1. INTRODUCTION
(1-2 pages)
This chapter contains some background information on the specific situation in the country that led to
proposing this SfP Project. Special attention should be given to the economic and industrial benefits
that are expected as a result of this Project as stated in the Criteria for Success Table, including its
contribution to strengthening the national scientific/technological infrastructure. This information
should be quantified, where possible.
The ecological devastation and economic impoverishment that have befallen the Aral Sea
communities are threats to the security of the region and to the health and well-being of the people.
There is an immediate need for the communities surrounding the Aral Sea to improve their quality of
life through economic opportunities. The Aral Sea ecological change has resulted in total obliteration
of the fish stocks upon which these communities formerly depended for their sustenance and
employment, and the contemporary East and West parts of the so-called ‘Big’ Aral are essentially
hypersaline water bodies with near-total elimination of species of fresh, brackishwater, or marine
origin. The likelihood of recovery of fish stocks to these parts of the Aral Sea is very low. Water
management alternatives, the introduction of drought tolerant crop strains, less wasteful irrigation
methods and other means of mitigating the economic losses and ecological demise of the region may
hold some promise for the Aral Sea, but they have been largely unsuccessful. Water management
construction works in the area of the so-called ‘Small’ Aral (in the north; Kazakh territory) have
resulted in a stabilization or even partial restoration of former salinity levels, but have virtually no
impact on the Big Aral. Similar, but even bigger-scaled hydrological engineering works are needed
for the stabilization or partial restoration of the Big Aral, but both funding and political consensus are
not available within the foreseeable future. Any means of economic recovery for the Aral Basin
communities would therefore be a most welcome and much needed benefit.
There is evidence of a recently colonizing population of the halophilic Artemia in the Aral Sea that
holds promise as a commercially viable resource. The development and prudent management of an
Aral Sea Artemia resource could provide some degree of economic and ecological recovery to the
region through sustainable management strategies. Artemia is an essential component of aquaculture
production worldwide. Global demand for Artemia biomass and cysts is on the order of approximately
2000 tonnes per year. Revenues from the sale of Artemia products are in the range of 55 to 95 million
USD per year. Over the course of the past two decades the demand for aquaculture products has
increased at a rate of 5% to 10% per year. The potential economic benefits gained through successful
development of the emerging Aral Sea Artemia resource could provide the people of the region with
employment and other opportunities that presently do not exist, contributing to local social security.
Additionally, secondary benefits arising out of demands for goods and services and an improved
infrastructure may follow.
However, it is insufficiently known whether the current hydrobiological and hydrochemical status and
primary productivity of the Aral Sea is sufficient to support a stable Artemia population. It is therefore
of crucial importance to implement an ecological monitoring program to determine the potential
suitability of the Aral Sea as an environment in which one could expect sustained Artemia population
growth and productivity. Furthermore, the potential for commercial exploitation will depend on
Artemia population dynamics: a consistent high annual rate of cyst production, coupled with high cyst
quality, and a low cost of production could certainly result in a marketable export product for the
region. Expert advice in harvesting, transport, handling, storage, and industrial production techniques
are additionally essential for the successful exploitation of the Artemia resource.
The research team for this project includes some of the world’s foremost experts in Artemia biology
and ecology. State-of-the-art methods for characterizing Artemia, conducting population studies,
water quality and phytoplankton assessments in use by scientists from NATO countries will be
introduced to partner country scientists. Well-established sustainable management practices
developed by the NATO project partners for Great Salt Lake Artemia, Utah, USA and successfully
applied for Artemia populations in Russia, Turkmenistan, Kazakhstan and China, will serve as a
model for an Aral Sea Artemia management strategy. Monitoring devices for sampling and water
1
quality determination, Artemia cyst harvesting, storage, processing and quality control, provide
significant advancements in research efficiency for partner country scientists. Population models of
the Great Salt Lake (GSL) Artemia have been developed and illustrate the economic and conservation
benefits of thorough research and well-formulated interpretation of the scientific data. Population
growth models of the GSL Artemia resource have proven to be reasonably accurate predictors of
harvest quantities over the past six years.
Harvestable quantities are an important aspect to evaluate the exploitation potential of a site, but
quality and application range of the marketable product also decisively determines the potential yield.
The average quality of cysts harvested from diverse geographical regions of the world varies
considerably. It is a function of the inherent quality of the cysts and, equally important, handling,
storage, and processing methods. At one extreme are the cysts from the GSL; skilled harvesters and
processors can produce dried cysts with an overall average quality of over 85% for all harvested cysts.
Cysts from other locations may have a maximum quality of 70% hatching percentage with the
majority of harvested cysts exhibiting a range of lower qualities. Quantities from other sources
(China, Russia, Central Asia) have shown improved consistency in quality and yields over the past 5
years and demand for alternative Artemia cysts will is directly related to the production at GSL. It is
unknown whether cysts from the Aral Sea will demonstrate high average quality. The market for
Central Asian cysts is generally well below that for alternative sources of cysts, resulting in a decrease
in the expected value. Buyers’ perception of quality plays a key role in the value of Artemia cysts.
Newly introduced cyst strains are often valued less than known sources, even with comparable quality
characteristics. It is also for this reason that exploitation and marketing of Aral Sea Artemia cysts
should be supported by the best science available.
2
2. SCOPE AND OBJECTIVES OF THE PROJECT
(1-3 pages)
This chapter contains a summary of the objectives to be achieved within the lifetime of the
Project.
- scientific goals
- participation by other national institutions/industries
- training
- enhancement of scientific infrastructure
- international co-operation
- etc.
Scientific goals
1. Description and characterization of the Aral Sea Artemia population: biology (incl. life history
studies), population dynamics, cyst quality characteristics and their potential for aquaculture
1.1. Field research
 hydrobiological and hydrochemical characterization of the Aral Sea: salinity; ion cmposition,
temperature
 phytoplankton density and species composition
 documenting and modeling Artemia population dynamics: age class composition; fecundity,
other zooplankton species: presence/absence studies of predatory or competitor zooplankton
1.2. Laboratory experiments
 phytoplankton: isolation, identification and growth of Aral Sea algal species at different
abiotic conditions: light, temperature, nutrient levels
 Artemia:
o strain characteristics and quality control: cyst biometrics, hatching quality, diapause
behaviour, nutritional characteristics (HUFA profile)
o life history and reproductive characteristics; effect of abiotic conditions temperature,
salinity, feeding
o predation on Artemia nauplii by other zooplankton organisms
2. Development of a population model for Aral Artemia resource. Population models of the Great
Salt Lake (GSL) Artemia have been developed and have proven to be reasonably accurate
predictors of harvest quantities. A similar model should be developed for the Aral Sea Artemia,
that can be used for resource managers and for determining the commercial viability.
3. Resource management recommendations: define sustainable management plan; sustainable is
defined in terms of consistently maintaining a commercial operation and striving to ensure that
harvesting efforts do not result in adverse impacts on cyst production. Determine commercial
viability and outline optimal means of exploitation of the Artemia resource. Through
documentation of Artemia population dynamics, Aral Sea hydrology and hydrochemistry, and
micro-algae analyses, potential production of cysts can be estimated. Therefore experimental
harvests can be conducted to provide information on:
 correlation between standing crop estimates and shoreline quantities available for harvesting;
 harvesting and processing yields;
 logistical consideration, especially transport time and constraints;
 cyst quality and optimal means of handling, storage, and processing.
Above information will be used in conjunction with Artemia market information to describe the
economic viability of the resource, and to issue recommendations towards end-users on optimal
utilisation of resource: site-specific strategies for harvest, transport, and processing, potential
economic benefit, estimation of employment generated by sustainable Artemia cyst industry, and
cost/benefit analysis.
3
Participation by other national institutions/industries
The project members envisage the practical use of the project’s realizations by the implementation of
the project’s results by the end-users. The latter consist of national, regional or local decision-making
authorities, and of institutes and other governnmental entities involved in activities related to some
aspect of the study and exploitation of the broad Aral Sea socio-economic and biological environment
(e.g. local employment, fisheries, water management, environmental issues…). These end-users have
expressed their interest through a Letter of Intent at the beginning of the project, and are involved in
the progress of the project through the organization of workshops in Tashkent by the project partners.
During these workshops, the end-users are informed about the project’s findings, and are invited to
provide feedback aiming at adjustment and optimization of the project’s activities. During the
Concluding Workshop, organized in the final phase of the project, they will be informed about the
overall results of the project.
Training
Transfer of know-how is an integral and essential part of this project; training of partner institutes in
all techniques needed to attain objectives 1-2-3, detailed above, and to realize a sustainable economic
exploitation of Aral Sea Artemia, will be organized. This implies:
 Training of Uzbek partners at NATO partner’s facilities
o hydrochemistry: methods of hydrochemical water analysis, with focus on nutrients
analysis in saline environment
o phytoplankton: methods of sampling, preservation and processing of phytoplanktonic
and microphytobenthic organisms; isolation, establishing and maintenance of
monoclonal cultures of microalgae; principles and methods of modern taxonomic
identification of aquatic microorganisms
o Artemia: biology, use in aquaculture, morphology and life cycle, ecology, cyst quality
control diapause termination, decapsulation, enrichment; culture tests to study life
cycle and reproductive characteristics at different abiotic and food conditions;
commercial harvesting, storage and processing of Artemia cysts; interpretation of
population dynamics data; population modeling.

Training of Uzbek partners in the field at Aral Sea: hydrochemical and hydrobiological sampling,
quantitative and qualitative sample analysis, data processing and interpretation
Enhancement of scientific infrastructure
The project aims at the enhancement of scientific infrastructure of the Uzbek partner institutes by
purchase, transport and installation of scientific equipment
 Laboratory equipment: all equipment (and consumables) needed to conduct the laborary work
described above; training will be provided in the operation and use of these equipment items;
 Field equipment: all equipment, infrastructure and vehicles (and consumables) needed to
conduct the field study (hydrochemistry, primary production, Artemia, zooplankton)
described above; i.e. material needed for sample-taking from the Aral Sea, field sample
analysis and transport of the samples to the laboratory.
International cooperation
Although international cooperation beyond the project partners is not a direct objective of the project,
the Uzbek partners will benefit from an increased visibility on the international forum (which may
provide leverage for faciliting international cooperation in the future) through their intense contacts
with the NATO country partners, which are thoroughly embedded in the international scientific
forum.
4
5
3. REALIZATION OF THE PROJECT
(1-3 pages)
An overview of the organisation of the Project, the management structure, and the participating
institutions and industries. If the Project is composed of several sub-projects, these items
should be treated for each sub-project.
Participating institutions
NATO countries:
 Laboratory of Aquaculture & Artemia Reference Center, Ghent University, Rozier 44, B9000
Gent, Belgium (ARC) (NPD: Prof. P. Sorgeloos; co-director Dr. G. Van Stappen)
 Laboratory of Protistology & Aquatic Ecology, Department of Biology, Ghent University,
Krijgslaan, 281-58, B9000 Gent, Belgium (LPAE) (co-director Prof. W. Vyverman)
 Utah Strategic Alliance (UtSA) (renamed into Great Salt Lake Artemia Cooperative in 2006),
5859; N. Cottonwood Canyon Rd. Mt. Green, UT, USA (co-director Th. Bosteels)
 INVE Technologies (INVE Tech), Artemia Task Force, PO Box 1306, 598 W Clark St.
