1

CHROMOSOME NUMBERS OF THREE OSTRACODA (CRUSTACEA)

SPECIES FROM KKEKMECE LAKE

Nerdin KUBAN, Diner GLEN

University of Istanbul, Faculty of Science, Department of Biology, Vezneciler
34459 Istanbul-Turkey

SUMMARY
In this study, three ostracoda species collected from Kkekmece
Lake have been examined. The chromosomes of Cypris bispinosa, Cypridopsis
vidua, Heterocypris salina were investigated in terms of their numbers. As a
result, it was observed that, Heterocypris salina have 21, Cypris bispinosa have
19 and Cypridopsis vidua have 24 chromosomes, average.

KKEKMECE GLNDEN OSTRAKOT (KRUSTASE) TRNN

KROMOZOM SAYILARI.
ZET
Bu almada Kkekmece Glnden toplanan ostrakot tr
aratrlmtr. Cypris bispinosa, Cypridopsis vidua, Heterocypris salina trlerinin
kromozomlar,

say

bakmndan

incelenmitir.

Sonu

olarak,

ortalama

Heterocypris salinann 21, Cypris bispinosann 19 ve Cypridopsis viduann da

24 kromozoma sahip olduu grlmtr.

INTRODUCTION
Ostracods are small crustaceans with evolved carapaces. They have
species that can live in fresh, salt and brackish water. Their sizes range from
0,5 mm to 2 mm. Since their carapaces contain calcium carbonate, they could
be traced as fossils, which enable the scientists, to have a high opinion about
the geographical features of region, diachronically.(Moore, 1961)
The aim of this research was to determine chromosome numbers of
three ostracoda species from Kkekmece Lake and to contribute to the
knowledge of Turkish Ostracoda species chromosome numbers.
The ability to reproduce asexually via unfertilized eggs in ostracods
requires several parallel mechanisms such as: the suppression of meiosis; the
suppression of the production of males and the ability to auto-initiative cleavage
of the diploid (unfertilized) eggs.
Parthenogenetic (asexual) species can be defined as: a set of all-female
generations that form a morphological and genetic cluster that is not known to
include any sexual generations. Species with mixed reproduction form a
genetical and morphological cluster of clonal and sexual lineages. Fully sexual
species are defined by the biological species concepts, they only consist of
sexual populations (K.Martens 1998). All types of reproduction can be seen in
different populations of some species, such as Eucypris virens that is
parthenogenetic in Europe, (Tetart 1978) but no male species were
encountered in North Africa, (Gauthier 1928-1937) makes this subject more
interesting.

Sex determination in non-marine ostracods is assumed to be genetic,

and this already considerably facilitates the suppression of the production of
males can be environmentally cued. For example; a possible exception, is the
suppression of the male phenotype by cytoplasmic, symbiont bacteria. The
presence of such bacteria in Heterocypris incongruens has been established
but the extent to which these bacteria indeed suppress males in non-marine
ostracods in general, remains yet to be determined (K Martens 1998). It is
possible that environmental aspects as well as genetic parameters can have an
effect on ostracods chromosome numbers too.

MATERIAL AND METHODS

Materials were collected from Kkekmece Lake in 12th Nov. 1999:
Cypris bispinosa, Cypridopsis vidua. (Sazldere river) and Heterocypris salina
(Altnehir) have parthenogenetic reproduction cycles. (Figure-1).
Collected materials by a net made of Mller fabric were placed in a
proper environment, prepared for reproduction, immediately. Later, temporary
preparations were made with lacto phenol for identification of morphological
characteristics of extremities and carapaces.
Egg cells of the species were blocked in metaphase, adding colchicine at
the final concentration of 0,01 mg/ml to the medium water of studied specimens.
Keeping period of ostracods in concentration varies respect to the species,
therefore each species period listed below as determined by try-error method
(Table- 1)

Table 1: Regional dispersion of species and keeping period in colchicine.

Name of Species
Cypris bispinosa
Cypridopsis vidua.
Heterocypris salina

Region
Kkekmece Lake
Sazldere
Kkekmece Lake

Period
25 Hours
20 Hours
27 Hours

After colchicine treatment, specimens were transferred into 6% F.T.S,

their ovaries were removed and after removal of the excess water with filter
paper the preparations were fixed with the 1/2 acid-alcohol for 10 minutes. Then
the same material was left in 5n HCl solution for 20 minutes, eventually,
preparations were left to dry. Dried preparations were stained after fixing in 0,1
strength Giemsa dye for 7 minutes and excess water left to dry in the air.
Conclusively, preparations were mounted with Canada balsam for permanent
usage. 20 cells were used to count chromosome numbers and the arithmetical
average was taken as a basis (Table-2).

