www.biodiversityjournal.com Biodiversity Journal ISSN 2039-0394 (Print Edition) ISSN 2039-0408 (Online Edition) SEPTEMBER 2012, 3(3): 157-260 with the support of FOR NATURALISTIC RESEARCH AND ENVIRONMENTAL STUDIES Julodis andreae Marseul, 1865 - Turkey world biodiversity association o n I u s 1 Julodis armeniaca Marseul, 1865, Turkey, Bingol Yayla, 20. VII.2007. 2 .J.aristidis, Tunisia, Tozeur, 8.IV.2004. 3. J. lucasi , Tunisia, Gafsa, 11.IV.2006. Cover: J. andreae, Turkey, Adiyaman - Nemrut Dagi, 12.VI.2007 (photos- collectionM. Gigli). The genus Julodis Eschscholtz, 1829 (Coleoptera Bupreslidae). The genus Julodis is the type genus of the Subfamily Julodinae Lacordaire, 1857. Species in this group are quite unusual Buprestidae for their general appearance, but especially for the larval biology. They don't live in wood or other plant tissues like other larvae of this family, but they dig into the soil and feed on the roots of various plants. In contrast to larvae of other Buprestidae, they are covered by long bristles directed obliquely backwards, helping in the movement in the ground, just like in the larvae of Coleoptera Cetonidae. The genus Julodis is currently divided into two groups. One in South Africa (especially Cape Region) and Namibia, represented by 34 taxa (species and subspecies), and another group primarily Palaearctic, spread from Spain and North Africa to the steppes of Central Asia (up to the Chinese province of Gansu), with some species in the Oriental Region (Pakistan and India) and other taxa in Kenya (92 species and subspecies). Many species are extremely variable. The widespread polymorphism and the existence of many similar species has led to the description of many species, then failed into synonymy, and probably others will follow the same fate, especially in certain groups of Palaearctic species. The European Fauna includes six species and subspecies, in the Iberian Peninsula, South France, Italy (Lampedusa Is.) and Balkans. Julodis onopordi s.l. is one of the most polymorphic species. It, according to actual conception, is widespread in the range of Mediterranean climate in Spain and southern France, and in North Africa, from Morocco to Sinai, with many different populations, some of which are considered subspecies. It is the only species in the genus in the Italian Fauna (Lampedusa Is., only). Another very variable species, spread over extensive territories, is Julodis andreae s.l., who lives in all the lands between Turkey, Iran and Azerbaijan. On the contrary, there are species very localized, limited to areas with well-defined characteristics of the soil and climatic conditions. Among these, three species live in the vast steppe and pre-desert around the large salt lakes of Northern Africa: Julodis aristidis Lucas, 1860, Julodis chrysesthes Chevrolat, 1860, Julodis kerimi Fairmaire, 1875. They are differently distributed over the territory in consequence of the type of soil, the presence of salt, gypsum, and its granulometry, of great importance for the needs of the larvae. Adults mostly feed on leaves and young twigs of shrubs and small trees, but also on herbaceous plants. Maurizio Gigli. Via Monte Macereton. 13-00141 Roma, Italy; email: gigli.maurizio@alice.it Biodiversity Journal, 2012, 3 (3): 159-164 New data of the freshwater fish genera Laubuca Bleeker, 1 860 (Cypriniformes Cyprinidae) and Phenacostethus Myers, 1 928 (Atheriniformes Phallostethidae) in Thailand Sitthi Kulabtong 1 *, Siriwan Suksri 2 & Chirachai Nonpayom 3 'Save wild life volunteer Thailand, Wangnoi District, Ayuttaya Province 13170, Thailand; email: kulabtong201 l@hotmail.com Reference Collection Room, Inland Fisheries Resources Research and Development Institute, Department of Fisheries, Thailand 10900; email: Siriwan. suksri@gmail.com 3 534/26 Soi Phaholyothin 58 Phaholyothin Rd. Sai Mai, Bangkok, Thailand; email : sornl33@hotmail.com * Corresponding author ABSTRACT In the present paper are reported, for Thailand, additional records of the cyprinid fish Lau- buca siamensis Fowler, 1939 and priapium fish Phenacostethus smithi Myers, 1928 re- spectively from Mekong Basin, Meklong Basin, Southeast Basin and from upstream of Bangpakong Basin and Yom Basin. Description and distribution data of the two freshwater fishes are also provided. KEY WORDS Laubuca ; Phenacostethus ; Cyprinidae; Phallostethidae; Thailand. Received 02.03.2012; accepted 26.06.2012; printed 30.09.2012 INTRODUCTION Freshwater fishes of the genera Laubuca Blee- ker, 1860 and Phenacostethus Myers, 1928 are po- orly known in Thailand. Currently, for Laubuca genus two valid species have been recorded in Thailand, namely L. caeru- leostigmata and L. laubuca (Smith, 1931; 1945; Banarescu, 1971). The cyprinid fish Laubuca siamensis was de- scripted by Fowler (1939) and the distribution of this species is known only in waterfall at Trang Pro- vince, peninsular Thailand. In Thailand, Laubuca species taxonomic status is still unclear, especially as far as concerns L. lau- buca'. in 1971, L. siamensis was reported as a junior synonym of Chela laubuca by Banarescu (1971) and in 2009, the genus Chela Hamilton, 1822 in Thailand was considered a junior synonym of genus Laubuca based on a molecular study of phylogene- tic interrelationships (Fang et al., 2009). Phenacostethus are small priapium fishes, found in large rivers and river estuaries in Sou- theastern Asia. The genus is separated into three valid species, namely P. trewavasae Parenti, 1986 from Sarawak, Malaysia, P. posthon Roberts, 1971 from peninsu- lar Thailand (Satul Province, Pungah Province), Malaysia and Sumatra, Indonesia and P. smithi Myers 1928 widely distributed: Mekong Basin (Thailand and Cambodia); Chantaburi River, Sou- theast Basin; Lower Chaophaya Basin (Bangkok); Malay Peninsula, Sarawak, Borneo, Malaysia (Myers, 1928; Roberts, 1971; Parenti, 1986; Rain- both, 1996; Parenti & Lim, 2005). In the present paper, the authors re-examine all specimens of the cyprinid fishes Laubuca stored in Inland Fisheries Resources Research and Develop- ment Institute, Department of Fisheries, Thailand [NIFI]. The authors found that the specimens named as L. laubuca from Thailand are clearly dif- ferent from the original description of Hamilton 160 S. Kulabtong, S. Suksri & C. Nonpayom (1822) of L. laubuca by the combination of the fol- lowing characters: lateral line scales, body depth, number of anal fin rays, pelvic fin length, pectoral fin length and absence of tubercles on lower jaw. The authors suggest the valid name of these speci- mens to be L. siamensis Fowler, 1939. In addition, the authors report additional records on the distribu- tion of L. siamensis in Mekong Basin, Meklong Basin and Southeast Basin, Thailand. As far as concerns the project at Yom River and upstream of Bangpakong Basin, Kabin Buri District, Prachin Buri Province and Nakhon Nayok Province, Central Thailand, we found many specimens of P. smithi in both areas, an additional record of this spe- cies in Thailand. ACRONYMS. Standard length (SL); Head length (HL). RESULTS Order Cypriniformes Bleeker, 1859 Family Cyprinidae Cuvier, 1817 Laubuca siamensis Fowler, 193 9 Chela laubuca. Smith, 1945: Peninsular Thai- land); Sontirat et al., 2006: Southeast Basin, Thailand. Examined material. NIFI 0079, 2 specimens, Aun River, Sakon Nakhon Province, Northeast Thailand, 1.1967, legit Sopa Trirat. NIFI 1227, 1 specimen, Klong Sang, Chav Raan Reservoir, Surat Thani Province, South Thailand, IV. 1983, legit Kar- nasuta, J. NIFI 1968, 1 specimen, Tapi River Basin, Phrasaeng Suratthani Province, South Thailand, IV. 1985, legit Sonkphan, L. (Fig. 1). NIFI 1969, 4 specimens, same data of NIFI 1968. NIFI 2527, 2 specimens, Tanow Sri River, Sune Pung Distric, Ratchaburi Province, West Thailand, XII. 1993, legit Chavalit Vidthayanon (Fig. 2). NIFI 2966, 4 speci- mens, Klong Phraya W.S. Krabi Province, South Thailand, 1970, legit J.N. Description. L. siamensis is distinguished from other species of Laubuca genus by the com- bination of the following characters: lateral line scales complete, with 31-33 + 2 scales; transverse line scales on body with 6-7/ 1 / 2-4 A scales; body depth is 28.6-33.0%SL. Pectoral fin is short not extending to the anus; anal fin with 3 unbranched rays and 1 8 A - 2 HA branched rays; pelvic fin is long (93. 8-136. 6%HL) reaching beyond the anus; pectoral fin length is 34.6-39.4%SL; a clearly black blotch above the pectoral fin base; thin black longitudinal stripe along the body; caudal pedun- cle with clearly dark blotch; lower jaw smooth, no tubercles on skin. Particularly, L. siamensis is compressed, body depth is 28.6-33.0%SL. Body width is 9.6- 13.2%SL. Scales in lateral series are medium to large, lateral line scales complete, with 31-33 +2 scales, transverse line scales on body with 6-7/ 1 / 2- 4 A scales and 16-18 predorsal scales. Head length is 21.2-26.8%SL. Eye is large, eye diameter is 25.5-37.9%HL (6.5-8.6%SL). Post orbital length is 39.0-43.8 %HL (8.6-10. 1%SL), snout length is short, 25.9-3 1.9%HL (6.0-7. 1 %SL) and interorbital width is 46.9-50.4 % HL (10.9-11.8 % SL), shorter than postorbital width (48.3-54.9 %HL or 12.0-12.5 %SL). Dorsal fin origin coincides with posterior anal fin origin, predorsal fin length is 62.8- 71.8%SL, prepectoral fin length is 25.4-30.6 %SL, prepelvic fin length is 43.0-46.5 %SL and preanal fin length is 64.1-68.9 %SL. Caudal peduncle depth is 9.2-10.9 %SL; pec- toral fin is long but not reaching beyond the anus, the pectoral fin length is 34.6-3 9. 4%SL showing 11-12 branched fin rays. Pelvic fin is long rea- ching beyond the anus, the pelvic fin length is 93. 8-136. 6%HL or 20. 8-36. 6%SL with 5 branched fin rays. Anal fin base is longer than dorsal fin base, the anal fin base length is 26.0-28.5%SL, dorsal fin with 3 unbranched rays and 8 branched rays and anal fin with 3 unbranched rays and ISA- 21 A branched rays. The dorsal fin base length is 10.5-14.0 %SL. Distribution. This species is distributed in Me- kong Basin, Meklong Basin, Southeast Basin and peninsular Thailand. Comparative notes. Other examined material. L. caeruleo stigmata: NIFI 0041, 1 specimen, Poung Klong Nong Moa, Ayuttaya Province, Central Thai- land, XII. 1966, legit Theachareon, p. NIFI 2602, 23 specimens, Pasak, Lopburi Province, Central Thai- land, 1.1994, legit Chukajom T. (Fig. 3). L. siamensis was descripted by Fowler (1931) from waterfall at Trang Province, peninsular Thai- land, and “ siamensis ” is referring to Siam, the old name of Thailand, the type locality of this species; this species has been considered unit now a junior New data of the freshwater fish genera Laubuca and Phenacostethus in Thailand 161 1 2 3 4 5 Figure 1. Laubuca siamensis , 48 mm SL, Tapi River Basin, Phrasaeng, Suratthani Province, South Thailand. Figure 2. L. siamensis , 51 mm SL, NIFI 2527 , Tanow Sri River, Ratchaburi Province, West Thailand. Figure 3. L. caeruleostigmata , 61 mm SL, N1F1 2602, Pasak River, Lopburi Province, Central Thailand. Figures 4, 5. Phenacostethus smithi , 15-17 mm SL, male (above) and female (below), N1F1 4545, Yom Basin, VIII. 2011, Siriwan Suksri leg., Thailand. 162 S. Kulabtong, S. Suksri & C. Nonpayom synonym of Chela laubuca (Banarescu, 1971) and genus Chela in Thailand has been considered a ju- nior synonym of Laubuca (Fang et al., 2009). The current status of L. siamensis is L. lau- buca. L. laubuca was descripted by Hamilton (1822) from Northern parts of Bengal (NorthEast India; Bangladesh). From a comparison with docu- ments and specimens from Thailand, we strongly believe that, in Thailand, the valid name of L. lau- buca is L. siamensis. L. siamensis from Thailand is distinguished from the original description of Hamilton (1822) and the report of Gunther (1868), who re-examined the C. laubuca specimens of Dr. Bleeker’s Collec- tion from Bengal, by the combination of the follo- wing characters: lateral line scales are 31-33 + 2 (in L. laubuca they are 37), body depth is 3.00-3.50 times greater than SL (in L. laubuca is 2.75-2.80 times); transverse line scales is 6-7/ 1 / 2-4 Vt. scales (in L. laubuca is 71/2/1/4); pelvic fin shows 5 rays (in L. laubuca 7); pectoral fin is short not extending to the anus (in. L. laubuca it reaches the anus); pel- vic fin is long extending beyond the anus (in L. lau- buca is shorter, not reaching beyond the anus); first ray of pelvic fin appears like a filament (in L. lau- buca it is undivided from the other branched rays); a thin black longitudinal stripe along the body and a clearly dark blotch on caudal peduncle (which are absent in L. laubuca). L. siamensis is clearly different from L. cae- ruleostigmata of Thailand by many characters such as: body depth is 3. 0-3. 5 times greater than SL (in L. caeruleostigmata is 2.25 times), lateral line scales includes 31-33 scales (in L. caeruleo- stigmata 34-35). L. siamensis has one black blotch above the pectoral fin base, a thin black longitudi- nal stripe along the body and a clearly dark blotch on caudal peduncle (L. caeruleostigmata shows 4- 5 dark vertical stripes above pectoral fin base on both sides of the body) (Smith, 1931; Smith, 1945). Order Atheriniformes Rosen, 1966 Family Phallostethidae Regan, 1913 Phenacostethus smithi Myers 1928 Examined material. NIFI 4545, 8 specimens, Yom Basin, Pakpot Subdistrict, Moung District, Su- Figure 6. Collection area, floodplain of Yom Basin in Sukhothai Province, Thailand. New data of the freshwater fish genera Laubuca and Phenacostethus inThailand 163 khothai Province, Thailand, 25.VIII.2011, legit Si- riwan Suksri (Figs. 4-5). NIFI 4548, 3 specimens, upstream of Bangpakong Basin, Kabin Buri Di- strict, Prachin Buri Province, Thailand, 5.II.2011, legit Sitthi Kulabtong. Description. P. smithi has a slender and cylin- drical body. Body depth is 16.5-19.9%SL. Head is short, head length is 18.2-19.1 %SL. Eye is big, eye diameter is 33.5-37.6 %HL. Post orbital length is 33.5-37.7%HL, snout short (23.5-24.3 %HL) and mouth is upward. First dorsal fin is very small, but second dorsal fin is large. Pre-second dorsal fin length is 70.6-70.7 %SL and the second dorsal shows 6-7 rays. Dorsal fin origin is clearly posterior the anal fin origin. Pre-anal fin length is 52.3-52.9%SL and the anal fin comprises 14-17 rays. Second dorsal fin base is shorter (12.4-14.4%SL) than anal fin base (23.5-25. 1%SL). The body is translucent, with tiny scales. Head with membranous dome. Males have a priapium (reproductive organ) below the head at the base of pectoral fin. The priapium of P. smithi is ruffled and hence distinguished from that (i.e. smooth) of other Phe- nacostethus occurring in Thailand. Variability. Priapium, the reproductive organ is found in males only. Biology and Distribution. Phenacostethus smithi were found in several habitats of Yom Basin (Fig. 6); floodplain canal and mainstream, charac- terized by slow and turbid waters and muddy bot- tom. In each habitat, submerged or marginal plants, such as green algae, papyrus and grass, were found. Specimens from upstream of Bangpakong Basin were found in a small stream nearly the mountain, with shallow, turbid and slow waters. This species is known from Yom Basin, Ban- gpakong Basin, Lower Chaophaya Basin and Sou- theast Basin in Thailand; Mekong Basin in Thailand and Cambodia; Malay Peninsula, Sara- wak, Borneo in Malaysia. ACKNOWLEDGEMENTS A special thanks to reviewers for reviewing this manuscript. Authors wish to thank Dr. Rohan Pe- thiyagoda. Wildlife Heritage Trust in Sri Lanka and Dr. Sorin Stefanut, Institute of Biology Bucharest, Romanian Academy, Romania for providing the original description of many species of Laabuca genus. Finally we are grateful to all partners for their support. REFERENCES Banarescu P., 1971. Further studies on the systematics of Cultrinae with reidentification of 44 type specimens (Pisces, Cyprinidae). Revue Roumaine de Biologie, Serie de Zoologie, 16: 9-20. Fang F., Noren M., Liao T.-Y, Kallersjo M. & Kullander S.O., 2009. Molecular phylogenetic interrelationships of the south Asian cyprinid genera Dcmio, Devario and Microrasbora (Teleostei, Cyprinidae, Danioni- nae). Zoologica Scripta, 38: 237-256. Fowler H.W., 1939. Zoological results of the third De Schauensee Siamese Expedition. Part IX. Additional fishes obtained in 1936. Proceedings of the Academy of Natural Sciences of Philadelphia, 91: 39-76. Gunther A., 1868. Catalogue of the fishes in the British Museum, v. 7. Catalogue of the Physostomi, contai- ning the families Heteropygii, Cyprinidae, Gonor- hynchidae, Hyodontidae, Osteoglossidae, Clupeidae, Chirocetridae, Alepocephalidae, Notopteridae, Halo- sauridae, in the Collection of the British Museum. Taylor & Francis, London, 512 pp. Hamilton F., 1 822. An account of the fishes found in the river Ganges and its branches. A. Constable e Co., Edinburgh, 405 pp. Myers G.S., 1928. The systematic position of the phal- lostethid fishes, with diagnosis of a new genus from Siam. American Museum Novitates, 295: 1-12. Parenti L.R., 1986. Bilateral asymmetry in phallostethid fishes (Atherinomorpha) with description of a new species from Sarawak. Proceedings of the California Academy of Sciences (Series 4), 44: 225-236. Parenti L.R. & Lim K.K.P, 2005. Fishes of the Rajang Basin, Sarawak, Malaysia. The Raffles Bulletin of Zoology Suppl. 13: 175-208. Rainboth W.J., 1996. FAO species identification field guide for fishery purposes. Fishes of the Cambodian Mekong. Rome, 265 pp. Roberts T.R., 1971. The fishes of the Malaysian family Phallostethidae (Atheriniformes). Breviora, 374: 1-27. Smith H.M., 1931. Descriptions of new genera and spe- cies of Siamese fishes. Proceedings of the United States National Museum, 79: 1-48. Smith H.M., 1945. The freshwater fishes of Siam, or Thailand. Bulletin of the United States National Mu- seum, 188: 1-622. 164 S. Kulabtong, S. Suksri & C. Nonpayom Sontirat S., Tunchareon S. & Soothornkit Y., 2006. Fish species diversity in the areas of National Parks and Wildlife Sanctuaries in the five eastern provinces of Thailand. Proceedings of 44th Kasetsart University Annual Conference: Fisheries, Bangkok (Thailand), p. 60-67. Biodiversity Journal, 2012, 3 (3): 165-172 Evaluation of the toxicity of metal pollutants on embryonic development of the sea urchin Paracentrotus lividus (Lamarck, 1816) (Echinodermata Echinoidea) Saliha Dermeche *, Fa/gal Chahrour & Zitouni Boutiba Laboratoire Reseau de Surveillance Environnementale (LRSE), Departement de Biologie, Faculte des Sciences ,Universite d'Oran, Algerie; e-mail: salidermeche@yahoo.fr ' Corresponding author ABSTRACT Bioassays are frequently used to evaluate biological effects of pollutants on marine orga- nisms. The objective of such tests is the detection of toxic effects on populations that are representative of a given ecosystem. Sea urchin is a model organism employed in the field of environmental toxicology due to its sensitivity towards various pollutants, particularly heavy metals. Preliminary bioassay tests on embryos and/or larvae of Paracentrotus lividus (Lamark, 1816) from Madagh (Oran, Algeria) were used to assess the potential toxicity and determine the LC$q of four metal pollutants, Cadmium, Copper, Lead and Zinc. KEY WORDS Bioassays; LCyp; Madagh; Heavy metals; Paracentrotus lividus. Received 22.05.2012; accepted 06.07.2012; printed 30.09.2012 INTRODUCTION Inland aquatic and marine systems represent containers for virtually all contaminants, via direct and indirect contributions (Peijnenburg et al., 1997). Research on the action of heavy metals on the development of the sea urchin eggs represent an important contribution to the progress of kno- wledge in the field of embryonic determination. Sea urchin is a preferred model in such a rese- arch, due to a number of reasons including external growth of embryos, rapid cell division rate and cell transparency, thus being commonly employed in the field of Environmental Toxicology (Guillou & Michel, 1993; Quiniou et al., 1997). Bioassays or bio-tests frequently use several techniques to mea- sure, predict and control the effect of the release of toxic substances on marine organisms. Present study examines the impact of four heavy metals, Cadmium, Copper, Lead and Zinc, on the embryonic development of the sea urchin Paracentrotus lividus (Lamarck, 1816) (Echinoder- mata Echinoidea). MATERIALS AND METHODS We investigated the site of Madagh bay (Oran, Algeria: 35°37'952" N; 000°104'243" W) (Fig. 1), a non-impacted area, closed at its ends by two small caps reducing the action of winds. Moreo- ver, the proximity of the Habibas island, which is considered a marine protected area, could make this site a reference station for comparative stu- dies regarding the monitoring of pollution impacts in the coastal marine ecosystem of western Alge- ria, a site rich in algae and Posidonia meadows (Benghali, 2006). Collection of sea urchins was carried out during the period March- June 2010, when spawning is at its peak in this echinoid species. Spawning was in- duced by injection of 0.5 ml of 0.5 M KC1 into the 166 S. Dermeche, F. Chahrour & Z. Boutiba X, MAROC ALGERIE A T L A S OKm 100 Km Figure 1. Sampling site: Madagh bay, Oran, Algeria. coelomic cavity of the sea urchin (Harvey, 1940); the male sex products were recovered "dried" and stored in melting ice. Moreover, sperms of several males were pooled. Females were placed in a 250 ml Erlenmeyer flask containing filtered seawater (FSW) so that the genital pore was in contact with the surface of the water. After spawning, eggs were sieved with a 160 micron sieve and collected in a test tube. Volume was adjusted to 500 ml using FSW and homogenized. Subsequently, the first 100 ml of the solution (containing eggs) was removed and replaced by FSW. This operation was repeated a second time (Dinnel et al., 1988; Quiniou et al., 1999; Guillou et al., 2000). Once suspensions of gametes were obtained, eggs and sperms were recovered separately in 2 ml of FSW. Fertilization was performed in beakers containing 1500 to 2000 oocytes to which 250 pi of sperm was added. After one hour of contact, we checked the fertilization success under a light mi- croscope. Bioassays were carried out according to Coteur et al. (2003). A 15 well plate was used for each metal pollutant (Cd, Pb, Cu and Zn). Four different increasing concentrations (10 gg/1; 50 jLig/1; 100 gg/1; 200 gg/1) and a control so- lution (FSW), employed as blank, were used. Each well contained 10 ml of each solution, then 250-300 embryos were transferred to each well and incuba- ted for 72 hours at 21 ± 1 °C. At the end, larval development was stopped by adding neutral formalin (35%) and the percentage of anomalies was determined according to the cri- teria of Klockner et al. (1985). Number of mal- formed eggs/larvae, expressed in percentage, was assessed under the optical microscope by scan- ning slides containing about 100 eggs each. The number of dead cells was adjusted by the formula: % mortalities corrected = (Po - Pt)/( 100 - Pt), where Po is the percentage of mortalities observed and Pt is the percentage of mortalities in controls (Abbott, 1975). Five replicates were performed for each concen- tration and each metal. The statistical treatment of experimental data was performed by the probit me- thod (Bliss, 1935), which is useful for experiments with reduced number of animals and particularly suitable for research on marine invertebrates, as con- firmed by Bendimerad (2000). RESULTS The mean percentages of embryonic abnorma- lities observed in each heavy metal solution ± stan- dard deviation are shown in Table 1 . As shown in Table 1, concentrations of 10 gg/1 and 50 gg/1 determined minor negative effects on larval development. At 100 gg/1, the malformation percentage is about double or more (respect to 50 Evaluation of the toxicity of metal pollutants on embryonic development of the sea urchin Paracentrotus lividus 167 [pg /l] Metals 10 50 100 200 Cd 7.31 ± 1.84 18.26 ±2.97 79.32 ± 15.38 88.00 ± 14.60 Pb 5.00 ± 1.68 12.20 ±7.85 22.40 ±7.97 40.33 ±4.04 Cu 5.93 ± 1.76 28.93 ±4.52 41.91 ±4.13 52.66 ±24.61 Zn 10.60 ±2.70 17 ±3.98 36.73 ± 10.32 40.5 ±24.02 Table 1 . Mean percentages of abnormal embryos ± standard deviation observed in the sea urchin P. lividus from Ma- dagh bay (Oran, Algeria) treated with four heavy metal solutions at different concentrations. jug/1) depending on the metal, in particular, it resul- ted 79.32% for Cd, 41.91% for Cu, 22.40% for Pb and finally 36.73% for Zn (Figs. 2-5). Graphs show results we partly expected: the more the concentration increases, the more the per- centage of malformations is important, although, surprisingly, percentage of malformations observed after treatment with Cadmium at 200 pg/1 is higher (88%) (Fig. 2) than that detected with the other me- tals: 52.66% (Copper, Fig. 3), 40.33% (Lead, Fig. 4) and 40.50 % (Zinc, Fig. 5). LCjp, calculated according to the method of Bliss (1935), resulted 61.65 pg/1 for Cd, 158.48 pg/1 for Cu, 389.04 pg/1 for Zn and 446.68 pg/1 for Pb. DISCUSSION Geffard (2001), using Pb solutions at 10 pg/1 and 50 pg/1, obtained, as percentages of malforma- tions, 14.8 ± 6.7% and 17.2 ± 3.9%, respectively; whereas, 100% of larvae with abnormalities were observed at 1200 pg/1. The LCjp was 482.0 ± 101.0 pg/1. The LC$o value we found for Pb is 446.68 pg/1. When comparing the two values, they appear to be very similar. In a previous study, carried out on the same spe- cies and in the same biotope, Dermeche (2010) re- ported, for Cd and Pb solutions at 10 pg/1, percentages of malformations of 8.33 ± 0.47% and 10.66 ± 0.47%, respectively. At 200 pg/1 the per- centages passed to 82.33 ± 0.94% and 40.67 ± 0.94% with a LC$g of 69. 1 8 pg/1 for Cadmium and 436.51 pg/1 for Lead. Once again, these values are close to the values discussed in the present paper (61.65 pg/1 for Cd and 446.68 pg/1 for Pb). Many authors use Zinc and Copper to determine the LC$q by using the sea urchin larvae. According to Hall & Golding (1998), Ghirardini et al. (2005) reported that a concentration of 30 pg/1 of Copper shows a negligible effect, while it takes a concentration of 50 pg/1 to observe the first malformed larvae. These results are consistent with those obtained by His et al. (1999) who observed developmental defects after treatment with a Copper solution at 60 pg/1. Our study gives a percentage of 28.93 of mal- formations at a concentration of 50 pg/1, a result that remains consistent with results obtained by different authors. Bougis & Corre (1974) suggested that the effect of Copper varies depending on the quality of brood stock. This would explain different results obtained. It is likely that gametes of poor quality are more sensitive to a toxic agent. Although echinoderms are capable of removing accumulated contaminants, the residence time in the body and the principal mode of elimination appear to depend on the metal (Warnau et al., 1997; Man- naerts, 2007). According to Basuyaux et al. (2009) and Petinay et al. (2009), larvae can develop up to a Copper concentration of 90 pg/1 but malforma- tions start appearing at 50 pg/1. Copper leads to a significant reduction in growth at 30 pg/1 with lar- vae showing spicules reaching 464 ± 7 microns while normal ones generally are up to 495 ± 9 pm. Bielmyer et al. (2005) noted that abnormalities in larval development and, above all, the stop at plu- teus stage were manifested at concentrations ran- ging from 40 to 80 pg/1. According to Fernandez & Beiras (2001), Cd causes 100% of arrest of development of the 168 S. Dermeche, F. Chahrour & Z. Boutiba °q Mnlfm ittaHcins ■ taNonaal Development : %Ma|fonnnliO[is ■ ^Normal DevdopninJt Figure 2. Percentage of deformed and normal larvae of Paracentrosus Jividus observed after tretament with Cad- mium solutions. Figure 3. Percentage of deformed and normal larvae of Paracentrosus Jividus observed after tretament with Cop- per solutions. Figure 4. Percentage of deformed and normal larvae of Paracentrosus Jividus observed after treatment with Lead solutions. Figure 5. Percentage of deformed and normal larvae of Paracentrosus Jividus observed after treatment with Zinc solutions. pluteus at 16 jLtg/1 and the stop at blastula and ga- strula stage at concentrations from 32 to 64 jng/1, re- spectively. Several studies demonstrated sensitivity of sea urchin embryos to heavy metals solutions in the range of 0.01-0.1 mg/1 for Hg and Cu, and 0.1- 10 mg/1 for Cd and Pb (Waterman, 1937; Kobaya- shi, 1981; Carr, 1996; Warnau et al., 1996). In a Cu solution at 64 pg/1, embryonic develop- ment was arrested at gastrula stage, and at morula stage at 128 jug/1. The toxic effects of Zinc on the larval development of sea urchin were as follows: the highest concentration used, 480 pg/1, completely inhibited the embryonic development; at very low concentrations (7.2 jng/1) no inhibitory effects were observed at first cleavage or at pluteus formation; exogastrula and Apollo-like gastrula were observed at concentrations ranging from 14 to 58 jug/1. At hi- gher concentrations (120 to 240 jng/1 ), embryonic de- velopment and the elevation of the fertilization membrane showed signs of delay and even malfor- mations, as well as polyspermies, permanent bla- stula, or exogastrula (Kobayashi & Okamura, 2006). Other metals known to cause exogastrulation in echinoids are: sodium selenite, cobalt chloride, zinc chloride or acetate, nickel, mercury chloride or ace- tate, the trivalent chromium and manganese chlo- ride (Rulon, 1952; 1956; Timourian, 1968; Timourian & Watchmaker, 1970; Kobayashi, 1971; 1990; Murakami et al., 1976; Pagano et al., 1982; Evaluation of the toxicity of metal pollutants on embryonic development of the sea urchin Paracentrotus lividus 169 Species Cu Cd Pb Zn References Paracentrotus lividus <32 pg/1 > 11 pg/1 — — Pagano et al., 1986 — — — > 3 3 pg/1 Ramachandra et al.,1997 — — 0.21-0.26 pg/1 — Warnau & Pagano, 1994 — — 482.68 pg/1 — Geffard, 2001 158.48 pg/1 61.65 pg/1 446.68 pg/1 389.04 pg/1 present study Strongylocentrorus pur- puratus 6.3 pg/1 — <9.7 pg/1 — Dinnel, 1990 — 0.5 pg/1 — 23 pg/1 Ramachandra et al.,1997 Strongylocentror us inter me dius — 0.5 to 2.5 pg/1 — — Gnezdilova et al., 1985 Arbacia Iixula — — — 10-100 pg/1 Castagna et al., 1981 Table 2. Toxicity (ZCjp) of heavy metal (Cd, Cu, Pb, Zn) solutions at different concentrations for different sea urchin species from the Mediterranean. Mitsunaga & Yasumasu, 1984; Vaschenko et al., 1994); according to Lallier (1955) and Timourian (1968), skeletal malformations of pluteus were caused by Zinc whereas delay in skeletal develop- ment was always caused by Cadmium and Cobalt (Kobayashi, 1990; Mannaerts, 2007). King & Riddle (2001) showed that exposure of Sterechinus neumayeri embryos to various con- centrations of Copper caused significant damages to the development at different stages and, parti- cularly, at the stage of blastula; moreover, signi- ficant abnormalities were observed at a concentration of 4-5 pg/1. High mortality of em- bryos was estimated at a concentration of 16 pg/1, and abnormalities were observed at a concentra- tion of 32 pg/1; a Copper concentration of 11.4 pg/1 caused 50% of developmental abnormalities after 6 to 8 days of exposure. Radenac et al. (2001) reported, for Cu solutions at 50 pg/1, about 36.9% of malformations which approximates the results (28.93%) observed, for the same concentration, in this study; however, at 100 pg/1 this rate reached 99.80% which exceeds our result (41.91%); 100% of larval malformations were obtained at 250 pg/1 while our study showed 52.66% of malformations at 200 pg/1. For Pb so- lutions, at 250 pg/1, a high percentage (96.6%) of malformation was reported. On the contrary, in our study, at 200 pg/1, we obtained only 40.33% of malformations. These results seem to suggest a different sensitivity of different species to heavy pollutants; concerning Zn, our results are coherent with those of Radenac et al. (2001). CONCLUSIONS Considering different stages of development of P. lividus , embryos and larvae were found to be the most sensitive and best suited to study heavy metal toxicity. Moreover, they can be used when testing short (embryotoxicity) and long-term (lar- val growth) issues. 