version: SM SR ISO 5667-12:2007 Calitatea apei. Prelevare. Partea 12: Ghid general pentru
prelevarea sedimentelor de fund (in Romanian). Chiinu: MOLDOVA-STANDARD, 2007. 41 p.
19. http://www.envirotools.hu/english/benthic_grabs.html.
20. http://www.gserentals.co.uk/details.aspx?product=93
21. http://clu-in.org/programs/21m2/sediment
22. http://www.osil.co.uk/Products/RentalEquipment/tabid/59/agentType/View/PropertyID/314/
Default.aspx
23. http://www.gemplers.com/product/134572/Stainless-Steel-Sieve-Set
24. http://www.inglass.com/amberjars.html
25. http://www.waecofridges.co.uk/coolbox/mobicoolu26.htm

IV. PRELEVAREA BACTERIOPLANCTONULUI

Pentru analize microbiologice se utilizeaz recipiente de sticl cu perei groi, de exemplu, flacoane,
cu un volum minim de 300 ml. nainte de autoclavare, dopurile recipientelor se acoper cu hrtie kraft,
pentru a le proteja de contaminare n timpul manipulrii. Pregtirea recipientelor const n splarea cu
detergent neionic i cltirea de cel puin trei ori cu ap distilat sau deionizat nainte de autoclavare.
Recipientele noi necesit aceeai pregtire.
Alegerea modului i locului de colectare a probelor de ap pentru cercetarea bacterioplanctonului
depinde de scopul cercetrii i condiiile locale. Principalele cerine sunt [5, 26 - 27]:
Flacoanele, n care se recolteaz probele de ap, preventiv se sterilizeaz n etuv timp de o or la
180 C, sau n autoclav timp de 20 min la o presiune de 1 atm, ceea ce corepunde temperaturii de
121 C (Fig. 14-15).
42

 Probele din straturile de suprafa ale apei nu se colecteaz n apropierea unei surse poluante (cu
excepia cazurilor prevzute de planul de cercetare).
Cantitatea de ap necesar variaz de la 500 ml pn la 1000 ml, n funcie de tipul analizei efectuate.
Flacoanele pentru colectarea probelor urmeaz s fie deschise numai nainte de colectarea probelor.
Probele de suprafa se colecteaz de la adncimi de 15-30 cm.
La colectarea probelor din bazine acvatice mici stttoare este necesar de a preveni tulburarea sedimentelor.
Flacoanele sunt umplute cu ap n aa mod nct distana dintre ap i dop s fie de cel puin 2 cm;
n timpul recoltrii probelor se evit orice posibilitate de contaminare accidental a apei;
Dup prelevarea probei, flacoanele se eticheteaz, indicnd locul, data, ora i eventualele caracteristici ale locului de prelevare. Toate probele sunt introduse n registru.

Fig. 14. Etuv i sterilizator de laborator [28]

Fig. 15. Autoclav de

laborator cu ncrcare
vertical [29]

Fig. 16. Cutie pentru transportarea

probelor de bacterioplancton [30]

Dac ntrzierea dintre prelevarea probelor i analiza bacteriologic este mai mic de 2 ore, atunci
probele pur i simplu sunt pstrate ntr-un loc rcoros, ntunecat. Dac trec mai mult de 2 ore, atunci
probele se rcesc rapid pn la aproximativ 4 C prin plasarea lor ntr-o cutie de rcire, n care urmeaz
s rmn i n timpul transportrii (Fig. 16). Dac perioada dintre prelevare i analiz depeete 6 ore,
atunci raportul de analiz trebuie s includ detalii privind condiiile i durata transportrii probei.
Colectarea probelor din ape curgtoare se efectueaz n acelai mod ca i din apele de suprafa stttoare. n unele cazuri, n particular la colectarea probelor de ap din staiile de epurare a apelor, este
necesar utilizarea unui colector de probe special.
Analiza probelor de bacterioplancton trebuie efectuat ct mai curnd posibil (timp de 6 ore) dup
recoltrea lor. Dac probele nu pot fi aduse n laborator n acest interval de timp, atunci ele se pun la
pstrare n frigider cel mult pentru 24 de ore [26].
Bibliografie
26. Toderas I., Negru M., Ionica D., Nicolescu D., Simon-Gruita A. Ecology of aquatic microorganisms.
Original title: Todera I., Negru M., Ionic D., Nicolescu D., Simon-Grui A. Ecologia microorganismelor acvatice (in Romanian). Chiinu: tiina, 1999. 281 p.
27. Kopilov A., Kosolapov D. Bacterioplancton of reservoirs of Upper and Middle Volga. Moscow:
Original title: A., . (in Russian). : , 2008. 377 .
28. http://eprus.pl/farma-play-suszarki-sterylizatory/.
29. http://labimpex.com.ua/p42078351-sterilizator-parovoj-avtoklav.html.
30. http://www.who.int/water_sanitation_health/resourcesquality/wqmchap5.pdf.

V. PRELEVAREA FITOPLANCTONULUI
Fitoplanctonul este reprezentat de organismele microscopice, liber vieuitoare n ap, capabile s fotosintetizeze. Cercetrile fitoplanctonului, determinate de rolul lui principal ca productor al substanei
organice i ca factor al formrii calitii apei, au importan deosebit la elaborarea bazelor tiinifice de
dirijare a ecosistemelor acvatice, construire a hidrobiocenozelor nalt productive, relativ stabile n condiiile influenei factorilor antropici. Structura i particularitile funcionale ale fitoplanctonului n cea
43

mai mare msur determin caracterul bioproductivitii ecosistemelor acvatice.

Productivitatea fitoplanctonului, spre deosebire de hidrobionii ce se refer nivelurilor trofice mai
superioare, poate fi determinat fr analiza fitoplanctonului ca atare.
Metoda de colectare a eantioanelor de fitoplancton este relativ simpl, ns prelucrarea lor necesit
o calificare nalt a cercettorului.
Alegerea punctelor de prelevare a probelor
Alegerea punctelor de prelevare a probelor pe orizontal i frecvena colectrii lor n timp sunt determinate n corespundere cu scopul i sarcinile investigaiilor hidrobiologice. n toate cazurile colectarea
probelor de fitoplancton i determinarea produciei primare se efectueaz sincronic n aceleai puncte.
Alegerea punctelor de prelevare a probelor de fitoplancton pe vertical este n corespundere cu caracteristicile factorilor fizici, ce influeneaz dezvoltarea algelor (lumina, temperatura).
n majoritatea bazinelor acvatice, cu excepia celor foarte adnci, numrul punctelor de colectare a
probelor la diferite adncimi (h), necesare pentru evidena fitoplanctonului la diferite nivele, se poate
limita la ase [31]:
1. Suprafa - zona de iluminare i nclzire maximal a apei, supus n acelai timp influenei vntului;
2. Orizontul situat la jumtatea valorii transparenei;
3. Orizontul la adncimea transparenei- practic coincide cu mijlocul stratului trofogen;
4. Orizontul ce corespunde adncimii duble a transparenei - de regul, coincide cu limita zonei trofogene (aici se situeaz punctul de compensare, iar la stabilirea stratificrii estivale termoclinul);
5. Orizontulce corespunde jumtii zonei trofolitice - se determin ca jumtatea distanei de la fundul bazinului pn la valoarea dubl a transparenei;
6. Orizontul situat la 0,3-0,5 m de la substrat.
La prelevarea probelor pe vertical este necesar de a lua n consideraie c i n bazinele relativ mici au loc migraiile verticale ale
fitoplanctonului, influenate de schimbrile diurne i cele ale condiiilor meteorologice. Deoarece n ruri distribuirea vertical a fitoplanctonului este relativ omogen, colectarea probelor se efectueaz de la orizontul 0,2 1m.
Metode de prelevare i echipament
Metodele de prelevare i investigare a algelor sunt foarte diverse, fiind determinate de particularitile ecologo-morfologice ale
reprezentanilor diferitor filuFig. 17. Plas planctonic [32]
muri i grupri ecologice, ct i
de diversitatea scopurilor i modurilor de abordare a investigaiilor.
Nu vom descrie aici acele metode care sunt orientate la atingerea
unor scopuri speciale. Ne vom limita la metodele de prelevare i
investigare a algelor ecosistemelor acvatice continentale cu scopul
cercetrilor floristico-sistematice i hidrobiologice.
n scopul studierii componenei specifice a fitoplanctonului n
timpul dezvoltrii lui intense, este suficient de prelevat ap din bazin ntr-un flacon i mai apoi de analizat proba la microscop. Dar, n
Fig. 18. Prelevator
majoritatea cazurilor, sunt aplicate diferite metode de concentrare
Ruttner pentru prelevarea
prealabil. Una din acestea este filtrarea apei prin plase planctonice
vertical a planctonului [32]
de diferit construcie (Fig. 17).
Cu un vas, volumul cruia este de un litru, se ia ap din straturile superficiale (pn la 15 20 cm
adncime) i se toarn n plas, filtrnd, astfel, 50-100 l de ap. n rurile i lacurile mari probele planctonice se preleveaz din luntre. Se recomand de tras plasa planctonic cu o funie subire dup luntrea n
44