Grantsville, UT, USA (co-director B. Marden)
Uzbekistan:
 Laboratory of Ichthyology and Hydrobiology, Institute of Zoology, Uzbek Academy of Sciences,
Niyazov st. 1, Tashkent, 700095 (LIH) (PPD: Prof. I. Mirabdullayev)
 Institute of Bioecology of the Karakalpak Branch of the Uzbek Academy of Sciences; 41,
Berdakh prospect, 742000, Nukus (IB) (co-director Dr. I. Zholdasova)
Management structure
see below project organigram
6
Organisation of project
List of (categories of) persons involved in project, their affiliation, and a summary of their tasks and
responsibiilties
Identity
Patrick
(NPD)
Affiliation
Tasks in the project








Sorgeloos ARC
Iskandar
Mirabdullayev
(PPD)
LIH
Co-directors
NATO countries
(Brad Marden,
Gilbert Van
Stappen, Thomas
Bosteels, Wim
Vyverman)
Co-directors
partner country
(Iliya Zholdasova)
INVE Tech, ARC,
UtSA, LPAE (resp.)
IB




Other scientific
personnel (esp.
Uzbek young
scientists)
LIH, IB









Logistic support
personnel
INVE
External NATO
advisors
S. Bradt1, D. Naftz²,
R. Baskin²
End-users
1
Univ. New Hampshire,
USA; ²US Geological
Survey







general project coordination
feedback to milestones, deliverables, reports
coordination of NATO country project partners
co-organizer of workshops
end-user
feedback to milestones, deliverables, reports
organizer of workshops
coordination of partner country project
partners: field and lab work; organization of
training missions to NATO countries
daily management of project, partim LIH
daily management of project
organization of training of Uzbek scientists at
NATO partners institutes
field training of Uzbek scientists in Uzbekistan
co-reporting
participation to workshops
daily management of project
co-reporting
participation to workshops
interaction of project consortium with regional
(Karakalpakistan) and local (Moinaq) level
conduct laboratory tests, field sampling and
sample analysis
participation to workshops
contribution to reporting through compilation
of field and laboratory work reports
target of training
assistance in field sampling
translation of documents and interpreter during
workshops
participation to workshops
advise in specific scientific issues
feedback on reports
participation to workshops
 authorities at different  provide feedback to project results
levels
 participation to workshops
 institutes/organisations  where relevant, facilitate and promote project
involved in Aral Sea
activities
management & study
1
4..SCIENTIFIC RESULTS (up to 40 pages)
The scientific results are reported here in the nature of an article in a scientific periodical or as a
presentation at a scientific conference. As a consequence, this reporting is relatively detailed
and may be organised per sub-project. Special attention should be given to the need for further
R&D activities after conclusion of the Project.
PART 1. FIELD STUDIES
Asf
Jedeli
Vozrojdeniya
JA
W
Akt
E
E
a
s
t
Lazarevo
Moinak
Artemia of the Aral Sea, Uzbekistan. G. Van Stappen et al.
10th Int. Conf. on Salt Lake Research. Salt Lake City, USA, May 11-16, 2008
slide 17 of 28
1. HYDROCHEMICAL COMPOSITION OF THE ARAL SEA IN THE PERIOD 2005-2008
2
30
temperature (°C)
25
20
15
10
5
West Aral
0
Jan-04
Aug-04
Feb-05
Sep-05
Mar-06
East Aral
Oct-06
Apr-07
Nov-07
month-year
1.1. Ionic composition of East and West Aral surface waters
In the first half of the twentieth century, the salinity of water in the open part of the Aral Sea reached
about 10 ppt. In the period 1960-1980, the salinity increased at an average rate of 0.2-0.6 ppt per year
(Table 1.1). Since 1989, the Aral Sea split into the Large and Small Aral and the salinity started
growing at the rate of 1 ppt per year. In the 1990’s, the central range of islands (Vozrozhdeniye and
Lazarev Islands) merged into one island, which caused a separation of the Big Aral into the western
and eastern basins. In recent years, the salinity of the water in the western basin increased at an
average rate of 5-10 ppt per year, while in the shallow eastern basin the rate of salinity increase 5-20
ppt per year (Table 1.1).
In 2005-2007 the salinity of the West Aral (which is relatively deep) increased relatively fluently
reaching 116 ppt at the end of 2007 (Fig. 1.1). At the same time the salinity in the shallow East Aral
underwent sharp fluctuations, caused by fluctuations of inflow from the Amu Darya delta. So,
salinity in the East Aral reached 150 ppt in 2002, and then dropped to less then 100 ppt in 2003. In
August 2005 a lot of water came into the East Aral causing a decreasing salinity till 67 ppt. In the
relatively dry period 2006-2007 salinity quickly increased again, reaching 265 ppt in coastal areas of
the East Aral in May 2007 causing crystallization of sulphates (Fig. 1.2). Apart from this, nu
substantial changes in ion composition were observed over the period 2005-2007, nor between the
East and West Aral, as the precipitation point for salts had not been reached.
Table 1.1. Salinity in Aral Sea surface water in period 1960-2007
Year
1960
1970
1975
1980
1985
1990
1992
1995
1996
Salinity (ppt)
10
12
14
17
23
32
35
42
44
West Aral
3
East Aral
1997
49–51
50–52
1998
54
58
1999
56
2000
58-63
2001
63-68
108-112
2002
69-74
150
2003
74-80
90-110
2004
84-94
90-116
2005
82-92
67-113
2006
96-101
70-136
2007
95-116
(265)*
*As the East Aral was virtually inaccessible for regular sampling taking in 2007, this value
corresponds with highly saline coastal samples
140
120
salinity (g/l)
100
80
60
40
West Aral
East Aral
20
0
Jan-04
Aug-04
Feb-05
Sep-05
Mar-06
Oct-06
month-year
Fig.1.1. Salinity fluctuations in Aral Sea surface water in period 2004-2007
4
Apr-07
Nov-07
Dynamic of ion composition of water in East Aral
100%
80%
60%
40%
20%
2005
2006
HCO3
Cl
SO4 Aral
Ca++
East
Sea Mg++
June
2007
Na+K
Fig. 2.1. Dynamics of East Aral surface water ion composition in period 2005-2007
5
September
August, 2
part
August, 1
part
June
May
April
November
October
September
August
0%
Dynamic of ion composition of water in West Aral
100%
80%
60%
40%
20%
2005
2006
October-
September
August
July
June, 2
June, 1
May
September
August-2
August-1
June
May
April
November
October
September
August
July
0%
2007
West Aral Sea
HCO3
Cl
SO4
Ca++
Mg++
Na+K
Fig. 1.3. Dynamics of West Aral surface water ion composition in period 2005-2007
1.2. Nutrient composition of deep West Aral water layers
In view of the low overall phytoplankton concentrations observed in West and East Aral (see 2.)
nutrient analysis (N and P) was performed for water samples taken from various depths in the West
Aral, in order to verify if a chemocline existed for these compounds. In the positive case, the project
partners might consider the possibility of valorising nutrient-rich deep-layer water using air-water lifts
for increased primary (and hence secondary) production in superficial water layers.
Samples were taken on 3 sites (Aktumsuk- AKT, Jidely Bulak-JID, Asphalt Kulau-ASP) at the
western shore, at 3 and 4 km offshore (S1 and S2, resp.), at different depths: 1 m, 10 m, and nearbottom.
As shown in Table 1.2., N, P and Fe levels were largely homogeneous throughout the water column,
or if variations occurred, these were not consistent with depth.
6
Table 1.2. Nutrient levels in deep-water samples taken along West Aral western coast
Sampling Site
AKT
S1
Total N (mg/l)1
2.18
2.80
2.43
2.34
2.46
2.52
2.72
2.98
2.59
2.51
2.57
2.82
2.87
2.77
2.65
2.80
2.92
3.04
Depth (m)
1
10
23
S2
1
10
25
JID
S1
1
10
33
S2
1
10
37
ASP
S1
1
10
36
S2
1
10
32
1
detection limit 0.050 mg/l
²detection limit 0.002 mg/l
³detection limit 0.020 mg/l
Total P (mg/l)²
0.048
0.277
0.078
0.039
0.041
0.077
0.046
0.068
0.055
0.041
0.041
0.045
0.042
0.040
0.041
0.038
0.039
0.115
Iron (mg/l)³
0.046
0.298
0.261
<0.020
<0.020
0.060
0.030
0.110
0.057
<0.020
<0.020
0.028
<0.020
<0.020
<0.020
<0.020
0.050
0.065
2. EVOLUTION OF THE PHYTOPLANKTON DENSITY AND SPECIES COMPOSITION IN THE ARAL
SEA IN THE PERIOD 2005-2007
2.1. Introduction
Phytoplankton of the Aral Sea was studied since the beginning of the 20th century (Kiselev, 1927).
Later on data on phytoplankton were presented by Pichkily (1981), Оrlova et al. (1998) and
Mirabdullayev et al. (2004). During the progressive salinization of the Aral Sea, the diversity of
planktonic algae has continuously decreased, being in the last years about 4 times lower than 80 years
ago (Table 2.1).
Table 2.1. Number of phytoplankton species found in the Aral Sea during its transition from a
oligohaline to a hypersaline environment
Тахon
Сyanobacteria
41
1967 –
1974²
79
Bacillariophyta
210
104
115
41
49
56
Pyrrophyta
15
28
3
3
4
3
Euglenophyta
0
3
2
0
0
2
19251
1999-2002³
2005
2006
2007
30
14
15
15
7
Chlorophyta
109
60
9
12
11
14
Total number
375
306
159
70
of species
1
Kiselev (1927); ²Pichkily (1981), ³Mirabdullayev et al. (2004)
79
90
Due to the high water transparency and shallow depths in the Aral Sea, most organics have been
produced by phytobenthos, not phytoplankton. In general, the biomass of phytobenthos reached 90%,
while phytoplankton reached 10% (Karpevich, 1975). Charophytes (mainly) yielded ca. 75% and the
chlorophyte ca. 13% of the phytobenthos biomass. In the 1990’s common phytobenthos such as
Tolypella aralica, Vauscheria dichotoma, Cladophora gracilis, Polysiphonia violaceae and Zostera
(Karpevich, 1975) became extinct in the Aral Sea. Presently the only benthic macroscopic plants in
the Aral are Cladophora glomerata, C. fraсta and Vauscheria cf. dichotoma (Zavialov et al., 2006).
2.2. Material and methods
Phytoplankton samples were fixed by lugol, and postfixed a few minutes later with formaldehyde at
0.5%. Samples were filtered using Millipore filters (1,2 μm) and counted in a Najot chamber under a
Zeiss Axiostar microscope.
In 2005-2006 samples were collected along the western shore of the Eastern Aral (depths 1.0-2.5 m)
and the eastern shore of the West Aral (depths 1.0-5.0 m). In 2007 samples were only collected on the
western shore of the West Aral (depths 1.0-40.0 m). A total of 132 samples were collected and
analysed.
2.3. Results
During the project lifetime 90 species of algae were recorded in total in the Aral Sea plankton (Table
2.2). Most species recorded belong to the diatoms (Bacillariophyta) which constituted 57-64% of all
species (Fig. 2.1). Also important were such groups as Cyanobacteria (17-22%) and Chlorophyta (1318%). Other algal phyla were represented by single species. The species composition of
phytoplankton in West and East Aral was largely similar. Not all recorded algal species are truly
planktonic. As the collection sites were shallow (2-4 m), a significant number of algal species are
representatives of phytobenthos and periphyton.
Table 2.2. Phytoplankton species observed in West and East Aral in period 2005-2007
Taxon
CYANOBACTERIA
1.Microcystis aeruginosa Kutz
2.Oscillatoria sp.(limosa?)
3.O.. chlorina(Kutz) Gom.
4.O.. planctonica Wolocz.
5.O.. amphibia Ag.
6.Merismopedia glauca (Ehr.)Nag.
