THE RAFFLES BULLETIN of ZOOLOGY 2011 CONTENTS (Continued from Front Cover) New Myrmoteras Ants (Hymenoptera: Formicidae) from the Southeastern Philippines

THE RAFFLES BULLETIN OF ZOOLOGY 2011 CONTENTS (continued from front cover) New Myrmoteras ants (Hymenoptera: Formicidae) from the southeastern Philippines. Herbert Zettel and Daniela The Raffles Bulletin Magdalena Sorger ...... 61

Four new species of Trurlia Jałoszy ski from Sumatra and Borneo (Coleoptera: Staphylinidae: Scydmaeninae). of Zoology P. Jałoszy ski ...... 69

Brevibora cheeya, a new species of cyprinid ﬁ sh from Malay Peninsula and Sumatra. Te-Yu Liao and Heok Hui Tan An International Journal of Southeast Asian Zoology ...... 77

CONSERVATION AND ECOLOGY CONTENTS

Introduction of an Indochinese freshwater crab Sayamia germaini (Crustacea: Brachyura: Gecarcinucidae) to Taiwan: Editorial ...... i morphological and molecular evidence. Hsi-Te Shih, Jhy-Yun Shy, Tohru Naruse, Ho-Tian Hung, Darren C. J. Yeo Peter K. L. Ng and ...... 83 TAXONOMY AND SYSTEMATICS

Distribution of small cetaceans in the nearshore waters of Sarawak, East Malaysia. Gianna Minton, Cindy Peter and The composition, diversity and community dynamics of limnetic zooplankton in a tropical caldera lake (Lake Taal, Andrew Alek Tuen ...... 91 Philippines). Rey Donne S. Papa and Macrina T. Zafaralla ...... 1

Preliminary report on mitochondrial DNA variation in Macaca fascicularis from Singapore. Michael A. Schillaci, Sandy The Acropora humilis group (Scleractinia) of the Snellius Expedition (1929-30). Sancia E.T. van der Meij and Saravia, Benjamin P. Y.-H. Lee and Carney Matheson ...... 101 Rémon R. Visser ...... 9

Natural-licks used by orangutans and conservation of their habitats in bornean tropical production forest. Hisashi New species of Pheretima, Pithemera, and Polypheretima (Clitellata: Megascolecidae) from Kalbaryo, Luzon Island, Matsubayashi, Nobuhiko Wakamatsu, Etsuko Nakazono, Masaaki Takyu, Noreen Majalap, Peter Lagan and Jum Philippines. Yong Hong and Samuel W. James ...... 19 Raﬁ ah Abd. Sukor ...... 109

Large mangrove-dwelling Elysia species in Asia, with descriptions of two new species (Gastropoda: Opistobranchia: Sacoglossa). Cornelis (Kees) Swennen ...... 29

A new subspecies of Amphidromus (Amphidromus) atricallosus from Singapore (Mollusca: Gastropoda: Camaenidae). Siong Kiat Tan, Sow Yan Chan and Somsak Panha ...... 39

Evidence of the earliest freshwater decapod fossil from Southeast Asia (Crustacea: Decapoda: Brachyura). Sebastian Klaus, Madelaine Böhme, Simon Schneider, Jérôme Prieto and Boungnavong Phetsomphou ...... 47

Guide to the aquatic heteroptera of Singapore and Peninsular Malaysia. 6. Mesoveliidae, with description of a new Nereivelia species from Singapore. Yang Chang Man and Dennis H. Murphy ...... 53

(continues on back cover) VOL. 59(1) Published by the Department of Biological Sciences, National University of Singapore Volume 59 28 February 2011 Number 1

Articles appearing in this journal are indexed in: SCIENCE CITATION INDEX®; CURRENT CONTENTS®; AGRICULTURE, BIOLOGY & ENVIRONMENTAL SCIENCE; SCISEARCH®; RESEARCH ALERT®; BIOLOGICAL ABSTRACTS®; CAMBRIDGE SCIENTIFIC ABSTRACTS®; AQUATIC SCIENCES & FISHERIES ABSTRACTS

Layout by Photoplates Pte ltd The Raffl es Bulletin of Zoology | R B Z | rmbr.nus.edu.sg/rbz/ INSTRUCTIONS TO AUTHORS The Raffl es Bulletin of Zoology (RBZ) is an online, peer-reviewed journal which publishes high quality papers in Taxonomy, Systematics, Ecology and Conservation Biology of animals from Southeast Asia and its adjacent areas. Submission of manuscripts. – All manuscripts are to be submitted via email to the Managing Editor, Dr. Tan Heok Hui The Journal aims to build up quality information on the “animal diversity” of Southeast Asia in particular. Papers from ([email protected]). Hard copy manuscript submissions will not be considered. 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ISSN 0217-2445 Detailed instructions to authors are available at http://rmbr.nus.edu.sg/rbz/author.htm THE RAFFLES BULLETIN OF ZOOLOGY 2011 59(1): i Date of Publication: 28 Feb.2011 © National University of Singapore

EDITORIAL

Since my childhood, I have always been fascinated with the natural world; toying with market fi sh and prawns, grasshoppers from the fi eld, tadpoles from the clogged drain, and teasing mimosa leaves. As I grew older, interest in keeping aquarium fi sh became a lifelong affair. On entering NUS, I got into “fi shy business” seriously, under the guidance of Peter Ng and Maurice Kottelat. Besides encountering ichthyological gems like Paedocypris progenetica, I found that Southeast Asia holds key to immense biological diversity. My fi rst introduction to the Raffl es Bulletin of Zoology (RBZ) was a truly eye opening experience in 1992 when I was introduced to my fi rst scientifi c journal as an undergraduate. Now in 2011, I am truly fortunate to step in as RBZ’s sixth managing editor.

Tan Swee Hee, the immediate past managing editor, had put in tremendous efforts to transform RBZ into an online journal since 2005, and this has been a rollercoaster journey. As of 2009, the impact factor of RBZ stands at 1.36, a very respectable index for an international journal dealing primarily with the taxonomy of Southeast Asian fauna. Part of the rising impact factor comes from the fact that biodiversity research and conservation have now become major global concerns.

The current policy prevails: the Rafﬂ es Bulletin of Zoology will continue to accept manuscripts on taxonomy and ecology of fauna from within the geographic scope of Southeast Asia. Papers on fauna from areas immediately adjacent or relevant to Southeast Asia (e.g., southern parts of China such as Yunnan or Hainan) or from drainages that are shared with Southeast Asia (e.g., Salween or Mekong) may, at the discretion of the Editors, be considered although priority will always be given to articles on Southeast Asian fauna. Furthermore, RBZ will not publish short notes or papers consisting solely of redescriptions or new records unless the Editors and/or Referees regard them as having exceptional interest. This is unavoidable as the number of quality papers the Bulletin has received over the years has been overwhelming and tough choices need to be made. However, occasional supplements will continue to be published (depending on the availability of external funding) – 23 have been published to date. Some exciting upcoming supplements include the proceedings of the 2009 hornbill conference, a tome on Southeast Asian inland ﬁ sh systematics, and one on Singapore freshwater aquatic biology.

All published papers in the regular issues of RBZ are available for download by any person with internet access completely free-of-charge. This continues to be in line with our policy of the dissemination of scientiﬁ c information to as wide an audience as possible. A minimal number of hard copies will still be printed in accordance to Article 8 of the 1999 International Code of Zoological Nomenclature to satisfy the criterion of publication for new taxa.

A general meeting was conducted in October 2010 with Editor-in-Chief and the majority of the Associate Editors. Amongst the agenda discussed, methods to support and improve current RBZ’s standing, and a limited appointment term for Editorial Board members. These will be discussed further and more details will be revealed at a later date.

I would like to thank Matthew Lim, who is now pursuing an overseas postdoctoral programme, and Martyn Low, who is now pursuing a graduate degree in Japan; and also extend a warm welcome to two new Associate Editors: Kevin Conway (Texas A & M University) and Zeehan Jaafar (NUS), covering freshwater and marine ﬁ shes respectively. I am also very fortunate to have the RBZ editorial team, consisting of Tran Anh Duc (Hanoi University of Science), Tohru Naruse (University of Ryukyus), Tan Koh Siang (Tropical Marine Science Institute), Chua Keng Soon, Joelle Lai, Li Daiqin, Rudolf Meier, Peter Ng, Greasi Simon, Peter Todd and Darren Yeo (NUS); the RBZ editorial board with new member, Tan Swee Hee; and look forward to their continued support and assistance to carry RBZ into the next phase.

Tan Heok Hui Managing Editor

i THE RAFFLES BULLETIN OF ZOOLOGY 2011

THE RAFFLES BULLETIN OF ZOOLOGY 2011 59(1): 1–7 Date of Publication: 28 Feb.2011 © National University of Singapore

THE COMPOSITION, DIVERSITY AND COMMUNITY DYNAMICS OF LIMNETIC ZOOPLANKTON IN A TROPICAL CALDERA LAKE (LAKE TAAL, PHILIPPINES)

Rey Donne S. Papa Department of Biological Sciences, Graduate School and Research Center for the Natural Sciences University of Santo Tomas, Manila, Philippines Email: [email protected]; reypaps@yahoo,com (Corresponding Author)

Macrina T. Zafaralla Institute of Biological Sciences, University of the Philippines Los Baños, Laguna, Philippines

ABSTRACT. – Limnetic zooplankton serve as a major food source for pelagic ﬁ sh. This paper updates the species composition, diversity and community dynamics of limnetic zooplankton in the two basins of Lake Taal, an active caldera lake ecosystem. Samples obtained from January to December 2008 included 15 rotifer, six cladoceran and three copepod species. Eight species are new records for the lake. The zooplankton community is dominated by copepods, which contributed 64% to total abundance and 84% to total biomass. Diversity values for rotifers and cladocerans were low and similar in both lake basins. The composition, diversity and homogenous spatial distribution of zooplankton in Lake Taal is typical for tropical lakes with a high trophic state which may be a response to the prevailing conditions inﬂ uenced by its location, geological origin and meteorological factors.

KEY WORDS. – Sardinella tawilis, Pseudodiaptomus brehmi, Tropical Limnology, Volcanic lakes, Eutrophication.

INTRODUCTION livelihood until fi sh farming of Oreochromis spp. (tilapia) and Chanos chanos (milkfi sh) in net cages started in 1975 Lake Taal (formerly known as Lake Bombon) (13°55'- which became a lucrative business thereafter (Aypa et al., 14°05'N, 120°55'-121°105'E; Altitude: 2.5 masl) is the 2008). The under-regulated proliferation of aquaculture has third largest Philippine lake which covers an area of 268 unfortunately led to a decrease in water quality (Mamaril km2. It is one of the deepest lakes in the country at 198 m Sr., 2001a). maximum depth and occupies a collapsed caldera formed by volcanic explosions. The lake is fed by water from 37 Three major zooplankton groups dominate freshwater partly seasonal tributaries while the 8.2 km Pansipit River ecosystems – the Copepoda, Cladocera and Rotifera. They connects it to Balayan Bay (Mutia, 2001; Papa et al., 2008; vary in contribution to total abundance and biomass depending Perez et al., 2008; Ramos, 2002). There are two lake basins on trophic status and predation by zooplanktivorous fi sh separated by the largest island in the center known as Taal (Fernando, 2002). Zooplankton composition, distribution Volcano Island which is regarded as the worlds’ lowest and movement are also infl uenced by physical and chemical active volcano (Zlotnicki et al., 2009). Volcanism has led characteristics of the ecosystem (Pinto-Coelho et al., 2005). to the evolution of landlocked endemic species of marine In lakes, distinct differences between the composition and origin such as the freshwater sardine (Sardinella tawilis) community dynamics of littoral and limnetic zooplankton may and the Taal ‘sea’ snake (Hydrophis semperi) (Hargrove, occur which necessitate separate evaluations (Matsumara- 1991). The Philippine government has declared it as a Tundisi & Tundisi, 2005). conservation and biodiversity priority (Ong et al., 2002) and a research hotspot for freshwater fauna. Fish surveys by Herre Studies on Philippine freshwater zooplankton have yet to (1927) and Villadolid (1937) have listed 101 species from reach the sophistication and level of applied limnology of 32 families making it one of the most diverse freshwater North American and European institutions (Mamaril Sr., fi sh populations in the country. Local fi sher folk depended 1986) and appear to be subsumed under fi sheries (Mamaril on capture fi sheries as their primary source of food and Sr., 2001b). The most comprehensive taxonomic papers by

1 Papa and Zafaralla: Zooplankton of Lake Taal

Table 1. Zooplankton species lists from previous limnological surveys in Lake Taal.

Brehm, 1939 Mamaril, 2001 Schiemer et al. (eds.), 2008 Ceriodaphnia rigaudi Brachionus angularis Bosmina fatalis Diaphanosoma sarsi Brachionus calycifl orus Ceriodaphnia cornuta Latinopsis australis Brachionus diversicornis Diaphanosoma spp Moina dubia Brachionus falcatus Moina micrura Simocephalus vetula Brachionus forfi cula Microcyclops varicans Brachionus havanaensis Thermocyclops crassus Kiefer, 1939 Brachionus plicatilis Tropodiaptomus vicinus Thermocyclops hyalinus Brachionus quadridentatus Brachionus urceolaris Woltereck et al., 1941 Hexarthra fennica Brachionus angularis Keratella procurva Brachionus calycifl orus Keratella tropica Brachionus calycifl orus dorcas / dorcas spinosa Keratella valga valga Brachionus plicatilis Lecane luna Brachionus forfi cula Lecane patella Diurella dixon-nuttalli Lecane ungulata Keratella valga valga Testudinella patina Pedalia fennica Trichocerca dixon-nuttali Diaphanosoma sarsi Tripleuchlanis plicata Ceriodaphnia rigaudi Biapertura pseudoverrucosa Moina dubia Bosmina fatalis Simocephalus vetula Bosmina longirostris Thermocyclops hyalinus Ceriodaphnia cornuta Chydorus barroisi Chydorus ventricosus Hauer, 1941 Diaphanosoma sarsi Pedalia intermedia Dunhevedia crassa Pedalia fennica Ilyocryptus spinifer Latinopsis australis Ueno, 1966 Moinodaphnia macleayi Brachionus calycifl orus Simocephalus vetulus Pompholyx complanata Ectocyclops pharelatus Ceriodaphnia cornuta Eucyclops serrulatus Diaphanosoma sarsi Mesocyclops leuckarti Latinopsis australis Microcyclops varicans Moina dubia parva Thermocyclops crassus Mesocyclops cf. thermocyclopoides Tropodiaptomus vicinus

Mamaril Sr. and Fernando (1978) and Mamaril Sr. (1986) MATERIALS AND METHODS listed 125 species belonging to Rotifera (61 spp.), Cladocera (49 spp.) and Copepoda (9 spp.). Studies on Lake Taal Sampling. – Zooplankton were collected monthly from zooplankton were pioneered by the Wallacea Expedition January to December 2008 in six sampling sites representing of the 1930s (Table 1) (Brehm, 1939; Hauer, 1941; Kiefer, the north and south basins of the lake (Fig. 1). Collection 1939b; Woltereck et al., 1941). Follow up studies by Ueno was done by making two replicate vertical tows from 40 (1966) and Mamaril Sr. (2001b) have led to the enumeration m depth using an 80µm mesh size conical plankton net of 37 species in Lake Taal including both littoral and (diameter = 30 cm) (Amarasinghe et al., 2008; Vijverberg limnetic forms. et al., 2008).

We want to update the species composition of limnetic Data Analysis. – Identifi cation was done to species level based zooplankton in Lake Taal and provide additional insights into on descriptions, taxonomic keys and illustrations in Mamaril its community structure. Specifi cally, we want to compare Sr. & Fernando (1978), Mamaril Sr. (1986), Korovchinsky the present zooplankton composition with previous studies (1992), Dussart & Defaye (2001), Fernando (2002) and considering recent developments in zooplankton taxonomy Petersen (2009). Sorting, identifi cation and counting were and systematics. We would also like update aspects of done under dissecting and compound microscopes. Density zooplankton ecology that have not been presented in previous was determined by counting three 1 ml Sedgewick Rafter studies by comparing composition, biomass and diversity sub samples per replicate. Biomass was computed based in the two basins of Lake Taal based on monthly surveys on the methods of Amarasinghe et al. (2008) and Wetzel in the year 2008. & Likens (1991). Biomass was not determined for rare

2 THE RAFFLES BULLETIN OF ZOOLOGY 2011 species. Species diversity for rotifers and cladocerans was Table 2. Limnetic zooplankton species of Lake Taal collected in computed using the Shannon-Wiener Diversity Index (H’) the year 2008 (*denotes new record while #denotes rare species). and compared across sampling months using One-way ANOVA. Copepod diversity was not determined due to the Phylum Rotifera Class Monogononta low number of species. Biomass values in the six sampling Family Brachionidae sites which represent the north and south basins of the lake Anuraeopsis navicula (Rousselet, 1910)*# were log (x+ s / 2) (where s is the lowest biomass value) Lecane bulla (Gosse, 1851)*# transformed and compared across sampling months using Keratella tropica (Apstein, 1907) One-way ANOVA. Statistical analyses were performed using Brachionus forﬁ cula Wierzejski, 1891 PAST Software Version 1.81. Brachionus havanaensis Rousselet, 1911 Brachionus falcatus Zacharias, 1898# Brachionus quadridentatus Hermann, 1783# # RESULTS Brachionus angularis Gosse, 1851 Brachionus diversicornis (Daday, 1883)# Brachionus calyciﬂ orus (Pallas, 1766)# There were 15 rotifer, six cladoceran and three copepod Family Synchaetidae species in Lake Taal. Brachionus was represented by seven Polyarthra vulgaris Carlin, 1943* species followed by Diaphanosoma (3) and Filinia (2). All Family Hexarthridae other genera were represented by one species each. No Hexarthra intermedia (Wiszniewski, 1929) new species or Philippine records were found, although a Family Filinidae comparison with previous studies revealed that six rotifers, Filinia opoliensis Zacharias, 1898*# two cladocerans and one copepod are new records for the Filinia longiseta (Ehrenberg, 1834)*# lake (Table 2). Ten rotifer and one copepod species were Family Trichocercidae Trichocerca capucina (Wierzejski and Zacharias, 1893)*# Phylum Arthropoda Class Branchiopoda Order Cladocera Family Moinidae Moina micrura Kurz, 1874 Family Bosminidae Bosmina fatalis Burkhardt, 1924 Family Sididae Diaphanosoma excisum Sars, 1886* Diaphanosoma sarsi Richard, 1895 Diaphanosoma tropicum Korovchinsky, 1998 Family Daphniidae Ceriodaphnia cornuta Sars, 1885 Class Maxillopoda Fig. 1. Map of Lake Taal with the six sampling sites (NB – North Order Calanoida Basin, SB – South Basin). The insert shows the location of Lake Family Diaptomidae Taal and the other lakes mentioned in the text (P – Lake Paoay, Tropodiaptomus vicinus (Kiefer, 1933) Lb – Lake Laguna de Bay, N – Lake Naujan and Ln – Lake Family Pseudodiaptomidae Lanao). Pseudodiaptomus brehmi Kiefer, 1939*# Order Cyclopoida Family Cyclopidae Thermocyclops crassus (Fischer, 1853)

considered rare since they had low densities and only occurred for a few months.