Table 2: In terms of species, the chromosome numbers of 20 ovary cells.

Chromosome
ovarium cells
Cell Number Heterocypris
salina
1
20
2
21
3
19
4
20
5
21
6
23
7
24
8
22
9
21
10
20
11
23
12
21
13
21
14
22
15
20
16
19
17
21
18
21
19
20
20
21

Numbers

of

Species

Cypris
bispinosa
18
17
18
19
19
20
21
19
18
19
19
20
19
18
19
20
20
18
19
20

from

20

Cypridopsis
vidua
27
25
24
22
22
24
25
24
23
25
24
27
24
22
25
24
22
24
23
24

Chromosome counting was carried out under a light microscope with a

100X immersion objective and a 10X eyepiece. 20 cells were used for all
species under 1000X magnification.
Pictures of chromosomes were taken using a Carl Zeiss-no 61555
photomicroscope with a 100X objective and intermediate magnification being
stable at 1.25=4.
Magnification ratio of photos was calculated in terms of Enlarger
magnification x Objective magnification x Intermediate magnification.
As chromosomes were too small and location of centromeres couldnt be
revealed, types of chromosomes couldnt be identified.

Fig. 1: Densely populated area

RESULTS
Given systematic classification and chromosome numbers of three
ostracods were handled in this study.
Hartmann and Puri's ostracoda classification study accepted in 1974 was
used for defining systematic placing of species in this search (Hartmann and
Puri, 1974).
Familia

:CYPRIDIDAE

Baird, 1845

Subfamilia

:CYPRIDINAE

Baird, 1845

Genus

:Heterocypris

Claus, l892

Heterocypris salina (Brady, 1866) Sars, 1890

Material:
Material was collected from Kckekmece Lake in 12 th Dec., 1999.
Chromosome number:
Parthenogenetic females were found in the material collected from the place
mentioned above. This species, to be kept in colchicine for 27 hours, have 21
chromosomes average. (Figure-2a, 2b)

Fig. 2a- Chromosomes picture of Heterocypris salina, Giemsa, X 1920

Fig.2b-Chromosome number of Heterocypris

population)

salina. (Parthenogenetic

Genus

: Cypris Mller, 1776

Cypris bispinosa Lucas, 1849

Material:
Material was collected from the reed bed near Kkekmece Lake in 12 th
Dec., 1999.
Chromosome number:
Cypris bispinosa parthenogenetic females, to be kept in colchicine for 25 hours,
have 19 chromosomes average. (Figure-3a, 3b)

Fig. 3a- Chromosomes picture of Cypris bispinosa, Giemsa, X 1920

Fig. 3b- Cypris bispinosa, chromosome number (Parthenogenetic population)

Familia

: CYPRIDOPSIDAE Kaufmann, l900

Subfamilia

: CYPRIDOPSINAE Bronstein, 1947

Genus

: Cypridopsis Brady, 1867

Cypridopsis vidua (Mller, l776) Brady, l867 (Cypris sella Baird, l850)
Material:
Material was collected from reed bed near Sazldere in 12 th Dec., 1999.
Chromosome number:
Cypridopsis vidua parthenogenetic females to be kept in colchicine for 20 hours
have 24 chromosomes average. (Figure4a, 4b)

Fig. 4a- Chromosomes picture of Cypridopsis vidua, Giemsa, X 1920

Fig. 4b- Cypridopsis vidua, chromosome number (Parthenogenetic population)

As three species all have small chromosomes, centromer locations

couldn't be identified. Morphological data of the chromosomes isn't available.
Because of this the shapes and the numbers of chromosomes seen in photos
were reflected directly to the drawings and types of chromosomes were not
displayed in these drawings.

DISCUSSION
First ostracoda studies were carried out by Tetart (Tetart, 1975,1978).
Belentepe (Belentepe, 1991), followed by Sar (Sar, 1991,1997) and Kuban
(Kuban, N., 1995), pioneered the studies in Turkey.
Heterocypris salina, Cypris bispinosa and Cypridopsis vidua species
found in Kkekmece Lake are handled in the study. All three species
reproduce parthenogenetically.
In a parthenogenetic population only female individuals are present and
new members resemble their mother. Their phenotypes are the same even if
there are bisexual forms encountered in the world. In addition to this, there are
differences between chromosome numbers of different local populations, this
situation causes, translocation and deletion to occur at a very high rate during
mitosis. In case of sexual reproduction females produce haploid eggs through
meiosis, but in case of parthenogenetic reproduction, there is no meiosis and
eggs are at least diploid. Individual females are not known to switch between
either mode (although both types can occur within one species and even

population), neither cyclically nor erratically, although intraspecific hybridization