micare timp de 5-10 min. Pentru colectri verticale ale planctonului sunt utilizate plase cu construcie
deosebit (Fig. 18).
n bazinele mici probele planctonice pot fi prelevate de la mal, treptat intrnd n ap i filtrnd apa
din fa prin plas, sau se arunc plasa n ap i se trage cu atenie de sfoar. Dup ce au fost efectuate
prelevrile, plasa planctonic trebuie cltit bine, scufundnd-o de cteva ori n ap pn la cerc n scopul
splrii algelor rmase pe suprafaa intern a plasei.
Concentrat n aa mod, proba planctonic din phrelul plasei se toarn prin tubul de evacuare n
flaconul pregtit n prealabil. nainte i dup colectarea probelor plasa trebuie bine cltit, iar dup finalizarea prelevrilor uscat i pstrat ntr-o hus special. Probele planctonice pot fi analizate n stare vie
sau fixate.Conservarea probelor se face ct mai curnd posibil dup colectare, dar ntotdeauna ea trebuie
fcut pe uscat, avnd la dispoziie ap n caz de stropire cu reactivi sau vrsare a acestora pe mini.
Pentru estimarea indicilor cantitativi ai fitoplanctonului, se preleveaz probe cu un volum determinat. n acest scop, pot fi efectuate i prelevri cu plasa, cu condiia evidenei cantitii de ap filtrate prin
plas i volumului probei prelevate. Dar cea mai sigur metod de prelevare a probelor de fitoplancton
este metoda cu utilizarea colectorului. Probele prelevate cu colectorul pot fi utilizate att la aprecierea
structurii taxonomice a comunitilor de alge, ct i la estimarea indicilor cantitativi ai fitoplanctonului.
n bazinele n care fitoplanctonul este srac se colecteaz nu mai puin de un litru de ap, iar n cele
bogate n fitoplancton volumul probei colectate poate fi de 0,5-0,25 l.
Etichetarea probelor
Fiecare prob trebuie etichetat. Pe etichet se indic data colectrii, locul,
denumirea sau numrul staiei (Fig. 19). Etichetele se lipesc pe sticlue. n registrul, n care se duce evidena tuturor probelor colectate, pe lng datele menionate se nscriu i alte caracteristici: date despre temperatura aerului i apei,
transparen, pH, viteza apei, aprecierea vizual a strii apei, de exemplu, gradul
de dezvoltare al macrofitelor .a.
Fig. 19. Flacon Nalgene
etichetat

Bibliografie

31. Algae. Handbook / Vasser S., Condratieva N., Masiuc N. et al. Kiev: Naukova Dumka, 1989 (in
Russian). Original title: . / .., ..,
.. . : , 1989. 608 .
32. http://www.duncanandassociates.co.uk/equip.htm

VI. PRELEVAREA ZOOPLANCTONULUI

Zooplanctonul este constituit din organismele planctonice heterotrofe, care plutesc n oceane, mri i
bazine acvatice continentale cu ap dulce. Ca i fitoplanctonul, zooplanctonul este o component-cheie
a ecosistemelor marine i oceanice. Teritoriul Republicii Moldova nu include nici mri, nici oceane. Prin
urmare, programele de monitorizare naionale i cercetrile tiinifice au ca obiect comunitile zooplanctonice din ecosistemele continentale lotice i lentice. Prelevarea zooplanctonului n ruri i bazine
acvatice cu ap stttoare (lacuri, heleteie i lacuri de acumulare) urmeaz a fi descris n aceast parte
a ghidului.
Localizarea prelevrii
Staiune de prelevare: zona general n cadrul bazinului acvatic din care sunt prelevate probele.
Punct de prelevare: poziia exact n cadrul locaiei de prelevare din care se iau probele.
Zona de litoral: zona din apropierea malului, n care lumina ptrunde pn la fund i permite creterea macrofitelor (Fig. 20).
45

 Zona pelagic: grosul apei dincolo de zona litoral.

Zona limnetic: stratul de ap de la suprafa, bine iluminat, nconjurat de zona litoral, n care
planctonul domin (att fitoplanctonul, ct i zooplanctonul).
Ap stratificat: stratificarea termic a apei n bazinele acvatice cu ap dulce i srat. Este caracterizat prin gradientul vertical al densitii. Apa rece este mai dens ca apa cald i epilimnionul, n
ansamblu, este format din ap, care nu este att de dens precum apa din hipolimnion.
Termoclin (metalimnion): strat dintr-un
bazin acvatic stratificat termic, n care gradientul de temperatur are valoare maxim.
Epilimnion: strat de ap poziionat de-asupra
termoclinei ntr-un bazin acvatic stratificat.
Hipolimnion: strat de ap poziionat sub
termoclin ntr-un bazin acvatic stratificat.
Identificarea punctelor de prelevare n
cadrul reelei hidrologice staionare permite colectarea probelor comparative pentru
crearea bazelor de date multianuale. n cele
mai multe cazuri, punctele de prelevare pot
fi uor regsite prin raportare la particularitile morfologice i hidrologice ale bazinuFig. 20. Distribuia zonal a habitatelor n lac [33]
lui acvatic. Pentru a controla calitatea apei,
n mod normal, probele de zooplancton sunt prelevate din acelai punct n care se colecteaz i alte probe
hidrobiologice (precum i probe hidrochimice).
Selectarea punctelor de prelevare n cadrul programelor de monitoring operaional, n primul rnd,
depinde de localizarea surselor de poluare (ape uzate menajere i industriale, ferme zootehnice, terenuri
agricole etc.). Pentru aprecierea condiiilor de referin n staiunea de prelevare, punctele de prelevare
ar trebui stabilite n amontele sursei de poluare. Alte puncte ar trebui stabilite n zona de contaminare:
pentru rurile mici distana de la sursa de poluare trebuie s fie nu mai mic de 50 m, pentru rurile mari
de minimum 200 m [34].
Punctele de prelevare din cadrul programelor de cercetri tiinifice se stabilesc n conformitate cu
obiectivele cercetrii i caracteristicile morfologice ale bazinului acvatic. n general, punctele selectate
trebuie s fie reprezentative pentru zona studiat. Zooplanctonul prezint o distribuie neuniform n
cursurile de ap i chiar mai neuniform n lacuri, datorit migraiei hidrobionilor att pe orizontal, ct
i pe vertical.
n zona litoral probele de zooplancton sunt colectate, de regul, la adncimi nu mai mari de 1 m. Se
recomand prelevarea separat a probelor din zonele care conin vegetaie acvatic (maluri protejate) i
din zonele cu vegetaie redus (maluri expuse).
Probele pelagice sunt prelevate de la adncimi specifice sau din ntreaga coloan de ap, n funcie de
obiectivele studiului. Dac adncimea bazinului acvatic nu se cunoate, aceasta ar trebui estimat prin
utilizarea sonarului portabil sau al unui alt instrument de msurare a adncimii. n general, n bazinele
cu ap stratificat termic probele pelagice ale zooplanctonului se preleveaz la adncimea termoclinului,
deoarece majoritatea planctonului se regsete n epilimnion. Pentru studierea distribuiei verticale a
zooplanctonului n straturile de epilimnion i metalimnion, probele sunt colectate la intervale de adncime nu mai mari de 2 m. n hipolimnion intervalele de adncime sunt mai mari. Pentru a obine rezultate
fiabile, numrul minim de puncte de prelevare n fiecare zon a bazinului acvatic ar trebui s fie de cel
puin trei, inclusiv zona litoral cu macrofite [35].
Frecvena i perioada prelevrii
Periodicitatea anual a prelevrii depinde de tipul monitorizrii. Pentru evaluarea strii ecosistemelor
acvatice n cadrul programelor naionale ale monitoringului de supraveghere, este stabilit o frecven
46