The three major zooplankton taxa were present throughout the year. There were between 15 and 20 species co-existing each month. The copepods comprised 64% of the average zooplankton abundance and 84% of total biomass which included copepodites and nauplii (Fig. 2). The months of January and March had the highest copepod abundances. January also had the highest zooplankton abundance which may be attributed to the copepod nauplii. Cyclopoid copepods outnumbered calanoid copepods for ten months. The rotifers Fig. 2. Average abundance (Ind. / l) and number of species of were most abundant in June. Eleven of the13 common Copepoda, Cladocera and Rotifera from the upper 40 m in Lake zooplankton species had similar over-all biomass in both Taal in the year 2008. The Copepoda were grouped into nauplii, Calanoida and Cyclopoida. basins; however, some species were noticeably more abundant

3 Papa and Zafaralla: Zooplankton of Lake Taal

Table 3. One-way ANOVA test statistic (F) and probability (P) the fi rst three months of the year which became more similar for zooplankton biomass. Signifi cance was accepted if P < 0.02; with the biomass values in the south basin as the months degrees of freedom (d.f.) = 5. progressed. Tropodiaptomus vicinus had the highest recorded biomass in a single month (January). Except for Brachionus Taxa F P havanaensis, the other common species were present in C. cornuta 1.383 NS almost all months. Rotifer diversity (15 spp.) ranged from B. fatalis 0.647 NS 0.77 to 1.58 while cladoceran diversity (6 spp.) was between M. micrura 3.055 NS 0.78 and 1.62 (Fig. 4). Lowest and highest diversity was D. excisum 2.218 NS D. tropicum 0.153 NS observed in February and April, respectively. No differences D. sarsi 0.102 NS in H’ was found between the north and south basins for both T. vicinus Male 1.822 NS rotifers (F = 2.865; 1 d.f.; P > 0.02) and cladocerans (F = T. vicinus Female 1.025 NS 1.543; 1 d.f.; P > 0.02). T. crassus Male 0.679 NS T. crassus Female 0.608 NS K. tropica 2.913 P < 0.02 DISCUSSION H. intermedia 1.175 NS B. havanaensis 0.300 NS Limnetic and littoral zooplankton have to be treated separately B. forfi cula 0.258 NS P. vulgaris 6.232 P < 0.02 when assessing species richness due to differences in ecology and sampling methods used for these two areas (Matsumara- Tundisi & Tundisi, 2005). The 37 species previously listed for in the north basin for the fi rst three months (cladocerans and Lake Taal included both littoral and limnetic forms (Mamaril copepods) or for most of the year (rotifers) (Fig. 3). Keratella Sr., 2001b). We focused on the community structure and tropica and Polyarthra vulgaris had higher biomass in the dynamics of Lake Taal’s limnetic zooplankton because of north basin for the entire year based on the results of One- their importance in the diet of pelagic fi sh species such as way ANOVA (Table 3). Other rotifers including Hexarthra S. tawilis (Papa et al., 2008). intermedia and Brachionus havanaensis had higher biomass in the north basin for six and two months, respectively but A comparison with previous zooplankton surveys may help did not show signifi cant differences over-all. The cladocerans better understand the current set of data and update the and copepods had higher biomass in the north basin during taxonomy and systematics employed by previous authors

Fig. 3. Mean monthly biomass (μg / l) of common zooplankton species from the north and south basins of Lake Taal for the year 2008.

4 THE RAFFLES BULLETIN OF ZOOLOGY 2011

(Tables 1 & 2). Thermocyclops crassus was the only The FISHSTRAT survey from August 1998 to July 2000 cyclopoid copepod species we encountered. It was the only only included six zooplankton collections with analysis copepod listed in Kiefer (1939b) and Woltereck (1941) later limited to microcrustaceans (Vijverberg et al., 2008). while Mamaril Sr. (2001b) found it together with four other A more frequent (e.g. biweekly) sampling regime is usually cyclopoids. Amarasinghe et al. (2008) then reported T. recommended for more in-depth zooplankton ecology studies crassus together with Microcyclops varicans in the lake. In (Gliwicz, 1986). Unfortunately, financial and logistical the year 2001, a calanoid copepod, Tropodiaptomus vicinus constraints did not permit the conduct of biweekly surveys. was included in a list of Lake Taal zooplankton (Mamaril We observed similar composition in both lake basins for the Sr., 2001b) where it was stated that T. vicinus was included dominant zooplankton species (Fig. 3). Due to its tropical based on Zafaralla (1992). The FISHSTRAT zooplankton location, meteorological factors affect the physical conditions data also included T. vicinus (Schiemer et al., 2008). In this of the lake throughout the year. The watershed has a study, we found T. vicinus together with another calanoid pronounced dry (November to March) and wet season (July species, Pseudodiaptomus brehmi. This rare demersal to September) which can also be classifi ed based on monsoon species has only been reported from marine habitats after seasons: Northeast (December to March), Southeast (March it was fi rst described by Kiefer using specimens from Lake to April), and Southwest (May to September) (Guerrero III, Naujan (Mindoro Is.) (Kiefer, 1939a) which is less than 2002). Vijverberg et al (2008) added an Intermediate Season 100 km south of Lake Taal. This is the fi rst record of P. (October to November) characterized by low rainfall and brehmi in Lake Taal and its second recorded occurrence in low temperatures. These conditions affect lake mixing which a freshwater habitat. infl uences zooplankton distribution. Tropical zooplankton are more prone to wind dispersal due to their smaller size The cladocerans Bosmina fatalis, Ceriodaphnia cornuta, and slower swimming speeds compared to temperate species Moina micrura and Diaphanosoma sarsi and D. tropicum (Blukacz et al., 2009). It was possible that this infl uenced have been recorded in past surveys while D. excisum is a spatial homogeneity in distribution either after lake mixing new record for the lake. Bosmina and Moina were described (cladocerans and copepods) or for the entire year (rotifers). as rare in limnetic areas of Lake Taal compared to nearby The varying response of these two groups may be attributed Lake Laguna de Bay (Mamaril Sr., 2001b). to differences in tolerance for higher trophic states and levels of predation. Rotifers had higher biomass in the north basin Nine of the 15 rotifer species we found have been earlier due to their tolerance for more eutrophic environments; reported by Woltereck et al. (1941), Hauer (1941), Ueno excessive feeding of cage-cultured fi sh increase nutrient (1966) and Mamaril Sr. (2001b). The remaining six species levels in the north basin. This also serves as the origin of - A. navicula, L. bulla, P. vulgaris, F. opoliensis, F. longiseta excess nutrients that are transported by wind and currents and T. capucina are new records for the lake. We also to other locations in the lake. We observed low diversity found Brachionus diversicornis which Mamaril Sr. (2001b) and a lack of seasonality in occurrence (Fig. 4), which was previously listed as a new record for Lake Taal and the the general observation for zooplankton of tropical lakes Philippines. A comparison with rotifer species richness in (Fernando, 2002). The highest H’ values were 1.58 (rotifers) other Philippine lakes showed that Lake Taal had less rotifer and 1.62 (cladocerans) with H’max values for 15 rotifer and 6 species than Lake Laguna de Bay (19 spp.) (Mamaril Sr., cladoceran species at 2.7 and 1.8 respectively. Cladocerans 2001b) but have more than Lake Paoay (11 spp.) (Aquino and rotifers had low diversity due to the dominance of C. et al., 2008) and Lake Lanao (seven spp.) (Lewis Jr., 1979). cornuta and B. fatalis among cladocerans and K. tropica, B. Other Southeast Asian lakes have similar numbers of rotifer forfi cula and H. intermedia among rotifers. Low diversity species (Fernando, 1980). may be related to lake trophic status. Lake Taal is now classifi ed as meso- to eutrophic (Perez et al., 2008). Eutrophic There have been no routine monthly zooplankton surveys lakes are known to support fewer zooplankton species with in Lake Taal covering an entire year prior to this study. higher abundances (Matsumara-Tundisi & Tundisi, 2005) where cyclopoid copepods and cladocerans are dominant over calanoid copepods (Pinto-Coelho et al., 2005). Our data shows that the copepods contributed the highest abundance and biomass of the three major groups almost every month similar to results from Aquino & Nielsen (1983), Perez et al. (2008) and Zafaralla (1992). The cyclopoids and cladocerans were also more abundant compared to the calanoid copepods. S. tawilis prefer calanoid copepods over other zooplankton which may have also infl uenced zooplankton composition and abundance. Lowest calanoid densities were recorded during months when S. tawilis were previously found to consume high quantities of calanoid copepods (Papa et al., 2008). Eutrophication in Lake Taal may be attributed to natural and man-made nutrient inputs from volcanic sediments, Fig. 4. Monthly variations in Shannon-Wiener Diversity (H’) Index values of rotifers and cladocerans in the north and south basins increased mixing layer due to thermal vents, wash-offs from of Lake Taal. the watershed and feeding of caged fi sh (Amarasinghe et

5 Papa and Zafaralla: Zooplankton of Lake Taal al., 2008; Vista et al., 2006; White & San Diego-McGlone, Fernando, C.H., 2002. A Guide to Tropical Freshwater Zooplankton 2008). Our data shows that based on analysis of monthly -Identifi cation, Ecology and Impact on Fisheries. Backhuys samples collected for an entire year, the limnetic zooplankton Publishers, Leiden, the Netherlands. 291 pp. of Lake Taal is typical of tropical eutrophic lakes. This is Gliwicz, Z.M., 1986. A lunar cycle in zooplankton. Ecology, 67: exemplifi ed by high copepod abundance, the dominance of 883–897. cyclopoid copepods and cladocerans over calanoid copepods, Guerrero III, R.D.,2002. The Fisheries of Lake Taal and its the higher biomass of rotifers in the north basin as well Management for Sustainability. FISHSTRAT Users Meeting. as having low species diversity while being present all Bangkok, Thailand, p. 36. throughout the year. Hargrove, T.B., 1991. The mysteries of Taal. A Philippine volcano and lake, her sea life and lost towns. Bookmark, Manila. 244 pp. ACKNOWLEDGEMENTS Hauer, J., 1941. Rotatorien aus dem Zwischengebiet Wallacea Internationale Revue der Gesamten Hydrobiologie und We would like to thank Reiner Eckmann and Augustus C. Hydrogeographie, 41: 177–203. Mamaril Sr. for reviewing the manuscript, T. Chad Walter Herre, A.W., 1927. Four new fi shes from Lake Taal (Bombon). for verifying our identifi cation of P. brehmi and comments Philippine Journal of Science, 34: 273–279. on the manuscript. The German Academic Exchange Service Kiefer, F., 1939a. Bemerkungen zur Pseudodiaptomidenausbeute (DAAD), Tonolli Fund Postgraduate Fellowship of the der Wallacea-Expedition. Internationale Revue der gesamten International Society of Limnology (SIL), Commission on Hydrobiologie und Hydrographie, 38: 75–98. Higher Education of the Republic of the Philippines (CHED) Kiefer, F., 1939b. Die yon der Wallacea-Expedition gesarnmelten and the University of Santo Tomas (UST) provided funds Arten der Gattung Thermocyclops Kiefer Internationale to the fi rst author for this research. Revue der Gesamten Hydrobiologie und Hydrogeographie, 39: 54–74. Korovchinsky, N.M., 1992. Sididae and Holopediidae. SPB LITERATURE CITED Academic Publishing, 86 pp. Lewis Jr., W.M., 1979. Zooplankton Community Analysis: Studies on Amarasinghe, P.B., M.G. Ariyarantne, T. Chitapalapong & J. a Tropical Ecosystem Springer-Verlag New York. 163 pp. Vijverberg, 2008. Production, biomass and productivity of copepods and cladocerans in tropical Asian water bodies and Mamaril Sr., A.C., 1986. Zooplankton In: (ed.), Guide to Philippine the carrying capacity for zooplanktivorous fi sh. In: Schiemer, Flora and Fauna. NRMC and UP Diliman, Quezon City. Pp. F., D. Simon, U.S. Amarasinghe & J. Moreau, (ed.), Aquatic 268. ecosystems and development: comparative Asian perspectives. Mamaril Sr., A.C., 2001a. Translocation of the clupeid Sardinella Biology of inland waters series Backhuys Publishers Leiden, tawilis to another lake in the Philippines: A proposal and The Netherlands. Pp. 173–194. ecological considerations In: Santiago, C.B., M.L. Cuvin- Aquino, L.V. & B.H. Nielsen, 1983. Limnological survey of some Aralar & Z.U. Basiao, (ed.), Conservation and Ecological lakes in Luzon Island, Philippines for pen and cage culture Management of Philippine Lakes in relation to Fisheries and potentials. 65 pp. Aquaculture SEAFDEC; PCAMRD and BFAR, Philippines, Quezon City. Pp. 133–147. Aquino, R., C. Cho, M.A. Cruz, A. Saguiguit & R.D.S. Papa, 2008. Zooplankton composition and diversity in Paoay Lake, Mamaril Sr., A.C., 2001b. Zooplankton diversity in Philippine Lakes Luzon Is., Philippines. Philippine Journal of Science, 137: In: Santiago, C.B., M.L. Cuvin-Aralar & Z.U. Basiao, (ed.), 169–177. Conservation and Ecological Management of Philippine Lakes in relation to Fisheries and Aquaculture. SEAFDEC; PCAMRD Aypa, S.M., S. Ingthamjitr, U.S. Amarasinghe, D. Simon & C. de and BFAR, Philippines, Quezon City. Pp. 81–93 Jesus, 2008. Fish cage culture in Asian lakes and reservoirs: Potentials and constraints In: Schiemer, F., D. Simon, U.S. Mamaril Sr., A.C. & C.H. Fernando, 1978. Freshwater Zooplankton Amarasinghe & J. Moreau, (ed.), Aquatic ecosystems and of the Philippines: Rotifera, Cladocera and Copepoda Natural development: comparative Asian perspectives. Biology of inland and Applied Science Bulletin, 30: 109–221. waters series. Backhuys Publishers, Leiden, The Netherlands. Matsumara-Tundisi, T. & J.G. Tundisi, 2005. Plankton richness Pp. 305–338. in a eutrophic reservoir (Barra Bonita Reservoir, SP, Brazil). Blukacz, E.A., B.J. Shutter & W.G. Sprules, 2009. Towards Hydrobiologia, 542: 367–378. understanding the relationship between wind conditions and Mutia, M.T.M., 2001. Assessment of local government’s plankton patchiness. Limnology and Oceanography, 54: implementation of open access policy in Taal Lake, Philippines: 1530–1540. Effects on lake conservation and management In: Santiago, Brehm, V., 1939. Die Cladoceren der Wallacea-Expedition. C.B., M.L. Cuvin-Aralar & Z.U. Basiao, (ed.), Conservation Internationale Revue der Gesamten Hydrobiologie und and Ecological Management of Philippine Lakes in Relation to Hydrogeographie, 38: 99–124. Fisheries and Aquaculture. SEAFDEC; PCAMRD and BFAR, Philippines, Quezon City. Pp. 123–132 Dussart, B.H. & D. Defaye, 2001. Introduction to the Copepoda. Backhuys Publishers, Leiden. 344 pp. Ong, P.S., L.E. Afuang & R.G. Rosell-Ambal, 2002. Philippine Biodiversity Conservation Priorities: A Second Iteration of the Fernando, C.H., 1980. The species and size composition of tropical National Biodiversity Strategy and Action Plan. Department freshwater zooplankton, with special reference to the Oriental of Environment and Natural Resources- Protected Areas Internationale Revue der Gesamten Region (South East Asia). and Wildlife Bureau, Conservation International-Philippines, Hydrobiologie und Hydrogeographie, 65 : 411–425. Biodiversity Conservation Program-University of the Philippines

6 THE RAFFLES BULLETIN OF ZOOLOGY 2011

Center for Integrative and Development Studies, and Foundation Vijverberg, J., P.B. Amarasinghe, T. Chitapalapong, R.C. Pagulayan, for Philippine Environment, Quezon City, Philippines. 113 M.G. Ariyarantne, E.R.J. Pamanian, E.I.L. Silva & L.A.J. pp. Nagelkerke, 2008. Structure of microcrustacean zooplankton Papa, R.D.S., R.C. Pagulayan & A.E.J. Pagulayan, 2008. communities in fi ve tropical Asian water bodies. In: Schiemer, Zooplanktivory in the Endemic Freshwater Sardine, Sardinella F., D. Simon, U.S. Amarasinghe & J. Moreau, (ed.), Aquatic tawilis(Herre 1927) of Taal Lake, the Philippines. Zoological ecosystems and development: comparative Asian perspectives. Studies, 47: 535–543. Biology of inland waters series. Backhuys Publishers, Leiden, The Netherlands. Pp. 153–172 Perez, T., E.E. Enriquez, R.D. Guerrero III, D. Simon & F. Schiemer, 2008. Catchment characteristics, hydrology, limnology and Villadolid, D.V., 1937. The fi sheries of Lake Taal, Pansipit River socio-economic features of Lake Taal, Philippines. In: Schiemer, and Balayan Bay, Batangas Province, Luzon Philippine F., D. Simon, U.S. Amarasinghe & J. Moreau, (ed.), Aquatic Journal of Science, 63: 191–225. ecosystems and development: comparative Asian perspectives. Vista, A., P. Norris, F. Lupi & R. Bernsten, 2006. Nutrient loading Biology of inland waters series. Backhuys Publishers Leiden, and effi ciency of tilapia cage culture in Taal Lake, Philippines. The Netherlands. Pp. 63–80. Philippine Agricultural Scientist, 89: 48–57. Petersen, F., 2009. An Illustrated Key to the Philippine Freshwater Wetzel, R.B. & G.E. Likens, 1991. Limnological Analysis Springer- Zooplankton. Including some brackish water species from Verlag New York. 391 pp. Laguna de Bay. With ecological notes.http://www.dafnier. White, P. & L.M. San Diego-McGlone, 2008. Ecosystem-based dk/philippines/keyzooplankton/introduction/an_illustrated_key. approach to aquaculture management. Science Diliman, 20: htm 1–10. Pinto-Coelho, R., B. Pinel-Alloul, G. Méthot & K.E. Havens, 2005. Woltereck, R., W.S. Tressler & D.M. Bunag, 1941. Die Seen und Crustacean zooplankton in lakes and reservoirs of temperate Inseln der “Wallacea”-Zwishenregion und ihre endemische and tropical regions: variation with trophic status. Canadian Tierwelt. Zweiter Teil: Inseln und Seen der Philippinen. Journal of Fisheries and Aquatic Sciences, 62: 348–361. Internationale Revue der Gesamten Hydrobiologie und Ramos, E.G., 2002. Origin and geologic features of Taal Lake, Hydrogeographie, 30: 37–76. Philippines. Aquatic Ecosystem Health & Management, 5: Zafaralla, M.T., 1992. Limnological assessment of Taal Lake 281 155–162. pp. Schiemer, F., D. Simon, U.S. Amarasinghe & J. Moreau, 2008. Zlotnicki, J., Y. Sasai, J.P. Toutain, E.U. Villacorte, A. Bernard, Aquatic ecosystems and development: comparative Asian J.P. Sabit, J.M. Gordon, E.G. Corpuz, M. Harada & J.T. perspectives. Biology of inland waters series Backhuys Punongbayan, 2009. Combined electromagnetic, geochemical Publishers Leiden, The Netherlands. 508 pp. and thermal surveys of Taal volcano (Philippines) during the Ueno, M., 1966. Freshwater Zooplankton of Southeast Asia The period 2005–2006. Bulletin of Volcanology, 71: 29–47. Southeast Asian Studies, 3: 94–109.