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Cadmium bioconcentration in the Echinoid Pa- racentrotus lividus. influence of the Cadmium concentration in seawater. Marine Environmen- tal Research, 43: 303-331. Waterman A. J., 1937. Effects of salts of heavy metals on development of sea urchin, Arbacia punctu- lata. The Biological Bulletin, 73: 401-420. Biodiversity Journal, 2012, 3 (3): 173-178 Metoncholaimus sp.(Nematoda Oncholaimidae) pseudopara- site of Mullus surmuletus (Linnaeus, 1 758) (Perciniformes Mul- lidae) in the western Algerian Sea Maya Meriem Hassani',S. Ahmed Kerfouf '* & Nawel Amel BrahimTazi 2 ‘Laboratoire d'eco-developpement des espaces, Universite Djilali Liabes, Sidi Bel Abbes, Algerie; e-mails: mayahassani@live.com; kerfoufahmed@yahoo.fr 2 Laboratoire Reseau de Surveillance Environnementale, Universite Es-Senia Oran, Algerie; e-mail: meltazi@hotmail.com jjj ' Corresponding author ABSTRACT This study was carried out between October 2009 and July 2010 to determine nematode pa- rasites of the red stripped mullet, Mullus surmuletus (Linnaeus, 1758), in the gulf of Oran (western Algeria), located at 35°43’ N - 0°37’ W. A total of 100 fishes caught from the local fishermen by gill-net were investigated. Our investigation revealed the presence of three ne- matodes (one female and two males) located in the intestine of two infected fishes, these ne- matodes were alive and not attached to the mucosa of the fish host. The examination of the nematodes recovered showed that they belong to the genus Metoncholaimus Filipjew 1918 (Oncholaimidae Oncholaiminae). These nematodes are free living mostly in the coastal marine sediment; Mullus surmuletus might acquire them accidentally while either feeding on them or along with other food items taken from the bottom. KEY WORDS Mullus surmuletus ; Nematode; Pseudoparasites; Oran; Western Algeria. Received 04.05.2012; accepted 26.07.2012; printed 30.09.2012 INTRODUCTION The striped red mullet Mullus surmuletus (Lin- naeus, 1758) (Perciniformes Mullidae), a benthic perciform fish with a widely known distribution, is very common in the Algerian coasts and is a com- mercially important species (Figs. 1-3). On the other hand, its nematode parasites in the western Mediterranean sea are poorly known al- though having been studied since more than one century by numerous authors who considered the system Helminthes-Mw//ws as the richest one and the most diversified of the Mediterr anean sea (Ba- youmy et al., 2008; Ferrer et al., 2005; Neifar et al., 2007; Ternengo et al., 2009). In this regard, we began a helminthological study during which we faced the problem posed by pseudoparasitism by known species of free-living nematodes. MATERIALS AND METHODS Fish were collected by means of a trammel net at a mean depth of 15 m, according to the traditio- nal local small-scale fishery techniques. Individual body weight and size (total length), sex, and matu- rity stage were recorded. The range of fish size (total length) was 10.5-22 cm. The whole gastro-intestinal tract was removed immediately after capture and all portions (sto- mach, pyloric caeca and intestine) were opened by a longitudinal incision. Removal of contents was obtained by successive washes with a wash bottle in a Petri dish, the food material collected was examined under a dissecting microscope Zeiss Stemi 2000. Only helminthes infesting this tract were exami- ned. The parasites were hand sorted and placed ini- 174 M.M. Hassani, S.A. Kerfouf & N.A. BrahimTazi Figures 1, 2. Sampling site: Madagh bay, Oran, Algeria. Figure 3. Mullus surmuletus (Linnaeus, 1758). tially in 2% NaCl saline solution and then stored in 75% ethanol. The nematodes specimens were stai- ned in Acetic-Carmin, dehydrated and mounted in Canada balsam. The collected nematodes were cleared in glyce- rin for examination. Drawings were made with the aid of a Camera Lucida connected to a Wild bright field microscope. For the identification of nematodes, drawings were compared with those of specialists in parasitic nematodes of fishes (Anderson, 1992; Moravec, 1998). Identification of nematode pseudoparasites did require the consultation of specialist works on free-living nematodes (i.e. Hope & Murphy, 1972; Tarjan, 1980; Platt & Warwick, 1983). RESULTS Systematic position Phylum Nematoda Rudolphi, 1808 Class Adenophorea Linstow,1905 Order Enoplida Filipjev, 1929 Family Oncholaimidae Filipjev, 1916 Genus Metoncholaimus Filipjew, 1918 Metoncholaimus sp. Description (Figs. 4-7). Body elongated, somewhat tapering to both cephalic and caudal regions. Metoncholaimus sp. (Nematoda, Oncholaimidae) pseudoparasite of Mullus surmuletus in the western Algerian sea 175 Cuticle. Smooth without transverse striations, thick, particularly in the cervical and the caudal regions. Anterior extremity. The head bears a crown of tubular and non-segmented sensilla; mouth opening is spacious, roughly hexagonal, with narrow mem- branous margin provided with small papillae; lips are very distinct and developed; buccal capsule large, with well sclerotized walls, sub-terminal next to a simple muscular esophagus very long and so- mewhat expanded at its posterior part. Nerve ring encircling esophagus anteriorly, excretory pore not located. Outer sensory organs represented by a crown of well developed tubular and non-segmen- ted bristles, never seen beyond the nerve ring. Caudal region: the tail of males and females is conical, tapered and curved, short bristled mainly located in both preanal regions in both sexes. Genital equipment, males: testes initiating at a short distance below the end of the muscular esophagus; strong spicules, regularly pairs are ar- ched without individualized capitulum and the di- stal end is flared into two points. Solid gubernaculum, consisting of two distinct parts, central bulging and curved ventral apophysis. Nu- merous short bristles (8 to 10) are present in the cloacal opening region. Genital equipment, females: only one female was found, it was non-gravid with monodelphic uterus. Ovary anteriorly starting below esophagus end. Ge- nital opening situated in posterior end of the body. DISCUSSION The examination of stomach contents of Mullus surmuletus revealed the presence of several preys difficult to identify because of their advanced state of digestion. Generally, they are mainly composed of fragments of polychaetes, small fish or crustace- ans (N'Da, 1992). The stomach content analysis of Mullus surmu- letus revealed a wide food range that can explain the observed parasite richness (Klimpel et al., 2008). Only parasites resist to digestion which led us to believe that the three Metoncholaimus speci- mens found alive and intact were nematode parasi- tes of the fish. More precisely, these nematodes are pseudoparasites, i.e. Mullus surmuletus may acci- dentally ingest them from marine sediments, the diet of the latter being composed largely of benthic prey (Quero & Vayne, 1997) that the mullet har- vests from the sediment through its burrowing and tactile barbells (Bougis, 1949). Pseudoparasitism is a fairly rare phenomenon, but was nevertheless described; indeed, pseudopa- rasites were found alive and in perfect condition in the fish Haemulon sciurus (Shaw, 1803) in Brazil. On a total of 50 fish examined, 13 contained nema- tode pseudoparasites at the rate of 2 to 50 speci- mens per fish. These nematodes were identified as Metoncho- laimus amplus Filipjev, 1918, Oncholaimidae (Mo- ravec et al., 1990) that is a species of free living nematodes described for the first time by Hopper (1967). There are small differences between Meton- cholaimus amplus specimens which may be consi- dered to be within the limits of intraspecific variability. This species was recorded in the coast of Flo- rida, Bermuda Islands and also from the Suez canal (Riemann & Rachor, 1972). Comparison of our de- signs with those of Hopper (1967) did not allow us to assign our specimens to this species, however, we can assign it to the genus Metoncholaimus whose identification keys according to Keppner & Tarjan (1989) are: • Cuticle smooth without transverse striations. • Non-segmented cephalic setae located in one to three crowns before the nerve ring. • Simply muscular and cylindrical esophagus without vesicular cells. • Well-developed buccal capsule. • Tapered shank short and curved. • Spicules paired, fronted a gubernaculum. • Presence or no of ventral apophysis. • Short caudal setae concentrated primarily on the preanal region of male. From a comparison of Metoncholaimus amplus Hopper, 1967 with our specimens, a few differences came to our attention: (i) M. amplus has two crowns of cephalic setae, Metoncholaimus sp. just one; (ii) as far as concerns shape and size of the copulatory apparatus, M. amplus shows thin and slender spi- cules and lacks of a ventral apophysis, our speci- mens - on the contrary- had stronger spicules and a ventral apophysis; (iii) in M. amplus the nerve ring of esophagus is situated at the mid-length of eso- phagus while in Metoncholaimus sp. is located more anteriorly. 176 M.M. Hassani, S.A. Kerfouf & N.A. BrahimTazi 5 0 0 m 4 bucai capsule muscular esophagus 5 cephalic seta esophagus l o 0 M m spicules gubernaculum - apophysis anus 5 o o m 7 anus Figures 4-7. Metoncholaimus sp. pseudoparasite of Mullus surmuletus. Fig. 4: anterior end; Fig. 5: detail of the anterior end; Fig. 6: posterior end of male; Fig. 7: posterior end of female. Metoncholaimus sp. (Nematoda, Oncholaimidae) pseudoparasite of Mullus surmuletus in the western Algerian sea 177 It should be noted that M. ampins approximates our specimens particularly by the appearance of the head region, the esophageal structure, the shape of the buccal capsule, the measurements of the body and the implementation of the bristles in the caudal region. The genus Metoncholaimus has twelve species and, in the Mediterranean Sea, is mainly represen- ted by the species Metoncholaimus pristiurus Zur Strassen, 1894 (Gerlach & Riemann, 1973), with an abundance of 68.54% of the total population of free-living nematodes (Hedfi et al., 2010). CONCLUSIONS It is clear that Metoncholaimus sp. is not a real parasite of Mullus surmuletus. Indeed, setae on its cephalic and caudal regions indicate a free-living mode of life. Nevertheless, the mode of survival of pseudoparasites into the digestive tract and their resistance to digestive enzymes remain un- known although they are known to be biomonito- ring tools in coastal ecosystems due to their extreme sensibility to any environmental stress (Mahmoudi et al., 2002). Pseudoparasitism by known species of free li- ving nematodes is rare, however it is interesting to report it in order to attract the attention of some ichtyoparasitologists who would easily confuse free living nematodes with parasitic nematodes of fishes because, despite nematodes are the most fre- quent and the most important parasites of fishes constituting a significant part of the parasite fauna of these hosts in freshwater, brackish-water or even marine environments throughout the world, there is a world-wide shortage of specialists capable of identifying unknown helminthological materials (Moravec, 2007). REFERENCES Anderson R.C., 1992. Nematode Parasites of Vertebrates. 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Free-living marine nematodes. Part I. British Enoplids, Cambridge Uni- versity, Cambridge, 307 pp. Quero J.C. & Vayne J.J., 1997. Les poissons de mer des peches francaises. Delachaux etNiestle, Lausanne- Paris, 304 pp. Riemann T & Rachor P, 1972. Geographical distribu- tion of species of the genus Metoncholaimus (Filip- jew, 1918). Journal of Zoology, 21: 167-187. Tarjan A.C., 1980. An Illustrated guide to the marine ne- matodes. Institute of Food and Agricultural Sciences University of Florida, Gainesville, 135 pp. Temengo S., Levron C., Mouillot D. & Marchand B., 2009. Site influence in parasite distribution from fishes of the Bonifacio Strait Marine Reserve (Corsica Island, Medi- terranean Sea). Parasitology Research, 104: 1279-1287. Biodiversity Journal, 2012, 3 (3): 179-188 Biodiversity and conservation of Wildlife at the Wafra area n Kuwait Hanan AI-Khalifa'^Afaf Al-Nasser 1 , Mohammad Safar Abbas 2 &Jamal Dashti 1 'Kuwait Institute for Scientific Research, RO. Box 24885, 13109 Safat, Kuwait; email: hkhahfa@kisr.edu. kw 2 Joint Operation (JO)- Wafra, Kuwait Oil Company, Kuwait ’Corresponding author ABSTRACT In response to the increasing needs to conserve wildlife and to enhance biodiversity, the Joint Operations-Wafra (JO- Wafra) protected their natural environment by fencing and de- dicating areas for conservation of biological diversity. The main objective of this study is to conduct a wildlife baseline assessment in the oilfields of JO-Wafra and to identify po- tential habitats of endangered or threatened species that could occur on site. The wildlife survey covered the winter and early spring seasons. Although short and insufficient to pro- vide a detailed assessment, the field data collected indicated significant differences in the number of individuals and wildlife fauna species within the fenced and unfenced oilfields. It also showed that the fenced JO-Wafra has rich and diverse wildlife fauna species, an in- dication of ecological health. In addition to JO-Wafra oilfield, it is recommended to protect the South Umm Guddair (SUG) oilfields from livestock grazing and wildlife hunters. The protected area could, therefore, increase wildlife habitats and might harbor some endan- gered wildlife species. It is also recommended to connect the two oilfields with native shrubs and trees planted along the road, to serve as “green corridor”, shelter and additional source of food for the animals of both oilfields. KEY WORDS Endangered wildlife; Oil fields; Wildlife fauna; Wildlife habitat; Wildlife monitoring. Received 07.06.2012; accepted 02.08.2012; printed 30.09.2012 INTRODUCTION Biological diversity refers to the variety of life forms including the genes they contain, and the eco- systems they form. There are three different levels of biodiversity: genetic diversity which refers to the variety of ge- netic material contained in all the individuals, spe- cies diversity which refers to the variety of living species and ecosystem diversity which refers to the variety of habitats and ecological processes. In other words, it is reflected by the variety of all forms of life on earth, which provides the building blocks for human existence. The total number of species (defined as a popu- lation of organisms which are able to interbreed freely under natural conditions) is estimated to range from 5 million to 100 million globally; though less than 1.7 million have actually been de- scribed (BUDEST, 1993; FEPA, 2003; Maffi, 2005; Sarkar, 2006). Biodiversity conservation is of a major impor- tance internationally because humans derive their food, medicines and industrial products from bio- logical diversity of the wild and its domesticated components. It also gives future generations the op- portunity to enjoy nature. In addition, biodiversity is important for the recycling of essential elements, such as carbon, oxygen, and nitrogen. It is also responsible for mitigating pollution, protecting watersheds and combating soil erosion; accordingly, experiencing and increasing our kno- 180 H.Al-Khalifa,A.Al-Nasser, M.S. Abbas &J.Dashti wledge about biodiversity transforms our values and beliefs (McGregor, 1994; FEPA, 2003; Boro- kini et al., 2010). The state of Kuwait covers an area of about 17600 Km 2 at the north-eastern corner of the Ara- bian peninsula, between 28° 30’ N and 30° 05’ N in latitude and between 46° 33’ E and 48° 35’ E longi- tude. Kuwait's environment is characterized by scarcity of rainfall (100 mm/yr) and extreme tem- perature variations throughout the year. The summer temperature is very hot (over 40- 50°C), while the winter is cool to mild with a mean temperature of 12.7°C. The water resources are very limited in Kuwait. There is no fresh surface water supply and very limited renewable groun- dwater. The adverse climate conditions of the state of Kuwait affected its biological diversity and en- vironmental ecosystems (Omar et al., 2001). The desert of Kuwait has long been known as an im- portant source of food, livestock grazing and wil- dlife hunting. Desert plants were used for fuel and medication purposes. Urbanization, rapid increase in population, over- grazing, recreation usage, environmental factors, and destruction due to Iraqi invasion in 1990 are main contributing factors to the ecological degra- dation of the country (McGregor, 1994; Omar, 2000; Selby, 2005). Biodiversity conservation has become one of the challenging priorities for many countries, including the state of Kuwait, to combat species extinction. The State of Kuwait ratified the International Convention on Biodiversity and the National Strategy for Biodiversity Conservation was adopted in order to conserve and enhance bio- logical diversity in the country. Numerous wildlife research studies were conducted in the State of Ku- wait. From all these studies, information on flora and fauna of Kuwait has been collected. A list of desert animals has been prepared. Also, threatened desert animals and those to be extinct were identified such as Houbara Bustard ( Chlamy - dotis undulata macquennii ), Desert Monitor ( Vara - nus griseus). Ostrich ( Struthio camelus ) and Arabian Oryx ( Oryx leucoiyx ) (KISR., 1999; De- lima et al., 2005; Zaman et al., 2005). In December 1922, a partitioned neutral zone (PNZ) was established by agreement between the Kingdom of Saudi Arabia and the State of Kuwait, in order to allow tribesmen from both countries to use this favorite grazing ground (Chichester, 2000). In 1938-1940, oil was discovered from the Al- Burgan area, near PNZ. In 1948, a 60 year conces- sion was granted by the Kuwait government to the Aminoil, a small group of oil companies to ex- plore and exploit the Kuwait side of the PNZ. Si- milarly in 1949, the Getty Oil Company (formerly Pacific Western Oil Corporation) gained grants from the Kingdom of Saudi Arabia (KSA) to ex- plore the PNZ (KSA side). The Joint Operations (JO) was born in 1960 when the two oil companies formed a joint commit- tee to oversee and supervise their operations with the resultant productions divided equally to both parties: the Kuwait Oil Company (KOC), which operates the Kuwaiti concession and the Texaco In- corporated, Saudi Arabian Texaco (SAT) that ope- rates the Saudi Arabian concession. The 3,600 Km 2 partitioned neutral zone oil- fields were not spared during the Iraqi invasion of Kuwait in 1990. The oil wells were destroyed and burned contributing to the environmental catastro- phe man had ever known. In 1999-2002, the peri- meter fence around the JO-Wafra main oilfield was constructed to prevent livestock grazing and wil- dlife hunters from the area. This resulted to the gradual rehabilitation of the flora and fauna in the oilfield. Literature on the wildlife fauna of the Wafra area is very limited. Example is the “Insect Fauna of Kuwait” by Al- Houty (1989), when some insect samples were collected in the Wafra area. An environmental impact review prepared by Chichester (2000) described the fauna of the upland deserts and sabkha of the Partitioned Neu- tral Zone (PNZ) as “Over 220 species of birds have been observed in the PNZ in recent years... fauna of the upland deserts and sabkha includes common insects; ants and beetles; lizards and sna- kes; such as Sand Boa, Rat Snake, Blue-throated Agamid, Desert Monitor, and Dhub. Small, noc- turnal mammals include Jerboa, Jirds, Desert Fox, and Long-eared Hedgehog”. The current study involves fauna baseline asses- sment within the JO-Wafra territories with the fol- lowing main objectives: assess the wildlife fauna in JO-Wafra main area; and compare the quality of ha- bitat inside and outside the JO-Wafra main area. The work has been implemented between Kuwait Institute for Scientific Research (KISR) and Kuwait Gulf Oil Company (KGOC). Biodiversity and conservation of Wildlife at the Wafra area in Kuwait 181 MATERIALS AND METHODS RESULTS The wildlife fauna study commenced in De- cember 2005 and terminated in March 2006. This report covers only the study period (i.e. from Ja- nuary to March, 2006), wherein 15 field data col- lection exercises were performed with a total of 345 field data collected from 11 selected wildlife (fauna) study sites, within the fenced JO- Wafra main oilfield and the unfenced SUG (South Umm Guddair) oilfield. Selections of wildlife study sites at the JO- Wafra oilfield were performed during re- connaissance surveys. The criteria used in selecting the possible wildlife study sites were: (1) type of habitat that include soil and vegetation cover; and (2) location within the oilfield, disturbed or undi- sturbed (see Table 1). Several wildlife survey methodologies were im- plemented to study the wildlife biodiversity, na- Types of habitat Wildlife study sites number 1 .Habitat with good vegeta- tion cover 2, 3, 4, 8, 9 and 10 2. Habitat with poor vegeta tion cover 1 and 5 3. Windblown sand covered habitat 6 and 7 4. Overgrazed and unprotec ted habitat 11 Table 1. Different Wildlife study sites according to types of habitat. mely: 1) Line Transects (LT): of 5 km to record animals within a specific habitat type. 2) Pitfall Trapping (PFT): to catch ground crawling animals such as reptiles and invertebrates. 3) Baited Mam- mal Trap Line: is usually a one kilometer long trap line. The large mammal trap (MTL) is placed bet- ween two small mammal traps (MTS) at a distance of 250 meters. 4) Mark-Release-Recapture (MRR): to estimate the population dynamics of an area. MRR models were developed for field studies in which the count statistics are numbers of marked and unmarked animals caught (Nichols, 1992; Grenwood, 2000). From December 2005 to March 2006, 17 trips were made to the JO- Wafra main (fenced) and the unfenced South Umm Guddair (SUG) oilfields. A total of 15 field data collecting exercises were performed. This included line transects, baited mammal trappings and pitfall trappings, conduc- ted over at least three consecutive days (Table 2; Figs. 1-2). Line Transects (LT): a total of 49 LT exercises were performed, covering the winter and early spring periods of the country. It was noticeable that during the last days of line transect exercises, more birds were observed and even the shy Red fox ( Vul- pes vulpes) was recorded. This may be due to the rise in temperature and the pleasant spring weather in the air. More than 34 species of wildlife fauna were recorded from the line transects performed. These included 27 species of birds, one species of mammal, one species of reptile and more than six species of invertebrates (butterflies, dragonflies, flies and ground hoppers). Table 3 lists the species recorded during line transect exercises at the JO- Wafra oilfields. The list is not conclusive, as it was taken during winter and early spring seasons. No. of Field Trips Type of Trips Location of Trips Field orientations, recon- naissance, site selections and installation of field JO-Wafra main and 2 study equipment e.g., SUG oiltield mammal traps and pitfall (south Um Gud- traps with drift fences dair) Field data collection resul- ting to the following field data collected: 49 from line transects (LT) 11 wildlife (fauna) 148 from two (x2) small study sites within 15 mammal traps at each the fenced JO- study site Wafra and the un- 74 from one (xl) large fenced mammal trapping at each SUG oilfields study site 74 from pitfall traps in as- sociation with drift fences (1x5) at each study site Table 2. Trips made by Wildlife Survey Team to JO- Wafra Oilfields. 182 H.Al-Khalifa,A.Al-Nasser, M.S. Abbas &J.Dashti Wildlife fauna recorded during LT (January 2006 to March 2006) 30 25 4- ® m 20 ■g S 15 C Q. = W10 5 0 ~7 Inside JO Outside JO Location of LT Figure 1. Wildlife species recorded during line transect in- side and outside JO-Wafra oilfields Wildlife fauna recorded at JO-Wafra (January - March 2006) Herbivores 3% Fruitivores 6 % Granivores 24% Consumers 9% Predators 15% Insectivores 43% Figure 2. Different trophic levels in the wildlife fauna re- corded at JO-Wafra oilfields from January to March 2006. Ground crawlers trapped in PFT at JO-Wafra oilfields (January 2006 - March 2006) 20 S 15 H Oi a to 10 © 2 Lizards Insects Beetles Arachnids ■ Number of species 5 16 15 10 Class of Animals Animal Species Trapped in PFT at JO-Wafra (January 2006 -March 2006) Figure 3. Classes of animal species trapped in PFT at JO- Wafra oilfields. Figure 4. Animals recorded from PFT at JO-Wafra are shown according to their trophic levels. More animal species are expected to be recor- ded if the survey covers the four seasons, especially the two migration periods of the country. Except for the residents, such as the Black-crowned finch lark. Crested lark, House sparrow and the Feral pigeon, the birds might only be over wintering in the coun- try e.g., Tawny pipit, Short-eared owl, Blue rock thrush. Pied wheatear, Woodchat shrike, Great grey shrike, and Hoopoe lark among others. The list is only 7.7% from the total number of bird species recorded in the country. The Black- crowned finch larks ( Eremopterix nigriceps ) were recorded breeding at study area, while both the Cre- sted larks ( Galerida cristata) and the Isabelline wheatears ( Oe nan the isabellina) were also obser- ved displaying courting behaviors. Pitfall trappings: there were 64 pitfall trapping exercises performed at the fenced main JO-Wafra and the unfenced SUG oilfields. More than 46 ani- mal species were recorded during these exercises, including 10 species of arachnids; lizards (5 spe- cies); beetles (15 species); and 16 species of insects (Figs. 3-4 and Table 4). Baited mammal trappings: there were seven (x7) mammal trapping exercises performed at the JO-Wafra study sites, for the du- ration of the study period. Biodiversity and conservation of Wildlife at the Wafra area in Kuwait 183 Study Site SN Animal Species G SLT NLT G SUG 1 Black-crowned Finch Lark, Eremopterix nigriceps (Gould, 1839) X X X 2 Barn swallow, Hirundo rustica (Linnaeus, 1758) X X 3 Blue Rock-Thrush, Monticola solitaries (Linnaeus, 1758) X X 4 Lepidoptera sp. 1 X X X 5 Chiffchaff, Phylloscopus collybita (Vieillot, 1817) X 6 Cream-coloured courser, Cursorius cursor (Latham, 1787) X 7 Crested lark, Galerida cristata (Linnaeus, 1758) X X X X X 8 Arabian babbler, Turdoides squamiceps (Cretzschmar, 1 827) X X X 9 Desert wheatear, Oenanthe deserti (Temminck, 1825) X X X 10 Dhub, llromastyx microlepis (Blanford, 1874) X X X 11 European roller, Coracias garrulous (Linnaeus, 1758) X X X 12 Feral pigeon, Columba livia (J.F. Gmelin, 1789) X X X X X 13 Great grey shrike, Lanius excubitor (Linnaeus, 1758) X X X X X 14 Ground hopper, Tetri x undulata (Sowerby, 1 806) X 15 Floopoe, Up upa epops (Linnaeus, 1758) X 16 Greater Hoopoe lark, Alaemon alaudipes (Desfontaines, 1789) X X X 17 House sparrow, Passer domesticus (Linnaeus, 1758) X X X 18 Isabelline wheatear, Oenanthe isabellina (Temminck, 1829) X X X X X 19 Kestrel, Falco tinnunculus (C.L. Brehm, 1855) X X X 20 Northern wheatear, Oenanthe oenanthe (Linnaeus, 1758) X X X X 21 Olivaceous warbler, Iduna pallida (Hemprich et Ehrenberg, 1833) X X X 22 Pallid harrier, Circus macrourus (S.G. Gmelin, 1770) X X X 23 Pied wheatear, Oenanthe pleschanka (Lepechin, 1770) X X X X X 24 Red-backed shrike, Lanius collurio (Linnaeus, 1758) X X X X X 25 Red fox, Amphicoma vulpes vulpes (Fabricius, 1792) X 26 Sand martin, Riparia riparia (Linnaeus, 1758) X 27 Short-eared owl, Asio flammeus (Pontoppidan, 1763) X X 28 Short-toed eagle, Circaetus gallicus (Gmelin, 1788) X X X 29 Short-toed lark, Calandrella br achy dactyl al longipennis (Eversmann, 1848) X X X 30 Stable flies, Stomoxys calcitrans (Linnaeus, 1758) X X X X X 31 Sundevall jird, Meriones crassus (Sundevall, 1842) X 32 Swift, Apus barbatus (Sclater, 1866) X X 33 Tawny pipit, Anthus campestris (Linnaeus, 1758) X X X 34 Turtle dove, Steptopelia turtur (Linnaeus, 1758) X Table 3. Animal species recorded during line transect at the JO-Wafra Oilfield. Legend: G=General line transect. SLT=South LT. NLT=North LT. SUG= Unfenced JO-Wafra oilfield LT. 184 H.Al-Khalifa,A.Al-Nasser, M.S. Abbas &J.Dashti Study Site SN Animal species 1 2 3 4 5 6 7 8 9 10 11 1 Arabian darkling beetle, Pemelia arabica (Kaszab 1982) X X X X X X X X X X X 2 Brilliant ground weevil, Bembidion sp. X X X 3 Camel spider, GaJeodis arabs (Koch, 1842) X X X X X X X X X X X 4 Capsid bug, Eurydema ornatum (Linnaeus, 1758) X 5 Centipede, Craterostigma sp. X X 6 Churchyard beetle, BJaps kollari (Seidlitz G von, 1896) X X 7 Desert runner, Cataglyphis niger (Andre, 1981) X X X X X X X 8 Domino beetle, Anthia duodecimguttata (Bonelli, 1813) X X X X X X X X X X X 9 Elevated stalker, Adesmia stoeckleini (Koch, 1940) X X X X X X X X X X X 10 Giant black ant, Camponotus xerxes (Forel, 1904) X X X X X X X X X X 11 Golden-tipped ant, Camponotus sericeus (Forel, 1904) X X X X X X X X X X 12 Ground mantis, Eremiaphila braueri (Krauss, 1902) X 13 Hairy capsid bug, Tropinota squalida (Scopoli, 1763) X 14 Joker bee, Parachistus pulchellus (Greathead, 1980) X X 15 Jumping spider, Salticidae X X X 16 Fesser scarab, Mnematium sp. X X X X 17 Fesser yellow scorpion, Uroplectes alstoni (Purcell, 1901) X 18 Fong-legged spider, Pholcidae X X X X X X X X X X X 19 Mesopotamian beetle, Sepidium mesopotamicum (R.,1904) X X X X 20 Mosquito, Anopheles pharoensis (Theobald, 1901) X 21 Opossum beetle, Mesostena puncticollis (Sober, 1835) X X X X X X X X X X X 22 Orb-weaver spider, Araneidae X X X X X X X X X X 23 Pill bug, Armadillidium vulgare (Fatreille, 1804) X X X X 24 Pinstripped ground weevil, Ammoc/eonus aschabadensis (Ft., 1884) X X X 25 Pitted beetle, Adesmia cancell ata (Klug, 1830) X X X X X X X X X X 26 Meloe "Queen", Meloe omanicus (Kaszab, 1983) X X X 27 Rack beetle, Tentyrina palmeri (Crotch, 1872) X X X X X X X X X X X 28 Rock gecko, Bunopus tuberculatus (Blanford, 1874) X X X X X X X 29 Sand gecko, Stenodactylus doriae (Blanford, 1874) X 30 Saber-toothed beetle, Scarites guineensis (Dejean, 1831) X 31 Scorpion (Black), Androctonus crassicauda (Olivier, 1807) X X 32 Seville row beetle, Paraplatyope arabica (Koch, 1965) X 33 Short-nosed lizard, Mesalina brevirostris (Blanford, 1874) X X X X X X X X 34 Silverfish, Thermobia domestica ( Packard, 1837) X X X X X 35 Small black ant, Monomorium gracillimum (Smith, 1861) X X X X X X X X X X X Biodiversity and conservation of Wildlife at the Wafra area in Kuwait 185 Study Site SN Animal species 1 2 3 4 5 6 7 8 9 10 11 36 Small red ant, Monomorium pharaonis (Linnaeus, 1758) X X X X X X X X X X X 37 Scutte lizard, Acanthodactylus scutellatus (Audouin, 1827) X X X X X X X X 38 Schmidt lizard, Acanthodactylus schmidti (Haas, 1957) X X X 39 Tiger moth, Utetheisa pulchella (Linnaeus, 1758) X 40 Ugly trox, Scleron sulcatum (Kulzer, 1956) X 41 Variable stalker, Adesmia cothurnata (Forskal, 1775) X X X X X X X X X X X 42 Winged ant, Podalonia sp. X 43 Wolf spider, Lycosidae X X 44 unidentified moth X X X 45 unidentified larva X X X 46 Lepidoptera sp. 2 X X Table 4. List of animal species trapped at different PFT in JO-Wafra Oilfields. The twenty two (x22) MTS trapped 15 rodents from one species ( Meriones crassus). Whereas the collapsible Tomahawk traps (MTL) trapped one species (feral dog). There were seven rodent recap- tures and three of the feral dog, which is ‘trap happy’ because it was recaptured every trapping exercises. Table 5 shows the animal species trapped and caught during mammal trapping exercises at the JO-Wafra oilfields. Mark-release-recapture (MRR): the field data collection exercises were performed during the win- ter, when the temperature ranged from 3°C to 8°C and during early spring, when the temperatures star- ted to rise (14°C to 18°C). Therefore, the field data collected is not representative of the entire popula- tion of each study site but indicative only for the duration of the study period. There were 32 trapping exercises performed at the JO-Wafra oilfields. This includes eight exercises each for the PFT; baited MTS1; MTS2; and MTL. These trapping exercises caught a total of 74 wil- dlife fauna (including recaptures) from different study sites at JO-Wafra. Five (5) species of lizards were caught in the PFT, namely: Mesalina breviro- stris (27 individuals); Acanthodactylus scutellatus (x8); A. schmidtii (x2); and 22 geckos ( Stenodacty - lus slevini and B unopus tuberculatus). Nine recaptures were recorded for the lizards during pitfall trappings; the first three species of the above-mentioned lizards were recorded to be active in winter, while the two species of geckos were trapped only when the ambient temperature at the study sites were higher at around 20°C. The fringed-toed lizards i.e. A. scutellatus and A. schmidtii seemed to be concentrated only at the low lying soot covered habitat, with good vegeta- tion cover. The two species of geckos: S. slevini and B. tuberculatus were trapped in the PFT only at the start of spring, when the temperatures at the study sites were higher. These species were recor- ded during the last two exercises. Two (2) species of mammals (Meriones crassus and Canis dome- sticus ) were trapped during baited mammal trap- ping exercises. There were 19 Sundevall jirds ( Meriones cras- sus) captured by the small mammal traps from eight study sites within the JO-Wafra main oilfield. Seven re-captures were recorded. DISCUSSION AND CONCLUSIONS The current study was conducted to assess the wildlife fauna in JO-Wafra main area; and to com- pare the quality of habitat inside and outside the JO- Wafra main area. The work has been implemented between Kuwait Institute for Scientific Research (KISR) and Kuwait Gulf Oil Company (KGOC). 