between parthenogenetic females and sympatric males has been reported for a
number of species: it is believed that the resulting offspring consists of
parthenogenetic polyploid females with mixed genotype (Dowling & Secor
1997).
Heterocypris salina have an average of 21 chromosomes and Cypris
bispinosa have an average of 19 chromosomes, however their chromosome
numbers couldn't be compared due to the absence of a pre-study in Turkey.
Cypridopsis vidua have an average of 24 chromosomes. Belentepe
(Belentepe, 1991) fixed chromosome numbers of Cypridopsis vidua, collected
from Baltaliman Hydrobiology Institution pool, as 19. Sar (Sar, 1992) obtained
the same species with 21 chromosomes around Kkekmece Lake and
Kuban (Kuban, N., 1995) found Cypridopsis vidua with 24 chromosomes.
Lastly, Sar (Sar, 1997) defined 21 chromosomes in the same species that he
collected from Terkos Lake.
As seen before all three observations made from the local populations of
Cypridopsis vidua and different chromosome numbers were found as a result.
However these differences cannot be observed in phenotypes. As a result, we
can assume that this chromosomal difference has no effect on phenotypes.
Therefore, it can be considered that these differences probably are derived from
polyploidy, which may happen during parthenogenetic reproduction. These
extra chromosomes have no obvious function and they can be acquired or lost
without causing serious effects. Many normal chromosomes are intensified
heterochromatins smaller than their normal sizes and are not homologous. The

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assumption regarding to the origins of these chromosomes is that they are

constituted through deletion and translocation of heterochromatic segments of
normal karyotype. The effect of changes in ecological conditions over the event
is just another subject to be argued.
As a result, three more ostracoda species chromosome numbers were
observed. A comparison between different local populations of a species was
made. Through this comparison, morphological and numeral differences
between local communities were revealed. Because of small size of the
chromosomes their shapes couldnt be obtained.

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REFERENCES
BELENTEPE, S.: Chromosome numbers and shapes of Thrace ostracode
(Crustacea) Populations. Istanbul Univ. Fen Fak..Biyoloji Der., 55 (1991),
29-37.
DOWLING, T. E. and SECOR, C. L.: The role of hybridisation and introgression
in the diversification of animals. Annu. Rev. Ecol. Syst., 28 (1997), 593620.
GAUTHIER, H.: Ostracodes et Cloderes de L Afrique du Nord

(1 ere note).

Bull. Soc. Hist. Nat. Afrique Nord 19 (1928), 10-19.

GAUTHIER, H.: Ostracodes et Cloderes de L Afrique du Nord (4 e note) Bull.
Soc. Hist. Nat. Afrique Nord 28 (1937), 147-156.
HARTMANN, G., PURI, H.: Summary of Neontological and Paleontological
Classification of Ostracoda. Mitt. Hamburg. Zool. Mus. ns. 70 (1974), 773.
KUBAN, N.: Baz Ostrakod Populasyonlarnda Kromozom Says ve ekli,. ..
Fen Bilimleri Enstits (1995), Yksek Lisans tezi.
MARTENS, K. : Sex and Parthenogenesis. Sex and Ostracods: A new
Synthesis 17, Backhuys Publishers, Leiden, Netherlands Press (1998).
MOORE, R. C.: Treatise on Invertebrate Palaeontology. Part Q. Arthropoda 3,
Crustacea, Ostracoda. Geol. Soc. Am. Univ. Kansas Press (1961).
SARI, H., ULAKOLU, G.: Ostrakod populasyonlarnda kromozom saylar.
Istanbul Univ. Su rnleri Der ., 2 (1992), 43- 60.
SARI, H.: Kromozom Saylarna Dayanlarak Terkos Glndeki Ostrakod
Trlerinin Saptanmas,. .. Fen Bilimleri Enstits (1997), Doktora Tezi.

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TETART, PAR. J.: Recherches sur la reprduction et L ecologie quelques

Ostracodes Cyprididae. A. L Universite Scientifique et Medicale de
Grenhoble (1975), Le Grade de Docteur es Sciences.
TETART, PAR. J.: Les garnitures chromosomiques des Ostracodes leau
douce.Trav. Lab. Hydrobiol. Grenhoble 69-70 (1978), 113-140.

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Fig. 1: Densely populated area

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Fig. 2a- Choromosomes picture of Heterocypris salina, Giemsa, X 1920

Fig.

2b-

population)

Chromosome number of Heterocypris

salina. (Partenogenetic

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Fig. 3a- Chromosomes picture of Cypris bispinosa, Giemsa, X 1920

Fig. 3b- Cypris bispinosa, chromosome number (Partenogenetic population)

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Fig. 4a- Chromosomes picture of Cypridopsis vidua, Giemsa, X 1920

Fig. 4b- Cypridopsis vidua, chromosome number (Parthenogenetic population)