7 THE RAFFLES BULLETIN OF ZOOLOGY 2011

THE RAFFLES BULLETIN OF ZOOLOGY 2011 59(1): 9–17 Date of Publication: 28 Feb.2011 © National University of Singapore

THE ACROPORA HUMILIS GROUP (SCLERACTINIA) OF THE SNELLIUS EXPEDITION (1929-30)

Sancia E.T. van der Meij Department of Marine Zoology, Netherlands Centre for Biodiversity Naturalis Darwinweg 2, 2333 CR Leiden, The Netherlands E-mail: [email protected] (corresponding author)

Rémon R. Visser Department of Marine Zoology, Netherlands Centre for Biodiversity Naturalis Darwinweg 2, 2333 CR Leiden, The Netherlands E-mail: [email protected]

ABSTRACT. – During the Snellius expedition (1929-30) in eastern Indonesia and the Philippines, a large collection of Acropora corals was made. This collection is comprised of roughly 700 specimens, many of which remain unidentiﬁ ed. In this paper we discuss species of the Acropora humilis group collected during the expedition and compare the records with the known distribution ranges in Indonesia. New records are found, most notably for Acropora multiacuta, A. retusa and A. fastigata.

KEY WORDS. – Acroporidae; biogeography; Indonesia; new records; Philippines.

INTRODUCTION or animal groups. The scleractinian coral family Faviidae was treated in a series of articles (Wijsman-Best, 1974, 1976, The Dutch naval vessel H.M. Willebrord Snellius left for 1977, 1980), whilst the Fungiidae of the Snellius expedition Indonesia and the Philippines in March 1929, on what would were incorporated in a taxonomic revision (Hoeksema, later become known as the Snellius expedition (Kuenen, 1989). Many specimens of the hydrocoral genus Millepora 1941; Aken, 2005). The expedition lasted 18 months and collected during the expedition were also studied (Razak & the programme encompassed research in oceanography, Hoeksema, 2003), as well as specimens of the scleractinian geology, marine biology and meteorology. As part of the coral genera Heterocyathus and Heteropsammia (Hoeksema marine biology programme under supervision of Dr H. & Best, 1991). All these publications included new records Boschma, hundreds of coral specimens were collected for Indonesia. and eventually deposited in the collection of the former Rijksmuseum van Natuurlijke Historie in Leiden, the present The present paper deals with species of the genus Acropora Netherlands Centre for Biodiversity Naturalis. In 1984-85 a (Scleractinia: Acroporidae) collected during the Snellius large Indonesian – Dutch marine expedition was organized, expedition, specifi cally those belonging to the Acropora which was named Snellius II expedition (Best et al., 1989). humilis species group. The small Indonesian collections Furthermore, a large coral collection in Leiden was produced of Dr D. Brongersma and Dr J.H.F. Umbgrove are also through the fi eldwork by Moll (1983) at southwest Sulawesi included. After identifi cation, the data was compared with as a result of the Buginesia Programme. A smaller coral the currently known distribution range of Acropora, resulting collection from Indonesia was obtained though the Siboga in one new species record for Indonesia. expedition (1899-1900) (Aken, 2005), as well as some small collections made by Dr H. Boschma, Dr D. Brongersma, and Dr J.H.F. Umbgrove. These Dutch collections of Acropora MATERIALS AND METHODS corals (approx. 2250 specimens) were not included in the taxonomic revision of Acropora by Wallace (1999). This The Acropora collection of the Snellius expedition [unless paper takes a step toward redressing this issue. otherwise specifi ed, Snellius expedition is referring to the fi rst expedition only] deposited at Naturalis (collection coded as The results of the first Snellius expedition appeared in RMNH.Coel), is comprised of roughly 700 specimens. Many numerous publications dealing with certain families, genera of these corals remain unidentifi ed. Due to the number of

9 Van der Meij & Visser: Acropora humilis group of the Snellius expedition

Table 1. Species of the Acropora humilis group collected at Snellius stations. Locality numbers correspond with Fig. 1. Locality names according to recent spelling (with alternative spelling between in parentheses), numbers indicate the encountered number of specimens in the NCB Naturalis ‘Snellius’ collection. samoensis cf. A. retusa A. digitifera A. A. multiacuta A. humilis A. gemmifera A. monticulosa A. fastigata No. Locality Date at locality A. samoensis 1 Sipangkot (Sipankot) near Sibutu, Sulu Isl. Sept 10-14, 1929 1 3 2 2 Wotap, Tanimbar (Tenimber) Isl. Oct 20-23, 1929 1 1 3 Kera, north of Kupang, near Timor Nov 11-23, 1929 2 4 Sailus Besar (Sailoes Besar), Paternoster Islands Feb 9-10, 1930 1 5 Tanahjampea (Tanah Djampea), N of Flores Feb 21-23, 1930 10 3 1 4 6 Bonetambung near Makassar March 2, 1930 4 4 1 7 Batuata (Batoe Ata) March 6, 1930 2 8 Binongko, Tukangbesi (Toekang Besi) Isl. March 7-10, 1930 1 2 9 Tusa Isl. near Obilatu (Obi Latoe), N of Obi Major April 23-27, 1930 1 1 1 2 10 Karatung (Karaton), Nenusa (Nenoesa) Isl. May 20-21, 1930 1 2 1 11 Akeselaka (Ake Selaka), Kau (Kaoe) Bay May 28, 1930 1 6 12 Beo, Karakelong, Taulaud Isl. June 14-21, 1930 1 2 1 2 13 Reimis (Reinis), Karakelong, Talaud Isl. June 14-21, 1930 1 1 1 1 corals to be identifi ed, and the sheer richness of the genus, specimens identifi ed by Dr J. Verwey, Dr C.C. Wallace, and it was decided to start with one species group, the Acropora Dr H. Moll (Snellius-II expedition, Buginesia Programme). humilis group (sensu Wallace, 1999). For ambiguous specimens a character matrix was designed, including characters such as shape and length of colonies, The A. humilis group is comprised of species with sturdy, branches and branchlets, inner and outer diameter of axial digitate to corymbose colonies. Branches have a large, corallite, shape and distribution of radial corallites, and obvious axial corallite and short thickened tubular radial characteristics of the coenosteum. The character states corallites. First, a pre-selection of specimens likely to were studied with the help of a calliper and a stereo belong to the A. humilis species group was made. These microscope. corals were then identifi ed to species level with the help of various publications (Nemenzo, 1967, 1981, 1986; Veron and Data collection during the Snellius expedition. – The corals Wallace, 1984; Wallace & Wolstenholme, 1998; Wallace, were collected by Dr H. Boschma and Dr P.H. Kuenen, 1999; Veron, 2000). In case of disagreement between the mainly during low tide on the reefs and sometimes through aforementioned authors, the revision by Wallace (1999) was dredging and diving. Thus, the majority of the material comes followed. Furthermore, the ‘registered’ Acropora collection from shallow water (0.5–5 m), while helmet divers of the at Naturalis was used for comparison, especially those Dutch Royal Navy collected material from depths down to 15 m. Occasionally corals were dredged from depths of more than 30 m (Wijsman-Best, 1974). For the majority of the Acropora specimens no depth was recorded. Descriptions of the Snellius localities are described in Boschma (1936) and Wijsman-Best (1974).

RESULTS

A total of 69 Snellius corals were identiﬁ ed as belonging to eight species of the Acropora humilis group: A. humilis (Dana, 1846), A. gemmifera (Brook, 1892), A. monticulosa (Brüggemann, 1879), A. samoensis (Brook, 1891), A. digitifera (Dana, 1846), A. multiacuta Nemenzo, 1967, A. retusa (Dana, 1846) and A. fastigata Nemenzo, 1967. These species were found at 13 Snellius stations (Table 1). Fig. 1. Map showing the localities where Acropora corals were In addition, seven specimens collected by Dr Brongersma sampled during the ﬁ rst Snellius expedition, numbers correspond and Dr Umbgrove in the Togian Islands and Sorido (Biak, with Table 1.

10 THE RAFFLES BULLETIN OF ZOOLOGY 2011

Table 2. Distribution of the Snellius corals belonging to the A. humilis species group compared with the known distribution range in Indonesia, and new records resulting from the present comparison. Little is known about species’ distribution ranges in the Philippines.

Acropora species Specimens Known distribution in Indonesia New records A. humilis 17 Indonesia: widely distributed Indonesia: Wotap, Tanahjampea, Binongko, Akeselaka A. gemmifera 17 Indonesia: widely distributed Indonesia: Wotap, Kera, Tanahjampea Binongko A. monticulosa 1 Indonesia: N Sulawesi, Molucca Sea – A. samoensis 16 Indonesia: widely distributed Indonesia: Bonetambung, Akeselaka, Beo; Philippines: Sipangkot A. cf. samoensis 2 – Indonesia: Tusa Isl., Reimis A. digitifera 3 Indonesia: widely distributed Indonesia: Binongko, Reimis A. multiacuta 4 Indonesia: Togian Isl, Flores Indonesia: Tanahjampea A. retusa 6 none Indonesia: Batuata, Bonetambung, Karatung, Beo A. fastigata 3 Indonesia: Flores; Indonesia: Sailus Besar; Philippines: Calamian Isl. Philippines: Sipangkot

Papua) were identifi ed. The newly obtained records were ‘marine kazerne’ [marine barracks], Feb. 1955, coll. D. Brongersma compared with the known distribution in Indonesia (Moll, (RMNH Coel.39900); Philippines, Sulu Isl., Sipangkot near Sibutu 1983; Best et al., 1989; Wallace & Wolstenholme, 1998; (RMNH Coel.39587). Wallace, 1999, see Table 2). Short description. – Colony digitate to corymbose; branches tapering or slightly terete, 8–24 mm in diameter and up to 95 SYSTEMATIC ACCOUNT mm long; growth determinate. Axial corallite outer diameter 3.0–7.0 mm; radial corallites evenly distributed, with some Most species have already been described extensively rows visible, tubular with round to dimidiate openings and elsewhere, for references see the short-listed synonymies. thickened walls. Short species descriptions are provided. Biogeography. – This common species is found throughout the Indo-West Pacifi c (Wallace, 1999). Wallace & Wolstenholme Family Acroporidae Verrill, 1902 (1998) show an overall distribution in Indonesia, with most Genus Acropora Oken, 1815 records east of Sulawesi. The Snellius I expedition collected specimens of A. humilis at eight sites. The NCB Naturalis The Acropora humilis group coral collection also contains a specimen of A. humilis, collected by Dr Brongersma, from Sorido (Biak, Papua). Colony corymbose or digitate; diameter of branch dominated by axial corallite; radial corallites short thickened tubular with dimidiate openings, evenly sized or in two sizes; coenosteum Acropora gemmifera (Brook, 1892) reticulate to reticulo-costate; sometimes with small amount (Fig. 2c–d) of secondary branching. Madrepora gemmifera – Brook, 1892 p. 457; 1893 p. 142 pl. 21. Acropora gemmifera – Veron and Wallace, 1984: 170; Veron, 1986: Acropora humilis (Dana, 1846) 137; Veron, 2000: 324–325. (Fig. 2a–b) Acropora (Acropora) gemmifera – Wallace and Dai, 1997: 292; Wallace and Wolstenholme, 1998: 212; Wallace, 1999: 120–121. Madrepora humilis – Dana, 1846 p. 483, pl. 31 fi g. 4; pl. 41 fi g. 4. Acropora humilis – Moll, 1983: 29; Veron and Wallace, 1984: Material examined. – Indonesia, Tanimbar Isl., Wotap (RMNH 166; Veron, 1986: 136; Best et al., 1989: 109; Veron, 2000: Coel.39583); Kera, near Timor (RMNH Coel.39577–39578) 318–319. Tanahjampea, N of Flores (RMNH Coel.39580–39582); Acropora (Acropora) humilis – Wallace and Dai, 1997: 292; Wallace Bonetambung near Makassar (RMNH Coel.39571–39574); and Wolstenholme, 1998: 210; Wallace, 1999: 116–118. Tukangbesi Isl., Binongko (RMNH Coel.39569–39570); Nenusa Isl., Karatung (RMNH Coel.39575–39576); Talaud Isl., Beo, Material examined. – Indonesia, Tanimbar Isl., Wotap (RMNH Karakelong (RMNH Coel.39568, 39894); Talaud Isl., Reimis, Coel.39598); Tanahjampea, N of Flores (10 specimens, RMNH Karakelong (RMNH Coel.39579); Papua, Biak, Sorido, W of Coel.39588–39597); Tukangbesi Isl., Binongko (RMNH Kampung, reef at ‘marine kazerne’ [marine barracks], Feb. 1955, Coel.39585); Tusa Isl. near Obilatu, N of Obi Major (RMNH coll. D. Brongersma (RMNH Coel.39901). Coel.39892); Nenusa Isl., Karatung (RMNH Coel.39586); Kau Bay, Akeselaka (RMNH Coel.39893); Talaud Isl., Beo, Karakelong Short description. – Colony digitate to corymbose; branches (RMNH Coel.39584); Papua, Biak, Sorido, W of Kampung, reef at tapering, 10–25 mm in diameter and up to 80 mm long; growth

11 Van der Meij & Visser: Acropora humilis group of the Snellius expedition

Fig. 2a-h. A. humilis: a, complete coral (RMNH Coel.39585), b, branch (RMNH Coel.39591); A. gemmifera: c, complete coral (RMNH Coel.39581), d, branch (RMNH Coel.39578); A. monticulosa: e, complete coral, f, close-up (both RMNH Coel.39599); A. samoensis: g, complete coral (RMNH Coel.39612), h, close-up (RMNH Coel.39622). Scale bars 2 cm (a-c, e, g-h), 1.5 cm for d, f.

12 THE RAFFLES BULLETIN OF ZOOLOGY 2011 determinate. Axial corallite outer diameter 2.0–4.5 mm; radial also sampled by Moll (1983). Two specimens are provisionally corallites in two sizes, often in longitudinal rows, immersed identiﬁ ed as A. cf. samoensis (RMNH Coel.39562-39563), corallites present, tubular with round to dimidiate openings since they show characteristics of both A. humilis and and thickened walls. A. samoensis. If these specimens indeed prove to be A. samoensis, one more locality for this species can be noted Biogeography. – The distribution is very similar to that (Reimis, Karakelong, Talaud Isl.). The species was already of A. humilis, with a wide Indo-West Paciﬁ c range. The known from a large number of reefs in the south Banda Sea Snellius expedition collected specimens at eight sites. The (Wallace & Wolstenholme, 1998). coral collection also contains a specimen of A. gemmifera, collected by Dr Brongersma, from Sorido (Biak, Papua). Acropora digitifera (Dana, 1846) (Fig. 3a–b) Acropora monticulosa (Brüggemann, 1879) (Fig. 2e–f) Madrepora digitifera – Dana, 1846 p. 454. Acropora digitifera – Moll, 1983: 29; Veron and Wallace, 1985: Madrepora monticulosa – Brüggemann, 1879 p. 576. 180; Veron, 1986 : 140; Best et al., 1989: 109; Veron, 2000: Acropora monticulosa – Veron and Wallace, 1985: 174; Veron, 1986: 328–329. 138; Veron, 2000: 320–321. Acropora (Acropora) digitifera – Wallace and Dai, 1997: 293; Wallace Acropora (Acropora) monticulosa –Wallace and Wolstenholme, 1998: and Wolstenholme, 1998: 218; Wallace, 1999: 126–128. 214; Wallace, 1999: 122–123. Material examined. – Indonesia, Tusa Isl. near Obilatu, N of Obi Material examined. – Indonesia, Talaud Isl., Reimis, Karakelong Major (RMNH Coel.39564–39565); Talaud Isl., Reimis, Karakelong (RMNH Coel.39599). (RMNH Coel.39896); Sebesi (RMNH Coel.39902).