186 H.Al-Khalifa,A.Al-Nasser, M.S. Abbas &J.Dashti Seventeen field visits to the JO-Wafra oilfields were undertaken by the wildlife team, to perform 15 field data collection exercises that resulted in 444 field data collected. The oil fires that occurred during the Iraqi invasion of Kuwait were the main cause of diversity loss in the country. It had signi- ficant effect on species and ecosystem, this effect shifted species distribution and caused reductions in population size that could be due to reduction in survival and fecundate rates. This negative effect is well documented in other studies that investigated similar items (Da Fonseca et al., 2005; Parmesan, 2006; Fischlin et al., 2007). Conserving wildlife and biodiversity is increasingly recognized as an essential element of life. Its importance involves production of plants and animals for food, providing recreational re- sources, flood and pest control, providing chemi- cals for treatments. Accordingly, biodiversity conservation is strongly related to finance, eco- nomy and poverty degree in a society. Petts & Platt (1990) demonstrated that most of the benefits derived from wildlife and biodiversity conservation are potentially quantifiable and can significantly add to the economy of a society. Many worldwide studies in the literature relate biodiversity to social poverty (Adams et al., 2004; Treves et al., 2005; Fisher & Christopher, 2007). Consequently, wildlife and biodiversity conserva- tion became a priority in the world. The approach of protecting natural resources and increasing extent of protected areas has been described and used in other worldwide studies to conserve wildlife and biodiversity (McNeely & Schutyser, 2003; UNEP, 2006). In addition to re- source protection, the current study also applied the technique of monitoring wildlife populations. It involves collecting, analyzing, and interpreting ecosystem information. Such techniques develop wildlife and natural resource management ap- proaches. Monitoring wildlife and ecosystems provide information that managers and compa- nies such as Kuwait Oil Company can use to ad- just or modify their commercial activities so that they minimize negative effect on natural resour- ces. These techniques have been used worldwide to conserve wildlife and biodiversity (Adger et al., 2003; Fischlin et al., 2007). The field data collected showed that the fenced JO-Wafra main oilfield has rich and diverse wil- dlife fauna species, which suggests that ecological health in this area is significantly better than the unfenced and unprotected SUG oilfields. Although short and insufficient to provide a de- tailed assessment of the studied areas, the field data collected showed that the fenced JO-Wafra main area is rich and diverse in wildlife fauna, in- dicating significant ecological health compared to the unfenced and overgrazed South Umm Guddair oilfield, which is located approximately 29 kilo- meters northwest of the main oilfield. The field data collected also indicated that additional sur- veys and monitoring activities for the wildlife at the JO oilfields are necessary and conducted to cover the different climatic seasons and migration periods of the country. The field data collected showed that there are at least four types of wildlife habitats at the JO- Wafra oilfields: high and low lying habitat with good vegetation cover; high and low lying habitat with poor vegetation cover; windblown sand co- vered habitat; and over-grazed and unprotected ha- bitat. The first three types of habitats are located inside the fenced oilfield, while the latter is at the unfenced South Umm Guddair (SUG) oilfield. Be- cause of the perimeter fence constructed in 2000, the JO-Wafra main oilfield enjoyed protection from livestock grazing and wildlife hunters. This has brought to the gradual rehabilitation of the flora and fauna within the perimeter fence. There were more than 78 wildlife fauna species recorded at the JO-Wafra main oilfield. This in- cludes the 19 species (24.36%) of wildlife fauna recorded at the unfenced SUG oilfield. The ti- ming of the field data collection might have im- pact on the numbers of individuals and fauna species recorded from both study areas because desert animals tend to hibernate during winter and only come out during spring. It is suggested and recommended that the SUG oilfield be fen- ced and protected from livestock overgrazing and wildlife hunters. The two oilfields then could be connected with native shrubs and trees plan- ted along the road. The fenced SUG and the additional plants will augment and increase the possible areas for re- sting, feeding and even breeding of some threate- ned and endangered migrating fauna that pass through the country during their migration move- ments (examples are the Houbara bustard, Chla- Biodiversity and conservation of Wildlife at the Wafra area in Kuwait 187 mydotis undulata macquennii and the Imperial eagle, Aquila heliaca). The Houbara bustard be- cause of the size of its habitat requirement may be- come the "umbrella" species in the wildlife conservation program. In other words, the pre- sence of Houbara bustard bird in any habitat re- flects richness in biodiversity in that habitat (Gregory, 2005). Accordingly, it is important that wildlife conservation programs are oriented speci- fically at particular species of most concern such as Houbara bustard. Such programs should be im- plemented based on regional, national and interna- tional scope (Young, 1997; Mawdsley et al., 2009). The weather during the field data collection exercises (occurrences of rains and low tempera- ture) might have impact on the animals’ availabi- lity. Ectothermic (cold blooded) animals tend to hibernate during cold weather (winter) and come out only from hibernation when the temperature is favorable (spring). This could be true because the Sand gecko (, Stenodactylus s lev ini ) and the Rock gecko ( Bunopus tuberculatus) were trapped only during the PFT trapping exercises in March, when the temperature in the oilfields had risen to above 20°C, whereas most of the beetles, ants and the fringe-toed lizards, such as Acanthodactylus scu- tellatus ; A. schmidtif, and Mesalina brevirostris were trapped when the temperatures in the field ranged from 3° to 8°C. The large percentage of predators (51%) of ani- mals recorded from the PFT indicated good supply of prey or food resources i.e. consumers 47% and scavengers 2%. The low numbers of captures in the baited mammal trappings were expected be- cause of the timing of the field exercises, winter. Desert animals tend to hibernate during winter and come out during spring and summer (examples are hedgehog, gerbil, jerboa, etc.). More animal spe- cies are expected to be recorded if the survey was to cover the four seasons, especially the two di- stinct migration periods of Kuwait. Expanding the survey period and applying more conservation strategies and programs is recommen- ded in future work. No conservation program or strategy is optimal, some strategies have to be oriented to a specific target. Development of a set of strategies or approaches that complement each other is sometimes important to create useful con- servation tools and to fulfill requirements needed for an appropriate wildlife conservation approach. ACKNOWLEDGEMENTS The authors would like to thank Joint Opera- tion management for facilitating the research as- sessment in Al Wafra fenced area. Thanks are also extended to KISR staff for their effort in executing field work. REFERENCES Adams W.M., Aveling R., Brockington D., Dickson B., Elliott J., Hutton J., Roe D., Vira B. & Wolmer W., 2004. Biodiversity Conservation and the Eradication of Poverty. Science, 306: 1146-1149. Adger W.N., Huq S., Brown K., Conway D. & Hulme M., 2003. Adaptation to climate change in the deve- loping world Progress in Development Studies, 3: 179-195. Al-Houty W., 1989. Insect Fauna of Kuwait. University of Kuwait. Al-Marzouk press, 1 89 pp. Borokini T.I., Okere A., Giwa A.O., Daramola B.O. & Odofin W.T., 2010. 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(ed.), Cambridge University Press. Gregory G., 2005. The birds of the state of Kuwait. Cupit Print. England. KISR., 1999. Soil Survey for the State of Kuwait - Vo- lume 1 Executive Summary. AACM International, Adelaide, Australia. ISBN 09 5770 0303X. Maffi L., 2005. Linguistic, Cultural, and Biological Diversity. Annual Review of Anthropology, 34: 599-617. Mawdsley J.R., O’Malley R. & Ojima D.S., 2009. A Re- view of Climate-Change Adaptation Strategies for Wildlife Management and Biodiversity Conserva- tion. Conservation Biology, 23: 1080-1089. McGregor J., 1994. Climate change and involuntary mi- gration: implications for food security. Food Policy, 19: 120-132. McNeely J.A. & Schutyser F., 2003. Protected areas in 2023: scenarios for an uncertain Future. Internatio- nal Union for the Conservation of Nature, Gland, Switzerland . Nichols J.D., 1992. Capture-Recapture Models: Using marked animals to study population dynamics. Bio- science, 42: 94-102. Omar S., 2000. 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Biodiversity Journal, 2012, 3 (3): 189-190 On the presence of the Caddisflie Stenopsyche siamensis Marty- nov, 1 93 1 from Central Thailand (Trichoptera Stenopsychidae) Nidsaraporn Petsut 1 *, Sitthi Kulabtong 2 & Patinya Sreesamran 3 ’Department of Agricultural Technology, Faculty of Science, Ramkhamhaeng University, Bangkok 10240, Thailand; e-mail: nidsaraporn@ru.ac.th 2 Save wild life volunteer Thailand, Wangnoi District, Ayuttaya Province 13170, Thailand; e-mail: kulabtong2011@hotmail.com 3 53 Moo 3, Banyang Subdistrict, Muang District, Nakronpatom Province 73000, Thailand; e-mail: man_evo@hotmail.com ■"Corresponding author ABSTRACT In the present paper, caddisflie larvae and pupae of Stenopsyche siamensis Martynov, 193 1 (Trichoptera Stenopsychidae) are recorded from upstream of Maewong Basin, Central Thailand. KEY WORDS Caddisflies; Stenopsyche siamensis ; Stenopsychidae; Trichoptera. Received 08.07.2012; accepted 26.07.2012; printed 30.09.2012 INTRODUCTION Caddisflies are aquatic insects of the order Tri- choptera Kirby, 1813. This order of aquatic insects is a very large one comprising more than 10,000 spe- cies in the world (Sangpradub & BoonSoong, 2006). In tropical Asia, Caddisflies include about 28 fa- milies (Dudgeon, 1999) and, more particularly, 491 species were recorded in Thailand (Malicky & Chantaramongkol, 1999). In natural freshwater eco- systems Caddisflies are important and fundamental components. Caddisflies of the family Stenopsychidae Mar- tynov, 1924 are different from other families of Tri- choptera by the combination of the following morphological and ecological characters: a) head longer than 2 times as long as wide; b) mesopleuron not extended anteriorly; c) dorsum of abdominal segment 9 without sclerotized plate; d) larvae of the family building nests between large stones (San- gpradub & BoonSoong, 2006). In Thailand, Stenopsychidae comprise one genus, Stenopsyche McLachlan, 1868, and six spe- cies (Malicky & Chantaramongkol, 1999) including Stenopsyche siamensis Martynov, 1931; life cycle and feeding habits of S. siamensis in Thailand were studied by Laudee & Chantaramongkol (2003). Stenopsyche siamensis Martynov, 1931 This species is widely distributed in Thailand and Malaysia (Laudee & Chantaramongkol, 2003). In a survey project of aquatic ecology at upstream of Maewong Basin, Pangsira Thong District, Kam- phaeng Phet Province, Central Thailand, carried out in May 2012, we found many specimens of S. sia- mensis at larvae and pupae stage. This report is important for faunistic and eco- logical aspects, considering that all the Trichop- tera are good ecological indicators. Particularly, larvae and pupae of S. siamensis (Laudee & Chantaramongkol, 2003), were found in upstream mountainous areas or islets of Maewong Basin, where water is of high-quality, the stream is tran- sparent and running fast, on average about less than 1 m deep and the stream ground is made of rough sand and large stones. These larvae make protective cases by fibers between large stones and/or under stones, stay in- 190 N. Petsut, S. Kulabtong & P. Sreesamran Figures 1-4. Residence constructions under stones (Fig. 1), larva (Fig. 2) and pupa (Fig. 3, 4) of Stenopsyche siamensis from Maewong Basin, Thailand. side the construction and live in the stream since they get to pupa stage when, the small sticky fiber, reinforced by rough sand from the stream ground, is wrapped around the body of the insect (Figs. 1-4). Present paper reports on an additional record of the species from Central Thailand confirming indirectly the good water quality of upstream of Maewong Basin. ACKNOWLEDGEMENTS The authors are grateful to reviewers for revie- wing this manuscript. A special thank to Mr. Tha- napol Saranark and Mr. Wathunyu Kalumpuk, for supporting the field survey. REFERENCES Boileau Dudgeon D., 1999. Tropical Asian Stream: Zoo- benthos, Ecology and Conservation. Hong Kong Uni- versity Press. Hong Kong. Malicky H. & Chantaramongkol P., 1999. A preliminary survey of the caddisflies (Trichoptera) of Thailand (Study No. 26 on caddisflies of Thailand). Procee- dings of 9th International Symposium on Trichop- tera: 205-216. Laudee P. & Chantaramongkol P, 2003. Life history of Stenopsyche siamensis Martynov, 1931 (Trichop- tera: Stenopsychidae) from Doi Chiang Dao cat- chment area, Chiang Mai Province, Thailand. Braueria, 30: 23-25. Sangpradub N. & BoonSoong B., 2006. Identification of freshwater invertebrates of the Mekong River and its tributaries. Mekong River Commission, Vientiane. 274 pp. Biodiversity Journal, 2012, 3 (3): 191-200 A contribution to the knowledge of the terrestrial Mammalian fauna of Comino and its satellite islets (Maltese Archipelago) Arnold Sciberras 1 *, Jeffrey Sciberras 2 , Michael Sammut 3 & Gaetano Aloise 4 '33 ‘Arnest’, Arcade Str, Paola, Malta; email: bioislets@gmail.com 2 24 ‘Camilleri Court’ flat 5, il-Marlozz Str, Mellieha (Ghadira), Malta; email: wildalienplanet@gmail.com 3 11, Sqaq Rigu, Birkirkara, Malta; email: aquilarus@gmail.com 4 Museo di Storia Naturale della Calabria e Orto Botanico, University of Calabria, Via P. Bucci, s.n., 87036 Rende, Italy; email: aloise@unical.it ’Corresponding author ABSTRACT The present work aims to contribute to existing knowledge on Mammalia species occurring on Comino and its satellite islets and to provide additional records collected between the years 1 998-20 1 2. At the present state of knowledge, on the islands of the Maltese Archipelago there are 19 different species of terrestrial mammals (Erinaceomorpha: AteJerix olgirus. So- ricomorpha: Suncus etruscus, Crocidura sicula. Chiroptera: Rhinolophus ferrum-equinum, R. hipposideros, Miniopterus schreibersii, Myotis purlieus, Eptesicus serotinus, Nyctalus noctula, PipistreUus pygmaeus , P. kulii, Plecotus austriacus, Tadarida teniotis. Lagomorpha: Oricto- Jagus cuniculus. Rodentia: Apodemus sylvaticus, Rattus rattus, R. norvegicus, Mus musculus. Carnivora: Mustela nivalis), more than half of which are bats. Out of 1 3 species listed here, 8 species are represented as new to the islands while 3 species are confirmed and 2 species are listed as unlikely occurring. KEY WORDS Mammalia; Comino; Satellite islets; Maltese Islands. Received 08.07.2012; accepted 18.08.2012; printed 30.09.2012 INTRODUCTION Very little work is directed to Maltese Mam- malia, and even less is known about their distri- bution, especially with respect to the smaller islands. Some scattered notes provide some lite- rature of past records. The first data on the mammalian fauna of the Maltese Archipelago were reported by Gulia (1890). Busuttil & Borg (1925) were the first to attempt to list the mammalian fauna present on Comino Island. Unfortunately their list only constitutes of Mal- tese vernacular names and this often causes diffi- culty in determining the precise species as local names change through time and may refer to a num- ber of closely related species. Lanfranco (1969) repeats these records in his work; Savona- Ventura (1982) focuses on giving new data on the mammals living on Comino and Cominotto. Baldacchino & Schembri (2002) along with the life history of mammalia and herpeto fauna of the Maltese Islands, give also some localities where the latter species were recorded. Recently Aloise et al. (2011) represented the possibility that Crocidura sicula (Miller, 1900) is present also on Comino. This latter record is con- firmed in the present work. What follows is the current mammalian fauna known to occur or have occurred on the latter is- lands. In addition to bibliographic data available, data were collected through direct observations, by collection of dead specimens and through discus- sion with local people. 192 A. SCIBERRAS, J. SCIBERRAS, M. SAMMUT & G.ALOISE All non-flying mammals of the Maltese Archi- pelago, however, are thought to have been introdu- ced, at different times, by Men. MATERIALS AND METHODS Study area The Maltese Archipelago consists of three main islands, which are Malta, Gozo (Ghawdex) and Co- mino (Kemuna) and a number of minor islands, islets and rocks. Comino, the third largest island of the Archipelago, is surrounded by following satel- lite islands: Cominotto, Old Battery's Rock, Lan- tern Point Rock, Comino Cliff Face Rock/ Pigeon Rock, Small Blue Lagoon Rock, Large Blue La- goon Rock, Ghemieri Rocks (Fig. 1; Table 1). Comino (Kemuna) Comino is the third largest island of the Maltese Islands, with an area of 2.7 km 2 . Comino is also considered to be the smallest of the three main is- lands, but it can be referred as the mainland with respect to its satellite islets. However, due to its mono-geological component of Upper Coralline Limestone above sea-level, it has much less habi- tats than Gozo and Malta. Moreover with its small size, the range in topography is also limited, with a genaral tilt from South to North, identical incli- nation to the one of Marfa Ridge. Cliffs are only dominant on the eastern and sou- thwestern perimeters of the island, while only two considerably long valleys exist there. Sand-dunes are minimal and localised. With respect to vegeta- tion communites. Thyme garigue and Anthyllis- Teucrium and Euphorbia melitensis phryganas are most common, with dense Hypericum aegyptium garigues along the coast, followed by Pistacia len- tiscus pseudomaquis further inland and few Pinus halepensis woodland patches. Other phryganas are rare on Comino, but a con- siderable large population of Senecio bicolor exists on Comino, in some areas it is dense enough to contribute to its own phrygana. Elsewhere in Mal- tese Islands, this species seems common in Malta and Gozo, and rare on the islets, but always scatte- Name of the lslet/rock Code (fig-1 ) Rank by area Height (m) Width (m) W-E orientation Length (m) N-S orientation Distance from the mainland (m) Topogra- phical in- clination Surface Geology Soils Cominotto 0 3 >20 500 -200 125 South to North Upper Co- ralline Terra Rossa Large Blue La- goon Rock M 6 20 170 57 50 South to North Upper Co- ralline Terra Rossa Small Blue La- goon Rock N 11 18 27 70 110 West to East Upper Co- ralline Terra Rossa Pigeon Rock L 15 20 20 45 20 East to West Upper Co- ralline Inglin complex Battery Rock J 22 6-7 18 9 4 East to West Upper Co- ralline Terra Rossa Lantern Rock K 23 8 7 17 30 / Upper Co- ralline / Ghemieri Rocks (3 rocks) X 19 6,1,2 14,14,10 19, 20,38 10,82, 30 / Upper Co- ralline / Table 1. Characteristics of the Satellite islets of Comino (Maltese Archipelago). A contribution to the knowledge of the terrestrial Mammalian fauna of Comino and its satellite islets (Maltese Archipelago) 1 93 Comino Figure 1. Satellite islets of Comino (Maltese Archipelago): Old Battery's Rock (J), Lantern Point Rock (K), Comino Cliff Face Rock/ Pigeon Rock (L), Small Blue Lagoon Rock (M), Large Blue Lagoon Rock (N), Ghemieri Rocks (X), Comi- notto (O). red (not in dense clumps). Sub endemic rare plants have been noticed occuring sporadically on the is- land, such as the Pelagian-Maltese endemics Lina- ria pseudolaxiflora Lojacono and Daucus lopadusanus Tineo, while the other is Senecio pyg- maeus D.C, a Sicilian-Maltese endemic. Cominotto (Kemunett) By far, Cominotto is the largest islet near Co- mino, with 9.9 ha; it is the second largest uninhabi- ted island of Malta. The island has an S -shaped topographical orientation. Cominotto also has a similar altitude of the other nearby islets on the south east of the island. Cliffs dominate the south coast, while a peninsula, larger than Small Blue Lagoon Rock, is connected on the south east of Cominotto. Cominotto has three slopes, east to west from its highest point to its sandy beach and nearby coast, south to north and west to east from the hi- ghest point too. Thymbra cap data. Convolvulus oleifolius , Teucr turn fruticans , Br achy podium retusum and Anthyllis hermanniae are dominant species on the eastern side of the island, especially facing Co- mino. Euphorbia melitensis , and Pistacia lenti- scus are dominant on the highest point of the island. Darniella melitensis is dominant on the south cliffs. Euphorbia melitensis is also dominant on the northwest of the island. In winter, several annual species dominate the island, such as Convolvulus althaeoides , Anthyllis vulneraria, Linum strictum , Linum trigynum, and Galactites tomentosa in the disturbed patches of the island. Phagnalon graecum 194 A. SCIBERRAS, J. SCIBERAS, M. SAMMUT & G.ALOISE subsp. ginzbergeri is only present in the peninsula of the island (Sciberras & Sciberras, 2010). Large Blue Lagoon Rock (Il-Hagra Ta’ Bejn il-Kmiemen il-Kbira) Large Blue Lagoon Rock is the second largest islet of Comino. The islet has one slope, which is slightly steeper than that of Small Blue Lagoon Rock. The south perimeter is dominated by cliffs, except for a sizeable sea cave on its eastern side, which goes right through the islet. Its topographic landscape shows that it used to form part of the col- lapsed western valley of Comino. The upper half is inhabited by vegetation. Hy- pericium aegypticum is the dominant species all over the islet, while Daucus carota is dominant on the west side of the islet. Some patches are domi- nated by Convolvulus oleifolius (Sciberras & Sci- berras, 2010). Small Blue Lagoon Rock (Il-Hagra Ta’Bejn il-Kmiemen iz-Zghira) Situated between Large Blue Lagoon and Co- minotto, the Small Blue Lagoon Rock is the third largest islet of Comino. Most vegetation also oc- curs on its upper half. Arthrocnemum macro sta- chyum and Daucus carota are the dominant species all over the islet. Lygeum spartum entirely covers a small patch of soil. Lavatera arborea ( Malva dendromorpha ), with only four specimens, barely survives near the Lygeum spartum and is only present on this islet in Comino' s Archipelago (Sciberras & Sciberras, 2010). Comino Cliff Face Rock/Pigeon Rock/Ta’ Taht sl-Mazz Rock(ll-Gebla ta’ Taht il-Mazz) Ta’ Taht il-Mazz rock is the fourth largest islet near Comino. The islet is very steep, with vertical sheer cliffs on its east side facing the western cliffs of Comino, while its western is slightly less steep, but still sheer. The majority of species of plants occur on its west side and upper half of it, while only one spe- cies inhabits its east side. Inula crithmoides is a do- minant species on the eastern face of the islet, while Darniella melitensis is dominant on the we- stern cliff face of the islet. Daucus carota and Li- monium melitensis are also dominant, but on a les- ser extent. Anthyllis hermanniae and Pistacia lentiscus are rare on the islet. No soil exists on the rock, vegetation is gro- wing on the debris (the accumulating debris) and in rough weather the lower area is inundated by wave action (Sciberras & Sciberras, 2010). Old Battery’s Rock (Gebla ta' taht il -Bat- terija) This small rock is situated on the southeast of Comino under the old battery it is the second smal- lest rock of Comino. Remnant soil exists on the islet which is inundated by water wave action du- ring rough weather. Till 2010 the flora of the islet consisted of 22 individuals of Inula crithmoides (Sciberras & Sciberras, 2010). Lantern Point Rock (Gebla Tal-Ponta Rqiqa) Lantern Rock it is slightly smaller than Old Bat- tery’s Rock, it supports very little vegetation, only two species, Limonium melitense and Inula cri- thmoides. It is a large boulder of no more than 7 m high, with another small boulder lying on top of it (Sciberras & Sciberras, 2010). RESULTS Erinaceomorpha Gregory, 1910 Erinaceidae G. Fischer, 1814 Atelerix algirus (Lereboullet, 1842) Algerian Hedgehog Taxonomy of Hedgehog of Maltese Islands was uncertain for a long time. The Hedgehog was first recorded for the Malese Islands by Gulia (1858), which, however, erroneously considers this popu- lation belonging to Erinaceus europaeus. After the taxonomic revision of the Maltese specimens (Lan- franco, 1969), currently all the populations occur- ring in the Maltese Archipelago belong to the species Atelerix algirus. Busuttil & Borg (1925) mention that a species of hedgehog was imported to Comino from the nei- A contribution to the knowledge of the terrestrial Mammalian fauna of Comino and its satellite islets (Maltese Archipelago) 1 95 ghbouring islands of Malta or Gozo probably by the ornithologist A. Schembri, in the 19th Century, to control infestations of Blattaria sp. Lanfranco (1969) and Savona- Ventura (1982) seem not to re- spond to the presence of this species to the island while Baldacchino & Schembri (2002) state that this species is not present. Two of the authors (A. Sciberras and J. Sci- berras) have recorded this species 8 times over a time span of 14 years and in one occasion (23. XI. 2005) a nest was found containing 5 young. Specimens were encountered mostly in the centre of Comino Island, particularly in a lo- cation known as Ta' Caruana. Several naturalists informed the authors however that during the 1990's several specimens saved from Maltese roads, especially from Mgarr to Mizieb area, were released on the island for safety reasons. Both white and dark forms were encountered on the island. The status of the population present in Comino is unknown. The Algerian Hedgehog is present in Comino, Malta and Gozo, but is absent from all the other smaller islands. Soricomorpha Gregory, 1910 Soricidae G. Fischer, 1814 Crocidura sicula (Miller, 1900) Sicilian shrew In the Maltese Archipelago is confirmed the presence of two species of Soricidae: Suncus etru- scus (Savi, 1822) and Crocidura sicula. On the basis of current knowledge S. etruscus occurs only in Malta, and C. sicula , classified as C. suaveolens, is recorded only from the island of Gozo (Schem- bri & Schembri, 1979). Only recently has advan- ced the possibility that this species is present on Comino (Aloise et al., 2011). Two specimens were obtained from the analy- sis of Asio otus (Linnaeus, 1758) pellets found from beneath the nest of the latter. Although most likely the latter specimens were caught from Co- mino, there is also the possibility that the speci- mens were caught from Gozo by the predator and then expelled on the nesting site. Similar pellets were collected by A. Sciberras containing Rattus sp. remains and were retrieved where Asio flam- meus (Pontoppidan, 1763) was occasionally si- ghted; presumably these remains belong to the lat- ter species. On 18. III. 2012 a dead specimen of Crocidura sicula was found drowned in a bucket presumably full of rain water close to the Northern Coast, confirming the presence of a population of this species on the island. On 19.X.2011 a lower jaw bone of Crocidura