Treubia 44: 1–14, December 2017 THE MORPHOLOGY AND ONTOGENETIC OF TAREBIA GRANIFERA (LAMARCK, 1822) FROM INDONESIA (GASTROPODA: CERITHIOIDEA: THIARIDAE) Nur Rohmatin Isnaningsih*1,2, Adi Basukriadi1 and Ristiyanti Marsetiyowati Marwoto2 1 Department of Biology, Faculty of Mathematics and Natural Sciences, University of Indonesia 2 Zoology Division (Museum Zoologicum Bogoriense), Research Center for Biology, Indonesian Institute of Sciences, Jl. Raya Jakarta-Bogor Km 46, Cibinong, Bogor 16911, Indonesia *Corresponding author: [email protected] Received: 14 November 2016; Accepted: 2 November 2017 ABSTRACT The freshwater gastropod Tarebia H. Adams & A. Adams, 1854, are found in rivers, lakes, and other limnetic habitats. In Indonesia, Tarebia granifera (Lamarck, 1822) is the only species within the genus that has a wide distribution. The systematics and identity of this species are still doubtful due to high variation in shell morphology, especially shell height and ornamentation or sculpture of shell. To determine the identity of T. granifera from Lombok, Banten, and Maros, ontogenetic studies have been conducted. The results showed that T. granifera from Lombok produce the highest number of embryonic shells. The number of progeny in the brood pouch from a single individual of T. granifera can vary between 9 to 203 embryonic shells which are found in various stages of 0.22 mm to about 5 mm in size inside the brood pouch. Key words: embryonic shell, ontogeny, subhaemocoelic brood pouch, Tarebia granifera, Thiaridae INTRODUCTION Tarebia granifera (Lamarck, 1822) (Thiaridae) occurs in freshwater bodies, in lotic as well as lentic habitats. This species belongs to benthic fauna and lives attached to different kinds of substrate or on the banks of aquatic habitats. Besides living in natural aquatic habitats, T. granifera is also found in artificial water bodies such as reservoirs and irrigation channels. This species is able to live in temperatures ranging between 6–38oC (Appleton et al. 2009). Tarebia granifera is resistant to turbidity and pollution and it tolerates a small degree of salinity (Benthem-Jutting 1956, Kartayev et al. 2009). The spreading of T. granifera occurs actively by its own movement and also passively by being carried away by birds or water plants (Appleton et al. 2009). Tarebia granifera was first described by Lamarck (1816) without any locality given (Cowie 1995). The distribution of T. granifera includes India, Malay Archipelago, Philippines, Taiwan, Japan, and various Pacific islands (Abbot 1952, Benthem-Jutting 1956). Nowadays, this snail is widespread and has become an invasive species at many locations around the world (Appleton et al. 2009). In 1947, T. granifera has been introduced to Florida, US. In 1999, this species was reported from South Africa, and in the same year, it has also been reported to occur in Cuba and small islands in the Caribbean, i.e. Saint Lucia, Martinique (Morrison 1954, Pointier 1998, Appleton & 1 Treubia 44: 1–14, December 2017 Nadasan 2002). The spreading outside of the natural distribution is attributed through the aquarium and ornamental trades. In Indonesia, T. granifera fossils were found from the upper layers of the Pliocene in Central and East Java (Benthem-Jutting 1937). This species was reported to occur in Sumatra, from Danau Laut Tawar, Danau Singkarak, Danau Di Atas, Danau Di Bawah, Langkat River and Asahan River (Rensch 1934, Benthem-Jutting 1959). Meanwhile, in Java, T. granifera was found widely distributed from West to East Java, among others from the Ciliwung river, Sukabumi, Cisolok, Cianjur, Pekalongan, Kebumen, Rembang, the Brantas River, Kali Mas and many other locations (Benthem-Jutting 1956). This species is also reported from Bali, Lombok and eastern part of Indonesia like Manado, Ternate, Aru, Waigeo and Raja Ampat Islands, Sorong and Uriami River in Manokwari (Rensch 1932, Benthem-Jutting 1959, 1963). Specimens from various locations in Indonesia were identified as T. granifera, but the shell morphology showed that there was a high degree of variation. So far morphological characters can be used to distinguish species within the genus Tarebia. However, anatomical characters are also important and required to support the validity and species identity. According to Wagner (2001), shell morphology was strongly influenced by environmental conditions or habitats hence it became easily changed. On the other hand, soft body (anatomical) characters represent the basic adaptation of behaviour that was more stable. The anatomical characters of reproductive organs can be used to determine