de prelevare a probelor de plancton (bacterioplancton, fitoplancton i zooplancton) de 3-4 ori pe durata perioadei de vegetaie (aprilie septembrie) [36]. Perioada colectrii pe parcursul a 24 de ore este
determinat de migraia diurn pe vertical a celor mai multe specii de zooplancton. n mod normal,
zooplanctonul ar trebui prelevat ntre ora 10 i ora 16.
Investigaiile tiinifice, care au ca scop studierea proceselor funcionrii ecosistemelor acvatice (ofer
informaii privind componena speciilor, efectivul numeric al hidrobionilor i productivitatea individual), ar trebui s includ cel puin probe lunare pe durata perioadei de vegetaie. Un aspect important
suplimentar este i numrul de probe, necesar pentru a caracteriza zooplanctonul ntr-un aa mod, care
ar ine cont de variaia sa pe vertical, orizontal i n timp.
Procedurile generale de colectare a zooplanctonului n apele stttoare, n scopul evalurii calitii
apei, sunt descrise in ghidul european EN 15110:2006 [37]. Procedurile respective sunt recomandate
pentru prelevarea zooplanctonului, care populeaz zona pelagic i cea de litoral a lacurilor naturale,
lacurilor artificiale i heleteielor. Ele nu se potrivesc pentru colectarea speciilor, care triesc, mai cu
seam, pe suprafaa sedimentelor sau pe suprafaa plantelor acvatice.
Tehnicii de prelevare
n funcie de mrimea organismelor planctonice, se utilizeaz dou tipuri de tehnicii de prelevare:
filtrare i sedimentare. Principiul filtrrii (direct sau indirect) este utilizat n diverse tipuri de echipamente: plase (filee) planctonice, capcane planctonice, prelevatoare tubulare, prelevatoare cu pomp. Fiecare
dintre aceste dispozitive au avantaje i dezavantaje i alegerea depinde de particularitile bazinului acvatic (adncime, efectivul numeric al algelor, etc.) i zooplanctonul propriu-zis.
Hidrobionii mezo- i microplanctonului pot fi colectai prin filtrarea apei cu ajutorul diferitor modele de plase planctonice (Fig. 21). Plasa planctonic este un dispozitiv conic, confecionat din nailon fin,
care se trage prin ap vertical sau orizontal pe o anumit distan. Animalele sunt capturate de pereii
de nailon ai plasei i colectai n recipientul de jos. Exist variante ale plaselor planctonice: cu sistem
nenchis i sistem nchis. Cel de-al doilea este utilizat pentru colectarea unui strat specific din coloana
de ap. Avantajul de baz al utilizrii plaselor const n posibilitatea de a filtra rapid volume mari de ap.

a
b
c
d
Fig. 21: a colectarea zooplanctonului cu ajutorul plasei planctonice, b plas planctonic tip Wisconsin [38],
c plas planctonic de tip nchis [39], d plas dubl de tip Bongo combinat ca un prelevator [40]

Plasele sunt folosite cel mai frecvent, dei ele prezint limite considerabile n obinerea datelor cantitative
de ncredere, n special, n apele bogate n nutrieni i alge. Pentru prelevarea calitativ se recomand
utilizarea plasei planctonice cu diferite dimensiuni ale ochiurilor: de 45 m pentru rotifere, de 90 m
pentru majoritatea crustaceelor i de 150 m pentru speciile rpitoare de copepode.
Un prelevator tubular este confecionat, de regul, din PVC sau Tygon. Un tub se coboar n ap i
atunci cnd partea lui de jos ajunge la adncimea dorit, se trage de un fir pentru a nchide partea de jos
a tubului cu un dop de cauciuc sau alt dispozitiv. Tubul se ridic n barc i apa colectat se toarn printr-o plas, pentru a colecta zooplanctonul. Acest dispozitiv ofer un grad sporit de certitudine privind
47

volumul de ap prelevat, dar nu poate fi o modalitate eficient de colectare

a animalelor de talie mare, care au un efectiv numeric mic, sau a animalelor
care pot detecta i evita un dispozitiv de prelevare ngust.
Cea mai frecvent utilizat capcan planctonic este capcana
Schindler-Patalas (Fig. 22), care poart numele celor doi inventatori ai si.
Aceasta reprezint o cutie de plastic incolor, care se coboar n ap pn
la adncimea necesar, apoi se nchide repede (partea de sus i cea de jos)
prin tragerea ei n sus n ap. Zooplanctonul este captat n interiorul cutiFig. 22. Capcan planctonic [41]
ei. Atunci cnd dispozitivul este ridicat, este permis trecerea apei printr-o
plas mica prins de peretele cutiei i zooplanctonul este colectat ntr-un flacon de prelevare, care este
situat la captul acestei plase. Acest dispozitiv, la fel, ofer un grad nalt de certitudine privind volumul
de facto de ap prelevat.
Prelevatorul cu pomp const dintr-un manon de nailon uor interanjabil, care este montat pe o pomp de aspiraie subacvatic (Fig. 23). Dispozitivul este compact i rezistent, cu o plas protejat de deteriorri externe.
Debitmetrele, care intr n componena prelevatorului, sunt eseniale pentru
estimarea exact a volumului. Dei conceput iniial pentru a fi utilizat la prizele de ap i canalele de scurgere ale centralelor electrice, acest prelevator iar putea gsi o aplicare mai general. ncorporarea unei pompe submersibile
ocolete problemele asociate trecerii planctonului prin furtunul lung. Fiind
colectai n aa mod, mai puin de 3% din numrul de indivizi de zooplancton
au prezentat vtmri sau au pierit timp de 48 de ore din momentul capturrii.
Anexarea unei pompe de absorie la captul unei plase cu odgon rigide este
Fig. 23. Prelevator
propus drept o metod de depire a erorilor poteniale, dac efectivul nucu pomp [42]
meric al organismelor colectate este mai mic de 1-5 ind./m3.
Metoda de sedimentare se aplic la colectarea nanoplanctonului. n acest scop, proba se preleveaz
ntr-un recipient cu volum determinat de la un anumit orizont al bazinului acvatic, se fixeaz i se pune
la pstrat pentru 7 10 zile. Dup acest timp, apa de-asupra sedimentului se decanteaz printr-un sifon
(tub de cauciuc, captul inferior al cruia este nchis cu plas cu mrimea ochiului de 45 50 m). Sedimentul rmas se prelucreaz conform procedurii stabilite [34].
n funcie de obiectivul monitorizrii, modalitatea de prelevare poate fi calitativ sau cantitativ.
Pentru analiza cantitativ pot fi utilizate prelevatoare volumetrice, de exemplu, capcana planctonic
Schindler-Patalas, prelevatoarele cu pomp cu debitmetru ncorporat, prelevatoare tubulare i prelevatoare cu butelie. Fileul planctonic Student (Fig. 17) poate fi, de asemenea, folosit pentru colectarea
cantitativ n modul urmtor: 50-100 litri de ap sunt filtrai prin plas. Materialul colectat n recipientul
de jos este cantitativ transferat n flaconul pentru pstrarea probei.
Conservarea i pstrarea probelor
Cnd echipamentul de prelevare este scos din ap, tot materialul prelevat se transfer n recipientul
pentru prob (container din plastic sau flacoane cu capac). Este important ca recipientele pentru probe
s fie umplute aproape complet. Un volum de prob mai mic dect volumul recipientului poate face ca
anumite specii s rmn prinse pe pereii recipientului i s se usuce n timpul depozitrii.
Reactivul conservant poate fi adugat n prealabil n recipient. n caz contrar, el trebuie adugat imediat dup prelevare. Probele de zooplancton neconservate pot fi depozitate ntr-un loc ntunecos, la o
temperatur de 2 5C i cel mult pentru 48 de ore. Probele pentru analiza cantitativ se vor conserva
imediat. De regul, probele se fixeaz cu soluie de formol, sau alcool etilic, sau soluie Lugol [37].
Soluia Lugol se adaug ntr-un volum de 0,5-1,0 ml la 100 ml de prob. Dac proba conine cantiti
mari de organisme planctonice i alte materiale organice (n special, n cazul colectrii din zona litoral),
atunci soluia Lugol va avea un volum de 3-5 ml la 100 ml de prob. Proba trebuie s aib culoarea coniacului. Probele consevate cu soluie Lugol ntotdeauna se pstreaz ntr-un loc ntunecos, de preferin
48

rcit sub 5C, cu excepia probelor, care vor fi analizate ntr-o sptmn (n acest caz ele pot fi pstrate
n ntuneric la temperatura camerei). Pstrarea la ntuneric este necesar pentru a preveni foto-oxidarea.
Proba trebuie verificat regulat, pentru a stabili, dac este nevoie de a mai aduga reactiv conservant.
Dac pentru conservarea probelor este utilizat alcoolul etilic, atunci el trebuie s aib o concentraie
de cel puin 70%. n cazul cnd probele urmeaz fi utilizate pentru analiza genetic (bazat pe extracia
AND-ului), sau msurarea parametrilor de reproducere, se recomand un etanol cu concentraia de
90%. Concentraia nalt a etanolului ajut la meninerea formei corpului (cladocere) i, de asemenea,
previne eliberarea oulor. Fiind pstrate n mod adecvat, probele i menin proprietile pentru un timp
ndelungat. Probele pentru analiza genetic se pstreaz n frigider sau congelator biologic.
n cazul utilizrii formolului, concentraia lui n proba conservat trebuie s fie de aproximativ 4%.
De exemplu, 20 ml de formol de 20% se adaug la 100 ml de prob. Conservarea n soluie de formol-zaharoz, rcit pn la 6C, prentmpin eliberarea oulor. Formolul conserveaz animalele prin prevenirea descompunerii bacteriene. Pentru un rezultat mai bun, probele conservate trebuie pstrate n frigider
n recipiente etanate (vase mici de plastic Nalgene, Fig. 19). Fiind depozitate corespunztor, probele vor
rmne n stare bun pentru civa ani. De notat c fomolul, sau formalina (soluia aldehidei formice de
37%) este carcinogen i trebuie utilizat cu grij. Echipamentul adecvat de siguran include mnui i
ochelari de protecie din plastic.
Bibliografie

33. http://www.lakeaccess.org/ecology/lakeecologyprim9.html.
34. Guidelines on hydrobiological monitoring of freshwater ecosystems / ed. by Abakumov V.
St. Petersburg: Gidrometeoizdat, 1992 (in Russian). Original title:
/ . . : , 1992. 318 .
35. Zooplankton and its production. Methodical recommendations on collecting and processing of
materials in hydrobiological investigations on freshwater bodies. St. Petersburg: GosNIORH, 1984
(in Russian). Original title: .