Short description. – Colony digitate; branches tapering, Short description. – Colony corymbose; branches terete, conical, and up to 25 mm long. Axial corallite outer diameter 12–16 mm in diameter and up to 30 mm long. Axial corallite 2.7–3.0 mm; radial corallites evenly distributed, tubular with outer diameter 3.2–3.5 mm; radial corallites dimidiate with the round openings. appearance of a ﬂ aring lip.

Biogeography. – The species has an Indo-West Pacific Biogeography. – This species shows a typical Indo-West range. A single Snellius specimen was collected at Reimis, Pacifi c range. Three specimens from Obilatu and Reimis Karakelong, near the Talaud Islands, north of Sulawesi. were identifi ed as A. digitifera, previously known records for this species. The NCB Naturalis collection also contains a specimen of A. digitifera from Sebesi. Acropora samoensis (Brook, 1891) (Fig. 2g-h) Acropora multiacuta Nemenzo, 1967 Madrepora samoensis – Brook, 1891 p. 468; 1893 p. 143 pl. 31 fi g. (Fig. 3c–e) A pl. 6 fi g. C. Acropora samoensis – Moll, 1983: 29; Veron & Wallace, 1985: 178; Acropora multi-acuta Nemenzo, 1967: 133-134, pl. 39 fi gs 1–3. Veron, 1986: 139; Best et al., 1989: 109; Veron, 2000: 323. Acropora multiacuta – Nemenzo, 1981: 156, fi g. 166; Nemenzo, 1986: Acropora (Acropora) samoensis – Wallace and Dai, 1997: 293; Wallace 116, fi g. 117; Veron and Wallace, 1985: 184; Veron, 1986: 141; and Wolstenholme, 1998: 216; Wallace, 1999: 124–125. Best et al., 1989: 109; Veron, 2000: 332. Acropora (Acropora) multiacuta – Wallace and Wolstenholme, 1998: Material examined. – Indonesia, Tanahjampea, N of Flores (RMNH 220; Wallace, 1999: 128–129. Coel.39622); Bonetambung near Makassar (RMNH Coel.39615- 39618); Tusa Isl. near Obilatu, N of Obi Major (RMNH Coel.39895); Material examined. – Indonesia, Tanahjampea, N of Flores (RMNH Kau Bay, Akeselaka (RMNH Coel.39608–39613); Talaud Isl., Beo, Coel.39600–39603); Togian Islands, N Sulawesi, coll. J. H. F. Karakelong (RMNH Coel.39614); Philippines, Sulu Isl., Sipangkot Umbgrove (RMNH Coel.39903–39906). near Sibutu (RMNH Coel.39619–39621). Short description. – Colony caespitose-corymbose to digitate; Short description. – Colony caespitose-corymbose; branches branches slightly tapering, 7–14 mm in diameter and up to terete or slightly tapering, 7–16 mm in diameter and up to 80 65 mm long, incipient branchlets at base of branches. Axial mm long. Axial corallite outer diameter 2.8–5.5 mm; radial corallite very prominent, outer diameter 3.0–7.0 mm; radial corallites not touching, sometimes in rows, tubular with round corallites at base of branch, immersed corallites visible, tubular to oblique openings. to downwards nariform with round openings.

Biogeography. – This species shows an Indo-West Paciﬁ c Biogeography. – Only a few records are known from the range. The Snellius expedition collected specimens at six central Indo-Paciﬁ c (Veron and Wallace, 1984; Wallace and sites from within the aforementioned range. Sipangkot Wolstenholme, 1998; Wallace, 1999; Fenner, 2002, 2003). (Philippines) yields a new species record for A. samoensis, Mactan Isl. (Cebu, Philippines) is the type locality from whereas Bonetambung (southwest Sulawesi) was previously where the species was recorded as rare (Nemenzo, 1986). It

13 Van der Meij & Visser: Acropora humilis group of the Snellius expedition

Fig. 3a-j. A. digitifera: a, complete specimen, b, branch (both RMNH Coel.39564); A. multiacuta: c, complete specimen (RMNH Coel.39601), d, close-up (RMNH Coel.39904), e, branch (RMNH Coel.39603); A. retusa: f, complete specimen (RMNH Coel.39607), g, branch (RMNH Coel.39604); A. fastigata: h, branch (RMNH Coel.39566), i-j, close-ups (both RMNH Coel.39567). Scale bars 2 cm: left scale bar for a, c-d, f, h; right scale bar for b, e, g, i-j.

14 THE RAFFLES BULLETIN OF ZOOLOGY 2011 is recorded as extremely rare by Wallace and Wolstenholme Biogeography. – Wallace (1999) considered the validity of (1998). In the present study, four specimens were identifi ed as this species insuffi ciently known, but provisionally included A. multiacuta. All specimens are very similar in appearance, it in the A. humilis group. This species was originally but represent separate colonies collected at Tanahjampea. described by Nemenzo from Malotamban Isl., Palawan, This new locality for A. multiacuta is in accordance with where it was seen as rare (Nemenzo, 1967, 1986). On the the previously known distribution in Indonesia (Wallace, recent Red List coral assessment of the IUCN the species 1999; Table 2). The coral collections furthermore contain was listed as data defi cient (Carpenter et al., 2008). The four specimens of A. multiacuta, collected by Dr Umbgrove, Snellius specimens are strikingly similar to the A. fastigata from the Togian Islands. pictured and described by Nemenzo (1986), and therefore identifi ed as A. fastigata. The species was listed by Fenner (2001a) from Banggi Isl., Kudat (northern tip of Borneo), Acropora retusa (Dana, 1846) but no picture is available to confi rm this record. Similarly (Fig. 3f–g) the species was reported from the Togian Islands (Fenner, 2001b). Veron (2000) shows pictures of A. fastigata from Madrepora retusa – Dana, 1846 p. 462. the Calamian Isl. (just north of Palawan, north-western Acropora (Acropora) retusa – Wallace, 1999: 130–131. Philippines), and Flores (Indonesia). The new records for Acropora retusa – Veron, 2000: 322. A. fastigata at Sipangkot and the Paternoster Islands are in line with these records. Material examined. – Indonesia, Bonetambung near Makassar (RMNH Coel.39606); Batuata (RMNH Coel.39604–39605); Nenusa Isl., Karatung (RMNH Coel.39607); Talaud Isl., Beo, Karakelong DISCUSSION (RMNH Coel.39897–39898). Ambiguous specimens. – Species within the A. humilis group Short description. – Colony corymbose; branches terete, show a high level of intraspecifi c morphological variability, appearing broader at tip, 7–13 mm in diameter and up to 30 with boundaries between species appearing indistinct mm long, growth determinate. Axial corallite outer diameter (Wallace 1999; Wolstenholme, 2003). Two specimens were 2.4–3.5 mm; radial corallites unevenly distributed and in identifi ed as A. cf. samoensis, since they show characteristics different sizes, tubular with dimidiate openings, sometimes of both A. humilis and A. samoensis. These species differ appearing nariform. in the habitat they occupy, with A. humilis occurring intertidally or subtidally on reef tops and upper slopes, and A. Biogeography. – The species is represented in the Snellius samoensis occurring in slope and lagoonal habitats (Wallace, collection, despite the disjunctive range presented by Wallace 1999; Wolstenholme, 2004). For the Snellius material this (1999): western Indian Ocean and central Pacifi c Ocean but information is not available. According to Wolstenholme not in between. Acropora retusa is found to occur on four (2004) there are intermediate morphs between A. humilis, A. sites in Indonesia, ranging from Nenusa Isl. and Talaud Isl., gemmifera and A. samoensis, suggesting a close relationship north of Sulawesi, to Bonetambung and Batuata in south between these species. One of the common morphs is the Sulawesi. It has also been reported from the Solomon Islands samoensis-humilis morph (‘sam-hum’), which appears to (Veron and Turak, 2006). share closest morphological affi nity with A. samoensis. For this reason it seems justifi ed to leave the two ambiguous specimens identifi ed as A. cf. samoensis. Acropora fastigata Nemenzo, 1967 (Fig. 3h–j) Value of historical material. – The Snellius expedition collected amongst others some presumably rare species, Acropora fastigata Nemenzo, 1967: 134–135, pl. 37 fi g. 3, pl. 38 present only known to occur at very few sites, such as A. fi g. 1. multiacuta, A. retusa and A. fastigata. The data presented Acropora fastigata Nemenzo, 1981: 156–157, fi g. 167; Nemenzo, here is in accordance with the major revision of the genus 1986: 116-117, fi g. 133; Veron, 2000: 331. Acropora (Wallace, 1999), and the new records are additions to the currently known distributions. This study yields no Material examined. – Indonesia, Paternoster Islands, Sailus Besar (RMNH Coel.39899); Philippines, Sulu Isl., Sipangkot near Sibutu new records for A. globiceps for Indonesia, in accordance (RMNH Coel.39566–39567). with the known species distribution, which is limited to the West Pacifi c, east of Australia, including Guam, Micronesia, Short description. – Colony corymbose to digitate; branches Cook Isl., and the Pitcairn Isl. (Wallace 1999). An important markedly tapering to narrow, conical apex, 15–25 mm in new record for Indonesia is A. retusa. As discussed before, diameter and up to 110 mm long, often with incipient branchlets Wallace (1999) comments on the unusual distribution of this at base of branches, appearing like a tall axial corallite with few species. The new record fi lls in the gap between the western tubular radial corallites. Axial corallite very prominent, outer Indian Ocean and central Pacific Ocean. Possibly more diameter 2.1–4.0 mm; radial corallites unevenly distributed and species records will be added in the future. Veron (2000) in different sizes, incipient radial corallites at base of branch, refers to the species’ occurrence of A. retusa in Indonesia immersed corallites visible, tubular to downwards nariform and South Africa, but does not present exact localities or with round openings. information on collected specimens.

15 Van der Meij & Visser: Acropora humilis group of the Snellius expedition

Presumably rare species, or species with a disjunct Edgar, A.J. Edwards, D. Fenner, H.M. Guzmán, B.W. Hoeksema, distribution, might be represented in scientifi c collections G. Hodgson, O. Johan, W.Y. Licuanan, S.R. Livingstone, E.R. without being noticed. This confi rms the value of historical Lovell, J.A. Moore, D.O. Obura, D. Ochavillo, B.A. Polidoro, collection material for biogeographical research, since W.F. Precht, M.C. Quibilan, C. Reboton, Z.T. Richards, A.D. Rogers, J. Sanciangco, A. Sheppard, C. Sheppard, J. Smith, S. museum specimens may show that species display a greater Stuart, E. Turak, J.E.N. Veron, C.[C.] Wallace, E. Weil & E. distribution range than previously assumed. Additionally, Wood, 2008. One-third of reef-building corals face elevated such records provide a baseline, although patchy, for extinction risk from climate change and local impacts. Science, examining possible loss and change in species composition 321: 560–563 (supplementary material). due to environmental degradation (see also Hoeksema & Koh, Dana, J.D., 1846-1849. Zoophytes. U. S. Exploring Expedition. 2009; Meij et al., 2009, 2010; Thacker et al., 2010; Hoeksema 1838–1842, 7: 1–740, pl. 1-61. et al., in press). In particular, specimens collected during large Fenner, D., 2001a. Reef corals of Banggi area reefs, Sabah, expeditions in the late 1800s or early 1900s, such as the US Malaysia. Pulau Banggi Project for Coral Reef Biodiversity. Challenger expedition (1872-77), the Dutch Siboga expedition Greenforce: ReefBase, 13 pp. (1899-1900) or the Swedish Albatross expedition (1947-48), Fenner, D., 2001b. Reef corals of the Togian and Banggai Islands, may prove to be important for comparisons between previous Sulawesi, Indonesia. In: Allen, G.R. &S.A. McKenna (eds) A and currently known species distributions. Hence, scientifi c marine rapid assessment of the Togian and Banggai Islands, collections from such expeditions undertaken long ago can Sulawesi, Indonesia. RAP Bulletin of Biological Assessment provide important information about distribution ranges of 20, Conservation International, Washington DC, pp. 18–25, species and changes therein, possibly offering new insights 63–71. into global environmental change. Fenner, D., 2002. Reef corals of the Raja Ampat Islands, Papua Province, Indonesia. Part II. Comparison of individual survey sites. Appendix 2. Coral species recorded at individual sites in ACKNOWLEDGEMENTS the Raja Ampat Islands. In: McKenna, S.A., G.A. Allen & S. Suryadi (eds) A marine rapid assessment of the Raja Ampat Islands, Papua Province, Indonesia. RAP Bulletin of Biological The authors are grateful to Dr Bert W. Hoeksema, for Assessment 22. Conservation International, Washington DC, overall assistance with the project and improving an earlier pp. 29–36, 104–112. version of the manuscript, and to Mr Jack van Oyen and Mr Fenner, D., 2003. Corals of Milne Bay Province, Papua New Koos van Egmond for technical and collection assistance. Guinea. In: Allen, G.R., J.P. Kinch, S.A. McKenna & P. Seeto Ms Zarinah Waheed (Borneo Marine Research Institute, (eds) A rapid marine biodiversity assessment of Milne Bay Universiti Malaysia Sabah) provided background information Province, Papua New Guinea – Survey II (2000). RAP Bulletin on the presence of Acropora fastigata around Borneo. of Biological Assessment 29. Conservation International, Dr Carden C. Wallace and Mrs Barbara Done were very Washington DC, pp. 20–26. helpful during a collection visit to the Tropical Museum Hoeksema, B.W., 1989. Taxonomy, phylogeny and biogeography of Queensland in Townsville. The manuscript was greatly of mushroom corals (Scleractinia: Fungiidae). Zoologische improved by the comments of Dr Jackie K. Wolstenholme Verhandelingen, Leiden, 254: 1–295. and two anonymous reviewers. Hoeksema, B.W. & M.B. Best, 1991. New observations on scleractinian corals from Indonesia: 2. Sipunculan- associated species belonging to the genera Heterocyathus LITERATURE CITED and Heteropsammia. Zoologische Mededelingen, Leiden, 65: 221–245. Aken, H.M. van, 2005. Dutch oceanographic research in Indonesia Hoeksema, B.W. & E.G.L. Koh, 2009. Depauperation of the in colonial times. Oceanography, 18: 30–41. mushroom coral fauna (Fungiidae) of Singapore (1860s–2006) in changing reef conditions. Raffles Bulletin of Zoology, Best, M.B., B.W. Hoeksema, W. Moka, H. Moll, Suharsono & Supplement 22: 91–101. I.N. Sutarna, 1989. Recent scleractinian coral species collected during the Snellius-II Expedition. Netherlands Journal of Sea Hoeksema, B.W., J. van der Land, S.E.T. van der Meij, L.P. van Research, 23: 107–115 Ofwegen, B.T. Reijnen, R.W.M. van Soest & N.J. de Voogd. Unforeseen importance of historical collections as baselines to Boschma, H., 1936. Biological Data - The Snellius-Expedition in determine biotic change of coral reefs: the Saba Bank case. the eastern part of the Netherlands East-Indies 1929–1930, vol. Marine Ecology, in press. 6. Leiden: Brill, 29 pp. Kuenen, P. H., 1941. Kruistochten over de Indische Diepzeebekkens. Brook, G., 1891. Descriptions of new species of Madrepora in the Den Haag: H. P. Leopolds Uitgeverij N.V., 220 pp. collections of the British Museum. Annals and Magazine of Meij, S.E.T. van der, R.G. Moolenbeek & B.W. Hoeksema, 2009. Natural History, 8: 458–471. Decline of the Jakarta Bay molluscan fauna linked to human Brook, G., 1892. Preliminary descriptions of new species of impact. Marine Pollution Bulletin, 59: 101–107. Madrepora in the collection so f the British Museum. Part II. Meij, S.E.T. van der, Suharsono & B.W. Hoeksema, 2010. Annals and Magazine of Natural History, 10: 451–465. Long-term changes in coral assemblages under natural and Brook, G., 1893. The genus Madrepora. Catalogue of the anthropogenic stress in Jakarta Bay (1920-2005). Marine Madreporarian Corals in the British Museum (Natural History), Pollution Bulletin, 60: 1442–1454. 1: 1-212, pls 1–35. Moll, H., 1983. Zonation and diversity of Scleractinia on reefs off Carpenter K.E., M. Abrar, G. Aeby, R.B. Aronson, S. Banks, A. S. W. Sulawesi, Indonesia. PhD Thesis, Leiden University, Bruckner, A. Chiriboga, Cortés, J.C. Delbeek, L. Devantier, G.J. 107 pp.