the identity of thiarid species. It was shown that characters of the reproductive organs and embryonic shells are more informative at the level of genera (Abbott 1952, Glaubrecht 1999). The structure and form of the reproductive organs as well as ontogeny (embryonic development) can vary inside a genus (interspecies) or even within species (intraspecies). To assure the validity of Indonesian T. granifera status and identity, a comprehensive systematic study should be conducted to accompany the study on the shell morphology. This study focuses on the investigation of the intraspecific variation of biometrical data and compares the number and size classes of embryonic shells of T. granifera from three different locations in Indonesia. This preliminary studies is expected to provide supplementary data to complete the information status and identity of valid T. granifera. MATERIALS AND METHODS Materials The study is based on materials from the collection of the Museum Zoologicum Bogoriense (MZB). All specimens were preserved in 70% ethanol. For this study, we used 60 individuals of T. granifera from 3 different locations in Indonesia: 2 Isnaningsih et al.: The morphology and ontogenetic of Tarebia granifera (Lamarck, 1822)... Lempenge estuary, Dsn. Lempenge, Ds. Rempeg, district. Ganga, Kab. North Lombok, coll.date: 29 April 2014, coll: NR. Isnaningsih, n: 20 (MZB Gst. 17.551); river at Ds. Cijeruk, Pandeglang, Banten, coll.date: September 2015, coll: Heryanto, n: 20 (MZB Gst. 18.393); Bantimurung river, Maros, South Sulawesi, coll.date: July 13, 2006, coll: Aaron, n: 20 (MZB Gst. 15.698). Methods Specimens were investigated using a stereo microscope. Quantitative data were obtained by measuring the shell biometry which includes shell height (SH), shell width (SW), body whorls height (BWH), aperture height (AH), and aperture width (AW) (Fig. 1). These measurements were done using a caliper with 0.01 mm accuracy. Total embryonic shells retained in each specimen were counted. Those data were obtained by dissecting the subhaemocoelic brood pouch in the anterior part. Embryonic shells were separated into four ontogeny stages (size classes): (a) 0.1–1 mm, (b) 1.1–2 mm, (c) 2.1–3 mm, (d) more than 3 mm. Figure 1. Parameters used to measure the shell dimensions (SH: shell height, SW: shell width, BWH: body whorls height, AH: aperture height and AW: aperture width). 3 Treubia 44: 1–14, December 2017 For SEM examination, the embryonic shells were soaked and washed with 20% clorox solution then were coated with gold at 5–8 mÅ for 5 min. After coated, specimens were studied using a JEOL JSM5310LV scanning electron microscope (SEM) at an accelerating voltage of 20 kV. RESULTS Examination on 60 individuals of T. granifera are presented here, and some representative shells are shown (Fig. 2). Thiaridae Tarebia Adam H. & A. Adam, 1854 Tarebia granifera Lamarck, 1822 Taxonomy and systematics The first figure of T. granifera was published by Lamarck (1816) with the name Melania granifera without any description and location. Later Lamarck (1822) published following description "M. testa ouato-acutâ, striis Iransuersis crasslusculis granosis cincta, luteO'virescente; ullùno anfractu pentricoso; spirce ânfractibus planis; aperlurâ Alba ". In 1854, Adam & Adam reassigned granifera to the genus Tarebia. A number of authors still refered to Lamarck (1819) and used the name Melania granifera (see Brot 1874, Rensch 1934), while others used the name Thiara granifera according to Adam & Adam (1854), Benthem-Jutting (1937, 1941). Besides the two names, some other authors also used the name Melania lineata due to distinguishable dark lines coinciding with spiral ridges (Mousson 1848, Martens 1897). Based on this distinct characters, Rensch (1934) divided Melania granifera into two subspecies, namely M. granifera granifera and M. granifera lineata. Benthem-Jutting (1956) incorporated this species to the Melanoides granifera and wrote 19 synonyms. Furthermore, Glaubrecht (1996) returned the name to Tarebia granifera, so all the names that refered to the species Melania granifera described by Lamarck (1822) were considered as synonyms. Shell - The shells of T. granifera has a medium shell size of 13.40 mm – 32.15 mm. Their shape are elongated conic and rather thick. There are 9 – 11 spiral lines below the periphery. Axial lines are not visible. Spiral lines are as prominent as the axial lines so that granules are formed at the interceptions. The shell colour ranges from yellowish or light brown to dark brown or black. The apex is pointed and sometimes eroded. Shells of T. granifera usually have 6 to 8 whorls, with a high spire and regularly increasing size. The body whorl is evenly rounded and comprises about 0.5 4 Isnaningsih et al.: The morphology and ontogenetic of Tarebia granifera (Lamarck, 1822)... Figure 2. Shells of T. granifera from Indonesia: A-C. MZB Gst. 17.551, River Lempenge, Dsn. Lempenge, Ds. Rempeg, district. Ganga, Kab. Lombok Utara; D-F. MZB Gst. 18.393, Ds. Cijeruk, Pandeglang, Banten; G-I. MZB Gst. 15.698, Bantimurung rivers, Maros, South Sulawesi. 