. : , 1984.
36. Regulation on monitoring of surface and underground water state. Decision of the Government
of the Republic of Moldova no. 932 on 20.11.2013. Published on 29.11.2013 in the Official Monitor of the Republic of Moldova no. 276-280, art. no.1038 (in Romanian). Original title: Regulamentul privind monitorizarea strii apelor de suprafa i apelor subterane. Hotrrea Guvernului Republicii Moldova nr. 932 din 20.11.2013 n: Monitorul Oficial al Republicii Moldova Nr.
276 - 280, art. Nr. 1038.
37. EN 15110:2006 Water quality. Guidance standard for the sampling of zooplankton from standing
waters. Chiinu: INSM, 2012 (in Romanian). National version: SM SR EN 15110:2012 Calitatea
apei. Ghid pentru prelevarea zooplanctonului din ape stttoare. Chiinu: INSM, 2012. 24 p.
38. http://shop.sciencefirst.com/wildco/575-wisconsin-sampler.
39. http://shop.sciencefirst.com/wildco/466-birge-closing-nets.
40. http://www.kc-denmark.dk/products/plankton-nets/bongo-net-2-x-oe60-cm.aspx.
41. http://shop.sciencefirst.com/wildco/plankton-samplers-and-processing/6326-replacement-net-for-schindler-patalas-nitex-153m.html.
42. http://www.kc-denmark.dk/products/plankton-pumps.

49

VII. COLECTAREA ZOOBENTOSULUI

Nevertebratele bentonice sunt animale care vieuiesc la fundul bazinelor acvatice, n stratul de ap
bental i pe diferite tipuri de substraturi i, n temei, sunt reprezentate de urmtoarele grupuri taxonomice: oligochete, chironomide, molute, crustacee, tricoptere, efemeroptere, plecoptere, etc. n funcie
de dimensiunile organismelor, bentosul se clasific n microbentos (sub 0,5 mm), meiobentos (0,5-2,0
mm) i macrobentos (peste 2 mm).
Colectarea probelor de nevertebrate bentonice se efectueaz n corespundere cu standardele internaionale i naionale EN ISO5667-6:2011, EN ISO5667-14:2011, ISO 7828:1985, ISO 8265:1988, ISO
9391:1993, SM SR EN ISO 8689-1:2011, SM SR
EN ISO 8689-2:2011, SM SR EN ISO5667-3:2011,
SM SR EN ISO5667-4:2007 [3-4, 11, 43-44]. La fel,
se utilizeaz i metodele unanim acceptate n hidrobiologie [45-46].
Nevertebratele bentonice se ntlnesc n diverse
tipuri de bazine acvatice (curgtoare i stttoare, adnci i puin adnci), populeaz att diverse
tipuri de substraturi (pietroase, nisipoase, nmoa
b
loase, macrofite, perifiton), ct i stratul de ap
bental, de aceea, nu exist instrumente universale
Fig. 24. Ben Petersen (a) i Birge-Ekman (b) [47]
de prelevare a probelor de bentos.
n dependen de tipul de substrat i grupul de nevertebrate cercetat, se folosesc urmtoarele instrumente i metode de colectare unanim acceptate:
1. Bene de exemplu, Petersen i Birge- Ekman, cu o suprafa de captare de 0,025 m2 (Fig. 24). La
o staie de pe o suprafa de 0,025 m2 se colecteaz cel puin 2 probe, iar de pe o suprafa de 0,01
m2 respectiv, 4-5 probe. Benele sunt utilizate pentru cercetarea cantitativ (efectivul numeric
(exemplare/m2), biomasa (g/m2)), ct i calitativ (diversitatea taxonomic) a macrozoobentosului.
2. Traluri, drage - de exemplu, draga dreptunghiular cu o suprafa de captare de 8 m2, care este
utilizat de ctre cercettorii Institutului de Zoologie al AM pentru evaluarea cantitativ (mizide)
i calitativ a zoobentosului (Fig. 25, a, g). Nu se recomand utilizarea dragei pe faciesuri pietroase
i pe poriunile de ru cu resturi de plante lemnoase.

e
f
g
h
Fig. 25 Instrumente pentru prelevarea nevertebratelor bentonice: a drag trtoare triunghiular, b - fileu [48],
c ram, d - dispozitiv de tip sond (Air Lift Sampler) [49], e i f diverse tipuri de substraturi artificiale [50],
g - prelevarea nevertebratelor bentonice cu drag trtoare dreptiunghiular, h - prelevarea nevertebratelor bentonice
cu ajutorul ciorpacului limnologic semicircular

50

3. Rame sunt folosite pentru analiza cantitativ i calitativ a molutelor (Fig. 25, c).
4. Filee (plase de nailon 38) sunt utile n desiurile de macrofite submerse. n cazul recoltrii
probei prin metoda cositului, fileul este tras timp de 3 min n sens opus curentului de ap sau pe
o distan de 1 m (Fig. 25, b). Procedura se repet de 3 ori. n acest caz, la utilizarea unui fileu cu
deschiztur ptrat de 2525 cm suprafaa total recoltat este de 0,75 m2.
5. Ciorpacuri (plase de nailon 23) sunt potrivite pentru colectarea probelor de pe substraturi
dure, inclusiv construcii hidrotehnice (Fig. 25, h).
6. Substraturi artificiale (Fig. 25, e, f).
7. Dispozitive de prelevare de tip sond sunt utilizate n rurile mari, cu ape repezi i adncimi de
peste 7 m (Fig. 25, d).
8. Prelevarea manual este practicat pentru diferite tipuri de substraturi pentru analiza calitativ
i cantitativ a zoobentosului.
Dac recoltarea se efectueaz cu ajutorul fileului sau ciorpacului, atunci operatorul trebuie s se mite
lateral n sens opus curentului apei, tulburnd, totodat, cu piciorul substratul i innd instrumentul mai
jos pe cursul apei, comparativ cu poziia corpului su.
La colectarea hidrobionilor din desiurile de macrofite sau acumulrile de alge filamentoase, acele
buci de plante care au nimerit n fileu, nainte de a fi aruncate se cltesc bine (fr a fi scoase din fileu).
n cazul colectrii de pe pietre organismele sunt splate n interiorul fileului cu ajutorul unei perii sau
pinete. Pentru a aprecia suprafea pietrei, se face proiecia pietrei pe hrtie, dup care se cntresc separat
ntreaga proiecie pe hrtie i 1 cm2 de hrtie. Prin mprirea masei ntregii proiecii pietrii la masa unui
centimentru patrat se calculaz suprafaa total a pietrei.
Selectarea locului de colectare a probelor depinde de obiectivele concrete ale cercetrii. Monitorizarea
biologic necesit selectarea substraturilor cu cea mai variat faun bentonic, innd cont de caracterul
mozaic al distribuiei organismelor i afinitatea lor pentru diverse substraturi [45-46, 51-61].
n ruri bentosul este mai bogat faunistic n locurile de alternan a vadurilor i adncurilor [45], n
lacurile naturale i de acumulare n zonele litorale [2] i n cele de mare adncime i, de asemenea, pe
substraturile artificiale [11]. De regul, probele de nevertebrate bentonice se recolteaz la adncimi de
pn la 1,5 m. n ruri probele se colecteaz la mal, n albia rului, n desiurile de macrofite, la hotarul
dintre vaduri i adncuri. Segmentul de ru de-a lungul cruia se colecteaz probele trebuie s aib o lungime de cel puin 50 m n cazul rurilor mici i de cel puin 200 m a rurilor mari. Pentru rurile mari se
recomand recoltarea a cel puin 3-4 subprobe/staie, proporional ariei substraturilor prezente la staia
dat (pietroase, nisipoase, nmoloase, desiuri de macrofite), din care se alctuiete proba integral. De
amintit c colectarea se efectueaz n sens contrar curentului apei.