sp. was found on Cominotto (A. Sciberras, J. Sci- berras and L. Pisani, unpublished data) but the pre- sence of a population of Crocidura Wagler, 1832 on this islet is not confirmed. The Sicilian shrew is absent from all the other smaller islands. Chiroptera Blumenbach, 1774 Rhinolophidae Gray, 1825 Rhinolophus hipposideros minimus (Heu- glin, 1861) Lesser Horseshoe Bat This species was already more commonly known in Gozo and less in Malta, which is repor- ted by the early work on the Maltese Islands (see Borg et al., 1997 and references therein). On 12. III. 2004 a dead specimen was found in a location known as Il-Hazina on Comino Island by one of the authors (A. Sciberras). This species t is a new record for this island. Vespertilionidae Gray, 1821 My otis punicus Felten, Spitzenberger & Storch, 1977 Mediterranean Mouse-eared Bat Taxonomy and distribution of the Mouse-eared bat of the Maltese Islands have been debated for a long time, because of its morphometric peculia- rities. In the past reported as M. oxignatus (Lanza, 1959; Van den Brink, 1967; Lanfranco, 1969), was then reported to M. blyti punicus and studied in detail (Felten et al., 1977; Savona- Ventura, 1984a, 1984b; Borg, 1998; Borg & Cachia-Zam- mit, 1988, 1994; Borg et al., 1990; 1997; Borg, 1998). M. punicus actually up to now is found on Malta and Gozo. On 12.III.2004 and 22.XI.2005 a specimen of this species was found in a subterranean area close to the Comino tower. This is the first record for the island. On the Maltese Archipelago, this species was formerly common but has suffered immense 196 A. SCIBERRAS, J. SCIBERRAS, M. SAMMUT & G.ALOISE decrease in the 1980's and according to Baldac- chino & Schembri (2002) the current population consists of only around 250-300 specimens. Pipistrellus kuhli (Kuhl, 1817) Kuhl’s Pipistrelle This species was presumed rare in the Maltese Archipelago before 1969 (Gulia, 1890; Lanfranco, 1969) but according to a 1990 study (Borg et al., 1990, 1997) the latter was found to be more fre- quent and it had a distribution all over the three main islands: Malta, Gozo and Comino. Pipistrellus pygmaeus (Schreber, 1774) Soprano Pipistrelle Busuttil & Borg (1925) record the presence of two species of bats, noting that one preferred roo- sting in cracks while the other in caves. Lanfranco (1969) repeats this record. Savona- Ventura (1982) observed several specimens in flight and entering crevices at Santa Marija bay on VIII. 1977. None were captured, and so the records are only from di- rect observations. It was suggested to be similar to Pipistrellus pipistrellus (Schreber, 1774). Nowa- days it is being considered that Pipistrellus pyg- maeus (Leach, 1825) is the likely species to exist in the Maltese Archipelago (Baldacchino & Schembri, 2002). Several specimens were noted in flight by the three of the authors ( A. and J. Sciberras, M. Sam- mut) and from field observations they look to be a Pipistrellus sp., but none were ever recovered for taxonomic identification. Several specimens were also observed flying over Cominotto and landing in crevices of Comino Cliff Face Rock. Plecotus austriacus (Fischer, 1829) Grey Long-eared Bat Present on Malta, Gozo and Comino, the spe- cies was reported already by Gulia (1890, 1914), Lanza (1959) and Van den Brink (1967) as P. au- ritus (Linnaeus, 1758). Baldacchino & Schembri (2002) state that this species had a distribution all over the three main islands. Although the authors never observed this species on Comino a dead spe- cimen was found on Cominotto Beach by one of the authors (A. Sciberras) on 6.VII.2001. This is the first record for Cominotto island. Tadarida teniotis (Rafinesque, 1814) European free-tailed bat A skull of this species was retrieved from Larus sp. pellet from Large Blue Lagoon Rock (13.6.2011 A. and J. Sciberras leg.). Although the shape and size of the island suggest that, probably, the animal was preyed on a different site than where the remains were found, this is an intere- sting record because besides being new to the islet, this species is extremely rare and was repor- ted from the Maltese Islands twice before the di- scovery of this skull. The first time was in 1993 in a locality known as Cittadella on the island of Gozo where 2 speci- mens were observed hunting insects under street lights. The same specimens were noted till mid - May in the same locality. Another specimen was recorded flying close to the cliffs of Had - Dingli in Malta in November of 1996 (Baldacchino & Schembri, 2002). To add to the interest on the same islet where the skull was located, on 8. VIII. 20 10 the same authors observed gulls, identified as Larus michahellis (Naumann, 1884) by M. Sammut, hunting bat specimens that approa- ched the vicinity of a colony of 7 gulls that were resting on the islet. This was just before sunset. It could be that this species does occasionally predate on bats as it was noted to predate on other terrestrial species such as Podarcis filfolensis (Bedriaga, 1876). It could also be that the bat was caught away from this site even offshore and it was regurgitated locally. Carnivora Bowdich, 1821 Mustelidae Fischer, 1817 Mustela nivalis (Linnaeus, 1766) Weasel Baldacchino & Schembri (2002) state that this species locally is restricted in the Maltese Islands to the island of Malta. It is very scarce on the main island but it is widespread and observations of this species range from the North to the South of the main island of Malta. A contribution to the knowledge of the terrestrial Mammalian fauna of Comino and its satellite islets (Maltese Archipelago) 1 97 Whilst most observations of this species are of single individuals, a den with cubs was observed in Balluta area (Wardija Limits) (A. Sciberras, 5.IV.2002, unpublished data) and an adult with five cubs was observed at Mtahleb (M. Sammut & C. Cachia Zammit, 13. XI. 2010, unpublished data), although illegal, most of the specimens (from Qammieh and Ahrax headlands in Mellieha) have been killed for taxidermal purpo- ses. On 23.IV.2012, one of the authors (M. Sam- mut) saw an adult specimen of this species at a location known as Il-Hazina (Comino island) mo- ving swiftly from behind a rubble patch. Following that it climbed from behind a rub- ble wall and popped up on the rubble wall where it was seen very well and could be identified wi- thout any doubt. It disappeared again as swiftly as it had appeared. It was seen in the afternoon and though the author remained in the area for over an hour it was not seen again. An indication of the presence of species on the island is the finding of 3 dead specimens of Rattus norvegicus and a young specimen of Oryc- tolagus cuniculus near Comino' s pig farm (6.V.2004, A. Sciberras, unpublished data). These carcasses showed signs that they had been killed and dragged from the neck, and less than 50 cm away a den similar to the one observed at Balluta was discovered. No cubs or adults were observed despite the constant monitoring. Interestingly, people who have lived all their lives and constantly patrol the island of Comino have never seen the species on the island (S. Vella, pers. comm.). It is also worth noting that Mustela putorius furo (Linnaeus, 1758) was once observed on Comino in the 1970's (M. Psaila, pers. comm.). In the past feral specimens have escaped from rabbit hunters which visited the island (S. Vella, pers. comm.). One of the authors (A. Sciberras) also has reports on Mustela sp. being present in Gozo but after checking the site (an area known as Munxar) 3 feral specimens of M. putorius furo were noted roaming free. One of the authors (J. Sciberras) received reports by locals of Mustela sp. at San Bias Bay. After observing the site and description of the locals it was confirmed that a few specimens of M. putorius furo were present at the site. Lanfranco (1969) also records the latter feral species in Malta and it is suggested that these must be escapees from O. cuniculus hunting parties. Rodentia Bowdich, 1821 Muridae Illiger, 1815 Rattus norvegicus (Berkenhout, 1769) Brown rat The Brown Rat is considered common, but can be very common and, under certain conditions, can become particularly problematic. Its presence has been detected on all the major islands, Cominotto and many other smaller islands. The presence of rats on the Comino and Cominotto Islands is referred by Busuttil & Borg (1925), while Lanfranco (1969) mentions this species and Rattus rattus , Savona- Ventura (1982) observed a specimen of the species on Cominotto in IX. 1975. Baldacchino & Schembri (2002) also record the presence of the species on Comino. Presently including the period of observations stipulated above, this species is numerous on the islands and was recorded on the following islets (A. Sciberras, unpublished data): Manoel Island, Qawra Point or Ta’ Fra Ben islet, Selmunett Is- land, Large Blue Lagoon Rock, Haifa Rock, and Tac-Cawl Rock. On Fungus rock and Selmunett Island, it is known of its devastating impact on the Insular biodiversity (Baldacchino & Schembri, 2002; Sciberras, 2007; Sciberras & Schembri, 2008). Rattus rattus (Linnaeus, 1758) Black Rat The Black Rat is reported as present and common on Malta, Gozo, Comino and some of the satellite islets. Busuttil & Borg (1925) record the presence of rats on the Comino and Comi- notto Island while Lanfranco (1969); Savona- Ventura (1982) states in III. 1978 he found a dead specimen of this species. Baldacchino & Schembri (2002) and Aloise et al. (2011) also record the presence of this species on Comino including Malta and Gozo and state that it is common. The species is also known for Fungus rock (Borg & Sultana, 2003). From experience with data also collected from a local Pest control company, this species is nu- merous where it is present but it does have a re- 198 A. SCIBERRAS, J. SCIBERRAS, M. SAMMUT & G.ALOISE stricted distribution. Most records of specimens come from Valletta and neighbouring harbour ci- ties. In all the years of observations and data col- lection, this species was never encountered on Comino or its satellite islets. Mus musculus (Linnaeus, 1758) House mouse Widespread and very common, the House Mouse is abundand throughout. Present on all major islands, although not proven, its presence can be regarded as likely also on the islands of smaller size, because of its unique ecological characteristics. Busuttil & Borg (1925) reported the presence of a mouse on Comino and so did Savona- Ven- tura (1982) when he noted a specimen at Santa Marija Bay in 1977. Since it was not caught, it could not be certain whether it was this species or Apodemus syfvaticus (Linnaeus, 1758). Seve- ral specimens were caught annually on the north Coast of Comino as a part of pest control treat- ment. From 34 studied samples all specimens re- sulted in being M. musculus. Lagomorpha Brandt, 1855 Leporidae Fischer, 1817 Oryctolagus cuniculus (Linnaeus, 1758) Wild Rabbit Busuttil & Borg (1925) reported the presence of a rabbit on Comino and this was repeated by Lanfranco (1969). This species was very common at their time and it is presumed that this species was introduced around 1890' s on the latter Island. Prisoners and soldiers during 1914-1918 did short work on the rabbit population on Comino. In 1969 Comino was declared a protected area from hunters and the rabbit population must have benefited, however Savona- Ventura (1982) assu- med the survival of the animal through its scattered dropping and not so much on the sightings. He also recorded the species on Cominotto by finding droppings of the latter in IX. 1975. In the 1980's, Myxomatosis was introduced and the Maltese po- pulation was virtually wiped out as happened with the Comino population. A local resident introduced this species from Gozo again in several occasions as he did with other species (Sciberras, 2009). Today the Comino population is the largest in density when compared to the size with other Maltese Islands. The populations recorded in past literature on Cominotto and Selmunett are extinct (Sciberras unpublished data). It is interesting to note that in the Maltese Archipelago, two colour morphs occur: the brown form and the yellow form. The Maltese population consists of almost entirely the brown form with occasional yellow and hybrids with domestic rabbits. Domestic rabbits are gene- rally set free because of some kind of illness such as VHD (Viral Hemorrhagic Disease), Myxoma- tosis and the most commonly found Ear Cancer. If these survive they sometimes interbreed with the wild stock (Sciberras, 2006). The Gozo population has both forms in equal numbers, whilst the Comino population constitutes entirely of the yellow form and only on very rare occasions, slightly darker specimens are noted. CONCLUSIONS At the present state of knowledge, on the is- lands of the Maltese Archipelago are 19 different species of terrestrial mammals (Erinaceomorpha: Atelerix algirus ; Soricomorpha: Suncus etruscus , Crocidura sicula ; Rhinolophus ferrum-equinum , R. hipposideros , Miniopterus schreibersii, Myo- tis purlieus , Eptesicus serotinus , Nyctalus noc- tula , Pipistrellus pygmaeus , P. kulii , Plecotus austriacus , Tadarida teniotis', Lagomorpha: Orictolagus cuniculus ; Rodentia: Apodemus syl- vaticus , Rat t us rattus, R. norvegicus , Mus mu- sculus ; Carnivora: Mustela nivalis ), more than half of which are bats. As regards the island of Comino and its satel- lite islands, most of these species are present (68,4%), not being up to now verified the presence of S. etruscus and A. sylvaticus , among the non- flying, and R. ferrum-equinum, M. schreibersii , E. serotinus and N. noctula among the bats. With the species listed above, Busuttil & Borg (1925) reported that a certain Captain Stivala re- leased on the island of Comino a pair of Gazella sp. These bred successfully but were eradicated by prisoners during the First World War. A contribution to the knowledge of the terrestrial Mammalian fauna of Comino and its satellite islets (Maltese Archipelago) 1 99 A population of Felis silvestris catus (Schreber, 1777) introduced in the 1980' s was exceeding over 20 specimens around Comino hotel (Northern Coast) in the late 1990's and it was eradicated by environmentalists for the protection of the native wildlife of Comino. This update on the mammalian fauna was a result of observations, made indirectly while the authors were conducting other studies or surveys mostly entomological, herpetological or or- nithological. Upon further investigation, especially on spe- cies most critical (eg. Crocidura sicida and Chi- roptera), would be necessary to verily the status of the population, to guarantee their conservation in the Archipelago. ACKNOWLEDGEMENTS A. and J. Sciberras are in debt to Esther Sciber- ras, Romario Sciberras and Luca Pisani for their constant assistance in the field. Special thanks go to Professor Patrick J. Schembri, Alfred. E. Baldac- chino and Pietro Lo Cascio for providing useful contacts and some literature. Thanks also go to Mario Gauci for his hospitality during Gozo visits. Mark Psaila and Salvu Vella are acknowledged for sharing their observations. REFERENCES Aloise G., Baldacchino A.E. & Amori G., 2011. Croci- dura sicula Miller, 1900 (Mammalia, Soricidae): a possible new record from Comino island (Maltese Is- lands). Biodiversity Journal, 2011, 2: 145-148. Baldacchino A.E. & Schembri P.J., 2002. Amfibji, rettili, u mammiferi fil-gzejjer Maltin. Sensiela Kullana Kulturali, Nru. 39. Pubblikazzjonijiet Indipendenza, Il-Pieta, Malta, xii + 256 pp. Baldacchino A.E. & Azzopardi J., 2007. L-Ghasafar li jbejtu fl-ambjent naturali tal-gzejjer Maltin. Malta University Publishers Ltd., Msida, Malta. Borg J. J., 1 998. The lesser mouse-eared bat Myotis blythi punicus Felten, 1977 in Malta. Notes on status, mor- phometries, movements and diet (Chiroptera: Vesper- tilionidae). II Naturalista Siciliano, 22: 365-374. Borg J.J. & R. Cachia-Zammit, 1988. Avian, Chiropteran and other remains in Barn Owl Tyto alba pellets from Gozo. Il-Merill, 24: 12-13. Borg J.J. & R. Cachia-Zammit, 1994. Diet of the Bam Owl Tyto alba in a rural area in Gozo. Il-Merill, 28: 24-25. Borg J. J., Fiore M., Violani C. & Zava B., 1990. Obser- vations on the Chiropterofauna of Gozo, Maltese Is- lands. Bollettino Museo Regionale Scienze Naturali Torino, 8: 501-515. Borg J.J. & Sultana J., 2003. The presence of the black rat Rattus rattus on Fungus Rock (Maltese Islands). The Central Mediterranean Naturalist, 4: 105-106. Borg J.J., Violani C. & Zava B., 1997. The Bat Fauna of the Maltese Islands. Myotis, 35: 49-65. Busuttil V. & Borg T., 1925. Dizjunarju Enciklopediku. Vol.5, E. Lombardi, Malta, 4267-4268. Felten H., Spitzenberger F. & Storch G., 1977. Zur Klein- saugerfauna West-Anatoliens. Senckenbergiana bio- logica, 58: 1-44. Gulia G., 1858. Repertorio di StoriaNaturale (di Malta). Tip. Anglo-Maltese, Valletta, 245 pag. Gulia G., 1 890. Elenco dei Mammiferi Maltesi. II Natu- ralista Maltese, 1:2-3. Gulia G., 1914: Uno sguardo alia Zoologia delle "Isole Maltesi" . - IX. Congres International de Zoologie, Monaco, sect. 4: 545-555. Lanfranco G., 1969. Maltese Mammals- (Central Medi- terranean), Progress Press, Malta, 28 pp. Lanza B., 1959. Chiroptera, 187-473. In: Toschi A. & Lanza B., Mammalia: Generality Insectivora, Chirop- tera. Fauna d’ltalia, IV, Calderini, Bologna, 485 pp. Savona- Ventura C., 1982. The mammalian fauna of Co- mino and neighboring islets. Potamon, 1: 137-139. Savona- Ventura C., 1984a. Observations of the Genus Myotis in Maltese caves. Potamon, 1: 77-78. Savona- Ventura C., 1984b. A study of the genus Myotis Kaup (1829) in Malta (Mammalia, Chiroptera: Ve- spertilionidae). The Central Mediterranean Natura- list, 1 : 51-54. Schembri P.J. & Schembri S.P., 1979. On the occurrence of Crocidura suaveolens Pallas (Mammalia, Insecti- vora) in the Maltese Islands with notes on other Mal- tese shrews. The Central Mediterranean Naturalist, 1: 18-21. Sciberras A., 2006. Domestic rabbits in countryside cause for concern. (Press release) The Malta Independent. April 12th pg. 7. Sciberras A. & Lalov S.V., 2007. Notes on the impact of the black rat ( Rattus rattus L.) on the flora and fauna of Fungus Rock (Maltese Islands). The Central Mediterranean Naturalist, 4: 207-210. Sciberras A. & Schembri P.J., 2008. Conservation Sta- tus of St Paul' s Island Wall Lizard (Podarcis fdfo- lensis kieselbachi). Herpetological Bulletin, 105: 28-34. Sciberras A., 2009. Short notes on the introduced Avi- fauna of Comino Island and some of their interactions 200 A. SCIBERRAS, J. SCIBERRAS, M. SAMMUT & G.ALOISE to local Herpetofauna. The Central Mediterranean Naturalist, 5: 46-49. Sciberras J. & Sciberras A., 2010. Topography and Flora of the Satellite islets surrounding the Maltese Archipelago. The Central Mediterranean Naturalist, 5: 31-42. Van Den Brink F.H., 1 967. A field guide to the mammals of Britain and Europe. Collins. London, 221 pp. Biodiversity Journal, 2012, 3 (3): 201-228 New and little known land snails from Sicily (Mollusca Gastropoda) Fabio Liberto 1 , Salvatore Giglio 2 , Maria Stella Colomba 3 * & Ignazio Sparacio 4 'Strada Provinciale Cefalu-Gibilmanna n. 93, 90015 Cefalu, Italy; email: fabioliberto@alice.it 2 Contrada Settefrati, 90015 Cefalu, Italy; email: hallucigenia@tiscali.it TJniversita di Urbino, Dept, of Biomolecular Sciences, via Maggetti n. 22, 61029 Urbino, Italy; email: mariastella.colomba@uniurb.it 4 Via E. Notarbartolo n.54 int. 13, 90145 Palermo, Italy; e-mail: isparacio@inwind.it ’Corresponding author ABSTRACT In the present paper are reported new and little known land snails from Sicily (Mollusca Gastropoda). Particularly, Platyla similis (Reinhardt, 1880) (Aciculidae) and Rumina sa- harica Pallary, 1901 (Subulinidae) are first recorded in the island; new taxonomic data, useful for a better systematic classification, are provided on two little-known taxa, Lam- pedusa lopadusae nodulosa Monterosato, 1892 (Clausiliidae) and Cernuella (Cernuella) tineana (Benoit, 1862) (Hygromiidae); and finally, a new species of slug, Tandonia mari- nellii n. sp. (Milacidae), currently known from North-Western Sicily, is described. KEY WORDS Mollusca; land snails; Sicily; taxonomy; new species. Received 10.07.2012; accepted 26.08.2012; printed 30.09.2012 INTRODUCTION During 19 th century several taxa of terrestrial molluscs of Sicily were established, many of which are still little known and of uncertain taxo- nomic status, nevertheless some genera or families of molluscs, including slugs, have been neglected for a long time. The study of terrestrial molluscs of Sicily undertaken by present authors in the last decade, despite the complexity of the subject, con- tinues with this further contribution in which some new faunal reports are provided, little known taxa are examined and, in addition, a new species of slug is described. ACRONYMS. APP = anterior portion of palatal plica; BC = bursa copulatrix; BCD = diverticulum of bursa copulatrix; CL = columellar lamella; DE = distal epiphallus; DG = digitiform glands; DSC = dart sac complex; DBC = duct of the bursa copulatrix; DCP = distal caviti of the penis; DGS = dart gun; DG = digitiform glands; DSC = dart sac complex; DSO = dart sac opening; E = epiphallus; F = flagel- lum; FO = free ovidutto; FR = frenula; G = penial pa- pilla; GA = genital atrium; ISO = inner stylophore opening; F = lunella; OSO = outer stylophore opening; P = penis; PCP = proximal caviti of the penis; PD = penial diverticulum; PE = proximal epiphallus; PE = parietal lamella; PEE = parallel lamella; PP = principal plica; PPP = posterior portion of (upper) palatal plica; PR = penial retractor muscle; SCL = sub- columellar lamella; SL = spiral lamella; SP = sutural plica/plicae; V= vagina; VAG = vaginal accessory gland; VC = vaginal chambre; VD = vas deferens; VP = vaginal pleats. The materials used for this study are deposited in the following Museums and private collections: M. Bodon collection, Italy, Genova (CB); D.P Cilia collection, Santa Venera, Malta (CC); S. Giglio col- lection, Cefalu, Italy (CG); Laboratory of Cytoge- netics and Molecular Biology, University of Urbino, Italy (LCMBU); F. Liberto collection, Cefalu, Italy (CL); Museo Civico di Storia naturale di Comiso, 202 F. Liberto, S. Giglio, M.S. Colomba & I. Sparacio Italy (MCSNC); Museo Civico di Storia Naturale di Genova “G. Doria”, Italy (MSNG); G. Nardi col- lection, Nuvolera, Italy (CN), A. Reitano collection, Tremestieri Etneo, Italy (CR); I. Sparacio collection, Palermo, Italy (CS); The Steinhardt National Col- lections of Natural History, Zoological department, Tel- Aviv University, Israel (TAU). MATERIALS AND METHODS All specimens were collected by sight on the soil and under the rocks or by sieving litter and soil. Observations on ecology of these organisms and their feeding behavior were made directly in the field. Dry shells have been studied as regards size, colour, morphology, sculpture, aperture, plicae and lamellae, lunella and clausilium. Photographs were taken with a digital camera. In order to study and illustrate genital organs, the specimens were drow- ned in water and fixed in 75% ethanol. Reproductive apparatus was extracted by means of scalpel, scissors and needles. Illustrations of ge- nitalia were sketched using a camera lucida. Height and maximum diameter of the shell along with some parts of genitalia were measured (in millime- ters) by a digital gauge. Voucher specimens were stored in collections indicated below. Toponyms (place-names) are reported following the Portale Cartografico Nazionale (PCN, http://www.pcn.mi- nambiente.it /PCN/), Map IGM 1:25000. Each lo- cality and/or collection site is named in the original language (italian). The material used for the mole- cular analysis was collected on the field during Fe- bruary 2012. All the specimens were studied and observed at the steromicroscope (Leica MZ 7.5). Genetic study described in the present study was based on a comparative analysis of COI partial se- quences which are frequently used as markers in the investigation of evolutionary processes at the specific level. Briefly, the study was conducted by DNA iso- lation, PCR amplification, sequencing, alignment of the sequences and phylogenetic reconstructions using the Maximum Likelihood algorithm. For a de- tailed description see below. RESULTS Class Gastropoda Cuvier, 1795 Ordo Architaenioglossa Haller 1890 Family Aciculidae J.E.Gray, 1850 Platyla similis (Reinhardt, 1880) Examined material. Italy, Sicily, Cefalu, Rocca di San Nicola, 37°59 , 07”N 14°02’42”E, 600 m, 01.VI.2008, 6 specimens (CG); Cefalu, Cozzo Carca- rello, 37°59 , 29” N, 14°03’05”E; 320 m, 15.VI.2008, 8 specimens (CL); idem, 23.VIII.2009, 22 specimens (CL); idem, 23.IV.2012, 12 specimens (CL). Figures 1,2. Shell of Platyla similis from Cefalu, Cozzo Calcarello, h: 2.32 - D: 0.89. Figure 3. Geographic distribution of Platyla similis (circles) and P. subdiaphana (squares) in Sicily (personal data). New and little known land snails from Sicily (Mollusca Gastropoda) 203 Description. Morphologically, this Sicilian po- pulation of P. similis has typical characters of the species: shell conical (Figs. 1-2), slender, colorless, height 2.16-2.58 mm, width 0.80-0.98 mm, with 4- 5 whorls slightly convex, last portion of last whorl slightly ascending in relation to the penultimate, aperture slightly oblique, sinulus little depth, exter- nal peristomal rib consists of a weak thickening, not clearly defined anteriorly and posteriorly. Biology and Distribution. P. similis lives in woodland litter, on the soil, in the cracks and at the base of the calcareous rocks. P. similis has South- Oriental European distribution including Bulgaria, Croatia, Serbia, Greece, Romania, Kosovo, and Central and Southern Italy (Boeters et al., 1989; Bodon et al., 1995; Bodon & Cianfanelli, 2008; Bank, 2012). Remarks. We report the presence of P. similis for the first time in Sicily, from Nothern Madonie mountains, on the calcareous rocks named “Rocca di San Nicola” and “Cozzo Calcarello”. Shells were collected by sieving litter and soil, sampled in cracks of the calcareous rock, at the base of rocky walls or under boulders in the woods of oaks. In Sicily it was known until now only the en- demic species: P. subdiaphana (Bivona, 1839) (Boeters et al., 1989; Bodon et al., 1995; Bodon 6 Cianfanelli, 2008; Bank 2012) (Fig. 3). Boeters et al. (1989) distinguish all species of Platyla Mo- quin-Tandon, 1856 into three groups on the basis of the presence or absence and conformation of the external peristomal rib (see also Bodon & Cianfanelli, 2008). P. similis is inserted into the second group cha- racterized by an external peristomal rib not robust and not clearly delimited posteriorly. P. subdia- phana belongs to the third group characterized by a robust external peristomal rib bounded by a sharp line or by a large groove. P. subdiaphana , also, is distinguished from P. similis for bigger size (height 3.5-4.45 mm). Ordo Stylommatophora A. Schmidt, 1855 Family Milacidae Germain, 1930 Tandonia marinellii n. sp. Examined material. Holotypus: Italy, Sicily, Custonaci, Monte Sparagio, Pizzo Giacolamaro, 38°03 , 18”N 12°44’35”E, 665 m, 08.1.2012 (MSNG 56989). Paratypi: Italy, Sicily, Custonaci, Monte Co- fano, 38°06’H”N 12°40 , 39”E, 255 m, 14.IV.1991, 2 specimens (CS); San Vito lo Capo, Macari, Pizzo Castelluzzo, 38°07’28”N 12°44 , 41”E, 364 m, 11.2007, 3 specimens (CR); Erice, 38°02’24”N 12°35 , 34”E, 500 m, 03.X.2011, 1 specimen (CN); Calatafimi, Cozzo Gessi, 37°54’44”N 12 o 50’41”E, 264 m, 20.XI.20 11, 3 specimens (CL); idem, 2 spe- cimens (CG); Calatafimi, Monte Bernarco, 37°54’56”N 12°49’45”E, 370 m, 20.XI.2011, 6 spe- cimens (CL); Castellammare del Golfo, Fraginesi, 38°0r06”N 12°50 , 08”E, 180 m, 4.XII.2011, 1 spe- cimen (CS); Custonaci, Monte Sparagio, Pizzo Gia- colamaro, 38°03’18”N 12°44’35”E, 665 m, 8.1.2012, 1 specimen (CL); idem, 1 specimen (TAU 76575); idem, 1 specimen (MCSNC 4411); Custonaci, Monte Sparagio, Pizzo Giacolamaro, 38°03’17”N 12°44 , 57”E, 716 m, 4. II. 20 12, 8 specimens (CL); Trapani, contrada Chinea, near the Lago Rubino, 37°53’49”N 12°44 , 05”E, 260 m, 18.11.2012, 2 spe- cimens (CS); Custonaci, Muciara, 38°03 , 27”N 12°43’64”E, 542 m, 4.III.2012, 11 specimens (CS); idem, 2 specimens (MSNG 56990; MSNG 56991). Description of Holotypus. Slug medium- sized, length 55 mm after preservation (the speci- men is contracted). Clypeus about 1/3 of body length, superficially granulated, with horseshoe- shaped groove, and a hollow near keel; evident Ca- rina running from clypeus to posterior apex of body. Body and mantle brownish-yellow with blac- kish pigment forming irregular reticulation and dots which disappear toward the sole, keel orange. Foot sole tripartite, creamy-coloured, mucus thick, viscous, white-yellowish. Shell (limacella) nail- like, oval, well calcified, white, with apex posterior and situated on major axis, at the highest point, convex above, slightly concave ventrally (Figs. 18- 20); length: 7.5 mm; diameter: 4.9 mm. Genitalia (Figs. 4-7, 11). Vas deferens thin, en- ding laterally at proximal epiphallus tip. Epiphallus very long (20 mm), divided by a slight constriction in a narrow cylindrical proximal portion with thin walls (proximal epiphallus) and in an ample conic distal portion with thick walls (distal epiphallus). Internally, the proximal epiphallus is crossed by around 15 anular crests more evident in the central zone, and 5-6 longitudinal folds in the slight broa- dening apical knob, while the distal epiphallus has a very narrow duct; penial retractor muscle ending 204 F. Liberto, S. Giglio, M.S. Colomba & I. Sparacio Figures 4-7. Genitalia of T. marinellii n. sp., holotypus: whole genitalia (Fig. 4), internal structure of vagina (Fig. 5), in- ternal structure of penis (Fig. 6), internal structure of penis and epiphallus (Fig. 7). New and little known land snails from Sicily (Mollusca Gastropoda) 205 Figure 8. Spermatophore of the holotypus of T. marinellii n. sp. Figure 9. Anterior portion of spermatophore of T. ma- rinellii from Calatafimi. Figure 10. Spermatophore of T. sowerbyi from Novara di Sicilia, Rocca Novara, Sicily. Figures 11, 12. Scheme of genitalia of T. marinellii n.sp., (Fig. 11) and of T. sowerbyi by Wiktor (1987), modified (Fig. 12). 