16 THE RAFFLES BULLETIN OF ZOOLOGY 2011

Nemenzo, F., 1967. Systematic studies on Philippine shallow-water Wallace, C.C. & J. Wolstenholme, 1998. Revision of the coral scleractinians: VI. Suborder Astrocoeniida (Montipora and genus Acropora (Scleractinia: Astrocoeniina: Acroporidae) in Acropora). Natural and Applied Science Bulletin, 20: 1–223. Indonesia. Zoological Journal of the Linnean Society, 123: Nemenzo, F., 1981. Guide to Philippine fl ora and fauna, vol. 2. 199–384. Philippine corals. NRMC Ministry of Natural Resources, 328 Wijsman-Best, M., 1974. Biological results of the Snellius pp. expedition: XXV. Faviidae collected by the Snellius expedition. Nemenzo, F., 1986. Guide to Philippine fl ora and fauna, vol. 5. I. The genus Favia. Zoologische Mededelingen, Leiden, 48: Corals. NRMC Ministry of Natural Resources, 263 pp. 249–261. Razak, T.B. & B.W. Hoeksema, 2003. The hydrocoral genus Wijsman-Best, M., 1976. Biological results of the Snellius Millepora (Hydrozoa: Capitata: Milleporidae) in Indonesia. expedition: XXVII. Faviidae collected by the Snellius Zoologische Verhandelingen, Leiden, 345: 313–336. expedition. II. The genera Favites, Goniastrea, Platygyra, Oulophyllia, Leptoria, Hydnophora and Caulastrea. Zoologische Thacker, R. W., M.C. Díaz, N.J. de Voogd, R.W.M. van Soest, C.J. Mededelingen, Leiden, 50: 45–63. Freeman, A.S. Mobley, J. LaPietra, K. Cope & S. McKenna, 2010. Preliminary assessment of sponge biodiversity on Saba Wijsman-Best, M., 1977. Indo-pacifi c coral species belonging to the Bank, Netherlands Antilles. PLoS ONE, 5: e9622. subfamily Montastreinae Vaughan & Wells, 1943 (Scleractinea- Coelenterata): Part I. The genera Montastrea and Plesiastrea. Veron, J.E.N., 1986. Corals of Australia and the Indo-Pacifi c. Zoologische Mededelingen, Leiden, 52: 81–97. Angus and Robertson, Sydney, 644pp. Wijsman-Best, M., 1980. Indo-Pacifi c coral species belonging to the Veron, J.E.N., 2000. Corals of the World, vol 1. Australian Institute subfamily Montastreinae Vaughan & Wells, 1943 (Scleractinea- of Marine Science and CRR. Queensland, Australia, 463 pp. Coelenterata). Part II. The genera Cyphastrea, Leptastrea, Veron, J.E.N & E. Turak, 2006. Coral diversity. In: Green, A., Echinopora and Diploastrea. Zoologische Mededelingen, P. Lokani, W. Atu, P. Ramohia, P. Thomas, J. Almany (eds). Leiden, 55: 235–263. Solomon Islands Marine Assessment: Technical report of Wolstenholme, J.K., 2003. Species boundaries in scleractinian survey conducted May 13 to June 17, 2004. TNC Pacifi c Island corals: a case study of the Acropora humilis species group. Countries No 1/06. PhD-thesis, James Cook University. Veron, J.E.N. & C.C. Wallace, 1984. Scleractinia of Eastern Wolstenholme, J.K., 2004. Temporal reproductive isolation and Australia − Part V. Family Acroporidae. Monograph Series gametic compatibility are evolutionary mechanisms in the 6. Australian Institute of Marine Science: Australian National Acropora humilis species group (Cnidaria; Scleractinia). Marine University Press, 485 pp. Biology, 144: 567–582. Wallace, C.C., 1999. Staghorn corals of the world – a revision of the genus Acropora. Melbourne: CSIRO Publishing, 421 pp. Wallace, C.C. & C-F. Dai, 1997. Scleractinia of Taiwan. IV. Review of the coral genus Acropora from Taiwan. Zoological Studies, 36: 288–324.

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THE RAFFLES BULLETIN OF ZOOLOGY 2011 59(1): 19–28 Date of Publication: 28 Feb.2011 © National University of Singapore

NEW SPECIES OF PHERETIMA, PITHEMERA, AND POLYPHERETIMA (CLITELLATA: MEGASCOLECIDAE) FROM KALBARYO, LUZON ISLAND, PHILIPPINES

Yong Hong Department of Agricultural Biology, College of Agriculture & Life Science, Chonbuk National University, Jeonju 561-756, Republic of Korea Email: [email protected]

Samuel W. James Natural History Museum and Biodiversity Institute, University of Kansas, Lawrence, KS 66045, U.S.A. Email: [email protected] (Corresponding author)

ABSTRACT. – Eight new species of pheretimoid earthworms collected from Kalbaryo, Ilocos Norte Province, Luzon Island, the Philippines are described herein: four new species of Pheretima, three new species of Pithemera, and one new species of Polypheretima. All the Pheretima species lack genital markings. Pheretima margaritata, new species, Pheretima kalbaryoensis, new species, and Pheretima thaii, new species, have spermathecae in vi–ix, but Pheretima globosa, new species, has spermathecae in vii–ix. The distances between the male pores in Pheretima margaritata, new species, Pheretima kalbaryoensis, new species, Pheretima thaii, new species, and Pheretima globosa, new species, are 2.4 mm, 4.5 mm, 1.4 mm, and 1.7 mm, respectively. Pithemera glandis, new species, and Pithemera fusiformis, new species, have ﬁ ve pairs of spermathecae in v–ix. Pithemera levii, new species, has four pairs in v–viii. Pithemera glandis, new species, has no genital markings, but Pithemera fusiformis, new species, has paired genital markings in xix and xx. Pithemera levii, new species, has paired genital markings in xviii and xix. Polypheretima pagudpudensis, new species, has spermathecae in vi–vii, and genital markings paired in xvii, xix, and xx. Descriptions of the new species are provided, including illustrations of the ventral view, male pore region, and spermathecae.

KEY WORDS. – Taxonomy, Pheretima, Pithemera, Polypheretima, Megascolecidae, Clitellata, Philippines.

INTRODUCTION The earthworms collected belong to the Pheretima complex (Sims & Easton, 1972) and fall into three genera, Pheretima, This paper is one of a series of reports on the largely Pithemera, and Polypheretima. unknown earthworm fauna of the Philippines (James et al., 2004; James, 2004; Hong & James, 2004; Hong & James, The four new Pheretima species fall within two provisional 2008a, b). Beginning in early 2001, we surveyed numerous species groups designated by Sims & Easton (1972): three sites in the northern Philippines, one of which is covered species in the darnleiensis group and one species in the dubia here. The present site is Kalbaryo-Pagudpud, a somewhat group. The darnleiensis species group needs re-examination isolated forested mountain on the very northwest corner of in light of collections made at Kalbaryo and elsewhere in the Luzon, 560 km from Manila, in the province of Ilocos Norte. Philippines and Indonesia. Prior to our work it contained 15 Nearby towns are Bangui and Dumalneg. Kalbaryo is one of species names all synonymized by Sims and Easton (1972) the last remaining forests at the northern end of the Central as Pheretima darnleiensis (Fletcher, 1886) and encompassed Cordillera Range of Luzon. At the time of our survey it was a large geographic area (Torres Straits to Philippines) with being considered for protected status by the Department of considerable, perhaps excessive, morphological variations Environment and Natural Resources of the Government of (James, unpublished data). The species reported here the Philippines. Kalbaryo has a maximum elevation of about extend both the range and the morphological variation of 400 m on the mountains. Lower slopes are mainly secondary the species group. forest, with primary forest at higher elevations. There is a pronounced dry season from March to April. Our visit was Pithemera has fewer known species compared to most in early June, early in the rainy season. other genera in the Pheretima-complex, less than the small

19 Hong & James – New earthworms from Kalbaryo, Luzon, Philippines genera Ephemitra Sims & Easton, 1972, and Archipheretima along dorsal vessel; intestinal caeca simple, originating in Michaelsen, 1928. Pithemera contains 12 species in the xxvii, and extending anteriorly about to xxv, each consisting bicincta, pacifi ca, and sedgwicki species groups and one of a fi nger-shaped sac; typhlosole almost none. Hearts x–xiii species transferred to Amynthas. In this paper the three new esophageal; ix lateral, right side much larger. Pithemera species belong to the bicincta group in Sims & Easton (1972). Ovaries and funnels in xiii, spermathecae in vi–ix with nephridia on spermathecal ducts; spermatheca with small Holotypes and some paratypes are deposited in the National broad oval ampulla, fl attened by gizzard, duct shorter than Museum of the Philippines Annelid collection (NMA). The ampulla, iridescent diverticulum, seminal chamber egg- remaining paratypes are deposited in the Kansas University shaped, with stalk longer than ampulla; several diverticula Natural History Museum (KUNHM), USA, National Institute pass through septum to the anterior of the next segment. of Biological Resources, Korea (NIBR) and the Zoological Male sexual system holandric, testes and funnels in ventrally Reference Collection (ZRC) of the Raffles Museum of paired sacs in x, xi. Seminal vesicles two pairs in xi, xii with Biodiversity Research, Singapore. dorsal lobes. Prostates in xvii–xviii, with short muscular duct entering central medial face of copulatory bursae without coelomic glands; copulatory bursa openings fl anked TAXONOMY anteriorly and posteriorly by circular pads; penis with long slit opening on one side. Pheretima margaritata, new species (Figs. 1A–B)

Material examined. – Holotype: One clitellate (NMA 4385), Philippines, Ilocos Norte Province, Pagudpud, Kalbaryo (18°33.65'N 127°57.83'E), 239 m, litter layers in the forest, coll. Y. Hong, A. Castillo & M. Levi, 3 Jun.2001. Four paratypes: 1 clitellate (NMA 4393), 1 clitellate (KUNHM), 1 clitellate (NIBR), 1 clitellate (ZRC): same data as for holotype.

Other material: 10 clitellates, 4 aclitellates (KUNHM), Kalbaryo (18°30.94'N 120°54.52'E), 408 m, litter layer in the forest, 3 Jun.2001.

Diagnosis. – Four pairs of spermathecal pores in 5/6–8/9; 0.4 mm openings of copulatory bursae, spermathecal pores and male pores 0.25–0.28 and 0.26 circumference ventrally apart, respectively.

Etymology. – The name margaritata is Latin for pearl, here referring to the color of the spermathecal diverticulum.

Description. – Dark brown dorsal pigment, segmental equators unpigmented around setae. Dimensions 85–148 mm by 3.0–3.7 mm at segment x, 3.4–3.7 mm at xxx, 3.2–3.8 mm at clitellum, segments 83–96; body cylindrical in cross-section. Setae regularly distributed around segmental equators, numbering 24 at vii, 30 at xx; 4 between male pores, size, distance regular; setal formula AA:AB:YZ: ZZ = 4:2.5:4:7 at xiii. Clitellum annular in xiv–xvi; setae invisible externally.

First dorsal pore in 9/10, four pairs of spermathecal pores in 5/6–8/9, obvious bump inside segmental furrows, laterally placed, distance between spermathecal pores 2.8 mm (0.25– 0.28 circumference ventrally apart). Female pore single in xiv on 0.5 mm oval, 0.4 mm openings of copulatory bursae paired in xviii, distance between openings 2.4 mm (0.26 circumference ventrally apart). Genital markings absent.

Septa 5/6–7/8 thin, 8/9, 9/10 absent, 10/11, 11/12 slightly Fig. 1. Pheretima margaritata, new species: A, ventral view; B, muscular, 12/13, 13/14 thin. Gizzard in viii–x, intestine spermathecae and diverticulum. Scale bars = 5 mm (A), 1 mm enlarged from xv, medium paired lymph glands from xxviii (B).

20 THE RAFFLES BULLETIN OF ZOOLOGY 2011

Remarks. – Pheretima margaritata, new species, keys to First dorsal pore in 11/12, four pairs of spermathecal pores the darnleiensis group in Sims & Easton (1972), composed in 5/6–8/9, conspicuous, distance between spermathecal of fi fteen species, all of which were synonymized by Sims pores 4.5 mm (0.23 circumference ventrally apart). Female & Easton (1972). This species group is defi ned by having pore single in xiv, on 0.6 mm oval, 1.0 mm openings of either four or fi ve pairs of spermathecae, the last pair in copulatory bursae paired in xviii, distance between male segment ix, and the rare fi fth in segment v. Pheretima pores 3.6 mm (0.17 circumference ventrally apart). Genital margaritata, new species, differs from P. darnleiensis in markings absent. having fewer setae per segment, fewer segments, generally shorter length (vs. restricted sense of P. darnleiensis, 155 Septa 5/6–7/8 thick, 8/9, 9/10 very thin, 10/11–12/13 thick mm), and consistently dark brown dorsal pigment, rather than with muscle, 13/14 thin. Gizzard in viii, intestine enlarged sometimes unpigmented. The spermathecal diverticula are from xv, small paired lymph glands from xxviii along dorsal longer in Pheretima margaritata, new species, and the male vessel; intestinal caeca simple, originating in xxvii, and and spermathecal pores are wider apart. Our examination extending anteriorly about to xxiv or xxiii, each consisting of many of the species included in the synonymy of P. of a large fi nger-shaped sac; typhlosole low simple fold from darnleiensis suggests that species-level differences have xxvii, 32 longitudinal blood vessels in intestinal wall from been ignored or discounted against the large number of xxviii. Hearts x–xiii esophageal; ix lateral, left side large. spermathecae. The range possessed by P. darnleiensis sensu lato (Torres Strait to the Philippines; Borneo, Indonesia, Ovaries and funnels in xiii, spermathecae in vi–ix with Malaysia) could be either the result of an overly broad species nephridia on spermathecal ducts; spermathecae with small concept or the consequence of human transport. We believe ampulla, diverticulum sausage-shaped, as long as ampulla that this species was suspected of being a peregrine, at least or longer than ampulla. Male sexual system holandric, within Southeast Asia, and for that reason the variations testes and funnels in ventrally paired sacs in x, xi. Seminal in size, coloration, and other characters were discounted. Certainly the original record on Darnley Island is outside the known natural range of Pheretima, so the suspicion was correct for that site. Even if there is a regional peregrine species embedded within P. darnleiensis we feel confi dent that 1) it does not include all the synonymized names and, 2) our material obtained from remote forested areas is native, not introduced. In any case, resolving the questions about the species divisions within P. darnleiensis will require a separate paper.

Pheretima kalbaryoensis, new species (Figs. 2A–B)

Material examined. – Holotype: One clitellate (NMA 4386), Philippines, Ilocos Norte Province, Pagudpud, Kalbaryo (18°33.65'N 127°57.83'E), 239 m, litter layers in the forest, coll. Y. Hong, A. Castillo & M. Levi, 14 Mar.2001, 3 Jun.2001.

Other material: 2 aclitellates, same data as for holotype (NMA).

Diagnosis. – Four pairs of spermathecal pores in 5/6– 8/9; spermathecal pores and male pores 0.23 and 0.17 circumference ventrally apart, respectively, typhlosole present, septa 8/9/10 present.

Etymology. – The species is named after its type locality, Kalbaryo in Philippines.

Description. – Dark brown dorsal pigment,1/3 segment length white stripes on setal rings, narrowing dorsally. Dimensions 210 mm (aclitellate 145 mm) by 6.2 mm at segment x, 7.0 mm at xxx, 6.7 mm at clitellum, segments 93; body cylindrical in cross-section. Setae numbering 38 at vii, 49 at xx; 7 between male pores; setae slightly more widely spaced dorsally, setal formula AA:AB:YZ:ZZ = 6:2:3:6 at xiii. Clitellum annular Fig. 2. Pheretima kalbaryoensis, new species: A, ventral view; in xiv–xvi; setae invisible externally. B, spermathecae and diverticulum. Scale bars = 5 mm (A), 1 mm (B).

21 Hong & James – New earthworms from Kalbaryo, Luzon, Philippines vesicles two pairs in xi, xii with dorsal lobes. Prostates in ovate ampulla, duct shorter than ampulla, diverticulum xvii– xviii, two lobes, smooth duct entering center of the seminal chamber egg-shaped, iridescent, as long as ampulla, copulatory bursae without coelomic glands, penis small, male stalks long. Male sexual system holandric, testes and funnels pore just below tip; copulatory bursa openings fl anked by in ventrally paired sacs in x, xi. Seminal vesicles two anterior and posterior large fl at circular pads. pairs in xi, xii with dorsal lobes, vas deferens slender, not muscular. Prostates in xvii–xix, with short muscular duct Remarks. – The species also keys to the darnleiensis group entering center of the copulatory bursae without coelomic in Sims & Easton (1972). P. darnleiensis has fewer setae glands; copulatory bursa openings fl anked by anterior and per segment, shorter body length, ampulla shorter than posterior circular pads, penis long with vertical slit pore on diverticulum and first dorsal pore in 9/10. The present medial face. species is larger than Pheretima margaritata, new species, and has more setae per segment, more closely placed male Remarks. – Pheretima thaii, new species, has the same and spermathecal openings, septa 8/9/10 present, and a number of spermathecae as the previous two new species, sausage-shaped diverticulum rather than a diverticulum with an egg-shaped seminal chamber.

Pheretima thaii, new species (Figs. 3A–B)

Material examined. – Holotype: One clitellate (NMA 4387), Philippines, Ilocos Norte Province, Pagudpud, Kalbaryo (18°30.94'N 120°54.52'E), 408 m, litter layers in the forest, coll. Y. Hong, A. Castillo & M. Levi, 3 Jun.2001.

Diagnosis. – Four pairs of spermathecal pores in 5/6–8/9; spermathecal pores and male pores 0.5 and 0.2 circumference ventrally apart, respectively.

Etymology. – The species is named for Dr. Thai Tran Bai, a Vietnamese biologist who made many good contributions to the taxonomy of earthworms.

Description. – Dark brown dorsal and brown ventral pigment, intersegmental furrows and large circles around each seta unpigmented, these circles fuse where setae are closely spaced. Dimensions 60 mm by 2.1 mm at segment x, 2.3 mm at xxx, 2.5 mm at clitellum, segments 81; body cylindrical in cross-section. Setae more crowded ventrally in anterior segments, almost evenly distributed in posterior half, numbering 16 at vii, 24 at xx; 3 between male pores, size, distance irregular; setal formula AA:AB:YZ:ZZ = 1.5:1.5:4:7 at xiii. Clitellum annular in xiv–xvi; setae invisible externally.

First dorsal pore in 9/10, four pairs of spermathecal pores in 5/6–8/9, conspicuous, lateral (0.5 circumference). Female pore single in xiv, on 0.4 mm oval, 0.3 mm openings of copulatory bursae paired in xviii, 1.4 mm between male pores (0.2 circumference ventrally apart). Genital markings absent.

Septa 5/6–7/8 thin, 8/9, 9/10 absent, 10/11–13/14 thin. Gizzard in viii–x, intestine enlarged from xv, medium paired lymph glands from xxviii along dorsal vessel; intestinal caeca simple, originating in xxvi, and extending anteriorly about to xxiii, each consisting of a ﬁ nger-shaped sac; typhlosole almost none. Hearts x–xiii esophageal; ix lateral.

Ovaries and funnels in xiii, spermathecae in vi–ix with Fig. 3. Pheretima thaii, new species: A, ventral view; B, spermathecae nephridia on spermathecal ducts; spermatheca with small and diverticulum. Scale bars = 5 mm (A), 1 mm (B).