a
b
c
Fig. 26: Transferarea probei de bentos din ben ntr-un vas cu ap (a) [63], vas pentru transportarea
i pstrarea probelor de nevertebrate bentonice (b), splarea probei de ap cu ajutorul fileului (c)

51

n bazinele acvatice stagnante separat se colecteaz probe din zona litoral, din sectoarele adnci i n
desiurile macrofitelor [2].
n cazul habitatelor srace din punct de vedere faunistic se practic sporirea numrului de probe colectate de pn la 20, n scopul obinerii unui numr maxim de specii [11]. Pentru ca proba s permit
obinerea unui material hidrobiologic statistic veridic, ea trebuie s conin cel puin 700 de exemplare
[52]. O atenie deosebit necesit sectoarele n care locuiesc specii invazive. Speciile rare, relicte, la fel,
solicit o abordare deosebit, fiindc extragerea lor din bazinul acvatic poate avea o influen nefast
asupra populaiilor lor.
Frecvena colectrii probelor, de asemenea, depinde de obiectivele cercetrii i poate fi: anual (pentru macrobentos n septembrie sau octombrie), trimestrial, lunar, decadal. Se va ine cont i de
situaiile excepionale [62].
Probele bentonice, de regul, nu se colecteaz n perioada inundaiilor, secetei, n cazul unei aciuni
umane directe, de exemplu, n timpul desfurrii lucrrilor de adncire a rului n staia supus cercetrii
[52]. n scopul stabilirii dinamicii densitii populaiilor i a modificrilor naturale a structurii lor taxonomice, se recomand colectarea repetat a probelor pe parcursul unui an, timp de ctiva ani consecutivi.
Dup colectare proba de bentos se transfer ntr-un vas cu ap (Fig. 26); prin splare, utiliznd o sit
sau un fileu, se nltur fraciile grosiere de substrat. n scopul transportrii i pstrrii, proba se mut
ntr-un vas din sticl sau plastic cu un volum de 200-1000 ml i se conserveaz. Pentru conservare se
utilizeaz fie alcool etilic de 96%, fie formol de 37%, care se adaug n prob pn la realizarea unei concentraii finale de 70% i, respectiv, 3,7% [15, 51-52]. Dac proba nu se conserveaz, atunci ea urmeaz a
fi pstrat la o temperatur de 1-5 i prelucrat n primele 24 de ore dup prelevare [15].
Fiecare prob de bentos este etichetat, indicndu-se numrul probei, denumirea bazinului acvatic,
punctul i data colectrii, adncimea, tipul de substrat, numrul de replicri (subprobe).
Bibliografie
43. SM SR EN ISO 8689-1:2011 Water quality. Biological classification of rivers. Part 1: Guidance
on the interpretation of biological quality data from surveys of benthic macroinvertebrates (in
Romanian). National version:: SM SR EN ISO 8689-1:2011 Calitatea apei. Clasificarea biologica
a rurilor. Partea 1: Ghid pentru interpretarea datelor biologice de calitate obinute din studierea
macronevertebratelor bentonice.
44. SM SR EN ISO 8689-2:2011 Water quality. Biological classification of rivers. Part 2: Guidance
on the presentation of biological quality data from surveys of benthic macroinvertebrates (in
Romanian). National version: SM SR EN ISO 8689-2:2011 Calitatea apei. Clasificarea biologica
a rurilor. Partea 2: Ghid pentru prezentarea datelor biologice de calitate obinute din studierea
macronevertebratelor bentonice.
45. Guidelines on the methods of hydrobiological analysis of surface watersand bottom sediments
/ Abakumov V.A. (editor). Leningrad: Gidrometeoizdat, 1983. 240 p. (in Russian). Original title:
/. .. .: , 1983. 240 .
46. Jadin, V.I. Methods of hydrobiological investigations. Moscow: Vysshaya shkola, 1960. 190 p.
(in Russian). Original title: . .: ,
1960. 190 . . .: , 1960. 190 .
47. http://www.anhydre.eu/wildlife.html.
48. http://www.nhbs.com/browse/subject/516/aquatic-samplingnets.
49. http://www.uwitec.at/html/river_benthos.html
50. http://www.iad.gs/index.php?item=zooplankton_zoobenthos&PHPSESSID=bfd9bce62c0a64891c9902f73d9f61b9
51. Unified methods of water quality investigation. Part III: Methods of biological analysis of waters.
M.: SEV, 1983. 227 p. (in Russian). Original title:
. III: . .: , 1983. 227 .
52

52. Manual for the application of the AQEM system. A comprehensive method to assess European
streams using benthic macroinvertebrates, developed for the purpose of the Water Framework
Directive. Version 1.0,February 2002.
53. Jadin, V.I. Molluscs of fresh and brackish waters of the USSR. .-L. AN SSSR, 1952. 376 p. (in
Russian). Original title: .. . .; .:
- , 1952. 376 .
54. Unified methods of water quality investigation. Part III: Methods of biological analysis of waters.
M.: SEV, 1983. 227 p. (in Russian). Original title:
. III: . .: , 1983. 227 .
55. Identification guide into the freshwater invertebrates from the European parts of the USSR
(plankton and benthos) / Kutikova L.A., Starobogatov E.I. (editors). Leningrad: Gidrometeoizdat, 1977. 510 p. (in Russian). Original title:
( ) / . .., ..
: , 1977. 510 .
56. Identification guide into the freshwater invertebrates of Russia and adjacent countries, Vol. 1.
Lower invertebrates / Tsalolikhin S.J. (ed.). Saint-Petersburg: Nauca, 1994. 394 p. (in Russian).
Original title:
/ . .. 1. . .: , 1994. 394 c.
57. Identification guide into the freshwater invertebrates of Russia and adjacent countries, Vol. 2.
Crustaceans / Tsalolikhin S.J. (ed.). Saint-Petersburg: Nauca, 1995. 627 p. (in Russian). Original
title: /. .. 2. . .: , 1995. 627 c.
58. Identification guide into the freshwater invertebrates of Russia and adjacent countries, Vol. 3.
Arachnids / Tsalolikhin S.J. (ed.). Saint-Petersburg: Nauca, 1997. 439 p. (in Russian). Original
title: /. .. 3. . .: , 1997. 439 c.
59. Identification guide into the freshwater invertebrates of Russia and adjacent countries, Vol. 4.
Diptera / Tsalolikhin S.J. (ed.). Saint-Petersburg: Nauca, 2000. 997 p. (in Russian). Original title:
/. .. 4. . .: , 2000. 997 c.
60. Identification guide into the freshwater invertebrates of Russia and adjacent countries, Vol. 5.
Higher insects / Tsalolikhin S.J. (ed.). Saint-Petersburg: Nauca, 2001. 836 p. (in Russian). Original
title: / .
.. 5. . .: , 2001. 836 c.
61. Identification guide into the freshwater invertebrates of Russia and adjacent countries, Vol. 6.
Shellfish. Polychaete. Nemerteans. / Tsalolikhin S.J. (ed.). Saint-Petersburg: Nauca, 2004. 528 p.
(in Russian). Original title: / . .. 6: . . . .: ,2004. 528 c.
62. ISO 5667-14:1998 Water quality. Sampling. Part 14: Guidance on quality assurance of environmental water sampling and handling. National version: SMV ISO 5667-14:2011 Calitatea apei.
Prelevare. Partea 14: Ghid pentru asigurarea calitii la prelevarea i manipularea probelor de ap
natural (in English). Chiinu: INSM, 2011. 18 p.
63. http://shop.sciencefirst.com/wildco/551-standard-ekman-grab