206 F. Liberto, S. Giglio, M.S. Colomba & I. Sparacio laterally on penial complex where slight constric- tion separates distal epiphallus from proximal epi- phallus. Border between epiphallus and penis externally evident, inside the epiphallus protrudes into the penis with a semispheric extension, bearing at its apex a small cylindrical papilla. Penis irregu- lar, cylindrical, 1 1 mm long, approximately X A of length of epiphallus, with thin walls, inside with some striations and divided by a constriction in two cavities: a narrower, oblong proximal cavity (8 mm) and a short, wide distal cavity (2.2 mm). A thin she- ath envelopes the penial complex, keeping proximal epiphallus bent on the distal epiphallus and the penis contracted; wide and short genital atrium, with fine folds around the genital opening. Vagina long (9.8 mm), inside there are some di- scontinuous longitudinal folds. Vaginal accessory gland enters by means of thin canaliculi at about midway along vagina. An annular pad separates the real vagina from a small chamber (vaginal cham- ber) with thick, smooth wall, where the duct of bursa copulatrix and the free oviduct end. Duct of bursa copulatrix short and slender. Bursa copulatrix elongate with a narrow apical prolungation. Long and slender free oviduct, slightly wider near vagina. Spermatophore (Fig. 8) worm-like, glossy, gol- den. Anterior apex lacking (broken), the remaining anterior portion is regularly tubular and bare, only on the distal part there are some short spines (with 4-5 apexes), the posterior portion is covered with some short spines with bifurcate point (two apexes) or simple (one apex); posterior apex of the sperma- tophora, bigger, completely covered of bifurcate or simple spines. The spermatofore was found in the channel of the bursa copulatrix with the posterior apex contained into "vaginal chamber". Variability. Body colour variable (Figs. 13-16) from uniform orange with some spots hardly visible to orange-brown speckled with darker patches; keel clear, orange or cream; clypeus with blackish hor- seshoe-shaped groove and sometimes with a dark central line; genitalia: proximal epiphallus and di- stal epiphallus generally have the same length, but in some specimens the proximal portion is slightly shorter; the epiphallus protrudes into the penis with a semispheric or conic extension, penis length va- rying from 6 mm to 10 mm; vagina length varies from 2 mm to 9 mm. Spermatophore variable in size (Fig. 9) and color from red to yellow-gold; these were found in the channel of bursa copulatrix with the big apex contained into "vaginal chamber". Etimology. The new species is dedicated to Aldo Marinelli (Roma), as sign of appreciation for creating the forum “Natura Mediterraneo” (availa- ble at: http://www.naturamediterraneo.com/forum/). Biology and Distribution. Species rather common in natural environments with forests, me- diterranean maquis or garrigue, nocturnal, during the day specimens shelter under rocks and logs; sexual maturity occurs in winter. T. marinellii n. sp., at present, is known only from North-Western Sicily (Fig. 21). Comparative notes. T. sowerbyi (Ferussac, 1823) was the only known species in Sicily (Wa- gner, 1931, Giusti, 1973; Manganelli et al., 1995; Bank 2012) (Fig. 21). In this region it seems to be native as widely widespread, especially in natural environments, and having been reported since 1800 (Lessona & Pollonera, 1882 sub Amalia carinata and A. carinata var. oretea\ Mina Palumbo, 1883 sub Amalia carinata ; Pollonera, 1891 sub Amalia carinata). The populations of eastern Sicily exami- ned by us show the typical morphological charac- ters of T. sowerbyi (Wiktor, 1987, Giusti et al., 1995) (Figs. 10, 12). The epiphallus in T. sowerbyi is of medium length and cone-shaped, while in T. marinellii n. sp. the epiphallus is very long and equipped with an evident proximal portion comple- tely absent in I sowerbyi ; the penis retractor mu- scle in T. sowerbyi ends at about 2/3 of the length of the epiphallus, while in T. marinellii n. sp. ends at half the length of the epiphallus exactly where the distal portion ends and the proximal one begins; a pair of short supplementary muscles inserted at the distal end of epiphallus observed in T. sowerbyi , lack in the new species. The penis in T. sowerbyi is divided by a con- striction in a short distal portion followed by a lon- ger proximal chamber, while in T. marinellii a proximal long portion is followed by a short distal one, wide and sac-shaped; penial papilla in I so- werbyi is squished, of vestigial type, while in T. ma- rinellii is of cylindrical shape; the fold-like thickening (reduced "stimulator") present in T. so- werbyi , lacks in T. marinellii ; the spermatophore in T. sowerbyi has a posterior portion covered with very branched spines (Fig. 10), whereas in I mari- nellii is covered by scattered spines not branched or New and little known land snails from Sicily (Mollusca Gastropoda) 207 Figures 13-16. T. maninellii n.sp., Custonaci, Monte Sparagio, Giacolamaro, 08.1.2012: variability of the body colour. In a specimen (Fig. 16) is visible the white-yellowish mucus. 208 F. Liberto, S. Giglio, M.S. Colomba & I. Sparacio O p^n-orm li ;an i im 16800 D panOrrililanum' 16933 D panomnilanumi 10935' O p^normilfinum, 16801 D punorm it an unv 16822 D panormiilsniirn. 16802 D panormilanum 16934 D panormiiamjm — 10980 D panormiitanurn 16799 D panormilanifm 16797 D gdclv&ri 0795 D golcheri 6796 D gplcheri d_r ie ' Gil — 1C 54 60 I 2 D laews TOO L D taciwc 05944 D irtvodena 16920 D lnw4«n« - 1 5984 D iniAidcnr: 16927 D invadens L 1 0026 D inid^Kjpns •pg * 15035 D irtvatlusTs I 16919 D I modems ■ 1-403-8 D inwirirnr, ■ 16924 D irivadtuis 16925 O invadens 1 1692G O mwsdons 3 D reticulatum O reticuialumi 2 O rc-ticulutum 4 D reticulatum TO top tOO □e 2 T budepest 3 T budapcsl 1 t buGapest TCUS2 COi LCO 1490. scf TCUS4 COI LCO 1490. scf TCUS 1 COI LCO 1490 scf TMAD2 COI LCO 1490. scf 1 TMAD4 COI LCO 1490.6CT . TMAD3 COI LCO 1490 . scf 90 * TM ADS COI LCO 1490. scf — 506 Loh marginals 14 10 L flatAJs flavus flaws A L flaws fioj 1A1C LM 2 L fit I— 896 L IOC L 3 L flt 4 iL c L 1 S3 fluws . flaws — W0O2 L wnlilPorcidli m p 1162 L clfiaiwsnlg«r I* 14/8 L cmcreciciigcr 1 S44 L cinereoniger 1125 L cinsreoniger B6 < 1 178 L ci acrconigcir p 1379 L cinttre’omger |i 734 L cineraoniger 55 1 701 L cinerconigcr Li 365 L Bamansis L5O0 L sarnensis L979 L samcnsis L 984 L aamoosis L644 L sarpenrsis 70 1 L965 L sarnertsis ■ L890 L samensis -I L053 L sarncnsis * L970 L sarne-nsis 068 L woltsrstorffl 90 100 too 143 L minimus 126 L cf cewsfcu* 169 L corslcus 1 70 L corsicus 129 L cf. eofslcus 1010 L sancsrrsiij ■ 301 L cimineansis — 81 1 L lanninfi 04 35 S3 or 70S I Prandstcticri 991 L maxiniua i — 1 185 L maximum f- 903 L maxrmus 671 L maxlmus 16308 L max &44 L mnxiimus 992 L maxlmus Li 109 I maximurs Figure 17. Maximum Likelihood consensus tree (rooted with respect to the genus Limax) inferred from a dataset of 83 (seven sequences obtained in the present paper and 76 retrieved from GenBank database) mitochondrial COI gene partial sequences. Numbers above branches represent bootstrap values. New and little known land snails from Sicily (Mollusca Gastropoda) 209 at most bifurcate with wider base, and the posterior apex is bigger. Examined material of Tandonia sowerbvi. Italy, Emilia Romagna, Castiglione dei Pepoli, Roncobi- laccio, 44°06 , 59 ,, N 11°13 , 42 ,, E, 593 m, 5. XE2011, 2 specimens (CS). Italy, Sicily: Castel- buono, Cozzo Luminario, Piano Sempria, 37°54 , 18”N 14°03’59”E, 1192 m, X.1990, 4 spe- cimens (CS); Palermo, Parco della Favorita, Val- lone del Porco, 38°10 , 07”N 13°20 , 39 ,, E, 243 m, 13. XI. 1990, 4 specimens (CS); Monreale, Bosco Ficuzza, Diga Scanzano, 37°55 , 14”N 13°22 , 25”E, 536 m, l.XII. 1990, 6 specimens (CS); Palermo, Fiume Oreto, Ponte delle Grazie, 38°04’4”N 13°19 , 25”E, 95 m, 19.XI.1990, 5 specimens (CS); idem, 3.XII.1990, 2 specimens (CS); Isnello, Con- trada Montaspro, 37°54 , 42 ,, N 13°59’30”E, 857 m, 5. III. 1991, 1 specimen (CS); Collesano, Contrada Croce, 37°55’23”N 13°55’20”E, 511 m, 5.III.1991, 6 specimens (CS); Cammarata, Monte Cammarata, Cozzo Panepinto, 37°38 , 16”N 13°36 , 34”E, 984 m, III. 1992, 4 specimens (CS); idem, 2 specimens (CL); Petralia Sottana, Fiume Imera Meridionale, 37°48 , 26”N 14°05’01”E, 808 m, 2.XI. 1992, 3 spe- cimens (CS); San fratello, Monte Soro, Pizzo Muto, 37°56 , 16”N 14°38’16”E, 1410 m, X.2001, 1 specimen (CS); idem, 1 specimen (CL); Melilli, Riserva Naturale Integrale Grotta Palombara, 2008 (CR); Monreale, Ponte Arcera, 37°55’42”N 13°23 , 01”E, 470 m, 14.XI.2008, 2 specimens, (CL); Melilli, Riserva Naturale Integrale Grotta Pa- lombara, 2008 (CR); Vizzini, Contrada Rubala, near the F. Vizzini, 37°08 , 28 ,, N 14°44 , 15”E, 376 m, 11.1.2009, 5 specimens (CL); Prizzi, Fontana Grande, 37°42 , 53”N 13°25 , 43”E, 800 m, 15.XI.2009, 4 specimens (CL); Torrenova, Rocca Scovoni, Piano Scodoni, 38°05’38”N 14°41’26”E, 25 m, 06. XII.2009, 1 specimen (CL); Castelbuono, S. Gu- glielmo, near the creek San Calogero, 37°55’04”N 14°04 , 22”E, 670 m, 1.2010, 3 specimens (CL); No- vara di Sicilia, Rocca Novara, 37°59’35”N 15°08’25”E, 1000 m, 07.XI.2010, 4 specimens (CL); Itala, Piano Fattaredda, 38°02’48”N 15 o 25’09”E, 612 m, 12.XII.2010, 2 specimens (CL); Isnello, Vallone Montaspro, 37 0 54’18”N 13°58’55”E, 783 m, 26.11.2012, 2 specimens (CL). Tandonia rustica (Millet, 1843) has an Euro- pean central and southern distribution and is found in the northern regions of Italy and along the Apen- nines up to the central regions. This species is cha- racterized by a very long epiphallus externally si- milar to that of T. marinellii. However, T. rustica is characterized by its whitish or creamy colora- tion, somewhat violetish, with numerous, small black dots; the penial complex (epiphallus+penis) is smaller (around 10 mm) compared to T. mari- nellii (20-31 mm), and has a different structure: penis proportionally shorter and epiphallus sho- wing internally long longitudinal rows of papillae, penial papilla proportionally larger and much more ornate; the place where musculus retractor inserts is not constricted; atrium is narrow and tube-sha- ped while in T. marinellii is short and very broad. Examined material of Tandonia rustica. Italy, Emilia Romagna, Castiglione dei Pepoli, Ronco- bilaccio, 5.XI.2011, 1 specimen (CS); Italy, Lom- bardia, Brescia, Valvestino, Armo, 45°46’N 10 o 35’E, 666 m, 22.X.2000, 1 specimen (CN); idem, Anfo, S. Petronilla, 45°46’N 10 o 29’E, 524 m, 4.V.2008, 1 specimen (CN); idem, Ghedi, 45°24’N 10°16’E, 85 m, 20.IX.1996, 1 specimen (CN), idem, Marone, Velio, 45°45’N 10 o 05’E, 200 m, 01.V.2007, 1 specimen (CN). Molecular analysis. Seven Tandonia speci- mens, three from Custonaci (TP) and four from Madonie mountains (PA), labelled as TCUS and TMAD respectively, were analyzed. Samples were stored in 75% Ethanol at -20 °C in test tubes. For each individual, a piece of about 40-50 mg was used for total DNA extraction. Pieces of each spe- cimen were deposited as vouchers at University of Urbino, Lab. of cytogenetics and molecular bio- logy. COI amplicons (654 bp) were obtained by LCO1490/HCO2198 universal primers (5’- GGTC AAC AAATC ATAAAGATATTGG-3 75 ’ - TAAACTTC AGGGT GACC AAAAAAT C A-3 ’ ) as in Folmer et al. (1994) with a PCR cycle of 95 °C for 5 min; 95 °C for 1 min, 42 °C for 1 min, 72 °C for 1 min (37 cycles); 72 °C for 10 min. Se- quencing of the purified PCR products was carried out using automated DNA sequencers at Eurofins MWG Operon (Germany). Finally, sequence chro- matograms of each amplified fragment were brow- sed visually. Sequences generated in this study were analysed with additional seventy-six Lima- cidae COI sequences retrieved from GenBank (IDs: AF239733-34, AM259702-06, AM259712-14, EF128217, FJ481179, FJ481181, FJ606455-71, FJ606481, FJ606483, FJ606485, FJ606487, FJ606489, FJ606491, FJ606493, FJ606495, 210 F. Liberto, S. Giglio, M.S. Colomba & I. Sparacio FJ606497, FJ606499, GQ145509, GQ145523, GQ145525, GQ145527, GQ145538-39, GQ145553, GQ145572-75, JN248291-99, JN248300-15; see also Reise et al., 2011). Sequences were visualized with BioEdit Sequence Alignment Editor 7 (Hall, 1999), aligned with the ClustalW option included in this software and double checked by eye. Standard mea- sures of nucleotide polymorphism and phylogenetic analyses were conducted in MEGA 5.0.3 (Tamura et al., 2011). The best-fit evolution model of nucleo- tide substitution resulted GTR+G (General Time Reversible+Gamma). The evolutionary history was inferred by using the Maximum Likelihood me- thod; the bootstrap consensus tree was inferred from 500 replicates; a discrete Gamma distribution was used to model evolutionary rate differences among sites (5 categories; +G, parameter = 0.4467). Codon positions included were lst+2nd+3rd. All positions containing gaps and missing data were eli- minated. Divergence among TCUS and TMAD groups (Dxy), assessed as p distance, was 5.9%. Although genetic differences (p distance) are only indicative when assigning a group to a given taxonomic rank, the distance we assessed (5.9%) between COI sequences obtained from specimens collected in Madonie mountains and Custonaci not only is in line with values considered discrimina- tory at the specific level in Mollusca (i.e. Herbert et al., 2003; Pfenninger et al., 2006), but it is even greater than estimated distances separating entities accepted as distinct species ( L . corsicus/L. senensis, 2.3%; L. ciminensis/L. senensis , 4.2%; L. mini- mus/L. wolterstorffi , 4.4%). In conclusion, phylogenetic tree (Fig. 17) and genetic distance between TCUS and TMAD groups firmly support the hypothesis that specimens from Custonaci and Madonie mountains belong to two distinct Tandonia species. Remarks. The genus Tandonia Lessona & Pol- lonera, 1882 has European-Mediterranean distribu- tion extended to the Black Sea coasts (Wiktor, 1987, Giusti et al., 1995). In Italy it’s verified the presence of six species (Bank, 2012): T. nigra (C. Pfeiffer, 1849), T. bndapestensis (Hazay, 1880), T. robici (Simroth, 1884), T. rustica , T. simrothi (Hesse, 1923), T. sowerbyi. Among them, T. sower by i and T. rustica are morphologically the more similar to T. marinellii n. sp. that, on the other hand, sharply differs from these taxa for all the characters descri- bed above. Noteworthy, in bibliography about Si- cilian and surrounding geographical areas a few taxa of uncertain taxonomic value are reported. These taxa are examined below. Amalia marginata var. oretea Lessona & Pollonera, 1882 is a taxon described for Sicily (locus typicus: “Palermo presso il fiume Oreto”) only on external morphological characters: “Typica, clipeo tantum zonula nigra lon- gitudinali mediana instructo” (Lessona & Pollonera, 1882). The body coloration in Tandonia ( =Amalia Moquin-Tandon, 1855) is quite variable and topo- typic specimens studied by us can be traced back, due to the shape of genitalia, to T. sowerbyi (Figs. 10-12). A. marginata var. oretea is then confirmed as a synonym of T. sowerbyi (Giusti, 1973; Bank, 2012). It would have been critical examination of typical material, but the specimens described by Lessona & Pollonera (1882) are no longer available in their collection housed at the Museo Regionale di Scienze Naturali di Torino (E. Gavetti in litteris). Bourguignat (1877) established a new genus and a new species, Palizzolia monterosati, on a single Milacidae shell from Calatafimi with the following description: “Limacelle ovalaire, epaisse, tres-bom- bee (comme spherique) en dessous, caracterisee en dessus: 1° Par une surface plane, sur laquelle on di- stingue un cucleus median, circonscrit par une pro- fonde depression; 2° par une forte echancrure a sa partie mediane superieure” Lessona & Pollonera (1882) recognise in Paliz- zolia diagnostic characters of the shell of Milacidae (oval, medial nucleus) and put Palizzolia in homo- nymy with the genus Milax Gray, 1855 (sub Ama- lia). This choise was followed by Cockerell (1991), Kennard & Woodward (1926), Wiktor (1987) al- though with a question mark, Alzona (1971) and Barker (1999). However, the shell of Milacidae of- fers no morphological characters useful for a relia- ble classification at both genus and species level and therefore it is possible that Palizzolia could be an older synonym of Tandonia. The taxon monterosati was usually treated as doubtful species (Lessona & Pollonera, 1882; Mina Palumbo, 1883; Cockerell & Collinge, 1893; Wik- tor, 1987; Cockerell, 1991;) because the rear hol- low, the considerable thickening of the lower part of the shell and the groove around the apex are ab- normalities detectable in different species of both Milax and Tandonia genera; only Kennard & Wo- odward (1926) pose the taxon monterosati in syno- nymy with Milax gagates (Draparnaud, 1801). New and little known land snails from Sicily (Mollusca Gastropoda) 211 Figures 18-20. Shell (limacella) of T. marinellii n.sp., holotypus, h: 7.65 mm - D: 5.05 mm. Figure 21. Geographic distribution of Tandonia marinellii n. sp (circles) and T. sowerbyi (squares) in Sicily (personal data). Figures 22-24. Typus of Palizzolia monterosati (MHNG BGT 2385), h: 4.5 mm - D: 3.8 mm, thickness 2.2 mm. Figure 25. Original label of P. monterosati (MHNG BGT 2385). Figure 26. Four sequential visions of the ampoule rotated progressively so that to allow the overall vision of the label of P. monterosati (MHNG BGT 2385). Giusti et al. (1995) note that Tandonia shells are generally thicker and oval, however examination of the Palizzolia monterosati type (MHNG BGT 2385) (Figs. 22-26) does not provide indications for univocal taxonomic attribution. At Calatafimi, locus typicus of P. monterosati , we surveyed M. ni- gticans (Philippi, 1851) and T. marinellii n. sp., but the presence of M. gagates and T. sowerbyi cannot 212 F. Liberto, S. Giglio, M.S. Colomba & I. Sparacio be definitely ruled out. Hence, it appears that the taxon/binomial Palizzolia monterosati Bourgui- gnat 1877 is a nomen dubium, attributable with certainty at neither genus nor species level. In these cases, on the basis of article 75.5 of ICZN, the Commission may be asked, in order to settle all taxonomic doubts, to set up a neotype which, in our opinion, should be Milax gagates since Pa- lizzolia , as prevalent use, has been considered a sy- nonym of the genus Milax , and monterosati synonym of Milax gagates. For North Africa, particularly Tunisia and North Eastern Algeria, no species of the genus Tandonia was ever reported (Cockerell, 1891; Wiktor, 1987; Abbes et al., 2010). Milax gasulli Altena, 1974 and Amalia ater Collinge, 1895 are well known morphologically (genitalia) and con- sidered as valid species of the genus Milax. Wiktor (1987) based on specimens of Algeria (without ad- ditional indications) puts Umax scaptobius Bour- guignat, 1861 in synonymy with Milax gagates and both Amalia cabiliana (Pollonera, 1891) and A. gagates (var. or subsp.) mediterrana Cockerell, 1891 in synonymy with Milax nigricans. Amalia maculata Collinge, 1895, described for the surroundings of Algiers, was considered by Wiktor (1987) synonymy of T. sowerbyi, but the original description does not allow per se a certain assignment to the genus Tandonia. The taxon ma- culata Collinge 1895, however, is pre-occupied by Amalia maculata Koch & Heynemann, 1874 =Ly- topelte maculate (Koch & Heynemann, 1874) of the family Agriolimacidae (see Wiktor, 1987) and, for this reason, Hesse (1926) published, in its place, the new taxon Milax collingei. Limax ere- miophilus Bourguignat 1861 (locus typicus Al- giers, Algeria) was described only based on color and remains a taxon of uncertain allocation at both genus and species level. T. sowerbyi is reported for the regions of Southern Italy, attested with cer- tainty up to Basilicata (Ferreri et al., 2005). For Calabria two little-known taxa were de- scribed by Paulucci (1879), unreported even by Alzona (1971): Amalia mongianensis (locus ty- picus: Monte Pecoraro, Mongiana, Catanzaro) and A. fulva (locus typicus: “Monte Sant'Elia, Palmi”). The specimens from Calabria we could examine are to be considered as T. sowerbyi , al- though there are some morphological features that require further study. Family Subulinidae Thiele, 1931 Rumina saharica Pallary, 1901 Examined material. Rumina saharica. Italy, Sicily, Egadi Islands, Marettimo, admist Case Romane and Buccerie 200-250 m, 37°58’N, 12°03’E, 30.V.2010, 6 specimens, 1 shell (CL); idem, 18 shells (CS); idem VIII. 20 12, 2 speci- mens, 10 shells (CC). Description. Shell dextral (Figs. 27-30), whi- tish, truncated, height 30.5 mm, maximum dia- meter 10 mm, slender, sub-cylindrical, with slightly convex sides, the last whorl is wider than the penultimate whorl. Animal white. Genitalia (Figs. 31-32) characterized by vagina internally with longitudinal pleats and penis internally with some sparsely distributed papillae towards the proximal end. Biology and Distribution. R. saharica is a thermophilic and xeroresistant species. The genus Rumina Risso, 1826 has Mediterranean distribu- tion extending to Macaronesia, but it was disper- sed by man in some extra-Mediterranean countries (United States, Mexico, Cuba, Bermudas, China, Japan). Currently, R. saharica seems to prevail in the north African-East European area (Carr, 2002; Prevot et al, 2007). Remarks. Prevot et al. (2007) with molecular analyses demonstrated the presence in the Mediter- ranean area of two groups of species: R. decollata and R. saharica. They also showed the presence in R. decollata of two clades genetically distinct but morphologically similar. In addition, Mienis (2002) re-evaluates the validity of R. paviae (Lowe 1861) from Morocco, Algeria, and Tunisia. In Sicily, ac- tually, is known only R. decollata (Manganelli et al., 1995; Bank, 2012) (Figs. 23-24). Carr (2002) signals, in the collection of Na- tural History Museum of London, the presence of three shells similar to R. saharica collected in Sicily, however he points out that without data on the genitalia the classification of Rumina spe- cies is not certain. The population of Rumina from Marettimo Island (Western Sicily) which we examined shows the typical morphological characters of R. saharica (sensu Carr, 2002) with the exception of the duct of bursa copulatrix which is slightly longer. New and little known land snails from Sicily (Mollusca Gastropoda) 213 Figures 27, 28. Shell of Rumina saharica , Marettimo, h: 31.9 mm - D: 10.2 mm. Figures 29, 30. Idem, h: 26.4 mm - D: 9.8 mm. Figure 31. Genitalia of R. saharica, Marettimo. Figure 32. Idem, internal structure of penis and vagina. 214 F. Liberto, S. Giglio, M.S. Colomba & I. Sparacio Family Clausiliidae Morch, 1864 Lampedusa lopadusae nodulosa Montero- sato, 1892 Clausilia (Lopadusaria) nodulosa - Monterosato, 1892: 29 Clausilia (Lopedusaria) nodulosa - Kobelt, 1893: 303 Clausilia (Lopedusaria) nodulosa - Kobelt, 1897: 292 Clausilia lopadusae var. nodulosa - Westerlund, 1901: 105 Clausilia lampedusae var. - Giglioli, 1912: 217 Lampedusa lopadusae nodulosa - Alzona in Zavat- tari, 1961: 427 Delima (Lmpedusa) lopedusae - Alzona, 1971: 92 Lampedusa lopadusae , (synonym) nodidosa - Ho- lyoak, 1986: 217 Lampedusa lopadusae , (synonym) nodulosa - Beckmann, 1992: 22 Lampedusa lopadusae , (synonym) nodulosa - Cian- fanelli, 2002: 61, T. 9, f. 29 Lampedusa lopadusae , (synonym) nodulosa - Bank, 2012 Examined material. Italy, Sicily, Pelagian Is- lands, Lampione, 31.VIII.2009, 12 specimens, legit T. La Mantia (CS); idem, 09.IX.2009, 23 speci- mens, legit A. Corso (CL); idem, 23. VII. 2010, 5 specimens, legit T. La Mantia and S. Pasta (CS). Description. Shell sinistral (Figs. 33-36), me- dium-sized (height 13-18.7 mm; maximun diameter 3. 6-4.4 mm), fusiform, apex obtuse, elongated and inflated at half of its height, rather thick and robust, yellowish-brown in colour when fresh; external sur- face with oblique, thin and close ribs, 54-81 ribs on penultimate whorl. Spire with 9-10 convex whorls slowly and regularly growing, last whorl distinctly narrower than penultimate whorl and tapering dow- nwards, rather gibbous near umbilicus. Sutures deep, subcrenulated; umbilicus slit-like, internally closed; aperture about % of shell height (height 3.5- 4.4 mm; maximun diameter 2. 9-3. 7 mm), irregu- larly ovalar or sub-squared, peristome continuous, reflected, little thickenek. Aperture with 5 lamellae on parietum and columellar side and 3 or 4 plicae and lunella on palatum. On parietum (Figs. 41-42), starting from suture, there are: parallel lamella in the form of small relief, spiral lamella at centre of parietum, columellar lamella, a little subcolumellar lamella, and a tooth like parietal lamella (upper la- mella); only columellar lamella and parietal lamella are visible trough the opening (in apertural view). On the palatum (Fig. 43) there is a lateral lunella and, starting from suture: a long, well developed sutural plica; a second sutural plica variable in length: as long as the first one, shorter, or someti- mes absent; principal plica thin and raised; palatal plica showing a rear portion merged with the upper part of lunella, a central indistinct part and an an- terior part in the form of relief just visible. Clausi- lium elongated, plough-like (Fig. 44). Body. Animal oval-elongate, narrow, posteriorly pointed, white-yellowish; upper tentacles short, cy- lindro-conical, apically widened, with small black eyes; lower tentacle very short (Fig. 51; see also Cianfanelli, 2002 fig. 29). Genitalia. Anatomical organization of the geni- talia (Fig. 49) is similar to L. lopadusae (Soos, 1933; Pinter & Varga, 1984; Holy oak, 1986; Giusti et al., 1995) with penial complex consisting of flagellum, epiphallus, penis and penial diverticulum; flagellum short and slender; epiphallus divided by insertion of penial retractor muscle into proximal (1.5- 1.6 mm) and distal (1.2- 1.9 mm) portions; long, hook-like pe- nial diverticulum (1.5-1.88 mm) arising on border between epiphallus and penis; penis long (2. 6-3. 6 mm); on the inner wall there are 5 longitudinal crests which are parallel in the distal portion and rather in- distinct towards the penial diverticulum. Vagina long; short, wide copulatory duct branched in a short and slender duct of bursa copulatrix with small oval bursa copulatrix, and a short diverticulum of bursa copulatrix; short free oviduct. Biology and Distribution. At the base of ve- getation, at the soil, under stones (T. La Mantia in verbis). L. lopaduse nodidosa is endemic of the lit- tle isle of Lampione (Fig. 52), Pelagian Islands, bet- ween Sicily and Tunisia. Remarks. Monterosato (1892) described "Clau- silia (Lopadusaria) nodulosa" from the island of Lampione, with the following words: ”Conchiglia solida, striata quasi obliquamente (nella C. Lopa- dusae le coste sono perpendicolari ed esattamente lamellate); apertura a bordi ben rivoltati, porcella- niosi; colorazione bianchiccia; anfratti cochleae- formi, apice piu ottuso. Dimensione quasi la stessa.” [“Solid shell, ribbed almost sideways (in C. lopadusae the ribs are perpendicular and exactly la- mellated); opening with edges well turned, porce- lain-like; whitish colour; cochlea-shaped whorls, apex more obtuse. Almost the same size"]. New and little known land snails from Sicily (Mollusca Gastropoda) 215 Figures 33-36. Shell of L. lopadusae nodulosa , Lampione, h: 16.94 mm - D: 4.16 mm. Figures 37-40. Shell of L. lo- padusae lopadusae , Lampedusa, h: 17.11 mm - D: 4 mm. 216 F. Liberto, S. Giglio, M.S. Colomba & I. Sparacio Figures 41-44. L. lopadusae nodulosa , Lampione: palatum of two specimens (Figs. 41-42), parietum (Fig. 43) and clausi- lium (Fig. 44). Figures 45-48. L. lopadusae lopadusae , Lampedusa, palatum of two specimens (Figs. 45-46), parietum (Fig. 47) and clausilium (Fig. 48). New and little known land snails from Sicily (Mollusca Gastropoda) 217 Subsequently, this taxon is reported by Kobelt (1893; 1897) while Westerlund (1901) and Alzona (1961) consider it respectively as variety and sub- species of L. lopadusae. Alzona (1971), Holyoak (1986) and Beckmann (1992) put L. nodulosa in sy- nonymy with L. lopadusae. No news of this taxon is reported by Manganelli et al. (1995) and Cossi- gnani & Cossignani (1995). Cianfanelli (2002), de- spite considering it a synonym of L. lopadusae , reports that the population of the island of Lam- pione “. . . presenta dei caratteri piuttosto distinti sia nella conchiglia che nell’animale” ["... shows pretty distinct characters both in the shell and in animal"]. Nordsieck (2007) did not mention it in his catalog on the Clausiliidae of the world, Bank (2012) still considers it a synonym of L. lopadusae. L. nodulosa differs from L. lopadusae (Figs. 37-40. Figs. 45-48. Fig. 5 1) for shell less robust, darker in color i.e. yel- lowish-brown (yellowish-grey in L. lopadusae ), with deeper sutures, and whorls more convex so that the shell profile, in frontal view, appears less linear than L. lopadusae ; peristome is less develo- Figure 49. Genitalia of L. lopadusae nodulosa. Figure 50, 51. Body colour in L. lopadusae lopadusae (Fig. 50) and L. lo- padusae nodulosa (Fig. 51). Figure 52. Lampione island (photo P. Lo Cascio). 218 F. Liberto, S. Giglio, M.S. Colomba & I. Sparacio ped and calcified; ribs are more oblique, more nu- merous and less robust; in the internal structure of the shell, L. nodulosa mostly shows a second sutu- ral plica (rarely present in L. lopadusae). Genitalia of L. nodulosa differ from L. lopadusae for penial diverticulum slightly longer; the animal is lighter in color. As reported in the original description of Monterosato (1892), highlighted by Cianfanelli (2002) and confirmed by our observations, L. no- dulosa presents some morphological differentia- tions with respect to L. lopadusae and therefore we believe it is worthy of taxonomic reconsideration, at least at the sub-specific level, also in view of its peculiar geographical isolation. Indeed, the island of Lampione, where L. nodidosa lives, reaches its maximum altitude at 36 m above sea level and is approximately 17.5 Km far from Lampedusa, from which is separated by a stretch of sea -80 m deep. Despite its very little size, this islet harbours a very rich pool of plant and animal species, particularly some local endemics of high biogeographic interest (Lo Cascio & Pasta, in press.) Family Hygromiidae Tryon, 1866 Cernuella (Cernuella) tineana (Benoit 1862) Helix tineana - Benoit, 1862: 185-187, t. 4, fig. 24 (Calatafimi) Helix tineana - Pfeiffer, 1868: 487 (Sicilia, Cala- tafimi) Helix tineana - Benoit, 1875: 14 (Calatafimini) Helix Xerophila tineana - Kobelt, 1875: 18 (Ca- latafimi) Helix (Xerophila) Jacosta tineana -Westerlund, 1876: 104 (Sicilia) Helix (Xerophila) Jacosta tineana var. kobeltiana - Westerlund, 1876: 104 Helix tineana - Kobelt in Rossmassler, 1877: 103- 104, fig. 1452 (Sicilia, Calatafimi) Xerophila (Jacosta) tineana - Kobelt, 1881: 47 (Sicilien) Xerophila (Jacosta) tineana kobeltiana - Kobelt, 1881:47 (Sicilien) Helix tineana - Benoit, 1882: 37 (Calatafimini) Helix (Helicella) Jacosta tineana - Tryon, 1887: 253 pi. 62 fig. 92-94 (Sicily) Helix (Xerophila) Jacosta tineana - Westerlund, 1889: 318 (Sicilien, Calatafimini) Helix (Xerophila) Jacosta var. mista - Westerlund, 1889: 318-319 (Sicilien) Helix (Xerophila) Jacosta tineana var. kobeltiana - Westerlund, 1889: 319 (Sicilien) Helix (Xerophila) Jacosta tineana var. mista -We- sterlund, 1890: 61 (Sicilien) Helix (Xerophila) Jacosta tineana var. kobeltiana - Westerlund, 1890: 61 (Sicilien) Helicella Jacosta tineana - Pilsbry, 1894: 260 Helicella Jacosta tineana var. mista - Pylsbry, 1894: 260 Helicella Jacosta tineana var. kobeltiana - Pylsbry, 1894: 260 Helicella (Xerotropis) tineana - Alzona, 1971: 174 Helicella (Xerotropis) tineana mixta - Alzona, 1971: 174 Helicella (Xerotropis) tineana kobeltiana - Alzona, 1971: 174 Examined material. Italy, Sicily, Monte Cofano, Gorgo Cofano, 38°06’07”N 12°40’31”E, 228 m, 14.X. 1984, numerous specimens (CS); Italy, Sicily, Sciacca, Torre Macauda, 37°28 , 58”N 13°10’59”E, 59 m, l.VII/3 l.VII. 1986, numerous specimens (CS); Italy, Sicily, Monte Cofano, Gorgo Cofano, 38°06’07”N 12°40’31”E, 228 m, 14.IV. 1991, nume- rous specimens (CS); Italy, Sicily, Ribera, Contrada Castello, 37°30 , 18”N 13°15 , 04”E, 153 m, IX.2005, 3 specimens, 10 shells (CR); idem, 37°30’23”N 13°14’12”E, 144 m, IX.2005, 2 specimens, 12 shells (CR) ; Italy, Sicily, Cava a Nord di Ribera, 30.XII.2007, 3 specimens, M. and E. Bodon (CB); Italy, Sicily, Sciacca, Torre Macauda, 37°28 , 58”N 13 o 10’59”E, 59 m, 24.V.2008, numerous specimens (CS) ; idem 22.11.2009, 23 shells (CL); Italy, Sicily, Sciacca, Torre Macauda, 37°28 , 58” N 13°10 , 59” E, 60 m, 22.XI.2009, 1 specimen, 8 shells (CL); Italy, Sicily, Custonaci, Rio Forgia, 38 o 03’42”N 12°39 , 32”E, 56 m, 6.II.2011, 3 subfossil shells (Figs. 70-72); Italy, Sicily, Monte Cofano, Gorgo Cofano, 38°06 , 07”N 12°40’31”E, 228 m, 20.XI.2011, nume- rous specimens (CS); Italy, Sicily, Monte Cofano, Gorgo Cofano, 38°06 , 07”N 12°40’31”E, 228 m, 4.III.2012, numerous specimens (CS); Italy, Sicily, Custonaci, Baglio Cofano, 38°06 , 11”N 12°40’40”E; 250 m, 05.VIII.2012, 29 shells (CL). Description. Shell dextral (Figs. 53-55, 57-75), medium-sized (height: 7.2 mm, maximum diameter 17 mm), depressed, robust, whitish or greyish-yel- low in colour with brown band and dark apex; ex- ternal surface finely and regularly ribbed, opaque. New and little known land snails from Sicily (Mollusca Gastropoda) 219 Figures 53-56. “Helix” tineana , Calatafimi, Paulucci collection (MZUF GC/10825) (Figs. 53-55) and original label (Fig. 56), photos Saulo Bambi. Figures 57-60. C. tineana , Custonaci, Monte Cofano, Baglio Cofano, h: 5.55 mm - D: mm 11.10 mm. 220 F. Liberto, S. Giglio, M.S. Colomba & I. Sparacio Figures 61-63. CernueUa tineana , Sciacca, Torre Macauda, h: 6.90 mm - D: 12.92 mm. Figures. 