Garra Rufa) Inhabiting Kangal Balıklı Çermik Thermal Spring (Sivas, Turkey)
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Iranian Journal of Fisheries Sciences 15(4)1593-1608 2016 Otolith and scale morphologies of doctor fish (Garra rufa) inhabiting Kangal Balıklı Çermik thermal spring (Sivas, Turkey) Yedier S.1; Kontaş S.1; Bostancı D. 1*; Polat N.2 Received: December 2015 Accepted: July 2016 Abstract Garra rufa is one of the most popular therapeutic and commercially important fish in ichthyotherapy. Otolith and scale morphology provide new and useful information for fish identification and classification. Left-right asteriscus and lapillus otoliths from juvenile and adult doctor fish, and the scales from 6 different regions of the juvenile and adult fish body have been examined in Kangal Balıklı Çermik thermal spring (Sivas, Turkey). The otolith and scale morphological characters such as type, size, shape, mesial surface, lateral surface, antirostrum and rostrum shapes, focus position, circuli appearance, radii type and posterior and anterior margin shapes were distinguishable features for the juvenile and adult G. rufa samples. Three different otolith shapes were observed such as squared and discoidal otolith shapes for asteriscus pairs and oval to elliptic otolith shapes for lapillus pairs in the juvenile and adult doctor fish. Seven different scale shapes were described from six different body regions of the G. rufa in the current study. The graphical illustration of wavelets was used for both asteriscus and lapillus pairs of the fish to discover otolith variabilities. This is one of the first otolith and scale morphology studies for the juvenile and adult doctor fish. All these otolith and scale characters and their morphologies could be used for best alternative tools to identification, classification, phylogenetic relationships among the different freshwater and marine fish species, genera, populations or stocks. Keywords: Garra rufa, otolithology, Scale morphology, Shape indices 1-Department of Biology, Ordu University, Ordu, Turkey 2-Department of Biology, Ondokuz Mayis University, Samsun, Turkey *Corresponding author's Email: [email protected] 1594 Yedier et al., Otolith and scale morphologies of doctor fish (Garra rufa) inhabiting … Introduction psoriasis and eczema (Fackler and Otolith and scales are crucial and Augustin, 2001; De Groot and versatile research materials used in Conemans, 2004; Grassberger and ichthyological studies due to their Hoch, 2006). Nowadays, G. rufa is unique characters such as shape, becoming a much more popular fısh contour, size and shape indices species for therapeutic purposes and the (Esmaeili et al., 2014; Bostancı et al., doctor fish were listed as a locally and 2015). The otolith and scale commercially vulnerable fish species morphologies have recently increased for treatment and aquarium industry in in importance with the development of Turkey. image analysis systems and software Previous studies on G. rufa were programs using a Scanning Electron limited and some of them are focused Microscope (SEM) (Razak, 2014; on length-weight, length-length Bostancı et al., 2015). Previous studies relationship (Esmaeili and Ebrahimi, have indicated fish scale and otoliths 2006; Gerami et al., 2013; Hamidan were used successfully for and Britton, 2013; Hashemzadeh et al., identification and discrimination of fish 2015), karyotypes (Gorshkova et al., species and stocks (Jawad and Al- 2012), microbiological risks for human Jufaili, 2007; Esmaeili et al., 2014; (Heistinger et al., 2011), reproductive Bostancı et al., 2015), and they widely biology and age determination (Abedi used for age determination (Shimose et et al., 2011), and mortality (Majtán et al., 2015; Elzey et al., 2015), growth al., 2012). The uses of scales and (Romo-Curiel et al., 2015; Mayank et otoliths morphological and al., 2015), prey-predator (Esmaeili, morphometric characters have been 2001; Škeljo and Ferri, 2012) and shown as an important tool to genetic studies (Kumar et al., 2007; determinate and discriminate marine Annabi et al., 2013). and freshwater fish species (Esmaeili et The genus Garra is represented 73 al., 2014; Bostancı et al., 2015). There fish species in the family of Cyprinidae. is little information related to the scale G. rufa is one of them and is small morphologies of G. rufa in Iran. For tropical fish that live in the Syria, Iran, instance, Keivany et al., (2015) Iraq and Turkey (Esmaeili et al., 2009; examined 28 morphometric characters Coad, 2014). G. rufa is also known as (standard length, body depth, body doctor fish because of the regular width, head length, etc.), 25 ratios (head treatment specialties. The doctor fish is length to standard length, body depth to one of the therapeutic fish which is standard length, pectoral fin length to used in ichthyotherapy for therapeutic standard length, etc.) and 10 meristic purposes (Özcelik et al., 2000), their characters (lateral line scales, scale regular treatment specialties can greatly above the lateral line, scale below the improve skin treatment especially lateral line, predorsal scales, keel Iranian Journal of