53

VIII. COLECTAREA MATERIALULUI IHTIOLOGIC

Astazi n Europa sunt utilizate n principal urmtoarele metode de colectare:
electronarcoza reversibil, util n special n rurile mici, mijlocii i unele lacuri (n Republica
Moldova aceast metod este interzis prin lege);
uneltele clasice din plas, folosite mai ales n lacuri (nvod, voloc, setci instalate staionar sau lsate
n deriv, etc.);
iruri de crlige cu momeala (pripoane) folosite n completarea altor mijloace de studiu.
Pentru a fi siguri c datele despre abunden i structura pe vrste sunt valide, trebuie recoltat un
numr suficient de probe. Staiile supuse pescuitului tiinific trebuie s fie ct se poate de reprezentative
ca s reflecte starea ntregului ecosistem. n lacurile cu adncimi mari se va ine cont de existena stratificrii verticale (hipo-, meta- i epilimnion), astfel, vor fi utilizatesetci de suprafa, setci pentru masa
apei i setci de fund[64, 68, 70, 74-75].
Numrul de colectri trebuie s fie suficient de mare ca s asigure statistic rezultatele obinute - marja
de eroare nu trebuie s depeasc 20%. Lungimea total a setcilor folosite depinde de abundena diverselor specii i grupe dimensionale de captur. Durata de colectare va egal cu cel puin 12 ore sau 24 ore.
Randamentul de colectare se apreciaz prin determinarea efortului de pescuit (CPUE), care exprim
numrul exemplarelor sau biomasa petilor din captur (total sau pe specii) colectate la o suprafa de
plas (ex. 100 m2) n 12 ore sau 24 ore.
n cazul pescuitului prin electronarcoz determinarea stocului numeric (nr. ex./100 m2 sau nr. ex./
ha) i a stocului gravimetric (g/100 m2 sau kg/ha) este relativ simpl, ntruct se poate determina uor
suprafaa total colectat prin pescuit electric i apoi se poate exprima cantitatea colectat sau numrul
de exemplare la uniti convenionale de suprafa (100 m2, 1 ha, 100 m liniari de ru etc.).
Examinarea petilor se efectueaz uzual, prin numrarea, msurarea i cntrirea lor. Identificarea speciilor se realizeaz pe baza caracterelor morfologice, folosind chei de determinare pentru fiecare unitate
sistematic [67, 69, 71-72]. Pentru determinarea vrstei petilor se preleveaz solzi sau alte esuturi dure
(opercule, otolite, seciuni de radii osoase). La fel, se determinsexul i gradul de evoluie a gonadelor.
Pentru determinarea sexului petilor se studiaz gonadele. La stadiile mature de dezvoltare determinarea sexului este uor de realizat: la femelele adulte ovulele se disting uor n ovare, iar la masculi
testiculele sunt netede, albicioase, n aparen negranulate. Pentru a uura stabilirea diferitelor stadii ale
ciclului sexual, au fost ntocmite diferite scale ale gradului de maturare a gonadelor. La baza acestora st
evidenierea maturaiei gonadelor n diferite stadii ontogenetice [73].
n scopul evidenierii ritmului de cretere a speciilor de peti, se va efectua o serie de msurtori biometrice[70, 73-75].
Caracterele determinate prin studii de biometrie sunt:
caractere metrice: lungimea absolut sau standard a corpului, lungimea capului, grosimea, nlimea maxim i minim a corpului, distanele ntre nottoare, diametrul ochiului, .a.;
caractere gravimetrice: greutatea corpului sau a unor organe care se stabilete prin cntrire;
caractere meristice: numrul de solzi pe linia lateral, numrul rndurilor transversale de solzi n
regiunea cea mai nalt a corpului, numrul de radii, spini branhiali, formula dinilor faringieni, .a.
Toate aceste date pot fi supuse ulterior prelucrrilor statistice n vederea calculrii valorilor medii,
limitelor de variaie, existenei sau inexistenei unor corelaii, .a.
Pentru a descifra relaiile stabilite ntre diferite specii n cadrul ecosistemului, ierarhiile ce se stabilesc
n cadrul ihtiocenozelor, n ajutor ne vin un ansamblu de metode matematice cunoscute sub denumirea
generic de analiz sinecologic. Acest tip de analiz ne permite s identificm cu precizie speciile care
au ponderea cea mai mare n ecosistem sub aspectul schimbului energetic cu mediul, care sunt speciile
caracteristice unui biotop, sau speciile care au ajuns ntmpltor n zona cercetat; de asemenea, se pot
stabili cu destul precizie interelaiile dintre speciile ce alctuiesc biocenoza.
n funcie de modul n care acetia se calculeaz, avem de-a face cu dou categorii distincte: 1. indici
ecologici analitici (se opereaz cu datele brute colectate pe teren) i 2. indici ecologici sintetici (se opereaz
cu indicii analitici i se folosec pentru evidenierea interelaiilor dintre specii, comuniti sau cenoze).
54

Cel mai des sunt utilizai urmtorii indici ecologici: dominana (D), constana (C), indicele de semnificaie ecologic (W), indicele de similaritate specific (S), indicele de diversitate Shannon-Wiener, echitabilitatea (e), indicele de dominare Simpson (Is), .a. [64-66].
La evaluarea structurii de vrst a populaiilor este suficient de a supune msurtorilor ihtiologice un
eantion de minim 30 indivizi/specie, ns precizia de reprezentare a datelor crete cu majorarea numrului de msurri [64].
Determinarea stocului numeric i gravimetric ofer informaii corecte i comparabile asupra numrului i biomasei fiecrei populaii dintr-o specie i din ntreaga ihtiocenoz, n punctele de colectare. De
asemenea, aceti indici au o mare valoare i pentru c ofer informaii de maxim importan n cazul
reconstruciei ecologice.

Cu excepia specimenilor pentru colecii muzeale i a celor pstrate pentru identificare i prelucrare
n laborator (caractere confuze, analiz genetico-molecular, histologic, identificarea spectrului trofic,
.a.), toi petii vor fi eliberai imediat ce a fost ncheiat procesarea. Procesarea i eliberarea se va face
dup fiecare ton, lungime parcurs.
Detaliile despre particularitile fizice ale hidrobiotopului, echipa de pescuit, efortul de pescuit, obiective, echipament, captur, etc. vor fi nregistrate ntr-o fi tip. Aceasta va fi ntocmit i distribuit de
organizatorul pescuitului, avnd un aspect i coninut similar unic.
Bibliografie
64. Davideanu G. (2013) Methodological guide for monitoring the ichthyocenoses structure. Ed.
Performantica, Iasi, 57 p.
65. Dediu I. (2007) Treaty of theoretical ecology: monographic study of synthesis. Chisinau: Ed. Academia Nationala de Stiinte Ecologice, 557 p. (in Romanian). Original title: Dediu I. (2007) Tratat
de ecologie teoretic: Studiu monografic de sintez. Chiinu: Ed. Academia Naional de tiine
Ecologice, 557 p.
66. Gomoiu M.T., Skolka M. (2001). Ecology. Methodologies for ecological studies. Constanta: Ed.
Ovidius University Press, 173 p. (in Romanian). Original title: Gomoiu M.T., Skolka M. (2001)
Ecologie. Metodologii pentru studii ecologice. Ed. Ovidius University Press, Constana, 173 p.
67. Koblitskaia A.F. (1981) Identification guide of freshwater fish fingerlings. Moscow: Legkaia i piscevaia promishlennosti, 209 p. (in Russian). Original title: .. (1981)
. : . , 209 c.
68. Kotlear O.A. (2004) Methods of fish farming researches. Tutorial. Ribnoe, 180 p. (in Russian).
Original title: O.A. . .
, 2004, 180 .
69. Kottelat M., Freyhof J. (2007) Handbook of European Freshwater Fishes, Ed. Delemont, Switzerland, 646 p.
55

70. Navodaru I. et al. (2008) Assessment of fish stocks and fisheries. Methods of assessment and prognosis of fishery resources. Ed. Dobrogea, 298 p. (in Romanian). Original title: Nvodaru I. .a.
(2008) Estimarea stocurilor de peti i pescriilor. Metode de evaluare i prognoz a resurselor
pescreti. Editura Dobrogea, 295 p.
71. Otsel V. (2007) Atlas of fish from the Danube Delta Biosphere Reserve. Tulcea: Ed. Centrul de
informare tehnologica Delta Dunarii, 481 p. (in Romanian). Original title: Oel V. (2007) Atlasul
petilor din Rezervaia Biosferei Delta Dunrii. Tulcea: Ed. Centrul de informare tehnologic
Delta Dunrii, 481 p.
72. Popa L. L. (1977) Fish of Moldova. Reference book identification guide. Chisinau: Cartea Moldoveneasca, 200 p. (in Russian). Original title: . . (1977) .
. : , 200 c.
73. Pravdin I.F. (1966) Guidelines on fish study. Moscow: Piscevaia promishlennosti, 376 p. (in Russian). Original title: .. (1966) . :
, 1966, 376 .
74. Pricope F., Stoica I., Battes K. (2013) Secondary production of aquatic ecosystems. Bacau: Ed.
Alma Mater, 151 p. (in Romanian). Original title: Pricope F., Stoica I., Battes K. (2013) Producia
secundar a ecosistemelor acvatice. Bacu: Ed. Alma Mater, 151 p.
75. Shibaev S.V. (2007) Industrial ichthyology. Saint Petersburg: Prospect nauki, 2007, 399 p. (in
Russian). Original title: . . (2007) . -:
, 399 .