7.Gloeocapsa alpina Nag.em.Band
8. G. alpina f. lignicola (Rabenh.)
9.G. minima (Keissl.)
10. G. minor (Kutz)
11.G. turgida (Kutz.)
12.Gomphasphaeria aponina Kutz.
13.G.. lacustris Chod.
14.Synechococcus sp.(salina?)
2005
2006
2007
East Aral West Aral East Aral West Aral West Aral
+
+
+
+
+
+
+
+
+
+
+
+
+
-
8
+
+
+
+
+
+
+
+
+
+
+
+
+
+
+
+
+
+
+
+
+
+
+
15.Phormidium ambiguum Kissel
16.Phormidium sp.
17.Ph. papillaterminatum Kissel.
18.Lyngbya sp.
19.L. Kuetzingii (Kutz.) Schmidle
20.Spirulina sp.
21.Spirulina major Kutz.
22.Anabaena flos-aquae (Spreng)
BACILLARIOPHYTA
1.Cyclotella sp.
2.C. meneghiniana Kutz.
3.Melosira varians Ag.
4.Rhizosolea longiseta Zacharias
5.Synedra tabulata (Ag.) Kutz.
6.S. tabulata v. parva (Kutz) Grun.
7.S. tabulata v. fasciculata (Kutz.)
8.Synedra sp.
9.S. minuscula Grun.
10.Fragilaria construens (Ehr.) Grun
11.F. .construens v. venter (Ehr.) Grun.
12.Actinocyclops Ehrebergii
13.Achnanthes minutissima Kutz.
14.A. affinis Grun.
15.Achnanthes sp.
16.Chaetoceros sp.
17.Coscinodiscus sp.
18.Stephanodiscus sp.
19.Cocconeis sp.
20.Cocconeis placentula Ehr.
21.C. placentula v.euglypta (Ehr.) Cl.
22.Diploneis intterupta (Kutz.)
23.D. Smithii (Breb) Cl.
24.D. Smithii v .pumila (Grun.) Hust.
25.Entomoneis paludosa Reimer
26.Amphora holsatica Hust.
27.A. coffeaformis Ag.
28.A. commutata Grun.
29.A. ovalis Kutz.
30.A.veneta Kutz.
31.A .robusta Greg.
32.Amphora sp. (long)
33.Mastogloia baltica Grun.
34.M. Smithii Thw.
35.M. pumilla (Grun.) Cl.
36.Navicula cincta (Ehr.) Kutz.
37.Navicula sp.
38.N. cryptocephala Kutz.
39.N. cryptocephala v.veneta Kutz.
40.N. kolbei Poretz et Aniss
41.N. protracta v.capitata Woronich.
42.N. pygmae Kutz.
43. N. salinarum Grun.
+
+
+
+
-
+
+
+
+
+
+
+
+
+
+
+
+
+
+
+
+
+
+
+
+
+
+
-
+
+
+
+
+
+
+
+
+
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+
+
+
+
+
+
-
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+
+
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+
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+
+
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+
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+
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-
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+
+
+
+
9
44. N. spicula Hickie
45. N. gracilis Ehr.
46. N. halophila (Grun.) Cl.
47. N. microcephala Grun.
48.Hantzshia virgata v.capitellata Hust.
49.Nitzschia closterium (Ehr)W.Sm.
50.N. acicularis W.Sm.
51.N. lorenziana v.incurta Grun.
52.N. microcephala Grun.
53.N. obtusa W.Sm.
54.N. obtusa v. scalpeliformis Grun.
55.N. cf .sigma (Kutz.)
56.N. palea (Kutz) W.Sm.
57.N. vermicularis (Kutz.)
58.N. punctata v.aralensis Borszczow
59.N. hungarica Grun.
60.N. capitellata Hust.
61.Gyrosigma scalproides (Rabenh) Cl.
62. G. acuminatum (Kutz) Raben
63.G. spenseri (W.Sm.) Cl.
64.Pleurosigma elongatum W.Sm.
65.P. obtuscurum W.Sm.
66.Surirella ovata Kutz.
67.Surilla sp.(.crumenosa?)
68.Campylodiscus aralensis Kissl..
PYRROPHYTA
1.Glenodinium sp.
2.Peridinium sp.
CRYPTOHYTA
1.Cryptomonas sp.
EUGLENOPHYTA
1.Euglena sp.
2.Phacus sp.
CHLOROPHYTA
1.Chlorella sp.
2.Oocystis sp.(borgei Snow?)
3.O. marssonii Lemm.
4.Cosmarium formulosum Hoff.
5.Algae gen. sp. (Tetraselmis?)
6.Dunaliella sp.
7.Chlorocococcus turgida
8.Ankistrodesmus minutissima Korsch.
9.A. acicularis (Ag.Br.)
10.A. falcatus
11.Dactyosphaerium pulchellum Wood.
12.Tetraedron minimum (Ag.Br.) Hans.
13.Monas sp.
14.Carteria sp.
15.Chlorococcum sp.
16.Chlamidomonas sp.
17.Chromulina sp.
18.Polytoma sp.(ocellatum?)
+
+
+
+
+
+
+
+
+
+
+
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+
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-
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+
+
+
+
+
+
10
19.Scenedesmus quadricauda
20.Cladophora fracta (L.) Kutz.
Total number of species
56
+
47
West Aral 2005
70
+
54
+
69
East Aral 2005
West Aral 2006
East Aral 2006
West Aral 2007
Cyanobacteria
Bacillariophyta
Dinophyta
Euglenophyta
Cryptophyta
Chlorophyta
Fig. 2.1. Species diversity of phytoplankton in West and East Aral in period 2005-2007
Both the phytoplankton cell densities and biomass were normally higher in the West Aral than in the
East Aral; the highest occurrence of cell density and biomass was observed in summer months (Figs
2.2 and 2.3).
11
2000
cell density per ml
West Aral
East Aral
1500
1000
500
Fe
b
-0
Ap 5
r-0
Ju 5
n0
Au 5
g0
O 5
ct
D 05
ec
-0
Fe 5
b0
Ap 6
r-0
Ju 6
n0
Au 6
g0
O 6
ct
D 06
ec
-0
Fe 6
b0
Ap 7
r-0
Ju 7
n0
Au 7
g0
O 7
ct
-0
7
0
month-year
Fig. 2.2. Seasonal and annual fluctuations in phytoplankton cell density in East and West Aral in the
period 2005-2007
West Aral
East Aral
140
biomass (mg/l)
120
100
80
60
40
20
Oct-07
Aug-07
Jun-07
Apr-07
Feb-07
Dec-06
Oct-06
Aug-06
Jun-06
Apr-06
Feb-06
Dec-05
Oct-05
Aug-05
Jun-05
Apr-05
Feb-05
0
month-year
Fig. 2.3. Seasonal and annual fluctuations in phytoplankton biomass density in East and West Aral in
the period 2005-2007
References
Karpevich, A.F., 1975. Teoriya i praktika akklimatizatsii vodnikh organizmov. Pischevaya
Promyshlennost, Moscow. 432 pp. (in Russian).
12
Kiselev I.A. 1927. Novye dannye o vodoroslyakh Aralskogo Morya. – Depart. Appl. Ichthyol. and
Scient. Research. GIOA. 1927. V. 5. P. 1-64 (in Russian).
Mirabdullayev I.M., Joldasova I.M., Mustafaeva Z.A., Kazakhbaev S., Lyubimova S.A.,
Tashmukhamedov B.A. 2004. Succession of the ecosystems of the Aral Sea during its transition from
oligohaline to polyhaline waterbody. J. Marine Syst. V. 47. N 1-4. P. 101-107.
Orlova M.I., Aladin N.V., Filippov A.A., Plotnikov I.S., Smurov A.O., Rusakova O.M., Zhakova
L.V., Pirulin D.D. 1998. Living assiciationsof the northern part of the Aral Sea in 1993-1995. In:
UNESCO (Ed.), Aral Sea Project 1992-1996, Final Scientific Reports. Paris. P. 95-138.
Pichkily L.O. 1981. Fitoplankton Aralskogo morya v usloviyakh antropogennogo vozdeystviya
(1957-1980). Kiev, Naukova Dumka. 228 pp. (in Russian).
Zavialov P.O., Arashkevich E.G., Dikarev S.N., Kudyshkin T.V., Kurbaniyazov A.K., Ni A.A.,
Sapozhnikov F.V., Soloviev K.A., Khan V.M. 2006. Monitoring sostoyaniya fizicheskikh,
khimicheskikh i biologicheskikh sistem Aralskogo Morya v usloviyakh ekologicheskogo krizisa. In:
Sovremennye problemy aridnykh i semiaridnykh ekosistem yuga Rossii. Rostov-na-Donu. P. 529-562
(in Russian).
3. ARTEMIA POPULATION DYNAMICS IN THE PELAGIC ZONE OF THE ARAL SEA IN THE PERIOD
2005-2008
West females
West gravid females
East females
East gravid females
density (individuals/liter)
0.5
0.4
0.3
0.2
0.1
0
Aug-04
Feb-05
Sep-05
Mar-06
month-year
13
Oct-06
Apr-07
Nov-07
West Aral
East Aral
18
cyst count per liter
16
14
12
10
8
6
4
2
0
Aug-04
Feb-05
Sep-05
Mar-06
Oct-06
Apr-07
Nov-07
month-year
4. EVOLUTION OF NON-ARTEMIA ZOOPLANKTON IN THE ARAL SEA IN THE PERIOD 2005-2008
4.1.Introduction
Until the 1970’s, the composition of zooplankton in the Aral Sea was stable, comprising over 40
species in pelagial zones (Anonymous, 1974). The basis of the zooplankton fauna was formed by
Arctodiaptomus salinus, Ceriodaphnia reticulata, Moina salina, Podonidae. In the 1960’s a copepod
Calanipeda aquaedulcis Kritchagin was released into the Aral Sea. In the 1970-80’s this species was
dominant in the zooplankton of the Aral Sea, which resulted in the disappearance of the former
dominants, A. salinus, C. reticulata, M. salina (Aladin and Andreev, 1984).
A drop in the inflow of the rivers and progressive salinization of the waters of the Aral Sea produced
an adverse impact on the freshwater and brackish-water species, and they quickly vanished from the
fauna. A quick decrease in the biodiversity of zooplankton was recorded in the first half of the 1970’s.
By 1976, the species composition became stable as the average salinity of the Aral Sea reached 14 ppt
(Andreev, 1989). Later, a gradual decrease in the zooplankton diversity took place (Table 4.1). Since
1997, the former dominant, Calanipeda aquaedulcis, vanished from the plankton, which was
apparently the reason for the emergence of Moina salina and parthenogenetic Artemia in the plankton
(Mirabdullayev et al., 2004).
Data on Aral Sea zooplankton are abundant for the period till the 1980’s. As since 1981 fisheries
collapsed on the Big Aral, data on its ecology since this period are very fragmentary. However, fast
salinization of the water body over the last 20 years has caused fast changes in its biota.
The goal of this study is to present recent data on the abundance of zooplankton, most of which
species are food competitors for Artemia.
4.2. Material and Methods
Samples were collected with a conical plankton net (diameter 36 cm), hauled from the bottom to the
surface, and were fixed with formaldehyde until a final concentration of 2%. Zooplankton organisms
were counted in a Bogorov counting chamber under a stereozoom microscope. In totally 132 samples
were collected and processed.
14
Table 4.1. Succession of species composition of zooplankton of the Aral Sea during its transition
from oligohaline to a hypersaline state (+ = present; - = absent)
Taxa
1971
1981
1989
1994
2000
2005
2006
2007
Hexarthra fennica
+
+
+
–
+
+
+
–
Brachionus plicatilis
+
+
–
–
–
+
+
–
Nereis diversicolor
+
+
+
+
+
–
–
–
Cerastoderma isthmicum
+
+
+
+
+
–
–
–
Syndosmya segmentum
+
+
+
+
+
–
–
–
Artemia sp.