64-66. Idem, h: 6.30 mm - D: 12.46 mm. Figures 67-69. C. tineana , Sciacca, Monte San Calogero, coll. Paulucci (MZUF GC/41419), foto Saulo Bambi. Figures 70-72. C. tineana , Custonaci, Rio Forgia, subfossil. New and little known land snails from Sicily (Mollusca Gastropoda) 221 Figures 73-75. Cernuella tineana, Sciacca, Torre Macauda, h: 7.07 mm - D: 12.42 mm. Figures. 76-78. C. amanda, San Vito lo Capo, Salinelle, h: 12.18 mm - D: 7.12 mm. Figures 79-81. C. rugosa , Castelluzzo, Calette degli Agliarelli, h: 6.30 mm - D: 11.95 mm. Figures 82-84. C. cisafpina, Castellammare del Golfo, Fraginesi, h: 6.90 mm xD: 10.80 mm. 222 F. Liberto, S. Giglio, M.S. Colomba & I. Sparacio Figures 85-87. Genitalia of Cernuella tineana , Custonaci, M. Cofano, Gorgo Cofano (Fig. 85), C. cisalpina, Castellam- mare del Golfo, Fraginesi (Fig. 86) and C. rugosa , Castelluzzo, Golfo di Cofano (Fig. 87). New and little known land snails from Sicily (Mollusca Gastropoda) 223 l mm Figure 88. Genitalia of Cernuella tineana, Sciacca, Torre Macauda. Figure 89. Idem, internal structure of penis, dart sac and vagina. 224 F. Liberto, S. Giglio, M.S. Colomba & I. Sparacio spire more or less flat, with 5 regularly growing whorls, slightly convex; marked sutures; last whorl very convex below and keeled at its periphery; um- bilicus deep and wide, about 1/3 of maximum shell diameter; aperture oval and slightly angled, peri- stome simple, interrupted, with internal rib. Body. Animal whitish; dorsal region provided, more or less extensively, of dark spots. Genitalia. Short free oviduct, duct of bursa co- pulatrix of medium length, with large base, ending in a sac-like bursa copulatrix; vagina short (1-1.6 mm), 2 tufts of digitiform glands with 4-5 slender lobes and 8-12 apexes, disposed on opposite sides of proximal vagina. Dart-sac complex consisting of a pair of stylophores located on one side of va- gina; large outer stylophore containing dart. Penial complex composed of flagellum, epiphallus and penis; flagellum long (2-2.4 mm), ending where vas deferens enters penial complex; epiphallus long 3-5 times the length of the penis (4. 5-5. 5 mm), ending where penial retractor muscle con- tacts penial complex wall; penis short (1.5-2. 2 mm); penial papilla cylindrical, elongate, with api- cal opening, and base connected to penial walls by three small muscles (frenula). Biology and Distribution. C. tineana is found on the ground, usually on grass often under stones. It is endemic to Sicily, distributed with point popu- lations in coastal and low-hill territories ranging from Custonaci (Monte Cofano) in the province of Trapani to Ribera in the province of Agrigento (Fig. 90). Comparative notes. Anatomical character of "Helix" tineana suggest to ascribe this species to the genus Cernuella Schluter, 1838 sensu stricto: penial papilla with three basal frenula, two groups of digitiform glands on opposite sides of the vagina, proximal vagina short or absent, proximal portion of the duct of bursa copulatrix wide (Manganelli & Giusti, 1987; Manganelli et al., 1996a, b, 2001). Currently, five species of Cernuella s. str. are reco- gnized in Sicily (Bank, 2012): C. aradasi (Pirajno, 1842), C. metabola (Westerlund, 1889), C. cisal- pina (Rossmassler, 1837), C. virgata (Da Costa, 1778), C. rugosa (Lamark, 1822). C. aradasi is a dune-species with limited distri- bution to the dunes near the lighthouse in Messina (North-east Sicily) and neighbouring sandy soils. It’s distinguished from C. tineana for shell smaller, smooth, globose and without keel; genitalia cha- racterised by relatively large penis and by epiphal- lus twice as long as penis. C. metabola is an endemic species from Lam- pedusa island; it is distinguished from C. tineana by the shell with the globose shell with disconti- nuous, thick ribs and narrow umbilicus. A prelimi- nary study on the genitalia of this species seems to highlight significant differences from the other Cer- nuella species (unpublished data). C. cisalpina is a polymorphic species, with a Mediterranean distribution, for which several taxa of still difficult taxonomic interpretation were established. Shell of small-medium size, “small sized Cernuella ” sensu Manganelli & Giusti (1987), subglobose, with thin ribs, sometimes well raised, last whorl usually rounded or angled at its periphery (keel-like) (Figs. 82-84). Some popula- tions of C. cisalpina present a shell similar to that of C. tineana , but in addition to the morphological characteristics of C. tineana pointed out above, they are always distinguishable by their genitalia with epiphallus 2-3 times longer than penis, fla- gellum and penis proportionally shorter and digi- tiform glands lower, i.e. between the vagina and the inner dart sac (Fig. 86). C. virgata is a polymorphic species showing a European-wide distribution. In Sicily it is common at low and medium altitudes where specimens can be found on grass and shrubs. C. virgata is distin- guished from C. tineana for the shell which is smo- oth or with faint wrinkles, without keel and larger, "large sized Cernuella " sensu Manganelli & Giusti (1987); genitalia resembling those of C. cisalpina but with epiphallus longer and more numerous di- gitiform glands. C. rugosa , endemic of Western Sicily known only for two locations (Figs. 79-81, 90), is an ex- tremely vulnerable species deserving of protection. From the morphological point of view C. rugosa is distinguished from C. tineana for the shell with rai- sed, irregularly spaced ribs and a cordlike, crenula- ted keel at its periphery, and for the penial complex (Figure 87; Manganelli at al., 1996b, Fig. 16) with penis longer, epiphallus and flagellum shorter. C. tineana is morphologically well distinguisha- ble from other Cernuella species sensu stricto. Dif- ferential diagnosis problems may arise with the shell of Cernuella (Xeroamanda) amanda Rossmassler, 1838 (see also Benoit, 1862-1857) (Figs. 76-78). In the latter species the shell is as convex infe- riorly as in the upper part, the keel is less obtuse than C. tineana , opening more angled and the um- New and little known land snails from Sicily (Mollusca Gastropoda) 225 bilicus markedly funnel- shaped. However, an exa- mination of genitalia can easily allow to distinguish the two species that belong to distinct subgenera (Manganelli etal., 1996). Remarks. Helix tineana was described by Be- noit (1862) for the surroundings of Calatafimi, "Pizzo di grasso " and dedicated to the then Director of the Orto Botanico of Palermo, Vincenzo Tineo. Benoit (1857) provides, in addition to the detailed description of the shell, also a comparative analysis of “Helix” rugosa Lamark, 1822 and “Helix” amanda Rossmassler, 1838, and draws the three species in table IV, figs. 24, 25, 29. In his later works, Benoit (1875, 1882) reported this species ci- ting only the locus typicus. Other authors cited this species: Pfeiffer (1868), Kobelt (1875), Kobelt in Rossmassler (1877), Tryon (1887). Westerlund ( 1876) reports it indicating the locality "Sicilia" and describes the variety kobeltiana on the basis of spe- cimens received by Kobelt under the name " H. tinei Ben.". Subsequently, Westerlund (1889) re-de- scribes H. tineana from " Sicilien bei Calatafimi " and adds to the variety kobeltiana the new var. mista with "Sicilien" as locus typicus. Alzona (1971) ascribes “tineana” to the genus Helicella Ferussac, 1821 subgenus Xerotropis Mon- te rosa to, 1892 and considers the two varieties de- scribed by Westerlund (1889) as valid subspecies. Neither Cossignani & Cossignani (1995) and Man- ganelli et al. (1995) nor Bank (2012) report "Helix" tineana for, respectively, the Italian fauna and the European fauna. Despite repeated searches, we have not found this species in the locus typicus, Calatafimi. Howe- ver, in Paulucci collection we saw a shell determi- ned as “Helix tineana” (MZUF GC/10825), collected in Calatafimi by a Sicilian naturalist De Stefani, in 1868 (Figs. 53-56). This topopypic sam- ple corresponds with Benoit’s original description and even with the specimens we have sampled and studied on Monte Cofano. The more southerly po- Figure 90. Geographic distribution of C. tineana (circles) and C. rugosa (squares) in Sicily. 226 F. Liberto, S. Giglio, M.S. Colomba & I. Sparacio pulations (Sciacca, Ribera) have slightly larger di- mensions. To these populations we attribute, by comparison, also one sample from Paulucci collec- tion sub H. caficiniana (MZUF GC/41419) picked up at Sciacca, Monte San Calogero (South-Western Sicily) (Figs. 67-69). “Helix” caficii , described by Westerlund (1876) with locus typicus Sciacca, corresponds, in our view, with the populations of C. tineana of Sciacca. If the examination of the type in the Westerlund col- lection will confirm this assumption "Helix" caficii may be a synonym of C. tineana. ACKNOWLEDGEMENTS We wish to thank Luigi Barraco (Valderice, Tra- pani, Italy), Marco Bodon (Italy, Genova), Saulo Bambi and Simone Cianfanelli (Museo di Storia Naturale delFUniversita di Firenze sezione di zoo- logia de “La Specola”), David R Cilia (Santa Ve- nera, Malta), Andrea Corso (Siracusa, Italy), Giulio Cuccodoro and Yves Finet (Museum d’Histoire na- turelle, Geneve, Switzerland), Elena Gavetti (Museo Regionale di Scienze Naturali di Torino, Italy), Alessandro Hallgass (Roma, Italy), Tommaso la Mantia (Palermo, Italy), Pietro Lo Cascio (Lipari, Messina, Italy), Giuseppe Maraventano (Lampe- dusa, Agrigento, Italy), Gianbattista Nardi (Nuvo- lera, Brescia, Italy), Roberto Poggi (Museo Civico di Storia Naturale di Genova “G. Doria”, Italy), Agatino Reitano (Tremestieri Etneo, Catania, Italy), Andras Varga (Andras Varga, Matra Muzeum Gyongyos, Hungary), Francisco Welter-Schultes (Zoologisches Institut, Berliner, Germany). 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Synopsis molluscorum in re- gione palaearctica viventium ex typo Clausilia Drap. Memories de l’Academie imperiale des Sciences de St.-Petersbourg. 11: 1-203. WiktorA, 1987. Milacidae (Gastropoda: Pulmonata). A sy- stematic monograph. Annales Zoologigi, 41:1 53-3 19. Biodiversity Journal, 2012 , 3 ( 3 ): 229-236 A stability assessment on seasonal variation of seaweed beds n theTrat peninsula of Thailand Nidsaraporn Petsut'jAnong Chirapart 2 & Methee Keawnern 1 ’Department of Fishery Management, Faculty of Fisheries, Kasetsart University, Bangkok 10900, Thailand; email: nidsarapom@ru.ac.th department of Fishery Biology, Faculty of Fisheries, Kasetsart University, Bangkok 10900, Thailand ’Corresponding author ABSTRACT Species diversity, biomass and distribution pattern of seaweed beds in the Trat peninsula, east coast of Thailand, were investigated in relation to environmental conditions from Ja- nuary to December 2011. The macroalgal samples and environmental factors were collec- ted monthly; covering cool-dry (January-February, November-December), hot-dry (March-April) and rainy (May-October) seasons at four sampling stations; Ao Cho, Ao Lane, Laem Tien and Laem Sok. A total of 26 taxa of marine benthic algae were recorded, of which 16 species of red marine algae were the most diverse group. It was found that Ccitenella nipae , Gracilaria salicornia, Gelidium pusillum , Hydropuntia changii, Hypnea hamulosa, Kyrtutrix maculans, Laurencia decumbents, Lyngbya majuscula , Peyssonnelia rubra and Ulva clathrata were the most abundant throughout the sampling period. The hi- ghest number of marine flora species was obtained in March (25 species), whereas the lo- west in June (12 species). Algal biomass had a maximum value in April (59.50 g/m 2 dry weight) and minimum value in July (20. 14 g/m 2 dry weight). KEY WORDS Seasonal variation; benthic algae; seaweed beds; Conservation; Trat peninsula. Received 12.07.2012; accepted 03.09.2012; printed 30.09.2012 INTRODUCTION The seaweed or marine macrophytic algae, which are a large and diverse group of eukaryotic photo synthetic organisms occurring in marine en- vironment, are one of the major marine fishery re- sources in Thailand (Edwards et al., 1982). Seaweed beds are a common habitat in coastal in- shore communities consisting of large benthic ve- getation and distributed widely along coastline of Thailand (Lewmanomont, 1998; Prathep, 2005). They are highly valuable ecologically and eco- nomically and perform a variety of functions within marine coastal ecosystems (Lobban & Harrison, 1994; Stachowicz et al., 2008). Most seaweed beds are served as a vitally important food sources for fish and aquatic invertebrates and provide breeding area for several species of marine animals (Zhang et al., 2008). Additionally, seaweeds are used around the world as food and fertilizers and for the extraction of valuable commercial products (Sam- bamurty, 2006, Graham et al., 2009). Trat is one of the provinces located in the east of Thailand, and encloses the upper Gulf of Thai- land adjacent to the border between Thailand and Cambodia. Along the coastline of Trat peninsula, there are many different types of coastal ecosystems including estuaries, mangrove areas, sandy shores and mudflats. In addition to coastal environment, there is a considerable amount of nutrient supply variation in the response of wave exposure gradient. Because of the properties of coastal area features and environmental diversity, Trat peninsula has re- markably diverse marine fishery resource, espe- 230 N. Petsut, A. Chirapart & M. Keawnern cially macroalgal flora; wild populations of macro- algae are widely distributed in the intertidal and subtidal zones of Trat peninsula (Pirompug, 1976) and used as human food and for agar extraction (Edwards & Tam, 1984; Chirapart et al., 1992). Currently, the abundance and diversity of Thai seaweed has been vulnerable to decline because of over-harvesting of natural populations and the eco- logical deterioration of several inshore and estuary ecosystems (Lewmanomont, 1998). In Trat costal areas and adjacent waters, the rapid extension and development of fisheries acti- vities by local fishermen and conversion of man- grove areas into shrimp farms and urban areas threaten aquatic organisms (Menasveta, 1997) and this situation has a potentially negative impact on the coastal ecosystem (Doydee, 2005). Accordingly, the change in ecosystem and environmental condi- tions, mangrove deterioration and costal land-use activities would result in a decrease of macroalgal biodiversity and biomass in Trat peninsula. The need to promote a scenario of seaweed re- source management is therefore important for su- stainable conservation and restoration of coastal ecosystem. However, information and knowledge regarding to macroalgal assemblages and their ecology is very limited in this coastal region. In order to provide useful data for a possible preli- minary management strategy for conservation of seaweed resources in Trat peninsula we determi- ned seasonal variation in the species diversity, biomass and distribution pattern of macroalgae with reference to some environmental variables for better understanding of the recent situation in algal communities. MATERIALS AND METHODS Study area The study site is located at the coastal area of Trat province, east of Thailand. Four coastal areas of intertidal habitat, which are different in shape and environmental condition, were chosen as research station: Ao Cho (site 1), Ao Lane (site 2) Laem Tien (site 3) and Laem Sok (site 4). Among the stations, Ao Clio and Ao Lane are semi-exposed areas. Ao Cho is characterized by the formation of sandy beaches alternated with rocky shores and partly surrounded by mangroves, while natural habitat of Ao Lane is composed mainly by mudflat and this area is moderately occupied by in- digenous fisheries community. Laem Tien is a non-exposed area with muddy sand bottom fringed by mangroves, and some parts of this area are heavily exploited for aquaculture and shrimp farms. Additionally, Laem Sok, a fully exposed area with rocky shore habitat, is partially converted for commercial and industrial activities. Sample collection and laboratory analysis Species diversity, biomass and distribution Field sampling was carried out once a month at four stations for a year (from January to December 2011). Benthic marine macroalgae were sampled thoroughly by wading or snorkeling. Complete thalli of live specimens were uprooted by hand or with paint-scraper, placed in plastic bags, labeled by location and date of collection, and transported to laboratory. Algal samples were rinsed to remove sediment and debris, photographed, preserved as herbarium vouchers, or, on some occasions, preserved in 4% formalin-seawater solution, and deposited at the Algal Bioresources Research Center, Faculty of Fi- sheries, Kasetsart University. The species identifi- cation was based on gross morphology and internal features following Lewmanomont & Ogawa (1995); Huisman (2000) and Litter & Litter (2000; 2003). In order to determine algal biomass and di- stribution pattern, a quadrat method along a vertical transect line set across the intertidal zone perpendi- cularly to the coastal line was performed monthly throughout the study period. Once a month 25 replicates of sampling quadrat (50x50 cm) from research stations were collected for determination of algal biomass. Algal samples from each quadrat were carefully cleaned with fre- shwater to remove sand, silt, epiphytes and other de- bris before weighting. Dry weight of algal biomass was obtained by drying the samples in the oven at 105°C for 48 hours (Wong & Phang, 2004). Environmental parameters Environmental parameters, including physical and chemical factors, were recorded seasonally at the moment of each sampling. For physical varia- bles, seawater temperature, salinity, pH, turbidity and dissolved oxygen (DO) were measured in the A stability assessment on seasonal variation of seaweed beds in the Trat peninsula ofThailand 231 field. Seawater temperature and DO were determi- ned using a salinity compensated dissolved oxygen meter (YSI Model 57). Salinity was determined by hand refractome- ter; pH was measured using a pH meter (YSI Model 60) and water transparency was estimated by a turbidimeter (LaMotte Model 2020). Total rainfall of Trat peninsula was obtained from the Meteorological Department ofThailand. For che- mical variables, nutrient concentration in the sea- water was evaluated. Water quality was sampled from each study site and fixed in ice chests to examine alkalinity, har- dness, ammonia, nitrate, nitrite and phosphate, using the methods of Sasaki & Sawada (1980) and Strickland & Parsons (1972). Statistical analysis Data for statistical analysis were tested initially for normality and homogeneity (Zar, 1984). One- way analysis of variance (ANOVA) was employed to search for any significant difference among month, site and biomass data of each species. Statistical significance was set at p<0.05 and the stability of the estimate reflected by 95% confident intervals. All tests and analyses were performed with SPSS version 12.0 (SPSS, Inc., Chicago, IL). RESULTS Species diversity A total of 26 taxa were identified including 3 species of Cyanobacteria, 3 species of Chlorophyta, 4 species of Phaeophyta and 16 species of Rliodo- phyta (Tables 1,2). The number of species varied during the study period and ranged from 25 (March 2011) to 12 (June 2011). Catenella nipae , Gracilaria salicor- nia , Gelidium pusillum , Hydropuntia chang 'd. Hyp- nea hamulosa , Kyrtutrix maculans , Laurencia decumbents , Lyngbya majnscula , Peyssonnelia rubra and Ulva clathrata were found throughout the sampling period. On the other hand, Brachytrichia quoyi, Chae- tomorpha crassa , Cladophora sp., Dictyota dicho- toma, Hydroclathrus clathratus , Padina australis , P. sanctae-crucis, Acanthophora spicifera , Bostry- cia tenella , Centroceras clavulatum , Ceramium flaccidum , Erythrotrichia sp ., Gelidiopsis intrica- tum , Gracilariopsis irregularis, Neosiphonia sava- tieri and Palisada papillosa had only single occurrences in time. Biomass and distribution pattern of marine benthic algae in each season and site Total marine macroalgal biomass gradually increased from January to March, reached to the peak in April (hot-dry season) with 59.50 g/m 2 dry weight and dramatically decreased in July (rainy season) with 20.14 g/m 2 dry weight, and then steadily increased from August to Decem- ber, reaching to the peak again in November (cool-dry season) with 55.46 g/m 2 dry weight. The seasonality of macroalgae biomass at the site was less uniform. During both dry and wet sea- sons, Gracilaria salicornia and Hydropuntia changii attained the maximum biomass mean value at 29.51 g/m 2 dry weight and 14.82 g/m 2 dry weight, respectively. Acanthophora spicifera , Hydroclathrus clathra- tus and Padina sanctae-crucis had greatest biomass during the cool-dry season, and less biomass during the wet season. In contrast, Gracilariopsis irregu- ralis , Hypnea hamulosa, and Padina australis exhi- bited higher biomass during the hot-dry season than in the wet season. Most seaweed biomass exhibited greatest abun- dance in Ao Clio e.g. Gracilaria salicornia, Graci- lariopsis irreguralis, Hypnea hamulosa, Hydropuntia changii, Palisada papillosa and Pa- dina sanctae-crucis. Some species, e.g. Acantho- phora spicifera, Hydroclathrus clathratus , Hypnea hamulosa, Hydropuntia changii, Palisada papillosa and Padina australis, were common in Ao Lane. The brown algae, Padina australis and P. san- ctae-crucis, however, were common in Laem Tien. Some species, e.g., Gracilaria salicornia, Hypnea hamulosa, and Palisada papillosa were common in Laem Sok. In general, there were no patterns found in biomass of algae among different sites. Ten species were found throughout the entire study period. These included two species of cyano- bacteria, Kyrtutrix maculans, Lyngbya majnscula, one species of green algae, Ulva clathrata, seven species of red algae, Catenella nipae, Gracilaria salicornia, Gelidium pusillum, Hydropuntia chan- gii, Hypnea hamulosa, Laurencia decumbents and Peyssonnelia rubra. 232 N. Petsut, A. Chirapart & M. Keawnern Ao Cho Ao Lane TAXA 1 2 3 4 5 6 7 8 9 10 11 12 1 2 3 4 5 6 7 8 9 10 11 12 CYANOBACTERIA Brachytrichia quoyi (Agardh) Bomet et Flahault + + + Kyrtutrix maculans (Gomont) Ume zaki + + + + + + + + + + + + Lyngbya majuscula (Dillwyn) Harvey + + + + + + + + + + + + + + + + + + + + CHLOROPHYTA Chaetomorpha crassa (C. Agardh) Kiitzing + + + + + + + + + + Cladophora sp. Kiitzing + + + + + + + + + + + + Ulva clathrata (Roth) C. Agardh + + + + + + + + + + + + + + + + + + + + + + -i- + PHAEOPHYTA Dictyota dichotoma (Hudson) Lamouroux + + + + + Hydroclathrus clathratus (C.Agardh) M. A. Ho we + + + + + Padina australis Hauck + + Padina sanctae-crucis Borgesen + + + + + + + + RHODOPHYTA Acanthophora spicifera (M. Vahl) Borgesen Bostrycia tenella (J.V. Lamouroux) C.Agardh + + + + + + Catenella nipae Zanardini + + + + + + + + + + + + Centroceras clavulatum (C.Agardh) Montagne -i- + Ceramium flaccidum (Harvey ex Kiitzing) Ardissone + + + + Erythro trichi a sp. Areschoug + + + + + + + + + + + Gelidiopsis intricatum (C.Agardh) Vickers + + + + + + + + + + Gelidium pusillum (Stackhouse) Le Jolis + + + + + + + + + + + + Gracilaria salicornia (C.Agardh) E.Y. Dawson + + + + + + + + + + + + Gracilariopsis irregularis Abbott + + + + + Hydropuntia changii (Xia et Abbott)Wynne + + + + + + + + + + + + + + + + + + + + Hypnea hamulosa (Esper) J.V. Lamouroux + + + + + + + + + + + + + + + + + + + Laurencia decumbents Kiitzing + + + + + + + + + + + + + Neosiphonia savatieri (Hariot) M.S.Kim + + + + + + -!- + + + + -h + + + Palisada papillosa (C.Agardh) K.W.Nam + + + + + + + + + + + + + + + Peyssonnelia rubra (Greville) J. Agardh + + + + + + + + + + + + + + + + + + + + + Table 1. Seasonality of macroalgae at Trat Peninsula January (=1)/December (=12) 2011: Ao Cho and Ao Lane. A stability assessment on seasonal variation of seaweed beds in the Trat peninsula ofThailand 233 Laem Tien Laem Sok TAXA 1 2 3 4 5 6 7 8 9 10 11 12 1 2 3 4 5 6 7 8 9 10 11 12 CYANOBACTERIA Brachytrichia quoyi (Agardh) Bomet et Flahault Kyrtutrix maculans (Gomont) Ume zaki Lyngbya majuscula (Dillwyn) Harvey CHLOROPHYTA Chaetomorpha crassa (C.Agardh) Kiitzing Cladophora sp. Kiitzing Viva clathrata (Roth) C.Agardh PHAEOPHYTA Dictyota dichotorna (Hudson) Lamouroux Hydroclathrus clathratus (C Agardh) M. A. Howe Padina australis Hauck Padina sanctae-crucis Borgesen RHODOPHYTA Acanthophora spicifera (M. Vahl) Borgesen Bostry’cia tenella (J.V. Lamouroux) C.Agardh Catenella nipae Zanardini Centroceras clavulatum (C.Agardh) Montagne Ceramium flaccidum (Harvey ex Kiitzing) Ardissone Erythrotrichia sp. Areschoug Gelidiopsis intricatum (C.Agardh) Vickers Gelidium pusillum (Stackhouse) Le Jolis Gracilaria salicornia (C.Agardh) E.Y.Dawson Gracilariopsis irregularis Abbott Hydropuntia changii (Xia et Abbott)Wynne Hypnea hamulosa (Esper) J. V. Lamouroux Laurencia decumbents Kiitzing Neosiphonia savatieri (Hariot) MS. Kim Palisada papillosa (C.Agardh) K. W.Nam Peyssonnelia rubra (Greville) J.Agardh + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + ”h + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + 4 - + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + Table 2. Seasonality of macroalgae at Trat Peninsula January (=1)/December (=12) 2011: Laem Tien and Laem Sok. 234 N. Petsut, A. Chirapart & M. Keawnern Physical and chemical factors study There were insignificant variations in water temperature (p>0.05) among sites but significant variations in water temperature (p<0.05) among months. The water temperature was 27.5-33.5°C during the dry season and 25.5-3 1.5°C during the rainy season. The range was rather wide and was likely to influence the species diversity, biomass and distribution pattern of seaweed beds. In addi- tion, there were insignificant differences in salinity (p>0.05) among sites but significant differences in salinity (p<0.05) among months. The salinity du- ring the dry season was 32-37 %o and 15-27 %o du- ring the rainy season. Such a rather wide range was likely to in- fluence the marine macroalgae. There were insi- gnificant differences in turbidity (p>0.05) among sites but significant variations in turbidity (p<0.05) among months. The turbidity was 4.70-346.67 NTU during the dry season and 10.33-983.33 NTU during the rainy season. These differences could influence the species diversity, biomass and distri- bution pattern of marine algae ANOVA revealed that there were insignificant variations in P0 4 3 ' (p>0.05) and TIN (total inor- ganic nitrogen: NH 4 + + N0 3 ~ + N0 2 ) (p>0.05) among sites but significant variations in P0 4 3 ' (p<0.05) and TIN (p<0.05) among months. The phosphate during the dry season was 0.0004- 0.0391 mg/1 and 0.0030-0.0670 mg/1 during the rainy season. And the TIN was 0.0134-0.2642 mg/1 during the dry season and 0.0343-0.2800 mg/1 during the rainy season. These ranges were rather wide, and were likely to influence the ma- rine macroalgae. DISCUSSION Species diversity study A total of 25 genera and 26 species of marine benthic algae were recorded, of which 16 species of red marine algae (a characteristically abundant and diverse group in the tropics) were the most as- sorted. Red algae occupy a wide range of irra- diance environments, including high-latitude and high- intertidal habitats subjected to long periods of full sunlight (Graham et al., 2009). Twenty-four percent higher species richness of marine algae was found at our study site compared to the study of Laehyeb (2011), in which only 21 species were reported throughout Trat peninsula; only three field collections were made in that pre- vious study as compared to the four field collec- tions in this study. Thus, the number of visits for field collection as well as the collection efforts could be important for appraising species diversity more accurately. In addition, the differences we observed suggested that there was temporal varia- tion in species diversity of marine algae. Many marine macroalgae have posed ecologi- cal problems in some marine ecosystems, which is probably due to environmental changes linked to decreasing of mangrove forest, sewage discharges, shrimp culture, tourism and factories activities (Doydee, 2005; Thongroy et al., 2007; Laehyeb, 2011). These activities have had serious impact on coastal environments. Biomass and distribution pattern study Among all four study areas of Trat peninsula, marine macroalgal biomass and diversity were the highest at Ao Cho during the dry season. This area is semi-exposed, sandy bottom with rocks and partly surrounded by mangroves. This same phenomenon was observed in macroalgae at Si- rinart Marine National Park by Prathep (2005) in Thailand. Water motion is one of the most impor- tant variables influencing marine macroalgae, be- cause it regulates turbidity, light penetration and nutrient availability (Nishihara & Terada, 2010; Kang et al., 2011). At Ao Cho, marine macroalgae were exposed to intermediate levels of water motion, which al- lows the exchange of gases and uptake of nutrients (Lobban & Harrison, 1994; Kang et al., 2011). Ao Lane is a semi-exposed area as same as Ao Cho but is characterized by large mudflats and harbours a lot of fishery communities. This place is very sensitive to environmental changes due to human activity stressing the natural environment, such as sewage discharges of fishery communities, which can cause the decrease of the species diversity and of seaweed abundance. Laem Tien is a wave-shel- tered area covered by mangroves with a large muddy sand bottom. Some parts of this place are used for aquaculture. A stability assessment on seasonal variation of seaweed beds in the Trat peninsula ofThailand 235 This site showed the lowest biomass and spe- cies diversity. Taking into account that the area is rather sheltered from wave action, marine macro- algae may be exposed to some physiological stress due to limited circulation of nutrients and gas exchange (Prathep & Tantiprapas, 2006; Thongroy et al., 2007). In addition, this site is subjected to environ- mental changes because of water pollution from the shrimp farms. Laem Sok is a wave-exposed area with a large rocky shore habitat and many re- staurants nearby the harbor. This place shows a high level of water motion, which is likely to affect species diversity and abundance of macroalgae (Prathep, 2005; Kang et al., 2011). In addition, in this area mangrove forests decreased in the last few years (Doydee, 2005; Laehyeb, 2011). Generally speaking, mangroves are really im- portant since these plants indirectly participate to habitat complexity and diversity of fauna and flora, particularly marine macroalgae (Ashton et al., 2003; Ellison, 2008, Doydee & Buot, 2011). In fact by trapping nutrients and sediments from river runoffs from the uplands and transpor- ting them to coastal waters (Anongponyoskun & Doydee, 2006; Ellison, 2008; Doydee, 2009) they contribute to improve shoreline stability and water quality. Therefore, the decrease of man- grove forests is certainly another reason affecting negative changes in species diversity and abun- dance of seaweed. ACKNOWLEDGEMENTS We thank the Japan International Research Center for Agricultural Sciences for partial finan- cial support. Mr. Narongrit Muangmai for the in- valuable editorial advice and Mr. Jiraweath Petsut for assistance with the field work. REFERENCES Anongponyoskun M. & Doydee P., 2006. The change coastline in Loi Island, Chonburi Province during 1997 to 2004. Kasetsart Journal (Nat. Sci.), 40: 249-253. Ashton E.C., Macintosh D.J. & Hogarth P.J., 2003. A baseline study of the diversity and community eco- logy of crab and molluscan macrofauna in the Se- matan mangrove forest, Sarawak, Malaysia. Journal of Tropical Ecology, 19: 127-142. Chirapart A., Lewmanomont K. & Ohno M., 1992. Sea- sonal variation of reproductive states of the agar- producing seaweed, Gracilaria changii (Xia and Abbott) Abbott, Zhang and Xia in Thailand. 