IX. SURSE DE ERORI N PRELEVARE

Surse de erori asociate prelevrii
Erorile asociate prelevrii pot fi generate de diferii factori, cei mai importani fiind [62]:
Contaminarea. Contaminarea poate fi cauzat de materialele din care este fabricat echipamentul
de prelevare, condiiile de transportare sau pstrare necorespunztoare.
Instabilitatea probei. Tipul vaselor de colectare sau recipientelor poate afecta stabilitatea analiilor
n perioada dintre prelevare i analiz, datorit instabilitii inerente a nsi probei i a condiiilor
n care probele sunt pstrate i transportate.
Conservarea incorect. Alegerea opiunilor privind conservarea probelor, conform prevederilor
ISO 5667-3 [15], poate influena negativ concentraia diferitor componente ale apei.
Prelevarea incorect. Abaterea de la procedura de
prelevare sau prelevarea
din bazine acvatice neomogene poate constitui o
surs de eroare.
Tehnici de control al calitii
n colectarea probelor msurile de control al calitii au trei
obiective de baz: 1) a oferi o
cale de monitorizare i depistare a erorilor asociate prelevrii,
n scopul eliminrii datelor incorecte sau datelor care duc n
eroare; 2) a demonstra c erorile care nsoesc prelevarea sunt

Fig. 27. Controlul calitii privind precizia colectrii probelor

56

controlate n mod corespunztor;

3) a indica variabilitatea prelevrii i, prin urmare, a controla
acest aspect important al erorii.
Tehnicile principale de control al calitii sunt:
a) Colectarea replicatelor drept
verificare a preciziei prelevrii.
Aceast tehnic poate fi utilizat
pentru a explora un ir de abordri privind controlul calitii, n
scopul aprecierii erorii aleatorii,
care nsoete procesul de colectare. Relaia dintre diferite variaii ale prelevrii este prezentat
n Fig.27. Diferena dintre A1 i
Fig. 28. Controlul calitii echipamentului de prelevare
A2 permite estimarea variaiei
cu ajutorul apei deionizate martor
per ntregul proces de colectare
(prelevare, recipient, pstrare i analiz). Diferena dintre B1 i B2 permite aprecierea variaiei la etapa
analitic plus variaia n timpul colectrii (incluznd pstrarea, dar excluznd recipientul pentru prob).
b) Cltirea echipamentului. Aceast tehnic poate fi folosit pentru a identifica eventualele erori legate
de contaminarea dispozitivelor de prelevare a probelor i a procesului de prelevare, cauzate de curarea incomplet a vaselor de prelevare. n condiii de laborator se mparte o prob de ap deionizat n
dou pri: partea A este pstrat n laborator, iar partea B este transportat n cmp i mprit n dou
poriuni B1 i B2. Poriunea B1 se returneaz n laborator fr careva procesare n condiii de cmp.
Poriunea B2 se analizeaz ca i o
prob real (Fig. 28). Diferena
dintre rezultatele analizei prii
A i poriunii B1 scot n eviden
erorile aprute n timpul transportrii probei. Diferena dintre
rezultatele analizei prii A i
poriunii B2 identific erorile determinate de prelevare, procesare
i transportare.
c) Recuperarea filtrrii. Aceast tehnic poate fi folosit pentru
a identifica orice eroare legat de
contaminarea instrumentelor de
prelevare i a procesului de prelevare, care sunt nsoite de filtrare.
Fig. 29. Controlul calitii echipamentului de prelevare
n laborator o prob de ap deioi filtrare cu ap deionizat martor
nizat este separat n dou pri:
partea A, care este lsat n laborator, i partea B, care ulterior este transportat n cmp i divizat n
poriunile B1 i B2. Poriunea B1 se returneaz n laborator fr careva procesare n condiii de cmp.
Poriunea B2 este procesat cu ajutorul instrumentelor de colectare i a echipamentului de filtrare ca i o
prob real (Fig. 29). Diferena dintre rezultatele analizei prii A i poriunii B2 scot n eviden erorile,
care au loc pe durata prelevrii, filtrrii, procesrii i transportrii. Compararea rezultatelor poriunilor
B1 i B2 identific erorile, care se datoreaz contaminrii instrumentelor de prelevare i a procesului de
prelevare, nsoite de filtrare.
57

d) Probe sintetice (obinute prin metoda adaosului unei cantiti cunoscute de analit). Pentru aprecierea
erorii sistematice a proceselor de prelevare, care se datoreaz instabilitii probei (volatilizarea compuilor
organici, adsorbia microelementelor sau consumul nutrienilor de ctre bacterio- i fitoplancton), pot fi
utilizate probele sintetice printr-o tehnic similar.
Analiza i interpretarea datelor de control al calitii
Cea mai rspndit form de interpretare a datelor de control al calitii este fia de control Shewhart
[76]. n acest caz se utlizeaz o fi, pe care este reprezentat consecutiv diferena dintre determinrile
duplicate (d = R1 R2)1.Valorile msurate sunt comparate cu valorile de limit (limitele sus/jos de avertizare i limitele sus/jos de aciune) i rezultatele sunt analizate prin analiza vizual a fiei de control
(Fig. 30).

Fig. 30. Fia de control Shewhart pentru probele martor duplicate

n general, procedurile de asigurare a calitii/control al calitii (AC/CC) sunt necesare pentru asigurarea calitii activitilor de prelevare n cadrul programelor de monitorizare, inclusiv conservarea
integritii probei. Asigurarea calitii prelevrii, care include selectarea probei, tratarea prealabil, colectarea subprobelor, conservarea, pstrarea i transportarea este determinant pentru calitatea rezultatelor finale ale analizelor chimice. Efortul depus pentru controlul calitii prelevrii depinde de obiectivele programei, dar se recomand ca cel puin 2% din efortul analitic s fie dedicat controlului calitii
recoltrii probelor.
Biblografie
76. ISO 8258:1991 Shewhart control charts. This standard has been revised by: ISO 7870-2:2013
Control charts. Part 2: Shewhart control charts. National version: SM SR ISO 8258+C1:2012 Fie
de control Shewhart (in Romanian). Chiinu: INSM, 2012. 44 p.

R1 este rezultatul primei probe analizate i R2 este rezultatul celei de-a doua probe analizate.
58

ANEXA 1. Frecvenele prelevrii de probe pentru parametrii fizico-chimici i hidrobiologici [36]

Elemente
calitative

Ruri
(o dat n intervalul menionat)

Lacuri / lacuri de acumulare

(o dat n intervalul menionat)

Parametrii fizico-chimici ai calitii

Starea regimului termic

1 lun

1 lun

Starea regimului gazos

1 lun

1 lun

pH-ul i potenialul redox

1 lun

1 lun

Conductivitatea

1 lun

1 lun

Ionii principali i mineralizarea

Turbiditatea i cantitatea
suspensiilor
Culoarea i mirosul
Starea de eutrofizare:
substanele nutritive

1 lun

1 lun

1 lun

1 lun

1 lun

1 lun

1 lun

1 lun

1 lun

1 lun

1 lun

1 lun

Consumul chimic i biochimic

de oxigen (CCOCr and CBO5)
Starea acidificrii: alcalinitatea
Substane organice persistente
prioritare
Metale grele

1 lun

1 lun

1 lun

1 lun

Ali poluani

1 lun

1 lun

Parametrii biologici ai calitii

Bacterioplancton

De 3 ori pe an n perioada de
vegetaie (aprilie septembrie)

De 3 ori pe an n perioada de vegetaie

(aprilie septembrie)

Zooplancton

De 3 ori pe an n perioada de
vegetaie (aprilie septembrie)

De 3 ori pe an n perioada de vegetaie

(aprilie septembrie)

Fitoplancton

De 3 ori pe an n perioada de
vegetaie (aprilie septembrie)

De 3 ori pe an n perioada de vegetaie

(aprilie septembrie)

Clorofila A

De 3 ori pe an n perioada de
vegetaie (aprilie septembrie)

De 3 ori pe an n perioada de vegetaie

(aprilie septembrie)

Nevertebrate bentonice

De 2 ori pe an n perioada de
vegetaie (aprilie septembrie)

De 2 ori pe an n perioada de vegetaie

(aprilie septembrie)

Macrofite

De 2 ori pe an n perioada de
vegetaie (aprilie septembrie)

De 2 ori pe an n perioada de vegetaie

(aprilie septembrie)

De 2 ori pe an
(primvara i toamna)

De 2 ori pe an
(primvara i toamna)

Peti

59

ANEXA 2. Metodele recomandate de conservare i durata de pstrare maximum admisibil [15]

Elemente
calitative

Material recipientului
recomandat2

Tehnici
de conservare

Durata de pstrare
nainte de analiz

Aciditate i alcalinitate

P sau S

Rcire la 1 5C

24 h

Aluminiu

P sau SB

Acidulare la pH 1 2
cu HNO3

1 lun

Rcire la 1 5C
Acidulare la pH 1 2
cu H2SO4

24 h
21 zile

Amoniac liber i ionizat

Arsen

P sau S
splat cu acid

Filtrare nainte de conservare

Acidulare la pH 1 2
1 lun
cu HNO3

Consum biochimic de
oxigen (CBO)