5 Treubia 44: 1–14, December 2017 times of the total shell length. The suture of T. granifera is shallow. This species has oblique aperture with curved and uncontinuous apertural lip. The umbilicus is enclosed. Operculum - The type of operculum is paucispiral, nucleus located in the lower edge of the operculum. There are eight rings sculpted on the outer surface of operculum. Soft body- Animal is coiling in 3 whorls. The headfoot is black, while the mantle is white with black line in serrated marginal edge. The body has white colour. The snout is broad. Cephalic tentacle is about 2 mm in length. The location of eyes is at the base of tentacle. Ctenidium is about 7 times of osphradium. Embryonic shell - The average of embryonic shell size is 0.90–1.04 mm in shell height with the maximum of shell height up to 5.85 mm. Shell of embryo consists of 5 whorls. The apical whorl is smooth. Subsequence whorls decorated with strong ridge especially below the suture. Apical view of embryonic shell appears more wrinkle (Fig. 3). The biometrical measurements showed that shell height varies in T. granifera from three different locations. The shell height of T. granifera from Lombok, Banten and Maros is 24.10 to 29.68 mm, 13.40 to 25.37 mm and 10.01 to 32.15 mm, respectively (Fig. 4). Although T. granifera of three locations showed high variation in the character of the shell height, but in general all specimens studied have the same shell shape index. This was indicated by the ratio of height and width of the shell (index SH / SW) which depicting similar pattern (Fig. 5). From 20 individuals dissected, 17 individuals of T. granifera from Lombok retain embryonic shells (Fig. 6). Meanwhile, there are 9 individuals of T. granifera from Banten and 11 individuals from Maros that retain embryonic shells. The highest number of embryonic shells in brood pouch was found in T. granifera from Lombok. One individual from this location is able to retain 9 to 203 embryonic shells in their brood pouch, while 26 to 190 embryonic shells were found from one individual of T. granifera from Banten. Tarebia granifera from Maros bears 3 – 80 embryonic shells (Fig. 7). The progeny develops in the maternal brood pouch. Because of that, the progeny in the brood pouch is found in increasing size classes. Most of embryonic shells are remained in brood pouch in 0.1–1.0 mm then followed by 1.1–2.0 mm in size (Fig. 8). A few were found reaching sizes of 2.1–3.0 mm and only 5 shells were found with more than 3 mm in T. granifera from Lombok. 6 Isnaningsih et al.: The morphology and ontogenetic of Tarebia granifera (Lamarck, 1822)... A B Figure 3. Embryonic shell of T. granifera from Ds. Cijeruk, Pandeglang, Banten (MZB Gst. 18.393), A. Lat- eral view: elongated conical, consists of 5–6 whorls, B. Apical view: apical whorl smooth, subsequent whorls sculptured with strong axial ribs especially below the suture. 7 Treubia 44: 1–14, December 2017 Figure 4. Boxplot of adult shell height of T. granifera from three different locations. Values below and above boxplot indicate minimum and maximum shell height, respectively. Figure 5. Scatter plot comparing shell height vs. shell width of T. granifera from three locations in Indonesia. 8 Isnaningsih et al.: The morphology and ontogenetic of Tarebia granifera (Lamarck, 1822)... A B Figure 6. A. External view of the brood pouch located in neck part of the head and lying above the esophagus, B. Brood pouch filled with embryonic shells, one single individual could contain up to 203 embryos with the size ranging from 0.1 to 3 mm. 9 Treubia 44: 1–14, December 2017 Figure 7. Boxplots of the total number of embryonic shells found in T. granifera from three different locations (Lombok, Banten and Maros). Values above error lines indicated the maximum number of embryonic shells in the brood pouch of one single female. Figure 8. Number of embryonic shells of T. granifera found in four different ontogenetic stages (0.1 to higher than 3 mm) from three different locations. Numbers of total embryonic shells from 20 individuals dissected are given above the respective columns. 10