–
–
–
–
+
+
+
+
Moina salina
+
–
+
–
+
+
+
–
Podonevadne camptonyx
+
+
+
–
–
–
–
–
Apocyclops dengizicus
+
+
+
–
–
+
–
–
Cletocamptus retrogressus
+
+
+
+
+
+
+
+
Baeotendipes cf. noctivaga
+
+
+
+
+
+
+
+
Total number of species
43
18
13
9
8
7
6
3
salinity (g. l-1)
12
18
30
37
58-63
68113
70136
>100
4.3. Results
In May 2005 only 2 species of zooplankton were recorded in the West Aral, near Cape Kabanbay. In
August 2005 4 species were recorded in the West Aral, with the rotiferan Hexarthra fennica as
dominant speices, and 2 species in the East Aral with the copepod Apocyclops dengizicus as
dominant. Zooplankton biomass was relatively low in May and August 2005 (Table 4.2). In
September 2005 a big amount of water was discharged into the south part of the East Aral introducing
nutrients and resulting in a decreased salinity and an increased bioproductivity. As a consequence,
zooplankton species diversity and biomass increased sharply in the East Aral (Table 4.2). Brachionus
plicatilis and Moina salina became dominants, species which had not been recorded over the last
years.
Table 4.2. Species composition and density of zooplankton in the Aral Sea in 2005 (D = dominant
species)
West Aral
Turbellaria gen. sp.
Hexarthra fennica
Brachionus plicatilis
Clethocamptus retrogressus
Apocyclops dengizicus
Moina salina
Baeotendipes cf. noctivaga
number of individuals/m³
biomass (mg/m³)
May
+
D
+
3,024
1.67
15
July
September
+
D
D
+
+
+
D
+
+
+
+
33,294
2,877
1.61
3.64
East Aral
July
+
D
+
144
2.07
September
+
+
D
+
D
+
48,654
609.23
In 2006 species diversity and quantitative development of zooplankton in the West Aral was
extremely low, with the harpacticoid Cletocamptus retrogressus as the most constant representative
(Tables 4.3 and 4.4). In the East Aral in April and May 2006 the diversity of zooplankton was
comparatively high with Hexarthra fennica as a dominant species and with the appearance of the
calanoid Arctodiaptomus salinus, a species not recorded in the Big Aral over about 20 years (Table
4.1). However in June 2006 the diversity and development of zooplankton decreased sharply, which is
probably caused by the drop of inflow of water and the increased salinity.
Table 4.3. Species composition of zooplankton in the Aral Sea in 2006
Turbellaria gen. sp.
Brachionus plicatilis
Hexarthra fennica
Moina salina
Arctodiaptomus salinus
Cletocamptus retrogressus
Blaeotendipes cf.
noctivaga
West Aral
Apr May Jun Aug
+
+
+
+
+
+
+
+
Sep
+
+
Apr
+
+
D
+
+
+
+
May
+
+
D
+
+
+
-
East Aral
Jun
+
+
+
Aug
+
+
+
Sep
+
+
+
Table. 4.4. Zooplankton density in Aral Sea in 2006
April
number of individuals/m3
biomass (mg/m3)
0
0
number of individuals/m3
biomass (mg/m3)
53,250
18.56
May
West Aral
484
3.28
East Aral
52,157
24.44
June
August
September
97
11.11
7
7.1
2
0.05
37
1.10
136
136.5
15
0.76
In 2007 only 3 species of zooplankton were recorded in the West Aral. No zooplankton was recorded
in the East Aral due too high salinity (265 ppt). Zooplankton density was very low and less than in
2006 (Table 4.5).
Table 4.5. Species composition and density of zooplankton in the Aral Sea in 2007
Turbellaria
Cletocamptus retrogressus
Baeotendipes cf. noctivaga
number of individuals/m3
biomass (mg/m3)
June
+
+
11.2
0.37
July
+
+
12.35
0.39
August
+
+
+
1.77
0.06
September
+
+
0
0
4.4. Discussion
Over the last five years, four species have represented almost permanent elements of the Big Aral Sea
zooplankton: the rotiferan Hexarthra fennica, the branchiopod crustacean Artemia, the harpacticoid
copepod crustacean Cletocamptus retrogressus and larvae of the dipteran chironomid Baeotendipes
16
cf. noctivaga (Table 1). Surprisingly, in 2004-2005 the cyclopoid Apocyclops dengizicus was
recorded, a pecies which had not been observed since the mid-1990’s.
There is a strong overall tendency towards decreased zooplankton species diversity.. However, sharp
fluctuations in water inflow into the shallow East Big Aral in 2005-2006 resulted in considerable
changes in the structure and composition of the zooplankton fauna: a fast salinity drop in the East Big
Aral Sea (from 100 to 70 ppt) in autumn 2005 - spring 2006 and the inflow of nutrients resulted in a
marked increase in zooplankton diversity and biomass (Tables 4.2 and 4.3) and in the appearance of
such species as the rotiferan Brachionus plicatilis, the cladoceran Moina salina, and the calanoid
copepod Arctodiaptomus salinus, which had not been recorded in the Big Aral in previous few years
Later onwards biodiversity and biomass fell back on their previous low values (Table 4.5).
The overall biomass densities, recorded during this sampling campaig, suggest that the development
of non-Artemia zooplankton is too low to play a significant role in competition with the Artemia
population, which is in accordance with the opinion expressed by Zavialov et al. (2006).
Literature References
Aladin, N.V.;Andreev, N.I. 1984. Vliyanie solenosti Aralskogo morya na izmenenie sostava fauni
vetvistousikh rakoobraznikh. Gidrobiologicheskiy Zhurnal. V. 20. N3. P. 23-28 (in Russian).
Andreev, N.I. 1989. The zooplankton of the Aral Sea during its initial period of salinisation. Proceed.
Zool. Inst. USSR Acad. Sci. V. 199. P. 26-51 (in Russian).
Anonymous. 1974.. Atlas of the invertebrates of the Aral Sea. (Ed. F.D. Mordukhay-Boltovskoy). M.:
Pishevaya promyshlennost. 272 p. (in Russian).
Mirabdullayev I.M., Joldasova I.M., Mustafaeva Z.A., Kazakhbaev S., Lyubimova S.A.,
Tashmukhamedov B.A. 2004. Succession of the ecosystems of the Aral Sea during its transition
from oligohaline to polyhaline waterbody. J. Marine Syst. V. 47. N 1-4. P. 101-107.
Zavialov P.O., Arashkevich E.G., Dikarev S.N., Kudyshkin T.V., Kurbaniyazov A.K., Ni A.A.,
Sapozhnikov F.V., Soloviev K.A., Khan V.M. 2006. Monitoring sostoyaniya fizicheskikh,
khimicheskikh i biologicheskikh sistem Aralskogo Morya v usloviyakh ekologicheskogo
krizisa. In: Sovremennye problemy aridnykh i semiaridnykh ekosistem yuga Rossii. Rostov-
na-Donu.
17
PART 2. LABORATORY STUDIES
1. STUDY OF PHYTOPLANKTON SPECIES FROM THE ARAL SEA
SEM investigations of frustules morphology of numerically dominant diatom species
(Bacillariophyceae) from Aral Sea were performed Detailed SEM morphological analysis was carried
out for some Cyclotella, Actinocyclus, Melosira, Cocconeis, Amphora, Navicula, Nitzschia, Surirella,
and Campylodiscus.. A series of microalgal species were isolated from Aral samples available and
introduced in monoclonal cultures. Representatives of genera Oocystis, Tetraselmis, Navicula,
Nitzschia, Hantzschia, Mastogloia and Surirella proved to be cultivatable. Oocystis and Tetraselmis
cultures were used in Artemia feeding experiments (see 2).
Two diatom species (Fig. 2.1.) cultured were studied in more detail. Nitzschia sigma is one of the
Nitzschia spp. deposited in the diatom culture collection of the PAE UGEnt (Chepurnov et al., 2008).
The cell- and life-cycle of this diatom were studied experimentally; the mating system is heterothallic.
Sexual reproduction was induced experimentally in monoclonal cultures of Surirella fastuosa; in
addition this diatom exhibited interesting morphological feature as heterovalvy (a peculiar variation in
structure of the siliceous exoskeleton of a single cell) – this was also studied in detail with the aid of
scanning electron microscopy.
Stomach contents of adult Artemia specimens (collected in Aral) were investigated too; microscopical
analysis showed that Oocystis spp. were dominating in the diet of Artemia, but diatoms of the genera
Cocconeis and Nitzschia were present in the contents as well.
Fig. 2.1. Microscopic images of Diatoms (Bacillariophyceae) from the Aral Sea:
Nitzschia
o (A) Monoclonal culture
o (B) Reproductive stage (auxospore)
Surirella
o (C) Siliceous part of cell wall, the frustule: light microscopy
o (D) scanning electron microscope
18
2. STUDY OF THE CHARACTERISTICS OF THE PARTHENOGENETIC ARAL ARTEMIA STRAIN
2.1. Objectives
The strain characteristics of the parthenogenetic Aral Sea Artemia were studied in a series of
laboratory tests. These tests had a double objective:
a) to study the suitability of the strain for larviculture applications; for this purpose, biometrics
enrichment behaviour were studied; and
b) to study the Artemia biology (survival and fecundity) in standardized laboratory conditions, and
relate these to the observations on the population, made in the field.
2.2. Biometric characteristics
2.2.1. Material and Methods
The cyst diameter was obtained by incubating a small sample of full cysts in 10 ppt seawater, add 2%
lugol and keeping the sample overnight in darkness (Lavens and Sorgeloos, 1996). The cyst diameter
of 100 cysts was measured with a microscope, equipped with a calibrated eyepiece. For determination
of naupliar length, a small cyst sample was incubated in 35ppt seawater. After hatching, 100 (instar I)
nauplii were measured (from tip of head until tip of telson) under a microscope.
For determination of the hatching quality, a cyst sample of 1.6 g was incubated in 800 ml of 32 ppt
artificial seawater (Instant Ocean) under continuous illumination (2000lux) at 28°C in a
cylindroconical tube, provided with aeration to keep the cysts in suspension, and run in triplicate.
After 24 h of incubation 6 samples of 250µl were taken from each cone, fixed using lugol solution and
the nauplii and umbrellae were counted under a microscope. Unhatched cysts were decapsulated by
adding NaOCl to each sample. The unhatched embryos were counted and the hatching percentage (H
%) was calculated as follows: H% = (nauplii/nauplii+umbrellae+ embryos) * 100
2.2.2. Results
Cyst diameter of the various samples analysed ranged between 249.4.5 -260.0 µm, but was in the
order of 250-252 µm for most of the nine Aral samples analysed, a rather small value for a
parthenogenetic strain, but comparable to the diameter of the commercially important Great Salt Lake
Artemia (Vanhaecke and Sorgeloos, 1980). Instar I length varied between 493.6-499.5 µm. The
hatching percentage of the respective samples varied between 10.0 and 93.5 %. The quantities of the
samples were either too small, or the hatching was too high to allow a detailed study of the diapause
requirements of the Aral Sea strain.
2. 3. Enrichment of Aral Sea Artemia nauplii
2.3.1. Objectives
In order to evaluate the potential use of Aral Sea Artemia for larviculture applications, nauplii of the
sample with ARC code number 1638 were hatched in standard conditions (Lavens and Sorgeloos,
1996), and enriched with a standard enrichment emulsion, the metabolic fate of which was followed
during a subsequent starvation experiment.