22 THE RAFFLES BULLETIN OF ZOOLOGY 2011 but it differs from them and P. darnleiensis in the much more bursae without coelomic glands; copulatory bursa openings widely spaced spermathecal pores, much smaller body size ﬂ anked by anterior and posterior circular pads, penis only and presence of ventral pigment. Its spermathecal ampulla is small bump on copulatory bursa roof. as long as the diverticulum, unlike Pheretima kalbaryoensis, new species. The diverticulum shape is similar to that of Remarks. – Pheretima globosa, new species, keys to the dubia Pheretima margaritata, new species, but other characters group in Sims and Easton (1972), which is composed of ﬁ ve are different. species, P. philippina (Rosa, 1891), P. callosa Gates, 1937, P. dubia (Horst, 1893), P. korinchiana Cognetti, 1922, and P. poiana Michaelsen, 1913. The present species is small, Pheretima globosa, new species with body length 35–42 mm, and 71–80 segments, but P. (Figs. 4A–B) poiana and P. philippina are large worms, greater than 180 mm in length with more than 110 segments. Pheretima Material examined. – Holotype: One clitellate (NMA 4388), callosa is large bodied (330 by 16 mm), has at least twice as Philippines, Ilocos Norte Province, Pagudpud, Kalbaryo (18°33.59'N many setae per segment as Pheretima globosa, new species, 120°57.50'E), 175 m, litter layers in the forest, coll. Y. Hong, A. pseudovesicles in xiii and xiv, and thickly conical penes Castillo & M. Levi, 3 Jun.2001. Paratypes: 1 clitellate (NMA 4394), with deep cleft on the tips. Pheretima callosa is from the 1 clitellate (KUNHM), 1 clitellate (NIBR), 1 clitellate (ZRC): same data as for holotype.

Other material: 15 clitellates and 5 aclitellates, same data as for holotype (NMA).

Diagnosis. – Three pairs of spermathecal pores in 6/7– 8/9; spermathecal pores and male pores 0.32 and 0.22 circumference ventrally apart, respectively.

Etymology. – The name globosa is Latin for spherical, here referring to the shape of the spermathecal diverticula seminal chamber.

Description. – Light brown dorsal pigment. Dimensions 35–42 mm by 2.3–2.5 mm at segment x, 2.5–2.7 mm at xxx, 2.5–2.6 mm at clitellum, segments 71–80; body cylindrical in cross-section. Setae regularly distributed around segmental equators, numbering 29 at vii, 52 at xx; 9 between male pores; size, distance irregular; setal formula AA:AB:YZ: ZZ = 1:1:1:2 at xiii. Clitellum annular in xiv–xvi; setae invisible externally.

First dorsal pore in 11/12, three pairs of spermathecal pores in 6/7–8/9, very small, distance between spermathecal pores 2.4 mm (0.32 circumference ventrally apart). Female pore single in xiv, on 0.3 mm oval, 0.3 mm openings of copulatory bursae paired in xviii, distance between male pores 1.7 mm (0.22 circumference ventrally apart). Genital markings absent.

Septa 5/6–7/8 thick, 8/9, 9/10 absent, 10/11–13/14 thin. Gizzard in viii–x, intestine enlarged from xv, paired lymph glands from xxviii along dorsal vessel; intestinal caeca simple, originating in xxvii, and extending anteriorly about to xxvi, each consisting of a short ﬁ nger-shaped sac; typhlosole almost none. Hearts x–xiii esophageal; ix lateral, right side only.

Ovaries and funnels in xiii, spermatheca in vii–ix with nephridia on spermathecal ducts; spermatheca with spherical ampulla, duct shorter than ampulla, diverticulum seminal chamber egg-shaped. Male sexual system holandric, testes and funnels in ventrally paired sacs in x, xi. Seminal vesicles Fig. 4. Pheretima globosa, new species: A, ventral view; B, two pairs in xi, xii with dorsal lobes. Prostates in xvii–xviii, spermathecae and diverticulum. Scale bars = 3 mm (A), 1 mm with short muscular duct entering center of the copulatory (B).

23 Hong & James – New earthworms from Kalbaryo, Luzon, Philippines

Philippines, Luzon Island, Benguet Province at 2,130 m asl in Septa 5/6–7/8 thick, 8/9 absent, 9/10–12/13 muscular. oak forest, so is geographically the nearest neighbor among Gizzard in viii–ix, intestine enlarged from xv; intestinal the dubia group. Pheretima philippina is from Mindoro, a caeca simple, originating in xxii, and extending anteriorly more southerly island in the Philippines. Pheretima dubia about to xx, each consisting of a small triangle-shaped sac; is only twice the length of Pheretima globosa, new species, typhlosole medium depth simple fold from xxii. Hearts x–xii and has spirally twisted spermathecal diverticula. Pheretima esophageal; ix lateral, viii absent. korinchiana is also much larger (90–180mm), has violet- brown dorsal pigment, 12 setae between the male pores, Ovaries and funnels in xiii, spermathecae in v–ix; no laterally placed spermathecal pores, and a club-shaped nephridia on spermathecal ducts; spermatheca with an diverticulum on an undulating or spiral stalk. Ignoring for elongate acorn-shaped ampulla, ducts slender, as long as the moment the numbers and locations of spermathecae that deﬁ ne the dubia-group, the present species is otherwise very close to the overall morphological pattern of the three species described here for the darnleiensis-group. It seems reasonable to hypothesize that Pheretima globosa, new species is derived from a darnleiensis-group ancestor, by loss of the anterior pair of spermathecae. James (2005) presents preliminary molecular evidence that similar reductions from the darnleiensis-group spermathecal battery have taken place in other Philippine Pheretima species.

Pithemera glandis, new species (Figs. 5A–B)

Material examined. – Holotype: One clitellate (NMA 4389), Philippines, Ilocos Norte Province, Pagudpud, Kalbaryo (18°33.65'N 127°57.83'E), 239 m, litter layers in the forest, coll. Y. Hong, A. Castillo & M. Levi, 3 Jun.2001. Paratypes: 1 clitellate (NMA 4395), 1 clitellate (KUNHM), 1 clitellate (NIBR), 1 clitellate (ZRC): same data as for holotype.

Other material: 5 clitellates and 10 aclitellates, same data as for holotype (NMA).

Diagnosis. – Five pairs of very small spermathecal pores in 4/5–8/9; spermathecal pores and male pores 0.15 and 0.18 circumference ventrally apart, respectively. Genital markings absent.

Etymology. – The acorn shaped spermathecal ampulla is the source of the species name (glandis is Latin for acorn).

Description. – Brown dorsal pigment. Dimensions 62–108 mm (aclitellate 72 mm) by 4.1–4.5 mm at segment x, 4.2–4.6 mm at xxx, 4.3–4.6 mm at clitellum, segments 129–136; body cylindrical in cross-section. Setae regularly distributed around segmental equators, numbering 53 at vii, 61 at xx; 8–11 between male pores, size, distance regular; setal formula AA:AB:YZ:ZZ = 3.5:2:2:3.5 at xiii. Clitellum annular in xiv–1/2xvi; ventral setae of xvi visible externally.

First dorsal pore in 12/13, ﬁ ve pairs of minute spermathecal pores in 4/5–8/9, ventral, 2.0 mm between spermathecal pores (0.15 circumference ventrally apart). Female pores paired in xiv within 0.5 mm oval, male pores paired in xviii, 2.5 mm apart (0.18 circumference ventrally apart), slightly protuberant white spot on 0.3 mm porophores within 1.0 mm outer rings. Genital markings absent. Fig. 5. Pithemera glandis, new species: A, ventral view; B, spermathecae and diverticulum. Scale bars = 5 mm (A), 1 mm (B).

24 THE RAFFLES BULLETIN OF ZOOLOGY 2011 ampulla, diverticulum shorter than ampulla with elongate oval Septa 5/6–7/8 thin, 8/9 absent, 9/10–13/14 thin. Gizzard in chamber. Male sexual system holandric, testes and funnels viii–ix, intestine enlarged from xv; intestinal caeca simple, in inverted U-shaped sacs in x, xi, testes sacs enclosing originating in xxii, and extending anteriorly about to xxi, hearts x, xi, seminal vesicles xi. Seminal vesicles two pairs each consisting of a small sac, laterally placed; typhlosole in xi, xii with ﬁ ne bushy dorsal lobes, prostates in xvii–xix, none. Hearts xi–xiii esophageal; ix lateral. four small main lobes, ducts long slender, widening slightly towards body wall, vas deferens very thin. Ovaries and funnels in xiii, spermatheca in v–ix; no nephridia on spermathecal ducts; spermatheca with small ampulla, Remarks. – Pithemera glandis, new species, keys to the duct longer than ampulla, diverticulum sausage-shaped, bicincta group in Sims & Easton (1972), which is composed shorter than ampulla. Male sexual system holandric, testes of two species, Pi. bicincta (Perrier, 1875) and Pi. violacea and funnels in paired lateral sacs in x, xi, not connected. (Beddard, 1895). The species shares some characteristics of Seminal vesicles two pairs in xi, xii with dorsal lobes. the spermathecal pores in 4/5–8/9, and ampulla longer than Prostates in xvii–xix, one or two lobes; prostatic duct with diverticulum, with Pi. bicincta and Pi. violacea. Pithemera spindle-shaped muscular expansion in ectal half. glandis, new species, has no genital papillae, but Pi. bicincta has paired genital papillae in segments xviii and xix, and one Remarks. – The species also keys to the bicincta group in pair of genital papillae just behind the male pores at 18/19. Sims & Easton (1972), but Pi. bicincta and Pi. violacea Pithemera glandis, new species, differs from Pi. violacea in have the ampulla longer than the diverticulum and different having 8 or more setae between the male pores, and lacking locations of genital papillae. Pithemera fusiformis, new the genital papillae on 18/19 found in Pi. violacea. species is greatly smaller than Pithemera glandis, new species, (20–22 vs. 62–108 mm), has genital papillae and also lacks a typhlosole. Pithemera fusiformis, new species (Figs. 6A–B)

Material examined. – Holotype: One clitellate (NMA 4390), Philippines, Ilocos Norte Province, Pagudpud, Kalbaryo (18°33.59'N 120°57.50'E), 175 m, litter layers in the forest, coll. Y. Hong, A. Castillo & M. Levi, 3 Jun.2001. Paratypes: 1 clitellate (NMA 4396), 1 clitellate (NIBR), 1 clitellate (ZRC): Same data as for holotype.

Other material: 10 aclitellates, same data as for holotype (NMA).

Diagnosis. – Five pairs of spermathecae in v–ix; male pores invaginate white spots on 0.3 mm small round porophores, male pores 0.31 circumference ventrally apart. Genital markings paired in xix and xx.

Etymology. – This species is named for the spindle-like appearance of the prostatic ducts.

Description. – Brown dorsal pigment. Dimensions 20–22 mm by 1.7 mm at segment x, 1.8 mm at xxx, 1.6 mm at clitellum, segments 90; body cylindrical in cross-section. Setae regularly distributed around segmental equators, numbering 36 at vii, 47 at xx; 10 between male pores, size, distance regular; setal formula AA:AB:YZ:ZZ = 1.5:1:1:1.5 at xiii. Clitellum annular in xiv–1/2xvi; setae of xvi visible externally.

First dorsal pore in 12/13, ﬁ ve pairs of spermathecal pores in 4/5–8/9, unrecognizable externally. Female pore single in xiv, on 0.2 mm oval, male pores paired in xviii, invaginate white spots on 0.3 mm small round porophores, distance between male pores 1.8 mm (0.31 circumference ventrally apart), male pore region protuberant laterally. Genital markings paired in xix, xx. Fig. 6. Pithemera fusiformis, new species: A, ventral view; B, spermathecae and diverticulum. Scale bars = 3 mm (A), 1 mm (B).

25 Hong & James – New earthworms from Kalbaryo, Luzon, Philippines

Pithemera levii, new species Etymology. – The species is named for the collector of the (Figs. 7A–B) type material, Mr. Matthew Levi.

Material examined. – Holotype: One clitellate (NMA 4391), Description. – Brown dorsal pigment. Dimensions 50–65 mm Philippines, Ilocos Norte Province, Pagudpud, Kalbaryo (18°30.94'N by 2.3–2.6 mm at segment x, 2.5–2.7 mm at xxx, 2.5–2.8 120°54.52'E), 408 m, litter layers in the forest, coll. Y. Hong, A. mm at clitellum, segments 82–111; body cylindrical in Castillo & M. Levi, 3 Jun.2001. Paratypes: 1 clitellate (NMA 4397), cross-section. Setae regularly distributed around segmental 1 clitellate (KUNHM), 1 clitellate (NIBR), 1 clitellate (ZRC), same equators, numbering 54 at vii, 63 at xx; 6–7 between male data as for holotype. pores, size, distance regular; setal formula AA:AB:YZ:ZZ Other material: 7 clitellates and 13 aclitellates, same data as for = 1:1:1:1.5 at xiii. Clitellum annular in xiv–1/2xvi; ventral holotype; 20 clitellates and 4 aclitellates, Kalbaryo (18°33.65'N setae of xvi visible externally. 127°57.83'E), 239 m, colls. Y. Hong, A. Castillo & M. Levi, 3 Jun.2001. First dorsal pore in 12/13, four pairs of spermathecal pores in 4/5–7/8 (one worm 4/5, 5/6 only), very close to Diagnosis. – Four pairs of spermathecal pores in 4/5–7/8; medio-ventrum, 0.8 mm between spermathecal pores (0.11 spermathecal and male pores 0.11 and 0.13 circumference circumference ventrally apart). Female pores paired in xiv, ventrally apart, respectively. Large circular genital papillae on 0.3 mm oval, male pores paired in xviii, on 0.3 mm dark paired presetal in xviii and xix. oval, hardened porophores, 1.1 mm between male pores (0.13 circumference ventrally apart); large circular genital papillae paired, presetal in xviii, xix in line with male pores.

Septa 5/6–7/8 thick, 8/9 absent, 9/10–11/12 thick, 12/13, 13/14 thin. Gizzard usual in viii–ix, intestine enlarged from xv; intestinal caeca simple, originating in xxii, and extending anteriorly about to xxi, each consisting of a ﬁ ngernail-shaped sac; typhlosole medium depth simple fold from xxii. Hearts x–xii esophageal; ix lateral, viii absent. Vessel branches from lateral heart ix to body wall, extends posteriorly to xiv.

Ovaries and funnels in xiii, spermatheca in v–viii (one worm v, vii only); no nephridia on spermathecal ducts; spermatheca with elongate cylindrical ampulla slightly expanded knob at ental end , duct short, diverticulum stalk slender, longer than ampulla; chamber small ovate. Male sexual system holandric, testes and funnels in paired sacs in x, xi, not joined dorsally. Seminal vesicles two pairs in xi, xii without dorsal lobes. Prostates in xvii–xix, 2–3 or more main lobes, prostatic duct with spindle-shaped muscular expansion in ectal half. Glandular tissue on body wall anterior to genital markings in xviii, xix.

Remarks. – Pithemera levii, new species, has one pair of presetal genital papillae in each of xviii and xix, but Pi. bicincta has paired papillae in xviii extending from segments xviii to xix, and one pair of genital papillae just behind male pores at 18/19. Pithemera levii, new species, differs from Pi. violacea and Pi. bicincta in having only four pairs of spermathecae and no intersegmental genital papillae.

Polypheretima pagudpudensis, new species (Figs. 8A–C)

Material examined. – Holotype: One clitellate (NMA 4392), Philippines, Ilocos Norte Province, Pagudpud, Kalbaryo (18°33.65'N 127° 57.83'E), 239 m, litter layers in the forest, coll. Y. Hong, A. Castillo & M. Levi, 14 Mar.2001, 3 Jun.2001.

Other material: 3 aclitellates, same data as for holotype (NMA). Fig. 7. Pithemera levii, new species: A, ventral view; B, spermathecae and diverticulum. Scale bars = 5 mm (A), 1 mm (B).

26 THE RAFFLES BULLETIN OF ZOOLOGY 2011

Diagnosis. – Two pairs of spermathecal pores in 5/6, 6/7; Septa 5/6–7/8 thick, 8/9–13/14 thin. Gizzard in viii, intestine male pores xviii on extending to lateral margin of ventrum, enlarged from xv, intestinal caeca absent; typhlosole large, distance between male pores 1.5 mm (0.23 circumference 1/2 lumen diameter from xxii. Hearts x–xii esophageal; ix ventrally apart). Genital markings paired in xvii, xix and lateral. xx. Ovaries and funnels in xiii, spermatheca in vi, vii without Etymology. – The species is named for its type locality, nephridia on spermathecal ducts; spermatheca with elongate Pagudpud in Philippines. ampulla, diverticulum stalk shorter than ampulla, chamber small oval. Male sexual system holandric, testes and funnels Description. – Worm unpigmented. Dimensions 56–63 mm in dorsally united sacs in x, xi. Sacs do not enclose hearts. by 2.1 mm at segment x, 2.2 mm at xxx, 2.0 mm at clitellum, Seminal vesicles two pairs in xi, xii, very small. Prostates segments 126–136; body cylindrical in cross-section. Setae broad but thin, in xvii–xxii, ducts slender. regularly distributed around segmental equators, numbering 63 at vii, 27 at xx; 8 between male pores, size, distance Remarks. – The species shares the spermathecal pores in irregular; setal formula AA:AB:YZ:ZZ = 3:1.5:1:1 at xiii. 5/6, 6/7, and male pores on prominent porophores with Clitellum annular in xiv–xvi; setae invisible externally. Polypheretima sempolensis Easton, 1979. Polypheretima sempolensis has paired postsetal genital markings on xvii, First dorsal pore in 12/13, two pairs of spermathecal pores and paired presetal on xix, but the present species has paired in 5/6, 6/7. Female pore single in xiv on 0.3 mm oval, male genital markings covering the ventrum of xvii, xix, and xx. pores xviii on protuberant white spot at center of round Also, Polypheretima pagudpudensis, new species, has no porophores, distance between male pores 1.5 mm (0.23 genital markings in the spermathecal pore region, but Po. circumference ventrally apart). Genital markings paired in sempolensis has paired presetal genital markings on ix, xii xvii, xix, xx; all slightly medial to male porophores; genital and occasionally xiii, median to the line of spermathecal markings within white epidermal thickenings spanning greater pores, and the genital markings are exceptionally large, width than male porophores; small paired pre- and post-setal invading the setal lines and the intersegmental furrows. epidermal thickenings medial to porophores in xviii. Polypheretima sempolensis were recorded from East Java, at 1,400–1,500 m elevation.

