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Karyotypes of Southeastern Turkish Scorpions Hottentotta saulcyi and Buthacus macrocentrus (Scorpiones: Buthidae).

INTRODUCTION

Currently, 213 genera and 2433 scorpion species are classified under 17 families (1). Although scorpions are widely distributed in the tropics and subtropics and all types of terrestrial habitats all over the continents (except Antarctica) (2), our present knowledge of their karyotypes is still scarce. Chromosome data on 155 scorpions belonging to 11 families have thus far been determined. Among them, 91 species of Buthidae have been studied, limited to some geographic regions-especially Brazil and Africa (3). Karyotypes of these scorpions are composed of holocentric chromosomes without a localized centromere region (4).

Cytogenetic studies have been carried out on Leiurus abdullahbayrami Yagmur, Koc&Kunt, 2009 and Compsobuthus matthiesseni (Birula, 1905) present a karyotype with 2n=22 (5), Androctonus crassicauda (Olivier, 1807) has a karyotype of 2n=24 (6), Aegaeobuthus gibbosus (Brulle, 1832) shows 2n=28, Mesobuthus eupeus (C.L. Koch, 1839) has 2n=20 (Buthidae) and Euscorpius aladaglarensis Tropea&Yagmur, 2016 (7) shows 2n=88 (Euscorpiidae), which are distributed in Turkey.

The present knowledge of the cytogenetics of Turkish scorpions is scarce and fragmented. The aim of this paper is to report the first chromosomal data of two species (Hottentotta saulcyi and Buthacus macrocentrus) from Turkey. The genus Hottentotta Birula, 1908 is widespread throughout Africa, the Middle East and Asia (8, 9). This genus comprises almost 51 species (10). H. judaicus (Simon, 1872) (11), H. tamulus (Fabricius, 1798) (12-14) and H. trilineatus (Peters, 1861) (15) have been analyzed in cytogenetic studies up to present day. Hottentotta saulcyi (Simon, 1880) was firstly recorded in Mardin (10) and then reported in Batman, Sirnak, and Hakkari in Turkey (9). Meanwhile, the genus Buthacus Birula, 1908 is distributed across northern and western Africa, Israel, Palestine, Jordan, Syria, Turkey, the Arabian Peninsula, Iraq, Iran, Afghanistan, and Pakistan. 23 species belonging to Buthacus have thus far been described (1, 10, 16-19). In Turkey, Buthacus macrocentrus (Ehrenberg, 1828) is known only from Sanliurfa (10, 18, 20). Cytogenetic analyses have been performed for only one species namely Buthacus stockmanni Kovraik, Lowe&Stahlavsky, 2016 having 2n=20 (21).

MATERIALS AND METHODS

The scorpions were collected using a UV lamp during the night from Sirnak and Sanliurfa respectively (Figure 1). The collected specimens were transferred to the laboratory in individual plastic containers. In total, six specimens of Hottentotta saulcyi and seven of Buthacus macrocentrus were analyzed (for detailed information, see "Material examined" below).

The gonads were used from both males and females. The specimens were killed by ventral puncture to the prosomal area. Under a stereomicroscope, the gonads were removed by dissection in the presence of physiological salt solution for invertebrates. The gonads were then kept in a hypotonic solution (0.075 M KCl) for 20 min. The gonads were fixed in a freshly prepared fixative (3: 1, methanol: acetic acid) for 20 min. A few drops of 60% acetic acid were then dropped on a slide and then shredded with a tungsten needle. The drop on the slide was placed on the heating plate and spread with tungsten needles (22). The prepared slides were stained with 5% Giemsa in Sorensen's phosphate buffer. The chromosome preparations were analyzed under a Leica DM 500 microscope with a 100x objective. Images were taken with a Leica camera using Leica Application LAZ software. The measurements were analyzed using software ImageJ 1.47 (23) with the plugin Levan (24). The relative length of the chromosomes was calculated as a percentage of the diploid set and it based on seven mitotic metaphases in Hottentotta saulcyi and on seven postpachytene in Buthacus macrocentrus. The preparations were kept in a slide box and the remains of the specimens were fixed in a solution of 96% alcohol and stored in a refrigerator at 4 [degrees]C at the Zoological Museum of Sinop University, Turkey (ZMSU).

RESULTS

Family : Buthidae C.L. Koch, 1837

Genus-1 : Hottentotta Birula, 1908

Species-1 : Hottentotta saulcyi (Simon, 1828) (Figures 2 and 3)

Material examined: 2[male] 3[female] and 1[male] subadult, Sirnak, 2.5 km SW of Sirnak, 37[degrees]29'57.3"N; 42[degrees]26'32.7"E, 1024 m, 17.07.2014, leg. E. A. Yagmur (Figure 1).