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Bondarev Benthos Ecology Department, Institute of Biology of the Southern Seas (IBSS), Ukrainian National Academy of Sciences (NASU), Nakhimov av., 2, Sevastopol, 99011, Ukraine; email: igor.p.bondarev@gmail.com ABSTRACT Present paper reports on the possible existence of recent freshwater fauna in the Black Sea. Based on information available from malacology, ecology, paleontology, stratigraphy, hydrogeology and observations in situ, the presence of freshwater biota on the shelf and continental slope is discussed, including the existence of aerobic life forms in the Black sea deep-water cavity. KEY WORDS Aerobic; Biodiversity; Ecosystem; freshwater; submarine springs. Received 03.08.2012; accepted 03.09.2012; printed 30.09.2012 INTRODUCTION The Black Sea is a very specific marine basin with anaerobic water mass spreading from the ma- ximal bottom depth (more than -2200 m) to about - 200 m. This water thickness is valuated as about 85% of the whole Black Sea water volume (Zaitsev, 2006) and suggested as absolutely unfit for euka- ryotic life (Vinogradov, 1997). The Black Sea water salinity varies from about 17-18%o on the inner shelf till 23%o in the deepest basins’ parts (Sorokin, 1982) which excludes any possibilities for freshwater biota inhabitance. Any- way, the possibility of existence of aerobic life "oases" related to fresh groundwater springs in sul- fured hydrogen zone was hypothesized, taking into account a few unusual faunistic findings, signs "of island" speciation (Zaika, 2008). Additionally, on the Black Sea continental shelf bottom surface were repeatedly found out the shells of several gastropod mollusk species typical of fre- shwaters or brackish complexes (ITina, 1966; Go- likov & Starobogatov, 1972; Chukhchin, 1984). Most of these species - met nowhere else and de- scribed as extinct - supposedly re-deposited from ancient (Neoeuxinian) sediment layers formed in much more freshwater environmental conditions (Golikov & Starobogatov, 1972). However, among the above-mentioned gastropods, only one ( Theo - doxus pallasi Lindholm, 1924) is known for certain from Neoeuxinian sediment deposits. During the R/V «Maria S. Merian» (Leibniz In- stitute for Baltic Sea Research, Germany) expedi- tion, which was conducted in the Black Sea within the framework of the European project HYPOX in 2010, a live gastropod mollusk was sampled at the depth of 250 m (Sergeeva et al., 2011). If one be- lieves that its presence in anoxic zone may be not casual, then it is necessary to admit the existence of oxygen input into the sulfiired hydrogen water layer by sources which serve as freshwater springs. The finding of meiobenthic crustaceans in the deep Black Sea (around -2000 m) and at a depth of 174 m (Korovchinsky & Sergeeva, 2008) may be considered as one of the biological and ecolo- gical proofs of the existence of aerobic fauna in anaerobic zone. In 1986, during a test diving on the submarine inhabited vehicle of the USSR Navy at a depth of 600-640 m in the Yalta canyon at bottom, one "oasis" of aerobic life was found 238 Igor P. Bondarev out (Prof. Gevorkyan V.Kh. personal report); this information was under secret for a long time and therefore never published. The most credible reason for the existence of such an "oasis" is the occur- rence of a zone of stable and powerful submarine unloading of oxygen-rich groundwaters. Oceanological and geological researches confir- med the presence of submarine discharge of fresh water zones on the Black Sea continental shelf and slope (Trotsyuk et af, 1988; Shnyukov & Ziborov, 2004). Actually, these zones can create proper con- ditions for the existence of freshwater or brackish biota, both in aerobic and in anaerobic water masses of the Black Sea. The aim of the present research, based on geo- logical, biological and ecological data, is to investi- gate the existence of specific biocenoses in the Black Sea, related to the zones of the submarine un- loading of freshwaters. A discovery and description of such a biocenosis would extend knowledge about the Black Sea biodiversity and give new ideas about possible ways of the “oasis” fauna evolution. DISCUSSION Geological and oceanological evidence Submarine freshwater springs are known from many regions. Very recently, a Max Plank Institute science troop found rich bacterial life connected with freshwater springs even in the Dead Sea (Io- nescu et af, 2012). Black Sea is also suitable for se- arching submarine sources of freshwater. On geological terms it is possible to distinguish three basic types of submarine sources of groundwater unloading: artesian, karstic and waters of subriver- bed flow (Shnyukov & Ziborov, 2004). Data on the dynamics of the artesian pools’ groundwater opened toward the Black Sea, testily movements of different hydrogeological floors. Ac- tually these pools embrace all coastline of the Black Sea (Shnyukov & Ziborov, 2004). Submarine springs issued from the limestone massifs or other kinds of karsting rocks are widely spread in the Black Sea. Researches, managing with submarine inhabited vehicles, showed the presence of numerous rocky outcrops from the bottom silty-mud sediments cover on a narrow and steep continental shelf and slope of south Crimea (Bondarev, 2008; 2009). Oceanological researches in 37 th voyage R/V “Academician Vernadsky” showed submarine un- loading of karsting-crack waters in head parts of many Crimean submarine canyons. The hydroche- mical tests of near- bottom water allowed to set that in canyons salinity is lowered notably up to 12, 14, 15, 17%o (with reference to base-line va- lues of 21-22%o). Particularly, the desaltation of salt waters on the canyons bottom was found out on the continental slope of Turkey, north of Kef- ken Island and on the extreme north-west of the Turkish shelf (at 300-500 m below the sea level;13-17%o). Signs of desaltation were found also in the canyons of north-west Black Sea (Shnyukov & Ziborov, 2004). Use of impermeable water samplers, vacuum degassing and chromatographic analysis allowed to set the presence of solved oxygen in the benthic layer of deep-water part of the Black Sea. In the area adherent to the estuarine part of Danube, oxy- gen in benthic water decreases from 4.2 ml/1 in an off-shore zone practically to zero on the shelf edge. As recorded in eight near-bottom samples taken at different depths on the bottom and on the sides of a deep canyon up to 1340 m of depth, below the slope, up to the distal part of canyon fan, oxygen amount ranges from 0.3 to 1.6 ml/1. On the Anato- lian side (the cone of dejection of Kyzil-Irmak river) water-solved oxygen was found out in two deep-water stations with bathymetric marks -2064 m and -2003 m, respectively. Measured oxygen concentrations were 0.5 and 0.7 ml-1. At depths of over 2000 m, near-bottom waters contained oxygen, as revealed by tests from the districts of western ha- listaza zone, interhalistaza zone and the east hollow of the Black Sea (Trotsyuk et af, 1988). Major part of water samples containing oxygen was collected in submarine canyons or in the sur- roundings where the underground source of aerobic freshwaters binds to the submarine unloading. All these features do suggest the presence of specific freshwater aerobic biocenoses in the Black Sea is to be considered highly probable. Biological and ecological evidence Shells of gastropod molluscs of freshwater or brackish habitats were discovered on continental shelf and slope of the Black Sea (Golikov & Staro- Freshwater and brackish “oasis” fauna in the deep Black Sea 239 bogatov, 1972), where water salinity is 19-21%o. Their list (Table 1) was recently filled up by one specimen of living gastropod from the near-Bospo- rus region, where background water salinity exce- eds 22%o. Neritidae and Hydrobiidae mostly consist of marine species. ditions, these shells should be absent. In addition, these shells are found in the middle and external part of shelf and are absent in littoral zone. These circumstances and the sub-fossil state of seashells allowed to suppose they to origin from more ancient layers, maybe formed in a freshwater- Taxa Locality / inhabitance (after Kantor & Sysoev, 2006, others indicated) Fam. Neritidae: Theodoxus milachevitchi Golikov et Starobogatov, 1966 Theodoxus pallasi Lindholm, 1924 Crimea offshore/ recent mud, depth 20-60 m Black Sea Ne oeuxinian, Vityazean & Kalamitian layers, recent muds on the depth 18-158 m (Author’s data)/Aral & Caspian Seas, very freshened littoral spots in the Azov Sea (Golikov & Starobogatov, 1972); rivers of Ural Mountains and Armenia Fam. Flydrobiidae, Subfam. Pyrgulinae: Caspia valkanovi (Golikov et Starobogatov, 1966) Crimea offshore, silt on the 20 m depth Caspiohydrobia sp. Near-Bosporus region, mud on the depth of 250m (Author’s data) Turricaspia crimeana (Golikov et Starobogatov, 1966) Crimea, 15 m Turricaspia iljinae (Golikov et Starobogatov, 1966) Crimea offshore, mud on the 80- 180 m depth Turricaspia lirata marisnigri Starobogatov, in Alexeenko et Starobogatov, 1987 Crimea, Modiolula phaseolina (Philippi, 1844) contained mud. Turricaspia nevesskae (Golikov et Starobogatov, 1 966) Crimea offshore, mud on the 80- 180 m depth Table 1. List of freshwater and brackish water gastropods found on the shelf and continental slope of the Black Sea. However the genus Theodoxus Monfort, 1810 comprises species dwelling exceptionally in fresh or strongly refreshing (up to 5%o) waters of Eura- sia. This characteristic allows to use them as bioin- dicators of freshwater environments (Goodwin, 2006). The Subfamilia Pyrgulinae Brusina, 1882 also includes genera and species from freshwaters (springs, rivers and lakes) of Europe and front Asia (Anistratenko, 1998). A considerable part of these species inhabits brackish seas - Aral and Caspian. In the Azove- Black Sea basin the subfamily is represented by three genera whose species are usually encountered in the brackish zones of estuaries and rivers (Goli- kov & Starobogatov, 1972; Kantor & Sysoev, 2006). Hence, substantial differences in water sali- nity of sampling locations and in typical habitats of mollusk species led to hypothesize a possible intro- duced origin of shells from a freshwater environ- ment. But, in that case, in the southern part of Crimea littoral, where there are special habitat con- like environment (Golikov & Starobogatov, 1972). Such environmental conditions existed in a Neoeu- xinian period of the Black Sea evolution (IEina, 1966); Neoeuxinian layers on the shelf of Crimea are covered by more young ground sedimentations, from 1 to 4 m thick (Shcherbakov et al., 1978). Moving of shells through such a thick layer by means of natural processes is hardly plausible. But even if we assume such a possibility, then in Neo- euxinian deposits these species should be much more abundant than in recent sediments. However, species reported in Table 1 were found only in re- cent bottom deposits, with the exception of Theo- doxus pallasi. This species is the only one that is really cha- racteristic for Neoeuxinian layers whereas in later deposits is quite rare. It is has been reported that shells of T. pallasi from recent bottom deposits have better saved surface and color pattern compared to those from Neoeuxinian sediments (IEina, 1966; Golikov & Starobogatov, 1972). 240 Igor P. Bondarev In our samples there is one T. pallasi subfossil specimen from a depth of 158 m from the recent bottom sediment of near-Bosporus region (Fig. 1). Maintenance of colouring of the shell testifies its relatively recent fossilization. Available data allow to suppose that the shells of the freshwater species complex discovered on Black Sea shelf belong to recent living species inhabiting within the limits of the freshwaters biotopes. In 2010, during the Black Sea international ex- pedition of R/V "Maria S. Merian" (Leibniz Institute for Baltic Sea Research, Germany) in a near-Bospo- rus region on a depth of 250 m, a live specimen, be- longing to the genus Caspiohydrobia Starobogatov, 1970, was collected (Fig. 2). As at this depth in the Black Sea there are anoxic conditions, it was hypo- thesized that the animal had been rescued only by chance (Sergeeva et al., 2011). However if we admit the existence of specific aerobic freshwater fauna in Figure 1. Subfossil of Theodoxus pallasi , shell height: 5.8 mm, width: 7 mm, Bosporus region, depth 158 m, R/V "Maria S. Merian", 2010. Figure 2. Live-collected Caspiohydrobia sp. (soft parts co- lored with “Bengal rose”), shell height: 3.4 mm, Bosporus region, depth 250 m, R/V "Maria S. Merian", 2010. the depths of the Black Sea, then the location of this mollusk corresponds to the biotope formed by a sub- marine unloading of freshwaters. In 1986, during test dive of submarine inhabited vehicle of the USSR Navy in the Yalta canyon at a depth of 600-640 m. Dr. Gevorkyan (personal comm.) looked at one "oasis" of eukaryotic suppo- sedly aerobic life. The biotope was characterized by unusually clear (for the Black Sea) water because of the absence, in the water column, of the charac- teristic particles of organic suspended debris (known as "marine snow"). On the outcropped rocks macrobenthos forms did remind hydroids. The most notable detail of the biota was the occur- rence of fishes, exceeding 20 cm in size. The most likely hypothesis for the presence of such a biotope and a biocenosis is the possible existence of a po- werful submarine spring of aerobic waters, stably existing since a long time. Notably, generally speaking, strategy of repro- duction [laying eggs attached to the substratum and non pelagic development, (see Chukhchin, 1984; Anistratenko, 1998,] and early ontogenesis peculiarities of these mollusks allow them to exist within the limits of localized biotopes showing pa- rameters contrasting with those of surrounding en- vironments. Another important ecological characteristic for allowing species to inhabit the deep Black Sea is the resistance to hypoxia. Hy- drobiidae comprise mollusks adapted to the lack of oxygen (Chukhchin, 1984), and the specimen of Caspiohydrobia sp. discovered at a -250 m of depth belongs to this family. In 2002, during the international expedition on R/V Meteor (Ger- many) to two stations in the north-western part of the Black Sea at the depths of 1900 m and 2190 m, respectively, it was discovered a meiobenthic crustacean species unknown to science. The same species was found out in 2003 in an expedition on R/V “Yantar” (Russia) in north-eastern part of the Black Sea at a 171 m of depth. The specimen (Cla- docera: Ctenopoda) allowed to describe a new spe- cies, Pseudopenilia bathyalis Sergeeva, 2004 and a new respective genus taxon. Consequently, also the Pseudopenilidae family Korovchinsky & Ser- geeva, 2008 was described. Hydrochemical analyses of near-bottom water in a deep-water place of sampling of Pseudopenilia bathyalis showed the presence of sulfiired hydrogen in an amount of 4-12 ml/1 at a salinity of 22-23%o. Freshwater and brackish “oasis” fauna in the deep Black Sea 241 However, finding out this crustacean species at a depth of 171 m in the hypoxia zone allows to sup- pose that we deal with an aerobic organism adap- ted to oxygen-deficit conditions. Places of submarine water unloading are local phenomena in the Black Sea. Their spatial charac- teristics and stability in time could be substantially differentiated depending on sources regime. For example, water supplement varies seasonally and may lead to a temporary stop of unloading. Characters of water springs related to the arte- sian layers are more stable. In addition, salt and sol- ved gases composition may be different. Spatial structure of freshwater biotope can change as a re- sult of dynamic influences of the surrounding water masses. The innate structure of such biotopes can be non-homogeneous, thus including several biota that differently react to presence/absence of oxygen and of refreshing/salting waters. Hence biotopes and biocenoses of submarine unloading zones may be extremely heterogeneous, various and very vul- nerable ecosystems. CONCLUSIONS The existence of aerobic life in the deepest part of the Black Sea is traditionally contested and even denied (see Vinogradov, 1997). On the contrary, the present paper strongly supports the hypothesis reporting several evidence on the issue, including the occurrence of the endemic Pseudopenilidae fa- mily (Korovchinsky & Sergeeva, 2008) which suggests the evolutionally-long existence of spe- cific fauna in the deep Black Sea. Probably, a fre- shwater relict fauna could have existed not only from Neoeuxinian time (27-10 thousand years ago) but also earlier. In fact, the existence of submarine freshwaters springs is independent from the change of sea sa- linity levels and quantitative and quality composi- tion of this fauna may have undergone transformation during time, depending also on the changes of sea salinity. Oceanological and hydro- geological researches (Shnyukov & Ziborov, 2004; Trotsyuk et af, 1988) reported on desalted waters in near-bottom layers from tide-mark to deep- water cavity bottom of the Black Sea. The general volume of the submarine discharge in the Black Sea is only approximately estimated. However, di- scharge volumes calculated for single areas show that this volume is ecologically significant. For example, only for the Crimean coast from Bala- klava to Simheiz (less than 50 km), karst submarine springs were appraised as about 700 thousand m 3 /day (Shnyukov & Ziborov, 2004). The same au- thors assessed the volume of subriver-bed flow as 1/3 of the volume of river flow. At the moment, water unloading in submarine canyons, although being demonstrated as a fact (Shnyukov & Ziborov, 2004), has not been calculated yet (not even preli- minarily). The process of the submarine unloading can and must have ecological consequences. This paper aims at not only discussing and con- tributing to a deeper knowledge of the refinement of the Black Sea biodiversity and of the recent state of the ecosystem, but also encourages the re- vision (by several colleagues) of some other aspects of the natural history of this ecosystem. ACKNOWLEDGEMENTS The Author is thankful to the Dr Gevorkyan V.Kh. (Institute of Geological Sciences NASU, Kyiv) for his personal communications and to Dr. Sergeeva N.G. (IBSS, NASU, Sevastopol) for ha- ving allowed the examination of macrobenthos samples collected in the expedition of R/V "Maria S. Merian"(Leibniz Institute for Baltic Sea Rese- arch, Germany) in 2010. REFERENCES Anistratenko V.V., 1998. Handbook for identification of Pectinibranch Gastropods of the Ukrainian fauna. Vestnik zoologii, Part 1 . Marine and brackishwater. Supplement 8: 3-65, Part 2. Freshwater and land. Suppl.8: 67-124. Bondarev IP., 2008. Landscape features of the northern Black Sea continental slope as paleoceanological in- dicators. In: Yanko-Hombach V. and Gilbert A., eds., Extended Abstracts of the Fourth Plenary Meeting of Project IGSP 521: Black Sea - Mediterranean corridor during the last 30ky: Sea level change and human adaptation. - Bucharest, Romania October 4- 16, 2008, pp. 27-28. Bondarev I.P, 2009. Submarine Landscape of the North Black Sea continental shelf-slope transitional zone. In: Yanko-Hombach V. and Gilbert A., eds., Extended Abstracts of the Fifth Plenary Meeting and 242 Igor P. Bondarev Field Trip of IGCP 521 - INQUA 501 “Caspian-Black Sea - Mediterranean corridor during the last 30 ky: Sea level change and human adaptive strategies” (2005- 2009). Istanbul-Izmir-Canakkale, Turkey, pp. 34-35. Chukhchin V.D., 1984. Ecology of gastropod mollusks of the Black Sea. Naukova dumka, Kyiv, 1 76 pp. Golikov A N. & Starobogatov Ya.I., 1972. Classis gastro- pod mollusks - Gastropoda Cuvier, 1797. In: Vodya- nitsky V.A., Ed., Guide on the Black Sea and the Sea of Azove fauna. V.3, Free living invertebrates. Nau- kova dumka, Kyiv, 65-166. (In Russian). Goodwin D.R., 2006. The Use of Molluscs as Biological Indicators in Assessing Climate and Environmental Change. Visaya, March, 2006. www.conchology.be. Il’ina L.B., 1966. Natural History of Black Sea Gastro- poda. Nauka, Moscow, 228 pp. Ionescu D., Siebert C., Polerecky L., Munwes Y.Y., Lott C, Hausler S., Bizic-Ionescu M., Quast C., Peplies J., Glockner F.O., Ramette A., Rodiger T., Dittmar T., Oren A., Geyer S., Stark H.J., Sauter M., Licha T., Laronne J.B. & de Beer D., 2012. Microbial and che- mical characterization of underwater fresh water springs in the Dead Sea. PLoS One, 7 (6): e38319. doi: 10. 137 1/journal. pone. 00383 19 Kantor Yu. I. & Sysoev A.V., 2006. Marine and brackish water Gastropoda of Russia and adjacent countries: an illustrated catalogue. KMK Scientific Press Ltd., Moscow. 371 pp., 140 plates. Korovchinsky N. & Sergeeva N.G., 2008. A new family of the order Ctenopoda (Crustacea: Cladocera) from the depth of the Black Sea. Zootaxa, 1795: 57-66. Shcherbakov F., Kuprin P, Potapova L., Polyakov A. S., Zabelina E.K. & Sorokin V.M., 1978. Sedimentation on the Continental Shelf of the Black Sea. Nauka, Moscow, 211 pp. Sergeeva N.G., Zaika V.E. & Bondarev I.P, 2011. The lowest zoobenthos border in the Black Sea Near-Bo- sporus region. Marine Ecology Journal, 10: 65-72. Sorokin Yu. I., 1982. The Black Sea (Nature, resources). Nauka, Moscow, 217 pp. Trotsyuk V.Ya., Berlin Yu. M. & Bolshakov A. M., 1988. Oxygen in near-bottom water of the Black Sea. Ocea- nology, 302: 961- 964. Shnyukov E.F.& Ziborov A.P, 2004. Mineral resources of the Black Sea. Kiev, 277 pp. Vinogradov M.E., 1997. Influence of sulfured hydrogen on the live distribution in the Black Sea. Obschaya Biologia Journal, 58: 43-60. Zaika V.E., 2008. Is their animal life at the Black Sea great depth? Marine Ecology Journal, 7: 5-11. Zaitsev Yu.P, 2006. An introduction on the Black Sea Ecology. Odessa: “Even”, 222 pp. Biodiversity Journal, 2012 , 3 ( 3 ): 243-246 Description of two new species of Carabus Linnaeus, 1 758 from China (Coleoptera Carabidae) Ivan Rapuzzi Via Cialla n. 47 - 33040 Prepotto (UD), Italy; email: info@ronchidicialla.it ABSTRACT In the present paper two new species of Carabus Linnaeus, 1 758, subgenus Apotomopterus Hope, 1838, are described and figured: Carabus (Apotomopterus) francottei n. sp. and Ca- rabus (Apotomopterus) eccoptopteroides n. sp., comparative notes with the related taxa are provided. KEY WORDS Coleoptera; Carabidae; Carabus ; Apotomopterus , new species; China. Received 28.08.2012; accepted 14.09.2012; printed 30.09.2012 INTRODUCTION In terms of number of species the subgenus Apo- tomopterus Hope, 1838 is the largest subgenus of the genus Carabus L., 1758. The subgenus is wide- spread in Southeast China and adjacent Countries, in many places several species are sympatric (Deuve, 1997a, 1997b; Kleinfeld, 2009). In the last decades thanks to the investigation of new or less known areas a large number of new species and subspecies was described (Kraatz, 1894; Boileau, 1896; Breuning, 1931, 1932-1936, 1950; Hauser, 1932; Deuve, 1991, 1995, 1997a, 1997b, 2001, 2002; Brezina, 2003; Deuve, 2004; Lassalle, 2006; Deuve & Li, 2009; Kleinfeld, 2009; Deuve, 2012a, 2012b). The examination of some Apotomopterus speci- mens from North Guangdong and South Sichuan provinces in Southern China allowed to identify two new species described herein. Carabus (Apotomopterus) francottei n. sp. Examined material. Holotypus male (Fig. 1), China, North Guangdong province, Mts. Nanling Shan, VI. 2009. The holotypus is deposited in the author’s collection. Paratypus: 1 male, same data as holotypus. The paratypus is deposited in the au- thor’s collection. Description of Holotypus male. Length inclu- ding mandibles: 33 mm, elytral width: 10.7 mm. Color black with very few metallic luster brownish- copper on dorsum and pronotum, mat. Head moderately thickened. Frontal impres- sions deep and rugose, exceeding anterior margin of eyes; vertex slightly convex, surface of the ver- tex slightly punctured and rugulose; short neck. Surface faintly punctulate and rugulose. Mandi- bles moderately long, strong, and regularly cur- ved. Palpi long, penultimate segment of labial palp multisetose (3-4). Eyes very convex and pro- minent. Antennae very long extending of 5 V 2 an- tennomeres pronotal base and extending the second half of elytra. Pronotum very large and si- nuate, transverse, about 1.34 times as broad as long, slightly convex; sides of pronotum narrow margined, slightly bent upwards; hind angles sli- ghtly protruding behind its base; surface of pro- notum uniformly punctured; basal depressions large, roughly punctured. Elytra elongate, sub-parallel sides, slightly emarginated at apex, moderately convex, maximum width behind middle; shoulders rather large and 244 Ivan Rapuzzi rounded; sculpture triploid homodyname, intervals uniformly convex, only the primary interrupted in the row in quite long links by small foveae, not punctured striae. Male aedeagus (Figs. 2, 3) small, regular curved; apical half slightly thickened, a little sinuate on the ventral side; apex a little nar- rowed spatulate. Variability. No variability of paratypus. Body length 3 1 mm. Etimology. The new species is cordially dedi- cated to Dr. Auguste Francotte (Liege, Belgium) na- turalist and specialist of Coleoptera Cerambycidae, my friend from many years. Comparative notes. The new species is closely related with the sympatric C. (Apotomopterus) sau- teri nanlingensis Deuve et Tian, 1999, but easy to be distinguished by the following characteristics (Deuve & Tian, 1999): larger and more sinuate pro- notum; homodyname triploid sculpture of elytra with very regular intervals; larger and flat elytra; different color of elytra and pronotum; larger and curved median lobe of aedeagus. Carabus (Apotomopterus) eccoptopteroides n. sp. Examined material. Holotypus male (Fig. 4), China, South Sichuan province, Pu-Ge County, Lianxiang, Kakaliangzi, 1/11. VI. 2012. The holoty- pus is deposited in the author’s collection. Paratypi: 18 females, same data as holotypus. The paratypi are deposited in the author’s collection. Description of Holotype male. Length inclu- ding mandibles: 36 mm, elytral width: 11 mm. Color black with very faint cupper luster on dorsum and pronotum, mat. Legs, antennae and palpi black. Head of normal shape, neck quite narrow, eyes small and slightly prominent. Flat vertex with a raised polish trilobate plate, the rest of the surface of the vertex rugulose. Mandibles short and stout. Palpi thin and very long, labial palp bi or three se- tose. Antennae long and thin, extending with 5 A antennomeres beyond pronotal base and extending the apical half of elytra. Pronotum rounded, sli- ghtly transverse, about 1.18 times as broad as long; disc of pronotum slightly convex; sides of pronotum very narrow margined, not bent up- wards; hind angles very short, slightly protruding behind its base; surface of pronotum thin punctu- red; basal depressions small and not deep. Elytra very elongate, narrow, oval, moderately convex, maximum width just behind the middle; shoul- ders very narrow and rounded; sculpture triploid homodyname, intervals uniformly convex, with the primaries cut into quite long segments by small fovea; not punctured striae. Male aedeagus (Figs. 5, 6) very elongate, basal and median por- tion rectilinear and sub-cylindrical, apical portion strongly curved and very elongated; apex large and rounded. Variability. Only females: the length of the body ranges from 36 mm to 43 mm. Pronotum more or less transverse: from 1.16 to 1.26 times as broad as long. Elytra very long, very narrow, rather con- vex, with very strong preapical emargination, the posterior angles are acuminate and very protruding, forming a sharp tooth. The apical half of elytra is marginated and bent upwards. Etimology. The given name wants to indicate the morphological vicinity of the present new species with C. (Apotomopterus) eccoptopterus Kraatz, 1894. Comparative notes. C. (Apotomopterus) ec- coptopteroides n. sp. is related with several Apo- tomopterus species: C. (A.) aeneocupreus Hauser, 1932; C. (A.) benardi^QxQxxmng, 1931; C. (A.) eccoptopterus Kraatz, 1894; C. (A.) keithi Deuve, 1995; C. (A.) piriformis Deuve, 1997 but easy separable by the following characters (Kra- atz, 1894; Breuning, 1931; Hauser, 1932; Deuve, 1995, 1997b): eccoptopterus : the new species is similar by the very elongate shape of body but differs by the stron- gly punctured pronotum, stronger preapical emar- gination of females elytra and by the shape of aedeagus strongly curved at the apical portion. keithi'. the new species differs by the larger size, strongly punctured pronotum, stronger preapical emargination of females elytra and by the shape of aedeagus strongly curved at the apical portion. benardi : the new species is very different for the much more elongate body, the more convex elytra with very regular sculpture and the faintly punctured pronotum. piriformis', the new species differs for the more elongate body, the more regular sculpture of elytra, the faintly punctured pronotum and the shape of ae- deagus more strongly curved at the apical portion. 