Fisheries Sciences 15(4) 2016 1595 scales, total vertebrae, etc.) of G. rufa and 90% ethanol for 30 minutes in different basins of Iran. However, respectively and they were dried on there are no studies about otolith filter paper to avoid curling (Lippitsch, morphology; therefore, the scales and 1990). Non-damaged otolith and scale otolith morphologies are largely samples were stored for further unknown for G. rufa in many areas. examination. The current study presents for the first While the scales were viewed under time the comprehensive variation a stereo binocular microscope with information on scales and left-right transmitted light and magnification asteriscus and lapillus otoliths in the strength of 10×, the asteriscus and juvenile and adult G. rufa inhabiting lapillus otolith pairs were photographed Kangal Balıklı Çermik thermal spring using Leica S8APO brand microscope (Sivas, Turkey). with Leica Application Suit (Ver. 3.7.0) software. The otoliths and scales Materials and methods morphometric parameters such as The doctor fish specimens were length, width, perimeter, and area were collected from Kangal Balıklı Çermik measured using Digimizer image thermal spring in Sivas, Turkey (Fig. analysis software (Ver. 4.5.1) program. 1). Total length (TL), fork length (FL), The left-right asteriscus and lapillus and standard length (SL) of each doctor otolith shape indices such as form fish was measured to the nearest 0.1 factor (FF), roundness (RD), aspect cm, and their weight (W) was recorded ratio (AR), circularity (C), to the nearest 0.1 g. Schematic shape of rectangularity (R), and ellipticity (E) the Garra rufa and the six body regions were calculated to describe the otolith for scale collection in the doctor fish shape for the juvenile and adult fish were shown in Fig. 2. The fish were (Tuset et al., 2003; Ponton, 2006). dissected and examined as a mature or Mesial and distal surfaces of the left- immature and then they were right otoliths were photographed for considered as a juvenile or adult. The each sample. Eleven otolith characters fish scales were gently removed from such as width, depth, shape, rostrum- six different body regions of the fish. antirostrum size and shape, anterior- Asteriscus and lapillus otoliths pairs of posterior regions, mesial-lateral surfaces each fish were removed using the were determined for each G. rufa. otolith removal procedure (Secor et al., SHAPE software (Ver. 1.3) program 1991). The scales and otoliths were was used to extract the contours of the gently cleaned using a fine brush and G. rufa otolith outline (Iwata and Ukai, then each otolith and scale were rinsed 2002) and AFORO database system by distilled water (Chugunova, 1963; (Lombarte et al., 2006) was used to Campana, 2001). The cleaned scales of determine contour wavelets of the the fish were dehydrated in 30, 50, 70 otolith for the juvenile and adult fish. 1596 Yedier et al., Otolith and scale morphologies of doctor fish (Garra rufa) inhaabbiting … Figuure 1: Sampling location of the Garra rufa in Kangal Balıklı Çermik thermal spring (Sivas, Turkey). Figuure 2: Schematic shape of the Garra rufaa; showing the six body regions for scale collection (modified from Coad, 2014). Due to the determination of were presented in Fig. 3. The aasteriscus graphical illustration of wavelets in the and lapillus otolith pairs morphological left-right asteriscus and lapillus otolith characters such as overall otoliith shape, contours, a total of 512 equidistant otolith with, otolith depth, rostrum size cartesian coordinates of the left and and shape, antirostrum size and shape, right otolith were extracted and they mesial and lateral surfaces, anterior and were analyzed using the wavelet posterior regions were examined for transformed. each G. rufa. Due to the small body of General otolith (A) and scale (B) the fish, six body regions were selected features of the doctor fish and the for scale morphology of G. rufa. terminology used in the current study Iranian Journal of Fisheries Sciences 15(4) 1597 Figure 3: Otolith (A) and scale (B) features of the Garra rufa, and their terminologgy used in the current study. The scales morphological characters regions of the juvenile and adult doctor such as scale type, overall scale shape, fish are shown in Table 2 and also fork scale size, focus position, circuli length (FL), standard length (SL), W appearance, radii type, rostral and and TL are presented in Table 2. The caudal margins were determined in six left-right asteriscus and lapillus otoliths different body regions of the doctor morphological characters of the fish. juvenile and adult fish were separately determinated. In this study, 11 otolith Results morphological characters such as In this study, G. rufa samples weight overall otolith shape, otolith with, (W) and total length (TL) and the mean otolith depth, rostrum size and shape, values of their left-right asteriscus and antirostrum size and shape, mesial and lapillus otoliths measurements are lateral surfaces, anterior and posterior shown for the juvenile and adult fish regions were used, and the results are (Table 1). The mean values of otolith presented in Table 3.