2.3.2. Material and methods
For sake of comparison with literature data, the standard enrichment emulsion “ICES50” (50% total
n-3 HUFA on dry weight basis; DHA/EPA ratio of 0.87) was used. Temperature and salinity during
enrichment were 28°C and 32g l-1. The enrichment was performed in triplicate 1 liter cones (800 ml
water volume). The enrichment emulsion was administered at a dose of 0.2g l-1 at 0 and 12 h after
hatching, i.e. at t0 and t12. Samples were harvested at t0, t12, and t24 for FAME analysis. After 24 h
enrichment period the surviving Artemia nauplii were gently rinsed with seawater on a sieve to
remove all residual emulsion, and then transferred to cylindroconical tubes at a density of 125  10ml1
, kept in a water bath at 28°C, for a subsequent starvation during 36 h. Samples were then taken after
19
12, 24 and 36 h of starvation (t36, t48 and t60).
2.3.3. Results and Discussion
The EPA and DHA level of the ICES50 emulsion was 205.20 and 178.00 mg/g, respectively. The
HUFA levels of the the non-enriched nauplii (t0), the enriched (t12 and t24) and the starved metanauplii
(t36, t48 and t60) are shown in Figure 2.2.
Absolute changes in HUFA contents
following enrichment and starvation
mg/g dry weight of Artemia
40
30
EPA
DHA
20
> (n-3)
> (n-6)
10
0
0
12
24
36
48
60
time (h)
Fig. 2.2. Changes in HUFA contents (mg/g DW) in Aral Sea Artemia enriched
for 12 h with emulsion ICES50, and during a subsequent 36 h starvation period
As in Great Salt Lake A. franciscana (Han et al., 2001), parthenogenetic Aral Sea Artemia shows a
differential accumulation of fatty acids during the enrichment period, followed by a differential
breaking down during the starvation period. The exact levels of accumulation depend on various
factors such as a) initial concentrations in non-enriched nauplii; b) composition of enrichment
emulsion; c) specific conditions, such as temperature during enrichment and exact age of nauplii at the
onset of the enrichment period. Nevertheless, the results show that comparable HUFA levels can be
obtained with Aral Sea Artemia, as with the commercial GSL Artemia (Han et al., 2000; 2001).
2. 4. Effect of culture conditions on survival and fecundity of Aral Sea Artemia
2.4.1. Objectives
A series of laboratory culture tests was set up in order to evaluate the effect of different culture
conditions on the survival and reproduction of Aral Sea Artemia. The following culture parameters
were analyzed:
 culture salinity
 culture temperature
 phytoplankton species (monodiet) applied (in comparison with a reference phytoplankton
diet)
 density of phytoplankton cells applied: as the phytoplankton density of Aral Sea is low,
different microalgae cells were tested, lower than the reference density used for standard
Artemia culture (Lavens and Sorgeloos, 1996).
2.4.2. Material and methods
20
2.4.2.1. Cyst sample
The Aral cyst sample with ARC code number 1638 (harvesting date: February 2005) was used
throughouth the culture experiments. Its hatching percentage was 90.5 % ± 0.2 upon arrival at ARC
(March 2005).
2.4.2.2. Algae species
Oocystis sp. (Fig. 2.3) and Nautococcus sp. (Fig. 2.4) were isolated from Aral Sea water samples;
Tetraselmis suecica was applied as reference diet for sake of comparison with literature data
(Coutteau et al., 1992). The different algae were mass-cultured under standard conditions (Lavens and
Sorgeloos, 1996) and the algae suspension was harvested and centrifuged in the exponential growth
phase. The cell concentration of the algal suspension was counted using the Burker counting chamber,
and the suspension was stored at 4°C and used for a maximum of 5 days (after which a new culture
was ready for harvest). For cell size and FAME analysis of these 3 algal species, see Tables 2.1 and
2.2, respectively.
Fig. 2.3. Microscopical view of
Oocystis cells
Fig. 2.4. Microscopical view of Nautococcus cells
Table 2.1. Size of the individual algal cells (n=100)
Species
Length (µm)
Width (µm)
Tetraselmis suecica
7.31 ± 0.66
4.29 ± 0.72
Nautococcus sp.
12.05 ± 1.3
9.08 ± 1.16
Oocystis sp.
5.08 ± 1.07
2.54 ± 0.61
* determined according to Lavens and Sorgeloos (1996)
Table 2.2. FAME –analysis in mg/g DW (3 replicates) of algal cells
Tetraselmis
Nautococcus
16:0
23.6 ± 0.2
38.5 ± 1.0
16:1(n-7)
2.6 ± 0.0
10.2 ± 0.3
18:1(n-9)
8.3 ± 0.1
50.2 ± 1.0
18:1(n-7)
2.8 ± 0.0
3.9 ± 0.1
18:2(n-6)-c
5.1 ± 1.0
13.2 ± 0.3
18:3(n-3)
22.0 ± 0.3
9.3 ± 0.2
18:4(n-3)
11.2 ± 0.1
2.3 ± 0.1
20:5(n-3)
7.6 ± 0.1
2.8 ± 0.1
22:6(n-3)
0.0 ± 0.0
0.0 ± 0.0
Sum(n-3) >or= 20:3(n-3)
8.1 ± 0.1
3.0 ± 0.1
Sum(n-6) >or= 18:2(n-6)-t
7.2 ± 0.1
16.5 ± 1.5
21
Biomass*
(pg dry weight
/cell)
153.90 ± 7.88
229.53 ± 7.78
283.75 ± 7.17
Oocystis
30.0 ± 3.4
5.9 ± 0.0
9.1 ± 0.6
2.9 ± 0.3
11.4 ± 0.4
19.4 ± 1.9
11.7 ± 1.1
6.6 ± 0.6
0.0 ± 0.0
7.2 ± 0.6
13.0 ± 0.5
Total mg FAME/g DW
115.3 ± 1.3
160.4 ± 4.6
129.1 ± 10.2
2.4.2.3. Feeding regimes
Different feeding regimes (= number of algal cells supplied per Artemia individual) were applied,
based on the feeding regime described by Coutteau et al. (1992) for Tetraselmis suecica, which was
used as reference regime (“regime 1”) (Table 2.3). Other feeding regimes were a fraction (in the
range ½ - 1/16) of the reference regime.
Table 2.3. Feeding regimes applied (x 106 cells per Artemia individual)
age of Artemia (days)
1
1/2
1/4
1/8
1/16
1
0.250
0.125
0.065
0.033
0.016
2,3,4
0.500
0.250
0.125
0.065
0.031
5,6
0.750
0.380
0.190
0.095
0.048
7
0.990
0.500
0.250
0.125
0.063
8
1.260
0.630
0.315
0.158
1.079
9
2.040
1.020
0.510
0.255
0.128
10,11
2.400
1.200
0.600
0.300
0.150
12,13
3.000
1.500
0.750
0.375
0.188
14,15
3.600
1.800
0.900
0.450
0.225
16,17
4.200
2.100
1.050
0.525
0.263
18,19
5.100
2.550
1.275
0.638
0.319
from 20 onwards
6.000
3.000
1.500
0.750
0.375
2.4.2.4. Experimental procedure
The experimental procedure implied hatching of the cysts in standard hatching conditions (Lavens and
Sorgeloos, 1996) and harvesting the nauplii after 24 hours. For each treatment (see further), 3
replicates of 500 instar I nauplii, incubated in one-liter culture cones were set up. Culture temperature
was 22.0 or 28.0 ± 0.5°C. Culture medium was Instant Ocean  artificial seawater solution (renewed
3 times weekly) of 60 or 120 g/l. Survival was monitored at each water renewal. At sexual maturity
50 specimens were isolated individually in 50 ml Falcon tubes (kept at the same salinity, temperature,
feeding regime as before isolation). At each water renewal (3 times weekly) survival and fecundity
(offspring per brood, number of broods per female, total offspring per female, percentage encystment)
were observed until day 54 post-hatching.
2.4.2.5. Experimental treatments
For the combinations of culture salinity, temperature and feeding conditions (phytoplankton species +
feeding regime), see Table 2.4.
Table 2.4. Experimental treatments
Algal species Feeding regime
Tetraselmis
Nautococcus
Oocystis
1
1/2
1/4
1/8
1/16
1
1/2
1/4
1/8
1/16
1
60 g/l
x
x
-
culture temperature and salinity
22°C
28°C
120 g/l
60 g/l
x
x
x
x
x
x
x
x
x
x
x
x
x
22
120 g/l
x
x
x
x
x
x
x
x
1/2
1/4
1/8
1/16
-
x
-
x
x
x
x
x
-
2.4.3. Results and discussion
2.4.3.1. Survival
Figs. 2.5, 2.6 and 2.7 show the survival of the culture tests run with Tetraselmis, Nautococcus and
Oocystis, respectively. Results are shown for survival before (average of 3 cones) and after isolation
(Figures A and B, respectively); red lines correspond with 28°C; blue lines with 22°C; full lines with
120 g/l and dotted lines with 60 g/l; different symbols correspond with different cell densities.
Tetraselmis
100
Fig. 2.5.A. Tetraselmis before isolation
80
120 g/l - 1 - 28°C
survival (%)
120 g/l - ½ - 28°C
60
60 g/l - 1 - 28°C
60 g/l - ½ - 28°C
120 g/l - 1 - 22°C
40
120 g/l - ½ - 22°C
60 g/l - 1 - 22°C
60 g/l - ½ - 22°C
20
age (days)
0
0
5
10
15
23
20
100
Fig. 2.5.B.Tetraselmis after isolation
80
120 g/l - 1 - 28°C
120 g/l - ½ - 28°C
survival (%)
60 g/l - 1 - 28°C
60
60 g/l - ½ - 28°C
120 g/l - 1 - 22°C
120 g/l - ½ - 22°C
60 g/l - 1- 22°C
60 g/l - ½ - 22°C
40
age (days)
20
20
25
30
35
40
45
50
55
60
Fig. 2.5. Survival of Aral Sea Artemia on Tetraselmis diet, using different feeding rates and
culture temperature and salinity before (2.5.A) and after (2.5.B) isolation of the individuals in
separate culture vials
Prior to isolation, highest mortality occurred within the first 10 days of culture. Final survival was
higher at 120 g/l (range 20-29 %) than at 60 g/l (range 5-11 %). There were no clear trends for the
other parameters. After isolation, survival generally gradually decreased to values inbetween 76-78 %
(60 g/l – feeding ‘1’ – 22°C and 60 g/l – feeding ‘1/2’ – 28°C) and 54-56 % (120 g/l – ½ -22°C; 120
g/l – 1 – 28°C). Again no clear trends showed up.
Nautococcus
24
100
Fig. 2.6.A. Nautococcus before isolation
120 g/l – 1 - 28°C
80
120 g/l – ½ - 28°C
120 g/l – ¼ - 28°C
120 g/l – 1/8 - 28°C
survival (%)
60
120 g/l – 1/16 - 28°C
60 g/l - 1 - 28°C
60 g/l – ½ - 28°C
60 g/l – ¼ - 28°C
40
60 g/l – 1/8 - 28°C
60 g/l – 1/16 - 28°C
120 g/l – 1 - 22°C
20
120 g/l – ½ - 22°C
age (days)
0
0
5
10
15
20
100
Fig. 2.6.B. Nautococcus after isolation
120 g/l – 1 - 28°C
80
120 g/l – ½ - 28°C
120 g/l – ¼ - 28°C
survival (%)
120 g/l – 1/8 - 28°C
120 g/l – 1/16 - 28°C
60 g/l – 1 - 28°C
60
60 g/l – ½ - 28°C
60 g/l – ¼ - 28°C
60 g/l – 1/8 - 28°C
60 g/l – 1/16 - 28°C
40
120 g/l – 1 - 22°C
120 g/l – ½ - 22°C
age (days)
20
20
25
30
35
40
45
50
55
60
Fig. 2.6. Survival of Aral Sea Artemia on Nautococcus diet, using different feeding rates and culture
temperature and salinity before (2.6.A) and after (2.6.B) isolation of the individuals in separate culture
vials
Survival before isolation was often higher when Nautococcus had been fed, as compared to
Tetraselmis (survival values inbetween 12 and 73 %). There was a general trend for lower survival
when lower feeding rates were applied, both at 120 g/l and at 60 g/l. After isolation final survival
varied between 36 and 90 %. During this culture phase, however, lower feeding rates resulted in
higher final survival for 60 g/l, whereas there was no link between both factors at 120 g/l.. None of
25
both salinities performed better than the other.