Karyotype Investigation

The chromosome complement of Hottentotta saulcyi consisted of 14 chromosomes (Figure 4). We observed only the mitotic phases for this species. The relative chromosome length gradually decreased from 11.65 to 4.39% of the diploid set (Figure 4a).

Genus-2 : Buthacus Birula, 1908

Species-2 : Buthacus macrocentrus (Ehrenberg, 1828) (Figures 5 and 6)

Material examined: 4[male], Sanliurfa, Birecik District, 2 km S of Mezra Village, 36[degrees]56'50.1"N; 38[degrees]01'20.3"E, 375 m, 08.07.2013, leg. E. A. Yagmur. 3[female], Sanliurfa, Birecik District, 2 km S of Mezra Village, 36[degrees]57'35"N; 38[degrees]00'43"E, 387 m, 27.07.2014, leg. E. A. Yagmur (Figure 1).

Karyotype Investigation

The number of diploid chromosomes in all male and female Buthacus macrocentrus specimens examined was 28 (Figure 7a). The relative chromosome length of the first chromosome (6.31%) was slightly larger than the remaining chromosomes which gradually decreased from 5.14 to 1.95% of the diploid set (Figure 7a). Achiasmatic bivalents were detected in males during the first meiotic division. In the female, mitotic metaphases were obtained (Figure 7b). A distinct quadrivalent or hexavalent association of chromosomes were found in all individuals and observed in four males during meiosis (Figures 7c and d). The size of the detected multivalent chromosomes gradually reduced. The chromosomes forming quadrivalent or hexavalent were different in size (Figure 7a). During the first meiosis division phase (anaphase, pachytene, postpachytene, metaphase-I), no indication of crossing-over was observed. In the polar view of metaphase I (Figures 7b and c), the majority of these bivalents presented parallel-arranged homologous chromosomes. In pachytene (Figure 7e), bivalents were all strip-shaped.

DISCUSSION

The present study provides the first cytogenetic analysis of the Hottentotta saulcyi and Buthacus macrocentrus, species of the Buthidae family. The karyotypes of these species consist of 14 and 28 chromosomes (Table 1). 91 species of Buthidae have been cytogenetically studied thus far, and which show the lowest chromosome numbers within scorpions. This family has a diploid chromosome number varying from 2n=5 [Tityus bahiensis (Perty, 1833)] to 2n=36 [Barbaracurus somalicus (Hirst, 1907) and Parabuthus mossambicensis (Peters, 1861)], excluding dubious information (see 3). Buthidae family, as well as the entire order Scorpiones, is characterized by achiasmatic meiosis in males (25). In contrast to other scorpions, the examined species of buthids usually possess a relatively low chromosome number and all of them have holocentric organization (4).

Although there are several faunistic and taxonomic studies on the genus Hottentotta, there is a paucity of cytogenetic studies. Chromosomal data are known for only three of the total of 51 currently recognized species of the genus Hottentotta species (10), were included in cytogenetic studies (11-15). Hottentotta tamulus species possesses 2n=22, 24 or 20-28 chromosomes forming a continuous series (12-14). But, Venkatanarasimhiah and Rajasekarasetty (13) stated that the chromosome number of same species have 2n=23 stable chromosome number in India. Sharma et al. (12) observed the tetravalent formation which is very common in H. tamulus. The diploid number was given as 2n=24 for Hottentotta trilineatus by Newlands and Martindale (15). Qumsiyeh et al. (11) reported diploid chromosomes as 2n=16 for Hottentotta judaicus specimens from the Palestinian Territories. We obtained diploid chromosomes as 2n=14 for Hottentotta saulcyi. Our results supported that genus Hottentotta displays interspecific karyotype differences with 2n ranging from 14 (this study) to 24. However, we could not observe multivalent in H. saulcyi as previously documented in H. tamulus.

The genus Buthacus was studied cytogenetically for the first time by Kovarik et al. (21). The karyotype of B. stockmanni has 2n=20 chromosomes with holocentric and achiasmatic meiotic complement (21). According to cytological observation of B. stockmanni, the first pair of chromosomes are distinctively larger (13.41% of the diploid set) than the other chromosomes that gradually decrease from 5.84 % to 2.69 % of the diploid set. The cytogenetic analyses revealed that Buthacus macrocentrus consisted of 2n=28. The chromosomes are holocentric and achiasmatic meiotic complement as Kovarik et al. (21). The ideogram show that chromosomes of 1, 4, 9 and 22 are involving the arrangement of quadrivalent chromosome (Fig. a). These findings confirm the results of Shanahan and Hayman (26) who stated that multivalent formations involve during the achiasmate meiosis of buthid. The first chromosome is significantly longer (6.31%) than the remaining chromosomes that gradually decrease from 5.14% to 1.95% of the diploid set. In our present study, it is interesting to note that the presence of quadrivalents and hexavalents seems to be frequent in B. macrocentrus species.