P sau S

Rcire la 1 5C
24 h
Probele se pstreaz n absena luminii

Bromuri

P sau S

Rcire la 1 5C

Calciu

P sau S

Carbon:
dioxid de carbon

P sau S

1 lun

Rcire la 1 5C
7 zile
Acidulare la pH 1 2
1 lun
cu HNO3
Rcire la 1 5C
24 h
Analiza se realizeaz de preferin n situ

Carbon:
carbon organic total
(COT)

P sau S

Rcire la 1 5C
24 h
Acidulare la pH 1 2
7 zile
cu H2SO4
Dac se suspecteaz prezena compuilor organici
volatili nu se utilizeaz acidulare, analiza trebuie
efectuat n 8 h

Consum chimic de
oxigen (CCO)

P sau S

Acidulare la pH 1 2 cu
H2SO4, rcire la 1 5C

1 lun

1 lun

Rcire la 1 5C

24 h

Acidulare la pH 1 2
cu HNO3

1 6 luni

Rcire la 1 5C

24 h

Cloruri
Clorofil
Crom total
Crom (VI)
Culoare apei
Compui ai metalelor
(Cu, Pb, Zn, Ni, Cd, Co,
V, Mn, Se)
Conductivitate
2

P sau S

P sau S
P sau S
(culoare brun)
P sau S
splat cu acid
P sau S
splat cu acid

Rcire la 1 5C
5 zile
Probele se pstreaz n absena luminii

P sau S
P sau SB
splat cu acid

Acidulare la pH 1 2
cu HNO3

1 6 luni

Rcire la 1 5C
24 h
Analiza se realizeaz de preferin n situ

P sau SB

P polietilen (PTFE politetrafluoretilen, PVC policlorur de vinil, PET polietilen tereftalat),

S sticl, SB sticl borosilicat

60

Rcire la 1 5C
24 h
Alcalinizarea la
7 zile
pH > 12 cu NaOH
Probele se pstreaz n absena luminii

Cianuri:
cianuri totale

Cianocloruri

Rcire la 1 5C

24 h

Detergeni: ageni de
suprafaa anionici

S splat cu alcool
metilic

Acidulare la pH 1 2 cu
H2SO4, rcire la 1 5C

2 zile

Detergeni: ageni de
suprafaa cationici

S splat cu alcool
metilic

Rcire la 1 5C

2 zile

P dar nu PTFE

Rcire la 1 5C

7 zile

Fluoruri
Hidrocarburi

S splat cu solvent
Acidulare la pH 1 2 cu
utilizat pentru extracie
H2SO4 sau HCl

Fier (II)

P sau SB
splat cu acid

Fier total

P sau SB
splat cu acid

Magneziu

P
SB
splat cu acid

Mercur

SB
splat cu acid

1 lun (dac este

posibil, extracia se
realizeaz n situ)

Acidulare la pH 1 2 cu 7 zile (izolare de oxigenul

HCl
din aer)
Acidulare la pH 1 2
cu HNO3
Rcire la 1 5C
Acidulare la pH 1 2
cu HNO3
Acidulare la pH 1 2
cu HNO3 i adugarea
K2Cr2O7 la concentraia
final de 0,05 %
Rcire la 1 5C
Acidulare la pH 1 2
cu HCl

1 lun
7 zile
1 lun

1 lun

Azot:
azot de nitrai (azotat)

P sau S

Azot:
azot de nitrii (azotit)

P sau S

Rcire la 1 5C

24 h

P sau S

Rcire la 1 5C
Acidulare la pH 1 2 cu
H2SO4

24 h
1 lun

Uleiuri i grsimi

S splat cu solvent

Acidulare la pH 1 2
cu H2SO4 sau HCl

1 lun

Oxigen (Winkler)

P sau S

Azot:
azot total (Kjeldahl)

Pesticide carbamice

24 h
7 zile

Fixare n situ
4 zile
Probele se pstreaz n absena luminii
Rcire la 1 5C
14 zile
Congelare la 20C
1 lun
Dac proba este clorinat, se adaug 80 mg
Na2S2O35H2O pentru fiecare 1000 ml prob
n recipient nainte de analiz

S
P
splat cu solvent

61

Pesticide:
organoclorurate
organofosforice
organoazotate

Fenoli

pH

S cu capac de PTFE
splat cu solvent

SB brun
cu capac de PTFE
splat cu solvent

Extracie (n 24 h de la
5 zile
prelevare)
(durata de pstrare
Rcire la 1 5C
extractului)
Dac proba este clorinat, se adaug 80 mg
Na2S2O35H2O pentru fiecare 1000 ml prob n
recipient nainte de analiz
Acidulare la pH < 4 cu
H3PO4 sau H2SO4

3 sptmni

Dac proba este clorinat, se adaug 80 mg

Na2S2O35H2O pentru fiecare 1000 ml prob n
recipient nainte de analiz
Rcire la 1 5C
6h

P sau S: complet
umple recipientul Msurarea se realizeaz de preferin n situ imediat
pentru a elimina aerul
dup prelevare
S sau SB

Filtrare n situ
(filtre de 0,45 m)
Rcire la 1 5C

24 h

S sau SB

Filtrare n situ
(filtre de 0,45 m)
Acidulare la pH 1 2
cu H2SO4

1 lun

Potasiu

Rcire la 1 5C
Acidulare la pH 1 2
cu HNO3

7 zile
1 lun

Silicai dizolvai

Filtrare n situ
Rcire la 1 5C

7 zile

Silicai totali

Rcire la 1 5C

1 lun

Sodiu

Rcire la 1 5C

7 zile

Rcire la 1 5C

7 zile

Fosfor dizolvat

Fosfor total

Sulfuri

Sulfai

P sau S

Rcire la 1 5C

1 lun

Materii n suspensie

P sau S

Rcire la 1 5C

2 zile

Turbiditate

P sau S

Rcire la 1 5C

24 h

Probele se fixeaz imediat dup prelevarea

cu soluie de acetat de zinc 10%

Probele se pstreaz n absena luminii

62

ANEXA 3. Proceduri recomandate de splare pentru elemente selectate ale calitii apei [30]
Element de calitate

Procedur de splare

Culoare, turbiditate, pH, aciditate, alcaliniSe cltete de 3 ori cu ap de robinet 1 dat cu acid
tate, calciu, potasiu, sodiu, magneziu, carcromic2 3 ori cu ap de robinet 1 dat cu HNO3
bon organic total, azot (de amoniu, nitrit,
(1:1) 3 ori cu ap distilat
nitrat, total), fosfor, cloruri, fluoruri, sulfai
aluminiu, cadmiu, cobalt, cupru, fier, zinc, Se cltete de 3 ori cu ap de robinet 1 dat cu acid
nichel, plumb, molibden, mangan, hidrar- cromic 3 ori cu ap de robinet 1 dat cu HNO3 (1:1)
giu, selen, stroniu, vanad
3 ori cu ap distilat ultrapur3
Se cltete de 3 ori cu ap de robinet 1 dat cu acid
cromic 3 ori cu ap care nu conine compui organici
Pesticide fosforoorganice, cloroorganice, 1 dat cu aceton cu grad special de puritate4 de 2 ori
policlordifenili, pentaclorofenoli, rini fe- cu hexan cu un grad nalt de puritate privind coninutul
reziduurilor de pesticide se usuc (fr capac) ntr-un
nolice, erbicide din clasa fenoxi
cuptor cu aer fierbinte la 360C pentru minim 1 or

Fig. 31. a instalaie de distilare din sticl [78], b Millipore Super Q Water System [79]

Bibliografie
77. GOST 4517-87 Interstate standard. Reagents. Methods for preparation of accessory reagents and
solutions used for analysis (in Russian) Original title: 4517-87
. . ,
. M.,1988
78. http://www.indiamart.com/avm-scientific/scientific-laboratory-equipments.html
79. http://www.medicalexpo.com/prod/merck-millipore/product-70876-658188.htm
Acid cromic: soluia bicromatului de potasiu cu o concentraie procentual de mas de 5% n acid sulfuric [77].
Apa distilat ultrapur se obine prin trecerea apei distilate printr-o instalaie de distilare din sticl (Fig. 31 a) i apoi printun sistem Millipore Super Q Water System (Fig. 31 b), care conine un cartu prefiltrator, un cartu cu crbune activ i
un cartu cu pat mixt de schimb ionic.
5
Aceton cu grad special de puritate: grad de puritate privind coninutul reziduurilor de pesticide n cazul utilizrii
cromatografiei de gaze i grad de absorbie a razelor ultraviolete la aplicarea cromatografiei de lichide.
3
4

63

64