ACKNOWLEDGEMENTS

This study was supported by National Science Foundation grant DEB-0072764 to the second author. We are grateful for the cooperation of Philippine Department of Environment and Natural Resources personnel for assistance in the ﬁ eld and for providing collection permits. Several local families in the Kalbaryo area assisted the team with guidance, lodging and meals.

LITERATURE CITED

Beddard, F. E., 1895. A monograph of the order Oligochaeta. Oxford: Clarendon Press, i–xii + 769 pp. Cognetti de Martiis, L., 1922. Descrizione di tre nuovi megascolecini. Bollettino dei Musei di Zoologia et Anatomia comparata della Reale Universita di Torino, 37(744): 1–6. Easton, E. G., 1979. Acaecate earthworms of the Pheretima group (Megascolecidae: Oligochaeta): Archipheretima, Metapheretima, Planapheretima, Pleionogaster and Polypheretima. Bulletin of the British Museum of Natural History (Zoology), 35(1):1– 126. Fletcher, J. J., 1886. Notes on Australian Earthworms. Part III. Proceedings of the Linnean Society of NSW, (2)2: 377–402. Gates, G. E., 1937. Notes on some species of Drawida and Pheretima with descriptions of three new species of Pheretima. Bulletin of the Museum of Comparative Zoology, Harvard University, Cambridge, 80: 305–334. Fig. 8. Polypheretima pagudpudensis, new species: A, ventral view; Hong, Y. & James, S.W., 2004. New species of Amynthas Kinberg, B, spermathecae; C. malepore region in xviii. Scale bars = 3 mm 1867 from the Philippines (Oligochaeta: Megascolecidae). Revue (A), 2 mm (C), 1 mm (B). Suisse de Zoologie, 111(4): 729–741.

27 Hong & James – New earthworms from Kalbaryo, Luzon, Philippines

Hong, Y. & James, S.W., 2008a. Nine new species of earthworms James, S. W., Y. Hong & T. H. Kim, 2004. New earthworms (Oligochaeta: Megascolecidae) of the Banaue Rice Terraces of of Pheretima and Pithemera (Oligochaeta: Megascolecidae) the Philippines. Revue Suisse de Zoologie, 115(2): 341–354. from Mt. Arayat, Luzon Island, Philippines. Revue Suisse de Hong, Y. & James, S. W., 2008b. Two new earthworms of the Zoologie, 111 (1): 3–10. genus Pheretima (Oligochaeta: Megascolecidae) from Mt. Michaelsen, W., 1913. Die Oligochaten von Neu-Caledonien Isarog, Luzon Island, Philippines. The Journal of Natural und den benachbarten Inselgruppen. Nova Caledonia. 1: History, 42(23&24): 1565–1571. 173–280. Horst, R., 1893. Descriptions of earthworms. No. 7. On Malayan Michaelsen, W., 1928. Miscellania oligochaetologica. Arkiv für earthworms. Notes from the Leyden Museum, Leyden, 15: Zoologie Series A, 20: 1–15. 316–329. Perrier, E., 1875. Sur les vers de terre des iles Philippines et de la James, S. W. 2004. New species of Amynthas, Pheretima, Cochinchine. Comptes Rendus Hebdomadaires des Séances de Pleionogaster (Oligochaeta: Megascolecidae) of the Mt. l’Académie des Sciences, Paris, 81: 1043–1046. Kitanglad Range, Mindanao Island, Philippines. Rafﬂ es Bulletin Sims, R. W. & E. G. Easton, 1972. A numerical revision of the of Zoology, 52(2): 289–313. earthworm genus Pheretima auct. (Megascolecidae: Oligochaeta) James, S.W. 2005. Preliminary molecular phylogeny in the with the recognition of new genera and an appendix on the Pheretima group of genera (Crassiclitellata: Megascolecidae) earthworms collected by the Royal Society North Borneo using Bayesian analysis. In: V. V. Pop & Pop, A. A. (eds.), Expedition. The Biological Journal of the Linnean Society, Advances in Earthworm Taxonomy II. Cluj University Press, London, 4: 169–268. Cluj-Napoca, Romania. Pp. 129–142.

28 THE RAFFLES BULLETIN OF ZOOLOGY 2011

THE RAFFLES BULLETIN OF ZOOLOGY 2011 59(1): 29Ð37 Date of Publication: 28 Feb.2011 © National University of Singapore

LARGE MANGROVE-DWELLING ELYSIA SPECIES IN ASIA, WITH DESCRIPTIONS OF TWO NEW SPECIES (GASTROPODA: OPISTOBRANCHIA: SACOGLOSSA)

Cornelis (Kees) Swennen Faculty of Science and Technology, Prince of Songkla University, Pattani 94000, Thailand Email: [email protected]

ABSTRACT. − Two large Elysia species have been described as speciﬁ c for mangrove habitats over the last 20 years: E. leucolegnote Jensen, 1990 and E. bangtawaensis Swennen, 1998. They are usually found aggregated on soft mud without algae in shallow, shaded pools during low tide. Two more species are added in the present paper as E. singaporensis, new species, and E. bengalensis, new species. The four Elysia species are easily recognizable. External and internal characters of the four mangrove-associated Elysia spp. are depicted, together with some biological notes, suggesting two morphological groups similar in behaviour and habitat.

KEY WORDS. − Elysia, Mangrove, Plakobranchidae, Western and Central Indo-Paciﬁ c.

INTRODUCTION Photographs were made of the habitat, and live specimens were described in the laboratory. Although important external Sacoglossan opisthobranchs have no jaws and their radula characters can be seen in live specimens, details such as consists of a single, longitudinal row of teeth of which the apertures and dorsal vessels are more easily examined older teeth are kept during their life in an ascus (Pruvot-Fol, in carefully relaxed preserved specimens. For this, some 1954). The classifi cation of the Sacoglossa was most recently specimens were relaxed by placing them in a solution of reviewed by Jensen (1996). The Plakobranchidae (Elysiidae) 71 g MgCl2 in l,000 ml water meanwhile removing mucous form the largest family in the Sacoglossa with Elysia as the accumulation and then fi xing them in a 5% formaldehyde- most speciose genus. The most obvious character of Elysia, seawater solution. Pharynx, radular teeth, and main details of shared with Plakobranchus and Thuridilla, is the presence the digestive system were checked via dissections by hand of wing-like parapodia bordering the posterior part of the with fi ne needles. For permanent storage, some specimens foot. The species have neither shell nor oral tentacles. The were transferred into 70% ethanol. Specimens of the fourth majority of the Elysia species are smaller than 20 mm, but a species were collected and given to me by a colleague working few are larger and measure beween 25Ð50 mm. Several large in India. Types of the new species have been deposited in tropical species from reef habitats were described over 100 the Zoological Reference Collection of the Raffl es Museum years ago [E. grandifolia Kelaart, 1858; E. rufescens (Pease, of Biodiversity Research, National University of Singapore 1871); E. subornata Verrill, 1901]. However, several large (ZRC) and in the Zoological Museum of the University of species living in turbid water in mangrove forests have only Amsterdam (ZMA). been discovered during the last 20 years. The fi rst known large mangrove Elysia was found in Hong Kong and named E. leucolegnote by Jensen in 1990. It was followed by E. SPECIES ACCOUNT bangtawaensis Swennen, 1998, found in the Gulf of Thailand. Meanwhile, two other species have been found elsewhere. Elysia leucolegnote Jensen, 1990 All are named and depicted herein this paper. (Figs. 1Ð7)

Material examined. – South China Sea, Hong Kong, inland, MATERIALS AND METHODS south side Starling Inlet, about 22¡31'28"N, 114¡31'28"E, tens, 11 Mar.2004; Near Ting Kok, about 22¡28'14"N, 114¡13'01"E, Specimens of three species were collected by the author in tens, 14 Mar.2004; Hong Kong , Mai Po Nature Reserve, in mangrove forest along Deep Bay (22¡29'52"N, 114¡01'48"E), mangrove forests at different sites on separate occasions. tens, Nov.2007; Gulf of Thailand, south of Chumphon at about

29 Swennen Ð Large mangrove-dwelling Elysia species

Table 1. Live specimens of the four known mangrove Elysia species can easily been distinguished by the following characteristics:

1. Parapodia with white or yellowish border, triangular white-yellowish mark on head. No other colours on skin ...... E. leucolegnote Jensen, 1990. –. Parapodia with tens of coloured dots ...... 2.

2. Old rose specks over dorsal and ventral sides including foot sole, forming line over dorsal sides of rhinophores ...... E. singaporensis, new species. –. No old rose, but white dots on parapodia ...... 3.

3. Tips of rhinophores pale or orange. Parapodial border with several orange or reddish glandular dots ...... E. bangtawaensis Swennen, 1998. –. Tips of rhinophores dark. Mainly white dots on parapodia and body ...... E. bengalensis, new species

10¡22'20"N, 99¡10'26"E, tens, 27 Ð 29 Mar.2007; Near Don Sak reached four years then accidentally died. Food source is at about 9¡18'59"N, 99¡45'34"E, and 9¡18'10"N, 99¡48'08"E, about the tiny, siphonal green alga Boodleopsis c.f. pusilla. ten, 2 Oct.2007; Pak Phanang Bay, at about 8¡35'37"N, 99¡58'23"E and 8¡29'19"N, 100¡11'33"E, tens, 28-29 Sep.2007, 7, 13 Apr. and Discussion. – It may be the result of a different treatment 30 Sep.2008; Near Bang Tawa, about 06¡51'28"N, 101¡09'31"E, of the pharynx, but Jensen (1990) described the descending tens, 30 Sept.2008; same site, 15, 16 Oct.2009; Andaman Sea Thailand, Ko Libong in mangrove at about 07¡14'50"N, 99¡26'30"E, limb of the radula as attached over its whole length, while tens, 9 Oct.2007; mangrove east of Krabi at about 08¡03'17"N, my specimens showed the ascus on a partly free stalk 098¡53'57"E, tens, 3 Oct.2010. (Fig. 3A). Jensen (2003) reported that her specimens feed on Boodleopsis pusilla, Cladophora, Cladophoropsis and Live colouration. – Narrow white line along parapodial Enteromorpha in the laboratory. Only Boodleopsis was taken borders, often locally widened. Some have few white spots as food in my experiments. Cladophora and Enteromorpha on renopericardial prominence, rarely few white spots on were not touched even when the slugs ﬁ nally lost size and dorsal sides of parapodia. White triangle on head connected colour. They started feeding when Boodleopsis was given with white line over dorsal sides of rhinophores. Tips of and then became green again after a few days. rhinophores and often also posterior side of renocardial prominence lack tubules of digestive gland (Fig. 1). Individuals with digestive gland yellowish starting in the Elysia singaporensis, new species central parts have not fed for some months as conﬁ rmed (Figs. 1Ð7) in captivity. Material examined. – HOLOTYPE: 1 ex. (length alive 23 External morphology. – Length alive up to 41 mm. Length mm)(ZRC, MOL.2974), coll. Singapore, Western Johor Straits, of renopericardial prominence about three times width. Two old mangrove forest bordering east side of Sungei Buloh Wetland Park, about 1¡26'47"N, 103¼43'57"E, 21 May 2002. major dorsal vessels connected to posterior side (Fig. 2A). PARATYPES: 3 ex. (ZRC, MOL. 2974); 3 ex. (ZMA, Cat. nr. Renopore in vaginal aperture anterior right of renopericardinal Moll. 179181), same site and date as holotype. prominence on dorsum (Fig. 7A). OTHERS: 22 ex., length alive 20-31 mm, with egg strings, 21 May 2002; same site, 18 Oct.2009, 12 ex. Length alive 15-22 Internal characteristics. – Pharynx small, round, ascus on mm, with egg strings. stalk, length 330-350 µm, height without ascus stalk 280-320 µm (Fig. 3A). Radular teeth: ascending limb 4-6, descending Etymology. – The species name is derived from the island limb 7-9, ascus 8 to 18 (N= 5). Teeth blade-shaped with state of Singapore where the specimens were found. upwards curved tips, cutting edges denticulate (Fig. 4A). Tooth length up to 63 µm. Oesophagus about 2.8 times Live colouration. – Basic colour green from content of length of pharynx. Stomach small compared to intestine digestive gland. Ruddy specks over dorsal and ventral (Fig. 5A). Green, glandular tissue of digestive gland starts in sides including foot sole, forming line over dorsal sides main branches far from stomach (Fig. 5A). Penis unarmed, of rhinophores. Tubules of digestive gland lacking in tips conical with narrow extension, length 400-500 µm (Fig. 6A). of rhinophores and over posterior part of renopericardial Vas deferens convoluted just before entering penis. Gonadal prominence (Fig. 1B). follicles individually connected to ducts. External morphology of live specimens. – Length alive Biology. – Egg strings up to 55 cm long, irregularly curved, up to 31 mm. Eyes not conspicuous, wide apart behind but when substrate allows in wide spirals, with yellow string rhinophores; rhinophores with groove over whole length. of extra-capsular yolk in the jelly, single ovum per capsule, Lateral groove on right side from anterior border of right 11 March 2004 Starling Inlet, 13 April 2008 Pak Phanang parapodium to foot; transversal groove dividing foot into Bay. Some specimens were kept in captivity in separate two parts; anterior foot corners slightly extended, bluntly bottles for several years. One large specimen was collected pointed; frontal border can be kept notched, but also smoothly and stayed alive in captivity for 53 months; three others curved; foot behind transverse groove weakly developed.

30 THE RAFFLES BULLETIN OF ZOOLOGY 2011

Thin tributaries of digestive gland visible over whole body, Parapodia and body do not bear protuberances. Male aperture missing in tips of rhinophores, over eyes, and sometimes below right rhinophore; anus and oviducal apertures close partly above renopericardium. Renopericardial prominence together in upper part of lateral groove. long and narrow, tapering posteriorly, length three to four times width. Renal part posterior of cardial part, about 2/3 Internal features. – Pharynx small, ascus on stalk, length of total length, posterior with pair of major dorsal vessels 570-760 µm, height without ascus stalk 550 to 630 µm (three and on each lateral side zero to two vessels, varying in 25 mm long specimens) (Fig. 3B). Radula with 4 to 7 teeth number and position (Fig. 2B). Renopore in vaginal aperture including ghost teeth in ascending row, 6 to 9 in descending on dorsum at anterior right side of pericardium (Fig. 7B). row, few in ascus. Teeth narrow blade-shaped (Fig. 4B),

Fig. 1. The four mangrove Elysia species: A, E. leucolegnote Jensen, 1990; B, E. singaporensis, new species; C, E. bangtawaensis Swennen, 1998; D, E. bengalensis, new species. A and B photographed in-situ, respectively Pak Phanang Bay and Singapore, C and D photographed in captivity. Ab, Bb, Cb, Db group photographed in-situ, respectively Pak Phanang Bay, Singapore, Bangtawa, and Andhra Pradesh (India). D and Db photographed by Dr. S. Bouillon.

31 Swennen Ð Large mangrove-dwelling Elysia species used teeth often with slightly incurved tips, fi nely denticulate between Mar.1997 and Oct.2010; Inland ditch near Ban Di, about cutting edges. Tooth length 140-180 µm. Oesophagus short 6¡52'17"N, 101¡18'48"E, 5-10 during several visits between 1999 about as long as pharynx. Relatively massive intestine plicate and 2002 (site now converted into shrimp ponds); Inner Pattani Bay, near anus. Glandular tissue of digestive gland starts in about 6¡53'50"N, 101¡20'49"E, 2 to 30 individuals during several visits between 1997 and 2007; Straits of Malacca, Johor State, main branches far from stomach (Fig. 5B). Penis, unarmed, 01¡21'30"N, 103¡30'37"E, 3 specimens, 17 Oct.2009; Andaman tapered, (Fig. 6B). Sea, Thailand, mangrove east of Krabi at about 08¡03'17"N, 098 53'57"E, one, 3 Oct, 2010. Arabian Sea, estuary Mandovi River, Goa, Biology. – Egg strings have been found in May and October. India, material seen. [See photo: Jagtap et al., 2009]. Bay of Bengal, Strings had 3Ð4 ova per capsule, but older string had one Andhra Pradesh, about 20 km south of Kãkinãda, 16°51'40"N, veliger per capsule. Yellow extra-capsular yolk, in interrupted 82¡15'10"E, some specimens, 4 Dec.2002 [specimens sent to me line over whole length of string (Fig.7F, G). The strings were by Dr S. Bouillon]; South Pacifi c Ocean, Australia, Queensland, irregular on algae, but more or less in counter-clockwise, near Coolangatta, about 28¡10'S, 153¡32'E, 28 Jul.2007 (photos loose spirals on glass in captivity. The longest measured by G. Cobb in Coleman, 2008). string was 320 mm. Live colouration. – Prominent reddish to orange, glandular warts along parapodial borders. White spots of different Elysia bangtawaensis Swennen, 1998 sizes and fi ne reddish specks dispersed on dorsal and ventral (Figs. 1-7) sides, including foot sole. Green ductules of digestive gland cover renal part in most specimens; they do not reach tips Syn.: Elysia bangtawaensis Swennen, 1997. (Rudman, 2007; of rhinophores. Tips pale or coloured by white and orange Coleman, 2008). Incorrect year of publication due to a difference spots (Fig. 1C). Individuals with digestive gland yellowish of the year printed on the volume and the date of distribution starting in the central parts have not fed for some months of the last issue which is stated as 18 April 1998. as confi rmed in captivity.