CONCLUSION

Our analysis of karyotype data provides a first step towards understanding the chromosome numbers and the structure of chromosomes in one of the most important and dangerously venomous members of the family Buthidae. In general, our findings support that buthids have a low chromosome number with holocentric chromosomes. In the present study, the karyotype of two buthids, Hottentotta saulcyi and Buthacus macrocentrus, was identified for the first time as 2n=14 and 2n=28 respectively. The genus Hottentotta possess interspecific karyotype differences in a continuous series of the chromosomal number with 2n ranging from 14 to 24. As far as we know, this is the second karyological analysis of the genus Buthacus, thus our study is a significant contribution to the description of the chromosomal features of Buthacus macrocentrus. Our data highlights that the frequency of the multivalent is very high in this species. The chromosomes were arranged and numbered according to their total length in a gradually decreasing size order and their ideogram was developed (see Fig. 4a and Fig. 7a). Nevertheless, the results show that chromosome numbers are not a useful character in some buthids, and are therefore not effective for taxonomic purposes. Moreover, more detailed analysis of the karyotype of the buthid species is required for comparative cytotaxonomy of the Buthidae.

Peer-review: Externally peer-reviewed.

Author Contributions: Conception/Design of study: H.K.; Data Acquisition: H.K., E.A.Y.; Data Analysis/Interpretation: H.K., F. S.; Drafting Manuscript: H.K.; Critical Revision of Manuscript: H.K., F. S., E.A.Y.; Final Approval and Accountability: H.K.

Conflict of Interest: The authors declare that they have no conflicts of interest.

Financial Disclosure: The project was financially supported by the Sinop University BAP Unit (FEF 1901-13-03).

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Halil Koc (1) (iD), Ersen Aydin Yagmur (2) (iD), Frantisek Stahlavsky (3) (iD)

(1) Sinop University, Faculty of Arts and Science, Department of Biology, Sinop, Turkey

(2) Celal Bayar University, Alasehir Vocational School, Manisa, Turkey

(3) Charles University in Prague, Faculty of Science, Department of Zoology, Prague, Czech Republic

ORCID IDs of the authors: H.K 0000-0003-0429-2824; E.A.Y. 0000-0002-0396-3975; F.S. 0000-0002-8520-9166

Please cite this article as: Koc H, Yagmur EA, Stahlavsky F. Karyotypes of Southeastern Turkish Scorpions Hottentotta saulcyi and Buthacus macrocentrus (Scorpiones: Buthidae). Eur J Biol 2019; 78(2): 111-116. DOI: 10.26650/EurJBiol.2019.0008

Address for Correspondence: Halil Koc

E-mail: koc.halil@hotmail.com

Submitted: 16.05.2019

Revision Requested: 17.07.2019

Last Revision Received: 09.09.2019

Accepted: 17.09.2019

DOI: 10.26650/EurJBiol.2019.0008
Table 1: Number of diploid chromosomes in eight species of scorpions
from Turkey.

Taxon                                     2n  Sampling locality

Buthidae
Androctonus crassicauda (Olivier, 1807)   24  Turkey: Sanliurfa Province
Buthacus macrocentrus (Ehrenberg, 1828)   28  Turkey: Sanliurfa Province
Compsobuthus matthiesseni (Birula, 1905)  22  Turkey: Gaziantep Province
Hottentotta saulcyi (Simon, 1880)         14  Turkey: Sirnak Province
Leiurus abdullahbayrami Yagmur, Koc
& Kunt, 2009                              22  Turkey: Gaziantep Province
Mesobuthus eupeus (C. L. Koch, 1839)      20  Turkey: Nigde Province
Aegaeobuthus gibbosus (Schenkel, 1947)    28  Turkey: Nigde Province
Euscorpiidae
Euscorpius aladaglarensis Tropea &
Yagmur, 2016                              88  Turkey: Nigde Province

Taxon                                     Reference

Buthidae
Androctonus crassicauda (Olivier, 1807)   (6)
Buthacus macrocentrus (Ehrenberg, 1828)   the present study
Compsobuthus matthiesseni (Birula, 1905)  (5)
Hottentotta saulcyi (Simon, 1880)         the present study
Leiurus abdullahbayrami Yagmur, Koc
& Kunt, 2009                              (5)
Mesobuthus eupeus (C. L. Koch, 1839)      (7)
Aegaeobuthus gibbosus (Schenkel, 1947)    (7)
Euscorpiidae
Euscorpius aladaglarensis Tropea &
Yagmur, 2016                              (7)
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Title Annotation:RESEARCH ARTICLE
Author:Koc, Halil; Yagmur, Ersen Aydin; Stahlavsky, Frantisek
Publication:IUFS Journal of Biology
Date:Dec 1, 2019
Words:2673
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