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Reproduction Efficiency of the Medicinal Leech Hirudo Verbana Carena

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Reproduction Efficiency of the Medicinal Leech Hirudo Verbana Carena www.trjfas.org ISSN 1303-2712 Turkish Journal of Fisheries and Aquatic Sciences 15: 411-418 (2015) DOI: 10.4194/1303-2712-v15_2_27 Reproduction Efficiency of the Medicinal Leech Hirudo verbana Carena, 1820 Mustafa Ceylan1,*, Osman Çetinkaya2, Ramazan Küçükkara1, Ufuk Akçimen1 1 Fisheries Research Institute, Eğirdir, Isparta, Turkey. 2 University of Süleyman Demirel, Eğirdir Fisheries Faculty, Isparta, Turkey. * Corresponding Author: Tel.:+90.246 3133460; Fax:+90.246 3133463; Received 14 January 2015 E-mail: [email protected] Accepted 21 May 2015 Abstract The aim of the study was to investigate the reproduction efficiency of a medicinal leech Hirudo verbana Carena, 1820. Totally 25 gravid medicinal leeches weighted 5.70±3.22 g were used. The leeches deposited 3.20±1.87 cocoons by losing 23.46±5.42% in body weight in every cocooning at 9.67±3.68 days intervals. The cocoons, having a weight 0.93±0.38 g, diameter 13.81±2.26 mm and length 21.44±3.81 mm, had 12.29±5.14 offspring weighted 29.11±13.62 mg. It was determined the negative correlation between the adult leeches weight and the weight loss percentage after each cocoon deposition, between the adult leeches weight and the cocoon deposition frequency, and between the number of offspring and the mean weight of them. The positive correlation was found between the adult leech weights and the number of the laid cocoons, between the adult leech weight and the laid cocoon weight, and between the cocoon weight and the number of offspring. H. verbena shows strong reproduction effort, it can reproduce even loosing 70% their initial weights. Reproduction success of the medicinal leech is strictly correlated with the mature individual weight. Therefore, heavier individuals should be chosen and used in artificial production procedures. Collection of heavier specimens from natural habitats must be prohibited to conserve natural populations Keywords: Reproduction efficiency, medicinal leech, Hirudo verbana, cocoon, offspring size and number. Tıbbi Sülük Hirudo verbana Carena, 1820 Türünün Üreme Verimliliği Özet Bu çalışmada tıbbi sülük Hirudo verbana Carena, 1820 türünün üreme verimliliği araştırılmıştır. Çalışmada ağırlıkları 5.70±3.22 g olan 25 adet döllü sülük kullanılmıştır. Sülüklerden 9.67±3.68 gün arayla, her bir kokonlama sonrasında %23.46±5.42 oranında ağırlık kaybı ile 3.20±1.87 adet kokon elde edilmiştir. Kokonların 0.93±0.38 g ağırlık, 13.81±2.26 mm çap ve 21.44±3.81 mm uzunlukta oldukları ve 29.11±13.62 mg ağırlığa sahip 12.29±5.14 adet yavru birey içerdikleri belirlenmiştir. Anaç sülük ağırlığı ile her kokonlama sonrası yüzde ağırlık kaybı ve kokonlama frekansı arasında ve kokondan çıkan yavru sayısı ile yavru bireylerin ortalama ağırlıkları arasında negatif korelasyonlar bulunmuştur. Buna karşın, anaç sülük ağırlığı ile bırakılan kokon sayısı ve kokon büyüklüğü arasında ve kokon ağırlığı ile yavru birey sayısı arasında pozitif korelasyonlar bulunmuştur. H. verbana türünün başlangıç ağırlığının yaklaşık %70’ini kaybetmesine rağmen güçlü bir üreme eforu gösterdiği belirlenmiştir. Türün üreme verimliliğinin anaç ağırlığı ile güçlü şekilde bağlantılı olduğu görülmüştür. Bu nedenle yapay üretim koşullarında ağır bireylerin tercih edilmesi ve doğal popülasyonların korunması için ağır bireylerin toplanmasının engellenmesi önerilmektedir. Anahtar Kelimeler: Üreme verimliliği, tıbbi sülük, Hirudo verbana, kokon, yavru büyüklüğü ve sayısı. Introduction traditional and modern medicine which is known as medicinal leech. In the Western Palearctic (Caucasia, Europe, Anatolia, Northern Africa) there are just 4 Leeches play important roles in aquatic ecosystems as an invertebrate predator or a blood species from genus Hirudo (Sket and Trontelj, 2008; sucking ectoparasite. Most of the species live in Minelli et al., 2014). freshwater habitats and constitute a quite small group The genus Hirudo is represented generally with with a total of 680 species in phylum Annelida. There the species Hirudo medicinalis, however there are 4 is a small number of blood-sucking species, used in species reported in the genus according to detailed © Published by Central Fisheries Research Institute (CFRI) Trabzon, Turkey in cooperation with Japan International Cooperation Agency (JICA), Japan 412 M. Ceylan et al. / Turk. J. Fish. Aquat. Sci. 15: 411-418 (2015) species identifications and molecular genetics studies used in this study. The adult leeches were collected (Siddal et al., 2007; Utevsky et al., 2010). Hirudo from the wetlands around Lake Eğirdir according verbana (Carena, 1820) has a dispersal area extending Kasparek et al. (2000) and Elliott (2008). The leeches from Switzerland-Italy to Anatolia