Oocystis
100
Fig. 2.7.A. Oocystis before isolation
80
120 g/l - 1 - 28°C
120 g/l - ½ - 28°C
survival
(%)
60
60 g/l - 1 - 28°C
60 g/l - ½ - 28°C
60 g/l – ¼ - 28°C
60 g/l – 1/8 - 28°C
40
60 g/l – 1/16 - 28°C
120 g/l - 1 - 22°C
120 g/l - ½ - 22°C
20
age (days)
0
0
5
10
15
20
25
100
Fig. 2.7.B. Oocystis after isolation
80
120 g/l - 1 - 28°C
120 g/l - ½ - 28°C
survival (%)
60 g/l - 1 - 28°C
60 g/l - ½ - 28°C
60
60 g/l – ¼ - 28°C
60 g/l – 1/8 -28°C
60 g/l – 1/16 - 28°C
120 g/l - 1 - 22°C
40
120 g/l - ½ - 22°C
age (days)
20
20
25
30
35
40
45
50
55
60
Fig. 2.7.. Survival of Aral Sea Artemia on Oocystis diet, using different feeding rates and culture
temperature and salinity before (2.7.A) and after (2.7.B) isolation of the individuals in separate
culture vials
When Oocystis was fed, survival before isolation ranged between 73 (120 g/l – 1 – 22°C) and 32%
26
(120 g/l – 1 – 28°C) (and was thus generally better than when Tetraselmis was fed). Lower
temperature gave higher survival than higher salinity, and, as in Nautococcus lower feeding rates
resulted in lower survival. After isolation, final survival ranged between 58 (60 g/l – 1/16 – 28°C) and
34% (120 g.l – 1 – 22°C). In contrast to the pre-isolation culture phase, there was no decreasing
survival with decreasing feeding rate (as in Nautococcus, there was rather an opposite trend at 60 g/l).
2.4.3.2. Fecundity parameters
Fecundity was expressed as average brood size (Fig. 2.8), average number of broods per female (Fig.
2.9), average total individual offspring (Fig. 2.10) and percentage encystment (Fig. 2.11) for the 50
females, as recorded over the post-isolation period.
Fig. 2.8. average brood size
Tetraselmis
Nautococcus
Oocystis
160
140
120
100
80
60
40
20
0
C
C
C
C
C
C
C
C
C
C
C
C
°C 2°C
8° 28 ° 28 ° 2 8° 28°
2° 22 °
8 ° 28° 28° 28 ° 2 8°
22
2
-2
-2
–
–
1
1
8
6
6
1
8
½
¼
1
½
¼
/
½
/
1
1
1
1
½
l – /l –
l – /l –
l – /l –
1/
1/
l–
l–
l–
–
–
–
g/
g/
g/
–
–
g
g/ g/l
g/ g/l
g
g / g /l
g
l
l
0
0
/
0
/
0
0
0
6
g
g
60
60 60
60 60
12 12
12 12
12 12 0 2 0
60
1
-2
culture conditions
Fig. 2.8. Average brood size of reproducing Aral Sea Artemia, cultured until the age of 54 days in
different feeding, salinity and temperature conditions; bars (with standard error bars) represent
average of values of 50 individual females
27
Fig 2.9. average individual number of broods
Tetraselmis
Nautococcus
Oocystis
10
8
6
4
2
12
0
g
12 /l –
1
0
g/
l – 28
12
½ °C
0
g/
l – 28
12
0
¼ °C
g
12 /l – - 28
0
1/
°C
g/
8
l–
-2
8
1/
16 °C
-2
8°
C
60
g/
l
60 – 1
g/
l – 28
60
½ °C
-2
g/
60 l – ¼ 8°C
g/
60 l – 1 28
°C
/
g/
l– 81 / 28 °
16
C
-2
8°
12
C
0
g/
12 l –
1
0
g/
l – 22
½ °C
-2
2°
C
60
g/
60 l –
g/ 1 –
l–
2
½ 2 °C
–
22
°C
0
culture conditions
Fig. 2.9. Average individual number of broods of reproducing Aral Sea Artemia, cultured until the age
of 54 days in different feeding, salinity and temperature conditions; bars (with standard error bars)
represent average of values of 50 individual females
Fig. 2.10. average total individual offspring
Tetraselmis
Nautococcus
Oocystis
1200
1000
800
600
400
200
12
0
12
0
g/
l–
1
-2
g/
l
8
–
12
½ °C
0
g
12 /l – 28 °
C
0
¼
g
12 /l – - 28
°C
0
1/
g/
8
l–
-2
8
1/
16 °C
-2
8°
C
60
g/
l
60 – 1
g/
l – 28
60
½ °C
g/
l – 28°
60
C
¼
g/
l – - 28
60
°C
1/
g/
8
l–
-2
8
1/
1 6 °C
-2
8°
12
C
0
g/
12 l –
1
0
g/
l – 22
½ °C
-2
2°
C
60
g/
l
–
60
g/ 1 –
l–
2
½ 2 °C
–
22
°C
0
culture conditions
Fig. 2.10. Average (total individual offspring of reproducing Aral Sea Artemia, cultured until the age
of 54 days in different feeding, salinity and temperature conditions; bars (with standard error bars)
represent average of values of 50 individual females
28
Fig. 2.11. % encystment
Tetraselmis
Nautococcus
Oocystis
100
80
60
40
20
12
0
g
12 /l –
1
0
-2
g/
l
8
–
12
½ °C
0
g
12 /l – 28 °
C
0
¼
g
12 /l – - 28
°C
0
1/
g/
8
l–
-2
8
1/
16 °C
-2
8°
C
60
g/
l
60 – 1
g/
l – 28
60
½ °C
g/
l – 28°
60
C
¼
g/
l – - 28
60
°C
1/
g/
8
l–
-2
8
1/
1 6 °C
-2
8°
12
C
0
g/
12 l –
1
0
g/
l – 22
½ °C
-2
2°
C
60
g/
60 l – 1
g/
l – – 22
½
°C
–
22
°C
0
culture conditions
Fig. 2.11. Percentage encystment of reproducing Aral Sea Artemia, cultured until the age of 54 days
in different feeding, salinity and temperature conditions; bars represent overall value for 50 individual
females
The following general conclusions can be drawn from the fecundity tests (Figs. 2.8. to 2.11):

Effect of temperature: for those treatments where comparison is possible, culture at 22°C resulted
in a a signifcantly higher average brood size, but significantly lower number of broods (p <
0.001), as compared to 28°C. No significant difference between both temperatures was found for
the total number of offspring;

Effect of salinty: where comparison is possible, lower culture salinity results in signifcantly
higher (p< 0.001) fecundity (average brood size, number of broods and total number of offspring),
which can be explained by the higher metabolic (osmoregulatory) costs inherent to a higher
salinity environment;

Effect of algal species: when comparison is possible, higher fecundity values (quantified as
average brood size, individual number of broods and total individual offspring) are obtained by
feeding Nautococcus then by feeding Tetraselmis. Fecundity values obtained with Oocystis are
more variable, and can be superior to Nautococcus, inferior to Tetraselmis, or inbetween both
species depending on the culture conditions (temperature and salinity), feeding rate and parameter
studied;

Effect of feeding rate: fecundity is negatively affected by lower feeding rates; from feeding rate ¼
onwards, a negative effect is observed on average brood size, number of broods and total number
of offspring (p < 0.001) when feeding Nautococcus and Oocystis (low feeding rates were not
applied for Tetraselmis). This trend is thus observed in spite of the considerable size differences
between both algae species;

Oviparity: higher ambient salinity during reproduction results in higher values for oviparity
(which is generally linked with the degree of unstability or stressfulness of the environment).
Other factors (higher or lower feeding rates, higher or lower temperature) did not result in a clear
29
trend towards either increased or decreased oviparity.
The individual cellular dry weight (Table 2.1.) of the algal species used, allows to compare the
fecundity parameters based as a function of the algal biomass supplied to each Artemia individual,
where comparison is possible. At 60 g/l salinity and 28°C culture temperature (comparison for
Nautococcus and Oocystis for 5 different feeding rates) feeding results in a higher brood size for
Nautococcus (Fig. 2.12 A) and a similar number of broods (Fig. 2.12 B) than Oocystis, resulting in a
higher total individual offspring for the former (Fig. 2.12 C), with for both species decreased values at
the lower end of the feeding range. Percentage encystment shows no clear link with feeding rate or
algal species (Fig. 2.12 D).
Nautococcus
Oocystis
Nautococcus
120
80
40
0
0
0.5
1
1.5
8
6
4
2
0
2
0
feeding rate as a function of algal biomass
0.5
1
1.5
2
feeding rate as a function of algal biomass
A. average brood size
B. average number of broods
Nautococcus
Nautococcus
Oocystis
1200
70
1000
60
% encystment
total individual offspring
Oocystis
10
average number of broods
average brood size
160
800
600
400
200
Oocystis
50
40
30
20
10
0
0
0
0.5
1
1.5
2
0
feeding rate as a function of algal biomass
0.5
1
1.5
2
feeding rate as a function of algal biomass
C. total individual offspring
D. % encystment
Fig. 2.12. Fecundity parameters (A. average brood size, B. average number of broods, C. total
individual offpsring, D. % encystment) as a function of feeding rate, taking into account algal biomass
(feeding rates are presented relative to ‘1’, i.e. feeding rate ‘1’ using Tetraselmis)
Comparison for the three algal species is only possible for 2 feeding rates, at three salinitytemperature combinations (Fig. 2.13; only values for total individual offspring are shown).
Tetraselmis results in the lowest total individual offspring, whereas the combination 60 g/l-28°C gives
the highest fecundity of the salinity-temperature combinations where comparison is possible.
30
60 g/l 28°C
120 g/l - 28°C
1200
1200
1000
total individual offspring
total individual offspring
1000
800
Tetraselmis
600
Nautococcus
Oocystis
400
200
800
Tetraselmis
600
Nautococcus
Oocystis
400
200
0
0
0
0.5
1
1.5
2
0
feeding rate as a function of algal biomass
0.5
1
1.5
2
feeding rate as a function of algal biomass
A) 120 g/l-28°C
B) 60 g/l-28°C
120 g/l - 22°C
1200
total individual offspring
1000
800
Tetraselmis
600
Nautococcus
Oocystis
400
200
0
0
0.5
1
1.5
2
feeding rate as a function of algal biomass
C) 120 g/l-22°C
Fig. 2.13. Total individual offspring as a function of feeding rate, taking into account algal biomass
(feeding rates are presented relative to ‘1’, i.e. feeding rate ‘1’ using Tetraselmis) for the three algal
species tested, and at three salinity-temperature combinations: A) 120 g/l-28°C; B) 60 g/l-28°C; C)
120 g/l-22°C
2.4.4. Conclusions still to be worked out further once I have some insight into the field data
The objectives of these laboratory tests were
a) to provide information about the usability of Aral Sea Artemia strain for larviculture
applications;
b) to provide background information on the reproductive potential of the strain in controlled
laboratory conditions, and to mirror these to the data recorded in the field.