245 Description of two new species of Carabus Linnaeus, 1 758 from China (Coleoptera, Carabidae) 3 6 Figure. 1. Carabus (Apotomopterus) francottei n. sp. holotypus. Figures. 2, 3. idem, male edeagus lateral view (Fig. 2) and frontal view (Fig. 3). Figure. 4. C. (A.) eccoptopteroides n. sp. holotypus. Figures. 5, 6. idem, male edeagus lateral view (Fig. 5) and frontal view (Fig. 6). Figure. 7. C. (A.) eccoptopteroides n. sp. paratypus female. REFERENCES Boileau H., 1896. Description d’un Carabe nouveau. Le Naturaliste, 18: 203-204. Breuning S., 1931. Cinq nouvelles formes de Carabini. Buletin du Museum D’Histoire Naturelle de Paris. (2), III: 620-623. Breuning S., 1932-1936. Monographic der Gattung Ca- rabus L. Bestimmungs-Tabellen der europaischen Coleopteren. Troppau, 1610 pp. Breuning S., 1950. Einige neue Arten und Rassen der Gattungen Carabus und Cychrus aus Ostasien. Ento- mologische Arbeiten aus dem Museum G Frey Tut- zing Bei Muenchen, 1 : 198-201 . Brezina B., 2003. World Catalogue of the Genus Carabus L. Pensoft, Sofia-Moscow 1999: 170 pp. 246 Ivan Rapuzzi Deuve Th., 1991. Descriptions et diagnoses de nouve- aux Coleopteres Carabidae asiatiques. L’Entomolo- giste, 47: 13-27. Deuve Th., 1995. Contribution la connnaissance taxo- nomique des Geners Carabus et Cychrus en Asie. Revue frangaise d’Entomologie (NS), 17: 69-76. Deuve T., 1997a. Catalogue des Carabini et Cychrini de Chine. Memories de la Soc. Ent. France, 1 : 236 pp. Deuve T., 1997b. Nouveaux Carabus L. et Cychrus F. de la Chine, de l’Asie Centrale et de la Turquie d’Asie. Coleopteres, 3: 209-229. Deuve T. & Tian M., 1999. Diagnoses preliminaries de nouveaux Carabus L. de Chine meridionale. Coleopteres, 5: 139-147. Deuve T., 2001. Noveaux Carabus L. et Cychropsis Boileau de Chine, de Birmanie et d’lran. Coleopte- res, 7: 49-61. Deuve T., 2002. Noveaux Carabus L. de la Chine du Sud-Ouest, de l’lran et de la Coree. Coleopteres, 8: 219-231. Deuve T., 2004. Illustrated Catalogue of the Genus Ca- rabus of the World (Coleoptera, Carabidae). Pen- soft. Sofia-Moscow, 461 pp. Deuve T., 2012a. Une nouvelle classification du genre Carabus L., 1758. Liste Blumenthal 2011-2012. As- socation Magellanes - Andresy - France. Deuve T., 2012b. Description d’un nouvel Apotomop- terus du Sichuan (Coleoptera, Carabidae). Coleop- teres, 18: 13-16. Deuve T. & Li J., 2009. Nouveaux Carabus de Chine et de Coree et confirmation de la validite specifique de Carabus (Carabus) cartereti Deuve, 1982 (Cole- optera, Carabidae). Coleopteres, 15: 1-12. Hauser G., 1932. Zwei neue Arten der Untergattung Apotomopterus. Mitteilungen der deuthschen ento- mologischen Gesellschaft, 3: 75-77. Kleinfeld F., 2009. Apotomopterus . Monographische Uberisicht ttder das Subgenus Apotomopterus Hope, 1838 des Genus Carabus Linnee, 1758. Dr. Frank Kleinfeld, Uhlandstrasse 15, 90768 Fiirth, 281 pp. Kraatz G., 1894. Apotomopterus eccoptopterus Krtz n. sp. von China. Deutsche Entomologische Zeit- schrift, 38: 137-139. Lassalle B., 2006. Nouveaux Carabes d’lran, et de la Chine. Lambillionea, 106: 103-106. Biodiversity Journal, 2012, 3 (3): 247-250 Newly reported marine red alga, Neosiphonia savatieri (Ha- riot) M.S. Kim et I.K. Lee 1 999 (Rhodophyta Rhodomelaceae) from Thailand Narongrit Muangmai 1,2 , Sinchai Maneekat 2,3 , Nidsaraporn Petsut 4 & Chatcharee Keawsuralikhit 2,3 * 'School of Biological Sciences, Victoria University of Wellington, Kelburn, Wellington 6140 New Zealand 2 Biodiversity and Aquatic Environmental Research Unit, Center for Advanced Studies for Agriculture and Food, Kasetsart Uni- versity, Phaholyothin Road, Bangkok 10900 Thailand; e-mail: ffischs@ku.ac.th department of Fishery Biology, Faculty of Fisheries, Kasetsart University, Phaholyothin Road, Bangkok 10900 Thailand department of Agricultural Technology, Faculty of Science, Ramkhamhaeng University, Ramkhamhaeng Road, Bangkok 10240 Thailand ■"Corresponding author ABSTRACT Neosiphonia savatieri (Hariot) Myung Sook Kim et In Kyu Lee, 1999 is reported for the first time from Thailand based on specimens collected from the Gulf of Thailand and Andaman sea. We herein describe the vegetative and reproductive morphology of the specimens. Im- portant features for species identification include the thallus configuration, number of peri- central cells, cortication, branching pattern, origin of rhizoids, origin of branches, occurrence of trichoblasts and reproductive characteristics. Our results expand the known geographic di- stribution of this species and confirm its taxonomic features. KEY WORDS Marine red alga; Morphology; Neosiphonia savatieri ; Rhodomelaceae; Thailand. Received 01.09.2012; accepted 15.09.2012; printed 30.09.2012 INTRODUCTION The genus Neosiphonia Myung Sook Kim et In Kyu Lee, 1999 was segregated from Poly siphonia Greville, 1823 based on the generitype, N. flavima- rina M.S. Kim et I.K. Lee, 1999 from Bangpo on the western coast of Korea (Kim & Lee, 1999). Currently, there are 30 assigned species (Guiry & Guiry, 2012) of which 15 species have been recor- ded for South East Asia region (Ho, 1969; Silva et al., 1987; Abbott et al., 2002; Kim et al., 2008). N. savatieri (Hariot) M.S. Kim et I.K. Lee, 1999 was originally described based on collected material from Kanagawa Prefecture, Japan and subsequently it has been reported from Philippine, Korea, Malay- sia, Norfolk Island, Hawaiian Island and Samoan Archipelago (Silva et al., 1987; Abbott, 1999; Mil- lar, 1999; Masuda et al., 2001; Kim, 2005; Skelton & South, 2007; Kim et al., 2008). In Thailand, only generic level of the genus Neosiphonia has been currently reported (Coppe- jans et al., 2010). During the collections under the project of the biodiversity inventory and informa- tion management in biodiversity hotspots, the tuf- ting red alga, Neosiphonia sp. was collected in both Andaman Sea and the Gulf of Thailand. Eventually we identified those specimens as N. savatieri and confirmed the taxonomic features based on morphological and anatomical characteri- stics of vegetative and reproductive plants. MATERIALS AND METHODS Specimens examined were hand-collected du- ring October 2010 at sand dune area around the estuary of Pak Bara, Satun province, Andaman Sea (99°43 , 2 ,, E; 6°51’27”N). 248 N. Muangmai, S. Maneekat, N. Petsut & C. Keawsuralikhit Additional material examined was from Ao Len, Trat Province, Gulf of Thailand (102°32’57”E; 12°4’13”N). Algal samples for morphological in- vestigation were fixed and stored in 5% formalin/seawater or pressed onto herbarium sheets. Voucher specimens were deposited in the herbarium of Laboratory of Applied Research for Aquatic Plant and Plankton, Biodiversity and Aquatic Envi- ronmental Research Unit of Faculty of Fisheries, Kasetsart University, Bangkok, Thailand. Specimens were stained with 1% aniline blue, acidified with IN HC1 and mounted in a 40% Karo®corn syrup on glass microscope slides. Digi- tal images were photographed by microscope digi- tal camera Olympus DP20 (Olympus, Tokyo, Japan) and eventually edited using Photoshop Ele- ments 6 (Adobe, San Jose, CA, USA). Species iden- tification was based on the literatures of N. savatieri from Japan, Korea and Malaysia (Hariot, 1891; Ma- suda et al., 2001; Kim, 2005; Kim et al., 2008). RESULTS AND DISCUSSION Based on diligent observations on morphologi- cal features of gametophytic and tetrasporangial thalli, we conclude that this is the first record of N. savatieri (Figs. 1-13) from Thailand. Our Thai materials are in agreement with de- scriptions of previous studies of N. savatieri from other localities. The description below is based on the Thai materials. Neosiphonia savatieri (Hariot) Myung Sook Kim et In Kyu Lee, 1999 Basionym: Polysiphonia savatieri Hariot, 1891: 226-227. Habitat: Plants from Andaman Sea grew epi- phytically on Gracilaria minuta Lewmanomont, 1994 and G. salicornia (Agardh) Dawson, 1954, which inhabited the lower intertidal on the sand dune nearby the river mouth. The specimens from the Gulf of Thailand were found on the thallus of Hydropuntia changii (Xia et Abbott) Wynne, 1989 at 1 m depth (Fig. 1). The specimens of N. savatieri are erect, grow individually, and reach up to 5-10 mm high. Axes with 4 pericentral cells ecorticate, 90-180 pm in diameter (Figs. 2-3). Specimens pseudodichotomously branched, showing a Y-shaped ramification with an angle of approximately 50 degrees (Fig. 2). Rhizoids aggre- gated in tufts in the lower segment of the axes, cut off from pericentral cells (Fig. 4), they penetrate into the tissue of G. minuta. Trichoblasts or bran- ches are produced on successive segments (Fig. 5). Trichoblasts formed on every segment in a spiral arrangement and deciduous, leaving persistent scar cells (Fig. 6). Trichoblasts are abundant only at the apical part (Fig. 6). Lateral branches are exogenous, replacing trichoblasts (Fig. 5). Exogenous branches develop at various spots on the axis and grow from scar cells of the trichoblasts (Fig. 7). Tetrasporangia arranged in slightly spiral series on the upper branches (Fig. 8). A single tetraspo- rangium is formed in each segment (Fig. 9). Mature tetrasporangia are prominent, 60-80 pm in diameter. Spores are tetrahedrally organized. Spermatangial branches are formed at the first dichotomy of fertile trichoblasts (Fig. 10). Mature spermatangial bran- ches are lanceolate, 180-260 pm long and 60-80 pm wide, and lack sterile apical cells (Fig. 10). An initial of carpogonial branch is formed on the second segment of the fertile trichoblast near the tip of lateral branches (Fig. 11). Procarp consists of a three-celled carpogonial branch and two sterile cells borne on the supporting cell. After fertiliza- tion, the gonimoblast gradually develops from the auxiliary cell (Fig. 12). Mature cystocarps are sphe- rical with a slightly protruding ostiole, 180-220 pm in diameter (Fig. 13). Based on the specimen collected from Kana- gawa, Japan, Hariot (1891) originally assigned N. savatieri to the genus Polysiphonia , which is cha- racterized by its four pericentral cells, unicellular rhizoids cut off by a cross wall from the proximal end of the pericentral cells and spermatangial bran- ches arising as a primary branch of a trichoblast. Kim & Lee (1999) later assigned species with rhi- zoids cut off from pericentral cells, spiral arrange- ment of tetraspores, three-celled carpogonial branches and spermatangial branches on the pri- mary dichotomy of trichoblast filament to a new genus, Neosiphonia. The Thai specimens reported here agree well with the original description of N. savatieri and confirm the important identifying characteristics of this genus. Our Thai specimens were morphologi- cally very similar to N. savatieri described from the Newly reported marine red alga, Neosiphonia savatieri (Rhodophyta Rhodomelaceae) from Thailand 249 Figures 1-13 Neosiphonia savatieri from Thailand. Fig. 1 : plants epiphytic on Hydropimtia changii (arrows), Scale bar, 1 cm. Fig. 2: thalli pseudodichotomously branched with an angle of about 50 degree, Scale bar, 500 pm. Fig. 3: transverse section of the middle portion of a branch with 4 pericentral cells and a axial cell, Scale bar, 50 pm. Fig. 4: rhizoid (arrow) cut off from pericentral cells, Scale bar, 10 pm. Fig. 5: apical portion of a branch showing oblique divisions of apical cells (arrows), Scale bar, 100 pm. Fig. 6: trichoblasts (arrows) and scar cells of deciduous trichoblast arranged in spiral manner in the apical part of a cystocarpic plant, Scale bar, 100 pm. Fig. 7: exogenous branches (arrows) of a tetrasporic plant, Scale bar, 200 pm. Fig. 8: tetrasporic plants showing the spiral arrangement of tetrasporangia, Scale bar, 200 pm. Fig. 9: mature tetrasporangium bearing one per segment, Scale bar, 20 pm. Fig. 10: spermatangial branches arising on a branch of the fertile trichoblasts (arrow), Scale bar, 100 pm. Fig. 1 1 : procarp (arrow) developing on the second segment of a fertile trichoblasts at the tip of branches, Scale bar, 20 pm. Fig. 12: post-fertilization stage, Scale bar, 50 pm. Fig. 13: mature cystocarps, Scale bar, 70 pm. 250 N. Muangmai, S. Maneekat, N. Petsut & C. Keawsuralikhit Philippines, Hawaii, Malaysia, and Japan and Korea; all were relatively small epiphytic algae and have spermatangial branches formed on a branch of trichoblasts, spiraled tetrasporangia and an appro- ximately 50 degree angle in the Y-shaped branching pattern (Silva et al., 1987; Abbott, 1999; Masuda et al., 2001; Kim, 2005; Kim et al., 2008). Furthermore, Thai specimens of A. savatieri ap- pear to be prevalent in river deltas surrounded by mangroves and grow specifically on gracilarioid algae. Our study shows that N. savatieri occur in both marine and brackish waters and that the distri- bution of this species extends to Thailand. Additionally, Thai specimens of N. savatieri showed that young lateral branches are formed by replacing trichoblats, while Kim (2005) and Ma- suda et al. (2001) described branches that are not associated with trichoblasts in N. savatieri from Korea and Malaysia. According to Stuercke & Fre- shwater (2008), the origin of branches has been used as one of the important characteristics to se- parate species in Polysiphonia sensu lato. It is important to take into account whether the relationship of branches and trichoblasts or other specific morphological characters are useful for species delineation of N. savatieri. Additional mo- lecular and morphological analyses of N. savatieri will be needed in order to gain more insights into the species delimitation and differentiation among closely related species, especially in South East Asian region. ACKNOWLEDGEMENTS We thank those who helped us obtaining valua- ble specimens: Sunan Pattarajinda, Teerapong Duangdee and Wirayut Kuisorn. Sincere thanks are also due to Khanjanapaj Lewmanomont and John Bower for providing many useful suggestions and critical comments to the English. This research was partly funded by Office of Natural Resources and Environmental Policy and Training, Bangkok, Thailand. REFERENCES Abbot LA., 1999. Marine red algae of the Hawaiian Is- lands. Bishop Museum press, Hawaii, 465 pp. Abbott I. A., Fisher j. & McDermid K.J., 2002. Newly reported and revised marine algae from the vicinity of Nha Trang, Vietnam. In: Abbott I. A. & Mcder- mid K.J. 2002. Taxonomy of Economic Seaweeds with reference to some Pacific species. Vol. VIII, California Sea Grant College Program, California, 291-321. Coppejans E., Prathep A., Leliaert F., Lewmanomont K. & De Clerck O., 2010. Seaweeds of Mu Ko Tha Lae Tai (SE Thailand): Methodologies and field guide to the dominant species. Biodiversity Research and Training Program, Bangkok, 274 pp. Guiry M.D. & Guiry G.M., 2012. AlgaeBase. World- wide electronic publication, National University of Ireland, Galway, http://www.algaebase.org; searched on 15 March 2012. Hariot R, 1891. Liste des algues marines rapports de Yo- koska (Japon) par M. le Dr Savatier. Memoires de la Societe des Sciences Naturelles et Mathematiques de Cherbourg, 27:211-230. Ho P.H., 1969. Rong bien Vietnam - Marine algae of South Vietnam. Saigon, 558 pp. Kim M.S., 2005. Taxonomy of a poorly documented alga, Neosiphonia savatieri (Rhodomelaceae, Rho- dophyta) from Korea. Nova Hedwigia, 81: 163-175. Kim M.S. & Lee I.K., 1999. Neosiphonia flavimarina gen. et sp. nov. with a taxonomic reassessment of the genus Polysiphonia (Rhodomelaceae, Rhodophyta). Phycological Research, 47: 271-281. Kim M.S., Lim P.E. & Phang S.M., 2008. Taxonomic notes on Malaysian Neosiphonia and Polysiphonia (Rhodomelaceae, Rhodophyta). In: Phang S.M., Lewmanomont K. & Lim P.E., (eds.), Taxonomy of Southeast Asian Seaweeds. Monograph series 2. Uni- versity of Malaya, Kuala Lumpur, 33-44. Masuda M., Abe T., Kawaguchi S. & Phang S.M., 2001. Taxonomic notes on marine algae from Malaysia. VI. Five species of Ceramiales (Rhodophyceae). Bota- nica Marina, 44: 467-477. Millar A. J.K, 1999. Marine benthic algae of Norfolk Is- land, South Pacific. Australian Systematic Botany, 12: 479-547. Silva PC., Menez E.G. &MoeR.L., 1987. Catalog of the benthic marine algae of the Philippines. Smithsonian Contribution to the Marine Sciences, 27: 1-179. Skelton PA. & South G.R., 2007. The benthic marine algae of the Samoan Archipelago, with emphasis on the Apia District. Nova Hedwigia Beihefte, 132: 1-350. Stuercke B. & Freshwater D.W., 2008. Consistency of morphological characters used to delimit Polysipho- nia sensu lato species (Ceramiales, Florideophyceae): analyses of North Carolina, USA specimens. Phyco- logia, 47: 541-559. Biodiversity Journal, 2012, 3 (3): 251-258 Additional data on the genus Muticaria L ndholm, 1 925 with de- scription of a new species (Gastropoda Pulmonata Clausiliidae) Maria Stella Colomba'^Agatino Reitano 2 , Fabio Li berto 3 , Salvatore Giglio 4 , Armando Gregorini 1 & Ignazio Sparacio 5 'Umversita di Urbino, Dept, of Biomolecular Sciences, via Maggetti 22, 61029 Urbino, Italy.; email: mariastella.colomba@uniurb.it; armando. gregorini@uniurb . it 2 Via Gravina n. 7, 95030 Tremestieri Etneo, Italy; e-mail: tinohawk@yahoo.it 3 Strada Provinciale Cefalu-Gibilmanna n° 93, 90015 Cefalu, Italy; email: fabioliberto@alice.it 4 Contrada Settefrati, 90015 Cefalu, Italy; email: hallucigenia@tiscali.it 5 Via E. Notarbartolo 54 int. 13, 90145 Palermo, Italy; e-mail: isparacio@inwind.it ’Corresponding author ABSTRACT Morphological analysis and molecular genetic studies conducted on the genus Muticaria Lindholm, 1925 (Pulmonata Clausiliidae) in Sicily allowed to identify a new species which is described in the present paper. KEY WORDS Clausiliidae; Muticaria ; Sicily, new species. Received 01.09.2012; accepted 18.09.2012; printed 30.09.2012 INTRODUCTION The genus Muticaria Lindholm, 1925 has a di- stribution limited to South-East Sicily and Maltese Islands. Currently it includes three species: Mutica- ria syracusana (Philippi, 1836) and M. neuteboomi Beckmann, 1990 spread in southeastern Sicily and M. macrostoma endemic to the Maltese Islands, where it occurs with four subspecies: M. macro- stoma macrostoma (Cantraine, 1835), M. macro- stoma scalaris (L. Pfeiffer, 1850), M. macrostoma oscitans (Charpentier, 1852) and M. macro stoma mamotica (Gulia, 1861) (Beckmann, 1992; Giusti et al., 1995; Bank, 2012). A preliminary molecular study on 16S rDNA partial sequences (Gregorini et al., 2008) carried out on Sicilian Muticaria revealed the existence of si- gnificant genetic differences between populations attributed either to M. syracusana or M. neute- boomi , including the topotypic ones. Particularly, M. neuteboomi resulted the most widespread species with populations inhabiting inner areas of Iblean plateau (South Eastern Sicily), while M. syracusana resulted confined to a few coa- stal locality of Syracuse province. A second and more detailed molecular study (Colomba et al., 2010) was conducted on topotypic specimens of M. syracusana and M. neuteboomi with a comparative analysis of mitochondrial 16S rDNA and cytochrome oxidase I (COI) gene partial sequences. This study, besides confirming prelimi- nary data (Gregorini et al., 2008), strongly corrobo- rated the validity of the two species. As additional contribute to the research on the genus Muticaria in South Eastern Sicily and within the context of a wider and more detailed work, in the present paper the population of Muticaria from Spi- nagallo (Syracuse) is described as new species on the grounds of morphological and molecular data. ACRONYMS. BC = bursa copulatrix; BCD = diverticulum of bursa copulatrix; CL = co- lumellar lamella; DBC = duct of the bursa copula- trix; DE= distal epiphallus; FO = free oviduct; GA = genital atrium; L = lunella; P = penis; PD = di- verticulum of penis; PE= proximal epiphallus; PL = parietal lamella; PLL = parallel lamella; 252 M.S. Colomba, A. Reitano, F. Liberto, S. Giglio.A. Gregorini & I. Sparacio PP = principal plica; PR = penial retractor muscle; SL = spiral lamella; SP = sutural plica/plicae; V= vagina; VD = vas deferens. The materials used for this study are deposited in the following Museums and private collections: A. Brancato collection, Syracuse, Italy (CB); S. Gi- glio collection, Cefalii, Italy (CG); Laboratory of Cytogenetics and Molecular Biology, University of Urbino, Italy (LCMBU); F. Liberto collection, Ce- falu, Italy (CL); Museo Civico di Storia naturale di Comiso, Italy (MCSNC); Museo Civico di Storia Naturale di Genova “G. Doria”, Italy (MSNG); Museo Naturalistico F. Mina Palumbo, Castel- buono, Italy (MNMP ); A. Reitano collection, Tre- mestieri Etneo, Italy (CR); I. Sparacio collection, Palermo, Italy (CS). Muticaria brancatoi n. sp. Examined material. Holotypus: Italy, Sicily, Siracusa, Cugno Lungo, 37°00 , 25”N 15°10 , 47”E, 110 m, 02.IX.20 12, legit A. Brancato (MSNG 57016). Paratypi: Italy, Sicily, Siracusa, Contrada Spinagallo, 37°00'12”N 15°10'50 ,, E, 120 m, 12.III.2008, 5 specimens, 3 shells (CR); idem, 14 specimens, 30 shells (CR); Siracusa, V.ne Mosca- santi, 37°00'58”N 15 o 09'53”E, 130 m, 28.XIL2010, 2 shells (CR); Siracusa, Cugno Lungo, 37°00'53”N 15°10'11”E, 135 m, 28. XII. 20 10, 2 specimens, 3 shells (CR) Siracusa, Cugno Lungo, 37°00’25”N 15°10 , 41”E, 110 m, 01.IV.2012, 16 shells (CL); Si- racusa, Cugno Lungo, 37°00 , 27”N 15°10 , 48”E, 80 m, 01.IV.20 12, 8 specimens, 86 shells (CL); idem, 2 specimens, 2 shells, legit F. Liberto (MCSNC 4412); idem, 6 shells (CG); Siracusa, Cugno Lungo, 37°00 , 25”N 15°10 , 47”E, 110 m, 02.IX.2012, 8 shells (CB); idem, 20 specimens, 32 shells (CS); idem, 2 specimens, legit I. Sparacio (MNMP ). Description of holotypus. Shell sinistral (Figs. 1, 2, 9), dimensions: height: 12.30 mm; maximum diameter: 4.20 mm, medium, cylindrical-fusiform, decollate, rather robust, light yellowish-grey in co- lour; external surface with minute, raised, close ribs, 69 ribs on penultimate whorl; last whorl with robust, evident and very spaced ribs; spire slowly and regularly growing, with 4 whorls; last whorl ta- pering downwards, with a very elevated and curved cervical keel; suture moderately deep; umbilicus slit-like, aperture about 1/3 of shell height, square. with 5 lamellae (on parietum and columellar side) and lunella and 4-5 plicae (on palatum); on parietum (Figs. 7, 8), starting from suture, there are: long, well developed, non-emerging parallel lamella; short spi- ral lamella, deviating from centre of parietum to adhere to parallel lamella, (upper) parietal lamella tooth-like; non-emerging columellar lamella; subco- lumellar lamella internal; on palatum (Figs. 5, 6) there is an evident, lateral lunella and, starting from suture: two sutural plicae, the principal plica with a robust posterior portion, not fused to lunella apex, and a thin anterior portion, basal plica small, internally fused to base of lunella, very small sulcal lamella; clausilium triangular and slender (Figs. 3, 4), plough-like basal plate, apically pointed; peristome continuous, reflec- ted, distinct from the wall of the last whorl. Genitalia (Figs. 12-14). Genitalia are characteri- zed by: short vagina, very short free oviduct, well de- veloped ovispermiduct and a short copulatory duct ending in branched bursa copulatrix complex; one branch consisting of a short and wide diverticulum of the bursa copulatrix; other branch consisting of very short bursa copulatrix duct and oval and elongated bursa copulatrix. Penial complex consisting of flagel- lum, epiphallus, penial diverticulum and penis; epi- phallus divided by point insertion of robust penial retractor muscle into proximal and distal portions, the latter very short; wide and pointed penial diverticu- lum arising on border between distal epiphallus and penis; penis short (2.5 mm). Internal walls of penis show a long, wide and elevated pleat and two thin and less evident pleats; left ommatophore long and well developed. Variability. Dimensions in decollate specimens (4-5 whorls): height: 11.02-12.30 mm; maximum dia- meter: 4. 16-4.55 mm. The number of ribs on the pe- nultimate whorl of the shell ranges from 57 to 70 (on average, 67); in some specimens the principal plica is absent in its central portion. Etimology. The new species is dedicated to Aldo Brancato (Syracuse, Sicily), dear friend and esteemed naturalist. Biology and Distribution. This species lives on calcareous rock. It is found in cavities and under stone on stony soil. Endemic species to the South-Eastern Sicily, at the time known only for the locality of de- scription. Comparative notes. M. syracusana shows slen- der and conical-fusiform shell with ribs on penulti- Additional data on the genus Muticaria Lindholm, 1 925 with description of new species 253 Figure 1. Shell of Muticaria brancatoi n. sp., Siracusa, Cugno Lungo, h: 11.57 mm - D: 4.33 mm. Figure 2. idem, h: 12.27 mm - D: 4.29 mm. 254 M.S. Colomba, A. Reitano, F. Liberto, S. Giglio.A. Gregorini & I. Sparacio Figures 3-8. Muticaria brancatoi n. sp., Siracusa, Cugno Lungo, clausilium of two specimens (Figs. 3, 4), palatum (Figs. 5, 6) and parietum (Figs. 7, 8). Additional data on the genus Muticaria Lindholm, 1 925 with description of new species 255 Figures 9-11. Cervical keel in Muticaria brancatoi n. sp., Siracusa, Cugno Lungo (Fig. 9), M. syracusana, Siracusa, Tea- tro Romano (Fig. 10) and M. neuteboomi, Ragusa, Cava d’Ispica (Fig. 11). Figures 12-14. Genitalia of M. brancatoi n. sp., Siracusa, Cugno Lungo (Fig. 12) internal structure of penis (Fig. 13) and ommatophore (Fig. 14). 256 M.S. Colomba, A. Reitano, F. Liberto, S. Giglio.A. Gregorini & I. Sparacio 99 NEU1 COI LCO LCO 1490.scf NEU3 COI LCO LCO 1490.scf NEU2 COI LCO LCO 1490.scf NEU5 COI LCO LCO 1490.scf NEU4 COI LCO LCO 1490.scf SYR4 COI LCO LCO 1490. scf SYR1 COI LCO LCO 1490. scf SYR5 COI LCO LCO 1490. scf SYR2 COI LCO LCO 1490. scf SYR3 COI LCO LCO 1490. scf 100 62 M GARGANENSIS COI Albinaria caerulea COI 100 SPI3 BIS COI LCO 1490. scf SPI5 BIS COI LCO 1490. scf SPI6 BIS COI LCO 1490. scf SPI7 BIS COI LCO 1490. scf SPI4 BIS COI LCO LCO1490.scf 0.06 Figure 15. Maximum Likelihood consensus tree inferred from 500 replicates. The tree is drawn to scale, with branch lengths measured in the number of substitutions per site. Bootstrap values, i.e.the percentage of replicate trees in which the associated taxa clustered together in the bootstrap test are shown next to the branches. mate whorl more spaced and less numerous (27- 54); on palatum, the principal plica is very short and fused to upper palatal plica. M. neuteboomi is cha- racterized by fusiform shell, from slender to mode- rately ventricose, with more numerous ribs on penultimate whorl (56-97); on palatum, the principal plica is independent of upper palatal plica. All Muti- caria from Maltese islands are characterized for a principal plica independent of the upper palatal plica. Molecular Analysis. Five Muticaria speci- mens from C.da Spinagallo (Syracuse, SE Sicily), labelled as SPI, were analyzed. Samples were sto- red in 75% Ethanol at -20 °C in test tubes. For each individual, the entire animal was used for total DNA extraction (by Wizard Genomic DNA Purification Kit, Promega). Para-voucher specimens, sensu Groenenberg et al. (2011) i.e. different specimens than the ones used for DNA analysis, but from the same sample or po- pulation, were stored by MSC (University of Ur- bino). COI amplicons (644 bp) were obtained by LCO1490/HCO2198 universal primers (5’- GGTC AAC AAATC ATAAAGATATTGG-3 75 ’-TA- AACTT C AGGGT GACC AAAAAAT C A-3 ’ ) as in Folmer et al. (1994) with a PCR cycle of 95 °C for 5 min; 95 °C for 1 min, 42 °C for 1 min, 72 °C for 1 min (37 cycles); 72 °C for 10 min. Sequencing of the purified PCR products was carried out using auto- mated DNA sequencers at Euro fins MWG Operon (Germany). Finally, sequence chromatograms of each amplified fragment were browsed visually. Se- quences generated in this study were analysed with additional Muticaria syracusana (labelled as SYR) and M. neuteboomi (labelled as NEU) COI sequen- ces, previously deposited by us in GenBank (IDs: HQ696869 and HQ696867, see also Colomba et al., 2010) Medora garganensis (ID: AY425595) and Albinaria caerulea (ID: NC_001761) COI se- quences were employed as outgropus. Sequences were visualized with BioEdit Se- quence Alignment Editor 7 (Hall, 1999), aligned with the ClustalW option included in this software and double checked by eye. Standard measures of nucleotide polymorphism and phylogenetic analy- ses were conducted in MEGA 5.0.3 (Tamura et al., 2011). The best-fit evolution model of nucleotide substitution resulted T92+G (Tamura 3 -parameter + Gamma). The evolutionary history was inferred by using the Maximum Likelihood method; the bo- otstrap consensus tree was inferred from 500 repli- cates; a discrete Gamma distribution was used to model evolutionary rate differences among sites (5 categories; +G, parameter = 2.1279). Codon posi- tions included were lst+2nd+3rd. All positions con- Additional data on the genus Muticaria Lindholm, 1 925 with description of new species 257 Figure 16. Geographic distribution of Muticaria species genetically analysed in SE Sicily: Muticaria brancatoi n. sp. (star), M syracusana (square) and M. neuteboomi (dots). taining gaps and missing data were eliminated. Di- vergences between SPI/S YR and SPI/NEU groups (Dxy), assessed as p distance, were 27.5% and 27%, respectively. Hence, phylogenetic tree (Fig. 15) and genetic distance between groups support the hypothesis that specimens from Spinagallo may be ascribed to a distinct Muticaria species. Remarks. Muticaria brancatoi n. sp. appears well differentiated morphologically from nearby and strictly related species currently known. Mo- lecular data showed a good differentiation for Spinagallo populations already in preliminary studies conducted on 16S rDNA partial sequences (Gregorini et al., 2008), but with this survey, car- ried out by the analysis of cytochrome oxidase subunit I gene, p distance from the other species is considerably greater. Based on available data no evolutionary and/or paleobiogeographic hypothesis is possible, nevertheless, this work highlights a remarkable complexity (Fig. 16) and differentiation within the genus Muticaria in Sicily (Gregorini et al., 2008; Colomba et al., 2010), much greater than supposed until now. ACKNOWLEDGEMENTS We wish to thank Andrea Corso (Syracuse, Italy) REFERENCES Bank R.A., 2010. Fauna Europea: Gastropoda, Clausi- liidae. Fauna Europea version 1.1. http//www.fau- naeur.org. Last access: September 20th 2012. Beckmann K.H., 1992. Catalogue and bibliography of the land- and freshwater molluscs of the Maltese Is- lands, the Pelagi Islands and the Isle of Pantelleria. Heldia, 2: 1-60. 258 M.S. Colomba, A. Reitano, F. Liberto, S. Giglio.A. Gregorini & I. Sparacio Colomba M.S., Gregorini A., Liberto F., Reitano A., Gi- glio S. & Sparacio I., 2010. Molecular analysis of Muticaria syracusana and M. nenteboomi from Sou- theastern Sicily (Gastropoda, Pulmonata, Clausilii- dae). 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