and Uzbekistan were attracted by disturbing the water, and were (Siddal et al., 2007; Utevsky et al., 2010). However, collected by hand net, and then put into pet jars. there is not enough information on detailed ecology Totally 25 gravid leeches, weighing from 1.85 to and reproduction features of this species except 15.11 g averaging 5.70±3.22 (X±SD) g, were dispersal information (Neubert and Nesemann, 1999; obtained. The gravidity of mature leeches was Elliott and Kutschera, 2011). determined by discolouration and swelling of the Medicinal leeches prefer moist soil out of water clitellum region according to Fernández and Stent or places under structures at coastal regions to deposit (1982), Wilkin (1989) and Elliott (2008) (Figure 1). their cocoons (Sawyer, 1986). It has been established They were acclimated to the laboratory conditions for that H. verbana, kept in artificial ponds, move out of one week before the trial. the aquatic area to deposit their cocoons and any development does not occur in cocoons kept under Experiments water and terrestrial environment is necessity for development and survival of the young (Kutschera This study was designed in accordance with the and Roth, 2006). methods used by Davies and McLoughlin (1996), Hirudotherapy, having a long history of use in Utevskaya and Atramentova (2002), Petrauskienė et traditional medicine, has been observed to take its al. (2009) and Petrauskienė et al. (2011). The trial place in modern medicine recently. The US-FDA was started at 31/07/2013 at room temperature (U.S. Food and Drug Administration) approved the (24.0±0.4°C) and lasted 84 days (54 days for cocoon use of leeches in plastic and reconstructive surgery in deposition and 30 days for hatching all the cocoons) 2004 (Gödekmerdan et al., 2011; Mumcuoglu, 2014). (23/10/2013). The leeches were kept as single in the Legislation on “Traditional, Supplementary and 2-L pet jars half filled with moist peat and they were Alternative Medicine Practices”, including use of monitored to check cocoon deposition three times per leeches in medicine, went into effect in 2014 in week. They were weighted every week, and after each Turkey (OG. 2014). According to the regulation, cocoon deposition to determine the weight loss. The using only cultured leeches (H. medicinalis and H. collected cocoons were weighted; their length and verbana) are permitted in medical treatment. diameter were measured. The leeches were not fed Preventing use of nature collected leeches will during the trial. The cocoons were also kept increase the demand for artificially breed medicinal individually in 350-ml pet jars half filled with moist leech. Turkey has a major share (~80%) in the global peat during 30 days, and then they were opened in leech market (Sağlam, 2011), therefore reproduction order to count and weighat end of this period. The features of H. verbana should be investigated and moist peat also was checked to determine of the artificial breeding methods should be established. The cocoon leave hatchling’sproportion. obtained information will also provide a basis for The cocoon deposition frequency (day), the protection, control and management of natural leech leech weight loss after the each cocoon deposition, the populations. number, diameter, length and weight of the cocoons There is only limited information on per leech, offspring per cocoon, the weight of the reproduction features of H. verbana. In the studies offspring and cocoon leave hatchling proportion were carried out by Petrauskienė et al. (2009 and 2011), investigated. number of cocoons obtained from leeches, number of hatchlings per cocoons, hatchling weight and Statistical Analysis interbreeding abilities among H. medicinalis, H. The data was analyzed with SPSS 17.0 and verbana and Hirudo orientalis were investigated. Putting forth reproduction ecology of the species Excel 2007 software packages. Means ( ) and and determining artificial propagation methods need standard deviations (SD) of the data were given, and to explore the variables which have impact on data was analyzed with Kolmogorov-Smirnov test for reproduction efficiency, and the relationship between checking normal distribution. Correlations between these variables. In this study, reproduction features variables were investigated and the most suitable regression equations were selected (SPSS, 2007). and efficiency of the medicinal leech species H. verbana, being an economic and ecological species and having a dispersal area in many wetlands in Results Turkey was investigated. All values obtained in this study were normally Materials and Methods distributed (P>0.05). Therefore Pearson's Correlation analysis was used. The descriptive values of the trial Leech Sampling are given in Table 1, the correlation coefficients between the parameters are given in Table 2 and the The medicinal leech
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