The results show that the parthenogenetic Artema strain is comparable with the commercially
important A. franciscana Great Salt Lake strain in terms of cyst biometrics, HUFA levels and
accumulation of HUFA’s post-enrichment. As samples of diapausing cysts could not be obtained, no
information could be generated on the diapause characteristics (and techniques needed to terminate
diapause) for this strain.
The laboratory culture tests show a clear effect of the feeding conditions (phytoplankton species and
feeding rate) on fecundity. The situation in the field (where Artemia feeds on a variety of
phytoplankton species, on bacterial conglomerates and on organic detritus particles) is much more
complex than the highly simplified laboratory trophic tests. Nevertheless it is clear that two of the
common algal species in the Aral (Nautococcus sp. and Oocystis sp.), when fed as sole diet, result in a
reproductive output of the Artemia population surpassing the laboratory reference Tetraselmis diet. It
is equally clear that there is a pronounced feeding rate effect on fecundity. Comparing the labotory
data with the data from the field, it is clear that the Artemia population in the Aral Sea is living and
reproducing at concentrations of algal cells below the optimal concentration.
31
2.5. Predation by Aral Sea copepods on Artemia nauplii
2.5.1. Objectives
Two series experiments with Arctodiaptomus salinus (Copepoda, Calanoida) and Apocyclops
dengizicus (Copepoda, Cyclopoida) were conducted. Both species were found co-occuring with
Artemia in the Aral Sea. The aim of these tests was to evaluate the predation pressure exerted by these
copepods on the Artemia population.
2.5.2. Material and methods.
Artemia cysts (collected in the Aral Sea) were incubated in standard hatching conditions (Lavens and
Sorgeloos, 1996) for 24 h. Fifty instar I nauplii were introduced into a Petri dish filled with Aral Sea
water diluted to 35 g/l salinity. One adult female of Arctodiaptomus salinus or Apocyclops dengizicus
was added to each Petri dish. The experiments were run at a temperature of 24-26°C. The test was run
in 10 replicates for each copepod species; after 24 h all Petri dishes were checked for the presence of
Artemia individuals.
2.5.3. Results and conclusion
Apparently no consumption of Artemia nauplii was observed during these experiments. The used
methodology is identical with the methodology used by LIH for the estimation of the consumption of
larvae of Culex pipiens (Diptera, Culicidae). According to the (limited) set-up of this experiment, A.
salinus and A. dengizicus can thus not be considered as major predators on Artemia.
References
Chepurnov, V.A., Mann, D.G., von Dassow, P., Vanormelingen, P., Gillard, J., Inzé, D., Sabbe, K.,
Vyverman, W. 2008. In search of new tractable diatoms for experimental biology. BioEssays, in
press.
Coutteau, P., Brendonck, L., Lavens, P., Sorgeloos, P. 1992 The use of manipulated baker's yeast as
an algal substitute for laboratory culture of Anostraca. Hydrobiologia 234:25-32.
Han, K., Geurden, I., Sorgeloos, P. 2000. Comparison of docosahexaenoic acid (22:6n-3) levels in
various Artemia strains during enrichment and subsequent starvation. J. World Aq. Soc. 31: 469-475.
Han, K., Geurden, I.,, Sorgeloos, P. 2001. Fatty acid changes in enriched and subsequently starved
Artemia franciscana nauplii enriched with different essential fatty acids. Aquaculture 199: 93-105.
Lavens, P., Sorgeloos, P., 1996. Manual on the production and use of live food for aquaculture. FAO
Fisheries Technical Paper No. 361, 295 p.
Vanhaecke, P., Sorgeloos, P. 1980. International Study on Artemia. IV. The biometrics of Artemia
strains from different geographical origin: p. 393-405. In: The brine shrimp Artemia. Vol. 3. Ecology,
culturing, use in aquaculture. Persoone, G.; Sorgeloos, P.; Roels, 0.; Jaspers, E. (Eds). Universa Press,
Wetteren, Belgium, 456 p.
32
5. IMPLEMENTATION OF RESULTS
(2-4 pages)
This chapter gives a status of the implementation of the results at the end of the Project and of
the expectations for the immediate and longer term future. Special attention should be paid to
the economic and industrial benefits (short and long term) for the country. Wherever possible,
a cost-benefit analysis, based on the R&D findings, should be included (in EUR).
33
6. CONCLUSIONS
(1-2 pages)
Here, the overall conclusions of the SfP experience are given. The tangible consequences of
NATO's funding in EUR of the Project for the research team, the participating institutions and, if
appropriate, the nation, should be outlined.
34
ANNEXES
Annex 1: List of collaborators (internal and external); enumerate under separate headings the
collaborators who obtained an advanced degree (MSc, PhD) through co-operation
with the Project.
Project directors
Patrick Sorgeloos
(NPD)
Iskandar Mirabdullayev (PPD)
Co-directors
Thomas Bosteels
Brad Marden
Gilbert Van Stappen
Wim Vyverman
Iliya Zholdasova
Young Uzbek scientists
Lola Abdullayeva
Nodira Jumaniezova
Ablatdyin Musaev
Zuri Mustafaeva
Svetlana Lyubimova
Marina Oryol
Rahmetulla Temirbekov
Other NATO scientists involved
Victor Chepurnov
External NATO advisor
Shane Bradt
End users
Haitmurat Abdurakhmanov
A. Abdusattarov
Ubbiniyaz Ashirbekov
Polat Reimov
Jarilkap Tursinbekov
Rukhulla Kurbanov
Ubaidulla Nudirkhanov
 Laboratory of Aquaculture & Artemia Reference Center,
Ghent University, Rozier 44, B9000 Gent, Belgium (ARC)
 Laboratory of Ichthyology and Hydrobiology, Institute of
Zoology, Uzbek Academy of Sciences, Niyazov st. 1,
Tashkent, 700095, Uzbekistan (LIH)
 Great Salt Lake Artemia Cooperative, 5859
N. Cottonwood Canyon Rd. Mt. Green, UT, USA
 INVE Technologies, Artemia Task Force, PO Box 1306,
598 W Clark St. Grantsville, UT, USA
 Laboratory of Aquaculture & Artemia Reference Center,
Ghent University, Rozier 44, B9000 Gent, Belgium
 Laboratory of Protistology & Aquatic Ecology, Department
of Biology, Ghent University, Krijgslaan, 281-58, B9000
Gent, Belgium (LPAE)
 Institute of Bioecology of the Karakalpak Branch of the
Uzbek Academy of Sciences; 41, Berdakh prospect,
742000, Nukus, Uzbekistan (IB)
LIH
LIH
IB
LIH
IB
IB
IB
LPAE
University of New Hampshire, Durham, New Hampshire,
USA
 Ministry of Economy, Republic of Karakalpakstan, 97
Dosnazarov St., 97 Nukus, 742000, Uzbekistan
 Scientific Research Center for Development of Fisheries,
Chilanzar 10, 700123 Tashkent, Uzbekistan
 International Fund of Aral Rescue, Doslik guzari 111,
Nukus, 742000, Karakalpakstan, Uzbekistan
 Goskompriroda-State Committee for Nature Protection of
Karakalpakstan, 742000, Nukus Ul. Berdakh Gozary,
Uzbekistan
 Community of Moinaq, Hakimiyat of Moinaq district,
Republic Karakalpakstan, Moinaq 743500, Uzbekistan
 Scientific Research Center for Development of Fisheries,
Tashkent, Uzbekistan
 International Fund of the Aral Sea, GEF Agency, Tashkent,
35
Nikolai Gorelkin
Fakhritdin Shaunsiev
Nagmet Aimbetov
Bakhretdin Muradov
Vladislav Talskikh
Uzbekistan
 Hydrological Center, Cabinet of Ministers, Tashkent,
Uzbekistan
 UNDP Project on Aral Sea
 Academy of Science Uzbekistan, Karakalpak Branch,
Nukus
 Ministry of Economy Uzbekistan, Tashkent
 Hydrometeorological Center of Uzbekistan, Tashkent
36
Annex 2:
List of publications resulting from the Project. List of presentations given to the
scientific community during major conferences, as well as to the broader society via the press (attach
photocopies of articles) and other media..
Publications What tentative titles can we further list here ????
Chepurnov, V.A., Mann, D.G., von Dassow, P., Vanormelingen, P., Gillard, J., Inzé, D., Sabbe, K.,
Vyverman, W. 2008. In search of new tractable diatoms for experimental biology. BioEssays, in
press.
Mirabdullayev I.M., Musaev A., Jumaniezova N.I. 2006. New data on zooplankton of the Aral Sea. –
Proceedings of the Uzbekistan Research Centre on Fishery Development, Tashkent, Uzbekistan 110115 (in Russian)
Musaev, A., Hankuliev, K., Marden, B., Jumaniezova, N.I., Abdullaeva L.I.,, Mirabdullaev, I.M.
2005. Dynamics of the Artemia population in the Aral Sea. Biodiversity of Uzbekistan – monitoring
and exploitation (in Russian), p. 179-179.
Van Stappen, G., Marden, B., Abullaeva, L., Musaev, A., Mirabdullaev, I.M., Sorgeloos, P. Survival
and fecundity of parthenogenetic Artemia from the Aral Sea, Uzbekistan, as a function of culture
temperature and salinity and feeding conditions. In prep.
Presentations at conferences
Abdullayeva L.N., Khalilov S. 2006. Development of green algae Nautococcus grandis Korsh
(Chlorophyta) from the Aral Sea in laboratory culture. – In: Current problems in biology, ecology and
soil sciences. Conference of the National University of Uzbekistan, Tashkent, Uzbekistan, 17-18
November, Book of Abstracts p. 6-7. (in Russian).
Abdullayeva L.N., Mahieu C.., Van Stappen G.. 2006. Peculiarities of development of Artemia
parthenogenetica from the Aral Sea in experimental conditions. – In: Current problems in biology,
ecology and soil sciences. Conference of the National University of Uzbekistan, Tashkent,
Uzbekistan. Book of Abstracts p. 50-51 (in Russian).
Hankuliev, K. 2008. Comparative changes in the hydrochemistry of the Aral Sea, Uzbekistan (20022007) and Karabogaz-Gol, Turkmenistan (2000-2002) and th corresponding alterations in resident
biota. 10th International Confernce on Salt Lake Research. Salt Lake City, USA, May 12-14.
Jumaniezova N.I. 2006. Biological characteristics of Artemia parthenogenetica of the Aral Sea. –
International Conference on Problems of rational use and conservation of biological resources of
Southern Aral Sea area. Nukus, Uzbekistan, May 16-17. Book of Abstracts p. 115.
Jumaniezova N.I., Mirabdullayev I.M. 2006. Growth, development, survival and fecundity of Artemia
parthenogenetica of the Aral Sea – Proceedings of the Ferghana State University, Supplement, p. 83
(in Russian).
Mirabdullayev I., Abdullayeva L., Musaev A. 2007. Succession of zooplankton in the Aral Sea. –
Human and climate forcing of zooplankton populations. 4th International Zooplankton Symposium. 28
May-1 June, Hiroshima, Japan. Book of Abstracts, p. 216.
Mirabdullayev I.M., Taizhanov E.B., Kuzmetov A.R., Urazova R.S. 2006. Aral Sea and problems of
increasing of productivity in terminal irrigation lakes - In: Materials of the International Scientific
Practical Conference ‘Urgent problems of ecology and nature use in Kazakhstan and adjacent
territories”, Pavlodar, Kazakhstan, May 25-26, p. 345-347 (in Russian).
37
Mirabdullayev I.M., Urazova R.S., Taizhanov E.B. 2006. Use of Aral Sea hydrofauna for increasing
of bioproductivity of waterbodies in Central Asia. – In: International Conference on Problems of
rational use and conservation of biological resources of Southern Aral Sea area. Nukus, Uzbekistan,
May 16-17, Book of Abstracts p. 16 (in Russian).
Musaev A., Abdullayeva L.N. 2006. Influence of food concentration, salinity and temperature on
survival of Artemia from the Aral Sea. – In: International Conference on Problems of rational use and
conservation of biological resources of Southern Aral Sea area. Nukus, Uzbekistan, May 16-17, Book
of Abstracts p. 117 (in Russian).
Musaev A., Abdullayeva L.N., Jumaniezova N.I., Mirabdullayev I.M. 2007. Succession of
zooplankton of the Aral Sea. – In: Conference on Problems of rational use of natural resources of
Southern Aral Sea area. Nukus, Uzbekistan. Book of Abstracts p. 27 (in Russian).
Mustafaeva Z.A., Abdullayeva L.N. 2007. New data on phytoplankton of the Aral Sea. – In: Biology
of Inland waters. Reports of the XIII International Youth conference, Borok, Russia (in Russian), 2326 October, p. 45 (in Russian).
Mustafaeva Z.A., Chepurnov V.A. 2006. К вопросу о систематическом положении диатомей
родов Cymbella и Encyonema – In: Conference of the National University of Uzbekistan Current
problems in biology, ecology and soil sciences. Tashkent, Uzbekistan, 17-18 November, Book of
Abstracts p. 25 (in Russian).
Mustafaeva Z.A., Abdullayeva L.N., Chepurnov V.A., Mirabdullayev I.M. 2006. Changes in
plankton of the Aral Sea in conditions of progressing salinization – Proceedings of the Ferghana State
University. Supplement, p. 79-80 (in Russian).
Urazova R.S., Mirabdullayev I.M. Materials to revision of the genus Moina Baird, 1850 (Crustacea,
Cladocera) in Uzbekistan. 2006. Conference of the Mamun Academy. Khiva, Uzbekistan, p. 189-191
(in Russian).
Van Stappen, G. 2008. Artemia of the Aral Sea, Uzbekistan. Field survey of an emerging population
and perspectives for commercial exploitation. 10th International Confernce on Salt Lake Research.
Salt Lake City, USA, May 12-14.
MSc Thesis
Achom, H.O. 2008. Characterization of the parthenogenetic Artemia population of the Aral Sea,
Uzbekistan. MSc Thesis, Laboratory of Aquaculture & Artemia Reference Center, Ghent University,
Belgium
38
Annex 3:
Complete Inventory Record
Serial Number
Date of
Purchase
Cost
USD
Cost
EUR
Location
TRX 500 FM
TRX 500 FM
TRX 500 FM
TRX 500 FM
IHFTE310854000655
20-Jun-05
IHFTE310754002204
20-Jun-05
IHFTE310554000676
20-Jun-05
IHFTE310154001128
20-Jun-05
5944,00
5944,00
5944,00
5944,00
4755.20
4755.20
4755.20
4755.20
IB
IB
IB
IB
YSI
4JE-221297
still to be registered
27-Jul-05
1536,00
1228.80
IB
YSI
4JE-221298
still to be registered
18-Aug-05
1674,00
1339.20
IB
Meiji
A-4802-00
still to be registered
7-Jul-05
1995,00
1596.00
LIH
Meiji
A-48401-15
still to be registered
7-Jul-05
1910,00
1528.00
IB
2138.64
2138.64
IB
IB
Inventory
Label No.
No. A-0180
No. A-0181
No. A-0182
No. A-0183
Property Item
Honda ATV
Honda ATV
Honda ATV
Honda ATV
Manufacturer
Honda
Honda
Honda
Honda
No. A-0184
YSI Model 85 Oxygen, Salinity,
Conductivity
No. A-0185
& Temperature Meter (50 ft cable)
YSI Model 85 Oxygen, Salinity,
Conductivity
Model
Number
& Temperature Meter (100 ft Cable)
No. A-0186
Meiji Stereozoom Microscope System
(7x to 45x zoom)(230VAC)
No. A-0187
Meiji Professional Trinocular Compound
Microscope with achromatic objective
No. A-0188
No. A-0189
Caribe Rigid Inflatable Vessel
Caribe Rigid Inflatable Vessel
Caribe Inflatables
C12/Light Grey
EMD 12136G506
25-Jul-05
Caribe Inflatables
C12/Light Grey
25-Jul-05
No. A-0176
DELL Notebook Computer
DELL Computers
Latitude X1
1-Sep-05
1839,83
1471.86
LIH
No. A-0177
DELL Notebook Computer
DELL Computers
Latitude X1
1-Sep-05
1839,83
1471.86
LIH
No. A-0178
DELL Notebook Computer
DELL Computers
Latitude X1
31-Aug-05
1839,83
1471.86
IB
No. A-0179
DELL Notebook Computer
DELL Computers
Latitude X1
EMD 12137G506
CN-0P8056-3652158J-009A
CN-0P8056-3652158J-0060
CN-0P8056-3652158J-008D
CN-0P8056-3652158J-0099
2673,30
2673,30
31-Aug-05
1839,83
1471.86
IB
39
SUMMARY REPORT OF THE FINAL REPORT
NPD: Patrick Sorgeloos, Laboratory of Aquaculture & Artemia Reference Center, Ghent University,
Belgium
PPD: Iskandar Mirabdullayev, Laboratory of Ichthyology and Hydrobiology, Institute of Zoology,
Uzbek Academy of Sciences, Tashkent, Uzbekistan
1. Background and objectives
The ecological devastation and economic impoverishment of the Aral Sea communities are threats to
the security of the region and to the health and well-being of the people. There is an immediate need
for the communities surrounding the Aral Sea to improve their quality of life through economic
opportunities, to compensate for the loss of fish stocks upon which they formerly depended for their
sustenance and employment. There is evidence of a recently colonizing population of the halophilic
brine shrimp Artemia in the Aral Sea, that holds promise as a commercially viable resource: this
crustacean is an essential and highly priced live food item in marine aquaculture worldwide. The
development and prudent management of an Aral Sea Artemia resource could provide some degree of
economic and ecological recovery to the region through sustainable management strategies. However,
it is insufficiently known whether the current hydrobiological and hydrochemical status and primary
productivity of the Aral Sea are sufficient to support a stable Artemia population. Detailed ecological
information is needed in order to evaluate the feasibility and potential benefits of commercial
exploitation of Artemia. Expert advice in harvesting, transport, handling, storage, and industrial
production techniques are additionally essential for the successful exploitation of this resource.
This project consequently aimed at providing the partner institutes and end-users with information on
possible benefits and outcomes extending beyond the project’s duration: a) characterizing the Aral
Sea Artemia population in terms of life history, population dynamics (and ecological conditons
affecting them), cyst quality characteristics and potential for aquaculture; b) pending the outcome of
(a) development of a population model for Aral Artemia resource and issue recommendations on
optimal sustainable resource management; and c) transfer of technology (field and laboratory
equipment) and know-how (extensive training programme).
2. Methodology
2.1. Field research
 Design of appropriate sampling campaign covering East and West Aral: sampling sites located
along transsects; sampling frequency at least once a month in period April-October;
 Sampling for hydrochemistry (nutrients), other abiotic factors, phytoplankton composition and
density, Artemia population composition and density, other zooplanktonic organisms.
2.2. Laboratory studies
 phytoplankton:, isolation, determination and culture of Aral Sea algal species at different abiotic
conditions;
 Artemia: strain characteristics and cyst quality control: hatching and diapause behaviour,
biometrics, nutritional profile; life history (growth, survival) and reproductive characteristics:
effect of abiotic conditions temperature, salinity, feeding.
2.3. Training program for partner country (young) scientists
Terms of reference: a) Ghent University, Belgium: algal isolation, determination and culture;
hydrochemical analyis; Artemia biology and quality control; theory on harvesting and processing of
Artemia cysts; b) INVE Thailand, Bangkok: Artemia cyst processing and storage; use of Artemia
biomass in shrimp culture; c) Great Salt Lake Artemia Cooperative, Mountain Green, USA:
industrial-scale harvesting and processing of Artemia cysts; Artemia resources management and
modeling; d) at partner institutes in Uzbekistan: hands-on training in field sampling, sampling
analysis and data processing.
40
3. Results
3.1. Field research
The project was successful in covering the designed sampling programme (see picture) in the seasons
2005, 2006 and 2007, in the period March/April-October/November -depending on weather
conditions. An additional sampling
expedition was organized in December
2007 focusing on the nutrient contents of
West Aral in deeper water layers. As
compared to the start of the project,
salinity in the West Aral has slowly
increased from 70-80 to 80-90 ppt, and in
the East Aral (with some occasional
salinity drops due to increased freshwater
inflow) from 90-100 to 120-130 ppt. This
moderate salinity increase has, however,
major consequences for the East Aral in
terms of its potential for Artemia
exploitation. Though this water body
initially seemed more promising than the
West Aral, thanks to its higher salinity and
its relatively higher nutrient status (occasional Amu Darya water inflow), the further drop of the water
level in the period 2005-2007 and the retreat of the shoreline have turned nearly the entire East Aral’s
perimeter (on Uzbek territory and as far as publicly accessible) into extended mudflats practically
inaccessible for any major vehicle (such as vessels or trucks for harvesting resp. transport of cysts).
Moreover, the remaining water body, though still relatively large in surface, is basically a shallow
(1.5- 2 m), nutrient-poor environment with low densities of Artemia. The West Aral, on the other
hand, though more easily accessible (at least along its western shore) shows even lower nutrient
contents, including in deeper water layers, and consequently very low phytoplankton and Artemia
densities. The continuing uncertain situation regarding the future water management of the tributary
rivers (a complex problem with many stakeholders involved – also at the international level) is a
fundamental problem for any conceivable Aral Artemia exploitation plan. As there are no concrete
short or medium-term indications that these basic impediments for Artemia exploitation are likely to
alter in a positive sense, a commercially viable Artemia industry at West or East Aral thus seems
unlikely in the foreseeable future.
3.2. Laboratory studies
Laboratory studies were conducted as planned and show that the characteristics of the parthenogenetic
Aral Sea Artemia strain, relevant for larviculture application (e.g. cyst biometrics, nutritional
composition, diapause characteristics) are comparable to the Great Salt Lake strain. Reproduction
tests show that the reproduction in the field is clearly negatively affected by the ambient low food
levels. Nevertheless, even in optimal (laboratory) food conditions the reproductive capacity of the
Aral Artemia strain is lower than in the commercially used A. franciscana.
3.3. Training program
The training programme was executed as designed.
4. Implementation
The data generated by the project have general generic value for possible Artemia exploitation issues
and opportunities elsewhere in Uzbekistan. Further, implementation of the project’s results is ensured
through the following channels:
a) organization of workshops: 2 workshops for the end-users have been organized (i.e. one more
than foreseen in the project proposal. The first workshop, organized November 7-8, 2005,
Tashkent, aimed at informing the end-users about the goals and methodology of the project, and
41
b)
c)
d)
e)
at providing a forum for end-users feedback. The concluding workshop, organized November 5-6,
2007, sketched the overall results of the project to the end-users and drew outlines for future
Artemia-related activities in Uzbekistan;
development of critical mass of Artemia expertise and know-how at Uzbek partners institutes;
purchase, transfer and installation of laboratory and field equipment at Uzbek partner institutes;
Artemia- and aquaculture- related literature and documentation made available for Uzbek partner
institutes;
project-related publications: 13 publications in national or regional journals; XXX publications to
be submitted to international peer-reviewed journals.
42