Material examined. – Gulf of Thailand. Pak Phanang Bay about External characters. – Length alive up to 52 mm. 08¡36'10"N, 99¡58'16"E and 8¡29'19"N, 100¡10'58"E, 2 respectively Renopericardial prominence oval, length less than twice Elysia leucolegnote, 4 specimens near hundreds of 28-30 Sep.2007; width. Two major dorsal vessels on posterior side and one, Bang Tawa, about 06¡51'28"N, 101¡09'31"E, type locality, yearly

Fig. 2. Renopericardial bulbs and attachments of dorsal vessels of: A,: Elysia leucolegnote; B, E. singaporensis, new species; C, E. bangtawaensis; D, E. bengalensis, new species. The highest variation in number of vessels is shown by E. singaporensis. Figures in the bulbs of this species indicate the number of specimens with that pattern found in a sample of 28 individuals.

Fig. 3. Lateral view of the shape of the pharynx: A, Elysia leucolegnote; B, E. singaporensis, new species; C: E. bangtawaensis; D, E. bengalensis, new species.

Fig. 4. Lateral view of the radular teeth: A, Elysia leucolegnote; B, E. singaporensis, new species; C, E. bangtawaensis; D, E. bengalensis, new species.

32 THE RAFFLES BULLETIN OF ZOOLOGY 2011 sometimes two, thinner vessels on both lateral sides (Fig. Biology. – Copulation by lying with the right side of the 2C). Renopore on dorsal side of pericardium in posterior frontal part of the body against the partner. Penis goes into part between heart and renal part (Fig. 2C). Vaginal aperture the underside of the right parapodium. At the end they may on ventral side of right parapodium near anterior border form a ball when penises withdraw. Egg strings without (Fig. 7C). extra-capsular yolk, up to 720 mm long, irregularly coiled over substrate; in captivity irregular spiral on glass. Internal characteristics. – Pharynx small, length about 400 µm, dark pigment band over sides, ascus vestigial (Fig. During low tide, when there is no water current, the slugs 3C). Radula with 7 to 9 teeth in ascending limb and same were often found in dense aggregations resembling fallen number in descending limbs. Teeth blade-shaped, more than leaves on bare mud in shallow, water-fi lled depressions or 80 very fi ne denticles along cutting edge, tooth length in gullies in mangrove forests. Depression can be as small as specimens of 23 to 49 mm length alive similar, about 67 a footprint of a water buffalo or human. When a sudden µm (Fig. 4C). Oesophagus narrow, length about 2.5 times movement in the water occurred (e.g., a foot step), the slugs length of pharynx. Stomach wide, green, glandular tissue closed their parapodia and become detached from the fl oor. of digestive gland starts in stomach. Intestine pale, wide, Their feet clearly did not strongly adhere to any substrate. runs upward from anterior part stomach (Fig. 5C). Penis The slugs rolled around across the substrate usually to unarmed, conical (Fig. 6C). Male follicles more central, slightly deeper parts from where they can be easily swept larger and less numerous than female follicles. out by another foot step. I was unable to determine what

Fig. 5. Dorsal view of the main parts of the digestive systems: A, E. leucolegnote; B, E. singaporensis; C, E. bangtawaensis. Legend: dg Ð digestive gland; in Ð intestine; oe Ð oesophagus; op Ð oesophageal pouch; ph Ð pharynx; st Ð stomach. Note that two specimens per species were studied for making the drawings. They were collected from the same site and day. When the distribution of the coloured glandular parts is inﬂ uenced by the feeding condition then the differences do not indicate differences between species.

Fig. 6. Shape of the penis of: A, Elysia leucolegnote; B, E. singaporensis, new species; C, E. bangtawaensis; D, E. bengalensis, new species.

33 Swennen Ð Large mangrove-dwelling Elysia species happened when the tide moved in. The movements of the tidal kept the parapodia expanded also during the night. Closing currents are strong and longer lasting than a step; I would the parapodia over the body occurred when the tray was expect that they be dislodged from the substrate. However, touched, but also when exposed to direct sunlight that even when the fl ood came in, the water turbidity was high due to stimulated them to move to a shaded position such as below the heavy load of mud particles and visibility immediately other specimens (Swennen, 1998). This made it unlikely that decreased to zero. Although the slugs are likely dislocated resembling a leaf was procryptic. Feeding trials with algae by the tidal currents, I have no idea how they could select from outside the forest had no success. depressions that contain suffi cient water for survival during the next low tide. I noticed during a subsequent visit that tiny green algae were present on the dry mud surface between mangrove roots above When discovering E. bangtawaensis in March 1997, I had not mean high tide level. Mud and algae could not be separated dealt with sacoglossans since 1959 (Swennen, 1961) and had and were placed in-situ in the trays with the slugs. This time, missed the developments in knowledge since then. Therefore, the slugs were attracted by the algae and appeared to feed on I wondered what was the meaning of their behaviour and them. At my request, Mr. M. Lavaleye (Royal Netherlands what was their food, because there were no visible algae Institute for Sea Research, Texel) conducted phytopigment in the shallow pools with a soft mud bottom. Specimens analyses of an E. bangtawaensis and the algae. That showed were put in trays in the laboratory with the local mud and that the spectra of both organisms were almost identical. seawater. They soon became immobile and stretched out Mr. K. A. Sjollema (Laboratory for Electron Microscopy, their parapodia as seen in the fi eld. When undisturbed, they University of Groningen) made images of a piece of slug

Fig. 7. Position of vaginal apertures in: A, Elysia leucolegnote; B, E. singaporensis, new species; C: E. bangtawaensis; D, E. bengalensis, new species. E, part of an egg string of Elysia leucolegnote showing uninterrupted band of extra-capsular yolk; F, part of an egg string of E. singaporensis showing interrupted band of extra-capsular yolk and more than one ovum per capsule; G, E. singaporensis, new species, producing an egg string in algae in aquarium. Position of vaginal apertures indicated by arrow.

34 THE RAFFLES BULLETIN OF ZOOLOGY 2011

Table 2. Differences and similarities of position of vaginal aperture and shape of ascus and radular teeth in the four large mangrove Elysia species.

E. leucolegnote and E. singaporensis E. bangtawaensis and E. bengalensis Vaginal aperture On dorsal side of right parapodium anterior of On anterior ventral side of right parapodium renopericardium together with renal aperture Ascus Prominent on long stalk largely free of ventral side Vestigial on ventral side of pharynx of pharynx Radular teeth Narrow serrate blades Wide serrate blades tissue and the alga. They showed that in the cells of the PARATYPES: 2 ex. (ZRC.MOL.2973), same site and date as digestive gland of E. bangtawaensis several chloroplasts holotype. occurred that were similar to the ones in the algae. Then, OTHERS: about 7 specimens internally rotten, same site and date Dr. W. Stolte (Royal Netherlands Institute for Sea Research) as holotype. placed a live slug in a respirometer and measured the oxygen from incoming and outgoing seawater. The specimen had a Etymology. – The species name is derived from the Bay of length of 3.4 cm, and had no contact with any algae for 20 Bengal into which the Godovari River discharges. days. The space was too small for allowing the slug to fully expand its parapodia in the measuring chamber, but still the Live colouration. – Basic colour green from content of measurements in various light conditions showed a gross digestive gland. Irregular row of larger white spots along photosynthesis of 4.2 micromole O /h and a respiration of parapodial border. Conspicuous white specks irregularly 2 spaced all over dorsal and ventral sides, not on foot sole. 1.3 micromole O2/h illustrating that E. bangtawaensis can indeed act as a plant with real photosynthesis taking place Tips of rhinophores and vaginal aperture dark coloured. in dimmed light (117 micromole photons/m2/s) producing Dark remains visible on both areas in specimens stored in more oxygen than used. This is known under such names 70% ethanol or 6% formaldehyde for ﬁ ve years. as chloroplast retention, kleptoplasty, chloroplast symbiosis, solar-powered slugs. Further investigations were stopped due External appearance. – Length alive up to 50 mm. to lack of time and funding. Rhinophores with groove over whole length. Parapodia and body smooth. Anterior foot corners slightly extended, Discussion. – Specimens from India, Malaysia, Thailand bluntly pointed. Renocardial prominence oval, length less and Australia show the same colour pattern. than twice width; pair of major dorsal vessels on posterior side of prominence; one to four smaller vessels on each lateral side (Fig. 2D). Renopore could not be detected in Elysia bengalensis, new species available material. Male aperture below right rhinophore; (Figs. 1Ð7) anus and oviducal apertures close together in upper part of lateral groove. Vaginal aperture on ventral side of right Material examined. – HOLOTYPE: 1 ex. (length alive 43 mm) parapodium near anterior border as in E. bangtawaensis, (ZRC. MOL.2972), Bay of Bengal, Andhra Pradesh, delta of but black marked (Fig. D). Godovari River about 20 km south of Kãkinãda, between 16°43'N, 16¡52'N and 82¡15'E, 82¡22'E, May 2001, coll. Steven Bouillon. Internal characteristics. – Pharynx small, length about 380 µm, with strong dorsal septate muscle, ascus vestigial. Radula with 7 teeth in ascending row, 10Ð11 in descending row, and bundle of about 15 worn teeth in ascus. Length of radular teeth 68–80 µm (two individuals). Teeth blade- shaped, blade about 38% of total length, 20-35 denticles on cutting edge (Fig. 4D). Shape of blade slightly differs from E. bangtawaensis, but has fewer and larger denticles. Penis unarmed, conical (Fig. 6D).

DISCUSSION

The four large mangrove Elysia species can be divided into two groups of two species according to some anatomical differences and similarities (Table 2). The four species live in shaded mangrove habitats and are often found congregated Fig. 8. Map showing the known distribution of the four mangrove in pools. They are usually found in shallow pools and Elysia species discussed in this paper. The sites in the Gulf of drains high in the intertidal zone during low tide, but they Thailand and Hong Kong were in close proximity and one mark cannot live amphibiously. All show a similar behaviour as may indicate more than one collection site.

35 Swennen Ð Large mangrove-dwelling Elysia species mentioned in the description of E. bangtawaensis, thus can Crabs or birds may have damaged these specimens, but tests likely be grouped under the name solar-powered slugs. That in captivity showed that E. bangtawaensis and E. leucolegnote does not mean that they like direct sun light, they seem to are highly unpalatable for local predatory fi sh and shrimps. dislike strong light and prefer the shade (Swennen, 1998). Only the nudibranch Gymnodoris pattani Swennen, 1996, Tests showed that E. bangtawaensis and E. leucolegnote swallows them entirely, notwithstanding the Gymnodoris can keep their chloroplasts for months; E. bengalensis was usually much smaller than the Elysia. The discussed and E. singaporensis could not be tested. Long-term species have a long life span compared to the small Elysia functional chloroplast retention has only been found in the siamensis Swennen, 1998, that reach only the age of three Plakobranchoidea. However, several large Elysia species do months. Individuals of E. bangtawaensis were kept up to not have this quality (Händeler et al., 2009). 14 months in captivity, and of E. leucolegnote up to more than four years. The food alga of Elysia leucolegnote and E. bangtawaensis is Boodleopsis c.f. pusilla that grows in high mud between I am unable to indicate specific differences in habitat mangrove roots. Here it becomes not wetted during neap preferences among the species. E. bangtawaensis was found high tides in the summer months in the Gulf of Thailand, in the same area as E. bengalensis in the Bay of Bengal. when the water level is lower due to the monsoon wind. This Elysia bangtawaensis often occurs with E. leucolegnote in means that these slugs cannot reach their food at libitum, the same forest patch, and were regularly found mixed on but have to wait for spring high tides. Experiments showed several sites in the Gulf of Thailand. In the Pak Phanang that they can feed for many hours and can survive without Bay, only a few specimens of E. bangtawaensis were found food for two to three months. However, then they become in rather remote sites in the mangrove forest in September gradually smaller and their green colour changes into 2007, while at the same time E. leucolegnote was numerous yellowish. This may suggest that they not only need new over most of the area. Therefore, it was a surprise to see both chloroplasts, but also some additional substances from their species numerous in April and October 2008. No obvious food alga. The famished slugs regain their colour and size changes were noted in the habitats. Generally, water in which after they feed on algae. Specimens of Elysia bangtawaensis, the slugs were found was clear during low tide, but turbid E. leucolegnote, and E. singaporensis that are not green but during high tide. During low tide, the water quality was likely yellowish, are rarely found among normal specimens of these poor due to rotting wood and leaves, but the slugs tolerate species in the fi eld. They look similar as specimens that in it. Elysia leucolegnote has even been found in apparently captivity were deprived of food for more than two months. anaerobic water conditions with white colonies and borders It remains unclear if this is the results of feeding diffi culties of sulphur bacteria around them both in Starling Inlet (Hong or other problems. The noted differences in the extent of Kong) and near Chumphon (Thailand) where the high tides the glandular tissue in the main branches of the digestive does not reach these sites during neap tides. system (Fig. 5) has been based on the greenish colour in two specimens per species collected from the same site and The known distribution of the four discussed Elysia species day. In retrospect, it may be possible that the distribution is limited to the tropical West Pacific and East Indian of the coloured glandular parts has been infl uenced by the Oceans on both sides of the equator to about 22¡North and physical condition of the slugs and if so, would not indicate 28¡South (Fig. 8). Elysia bangtawaensis seems to have the differences among species. widest distribution. However, the ranges of the species are in fact nearly unknown as turbid intertidal and mangrove I did not study the reproductive organs in detail. The status habitats are generally poorly studied. Mangrove forests of the reproductive cycle seemed to induce large differences are notoriously diffi cult to explore due to lack of trails, in shape and size of albumen gland, bursae, and gonadal orientation marks, unobstructed views, and an inclement follicles. The reproductive periods seemed short and human environment that include deep soft mud, biting ants, remained unpredictable for me. The long egg strings were stinging mosquitoes, and skin-cutting oysters. The discoveries not always found in the fi eld nor deposited in captivity. The of Elysia leucolegnote in the Gulf of Thailand and Andaman egg strings of E. leucolegnote, E. singaporensis contain a Sea are the fi rst reports outside Hong Kong. I expect that band of yellowish extra-capsular yolk (Boucher, 1983) that the four species have a broader distribution than could be is interrupted in E. singaporensis. Three to four ova were reported here. found per capsule in fresh laid strings of the latter species, but only one veliger in older strings. Perhaps the other eggs Although there is still much potential mangrove habitat were eaten by the developing veliger as no remains were available where the four mangrove Elysia species can occur, detected, or the string was aberrant. The egg masses of E. these habitats are at increasing risk of reduction. I have bengalensis are still unknown. observed large-scale destruction of mangrove forests in SE Asia since my fi rst visit in 1984. For example the small Damaged specimens were frequently found. They lacked remnant of the mangrove forest near the village Bangtawa small or large parts along the border of the parapodia or had that was saved by the villagers from being converted into holes in the parapodia. These specimens behaved normally shrimp ponds, and is the type locality of Elysia bangtawaensis and small wounds completely healed when kept and fed in Swennen, 1998, Gascoignella nukuli Swennen, 2001, and captivity. Such damages seemed to occur more often in E. Costasiella coronata Swennen, 2007; has recently been bangtawaensis and E. singaporensis than in E. leucolegnote. damaged by mechanically digging a wide ditch with high

36 THE RAFFLES BULLETIN OF ZOOLOGY 2011 dikes along both sides. The site near Ban Di, type locality Jensen, K. R., 1990. Three new species of Ascoglossa (Mollusca, of Gascoignella jabae Swennen, 2001, was converted into Opisthobranchia) from Hong Kong, and a description of the shrimp ponds that have already been disused. Today, disused internal anatomy of Costasiella pallida Jensen, 1985. In: ponds are not restored to their natural condition and remain Morton, B. (ed.), Proceedings of the Second International Marine Biological Workshop: the Marine Flora and Fauna of empty and barren. Dikes, concrete sluices and other man- Hong Kong and Southern China, Hong Kong, 1986: 419Ð432. made constructions around the ponds block tidal fl ows that Hong Kong University Press. could aid in the natural recovery of these areas. On-going conversion of mangrove forests to shrimp ponds put these Jensen, K. R., 1996. Phylogenetic systematics and classifi cation of the Sacoglossa (Mollusca, Gastropoda, Opisthobranchia). habitats and associated species at great risk for species loss Philosophical Transactions of the Royal Society London B and possible extinction. 351: 91Ð122. Jensen, K. R., 2003. Distribution, diets and reproduction of Hong Kong Sacoglossa (Mollusca: Opisthobranchia): a summary of ACKNOWLEDGEMENTS data, 1980-2001. In: Morton, B. (ed.). Perspectives on Marine Environment Change in Hong Kong, 1977–2001: 347Ð365. I am grateful to Mr. M. Lavaleye (Royal Netherlands Institute Hong Kong University Press. for Sea Research, Texel), Mr. K. A. Sjollema (Laboratory Kelaart, E. F., 1858. Description of new and little known species for Electron Microscopy , University of Groningen), Dr. of Ceylon nudibranchiate molluscs and zoophytes. Journal of Jiro Tanaka (Tokyo University of Marine Science and the Royal Asiatic Society (Ceylon Branch), Colombo 3 (1): Technology) and Dr. W. Stolte (Royal Netherlands Institute 84Ð139. for Sea Research,Texel) for investigating and enlightening my Pease, W. H., 1871. Descriptions of nudibranchiate Mollusca understanding of food and behaviour of Elysia bangtawaensis. inhabiting Polynesia. American Journal of Conchology 4: I also like to thank Prof. Steven Bouillon of the Vrije 299Ð305. Universiteit Brussel for allowing me to study some of his Pruvot-Fol, A., 1954. Mollusques opisthobranches. Faune de Elysia specimens from India, Mr. Somsak Buatip for his France 58: 1Ð457. help to elucidate the distribution of the species in the Gulf Rudman, W. B., 2007 (August 8) Elysia bangtawaensis Swennen, of Thailand, Andaman Sea and Johor. Thanks are also due 1997. [In] Sea Slug Forum. Australian Museum, Sydney. to two anonymous referees for their numerous suggestions Available from http://www.seaslugforum.net/fi nd/elysbang for improving my manuscript. Swennen, C., 1961. On a collection of Opisthobranchia from Turkey. Zoölogische Mededelingen 38: 41Ð75. LITERATURE CITED Swennen, C., 1996. Gymnodoris pattani, a new dorid nudibranch from Pattani Bay, Gulf of Thailand (Gastropoda, Nudibranchia). Bulletin Zoölogisch Museum, University of Amsterdam 15 (6) Boucher, L. M., 1983. Extra-capsular yolk bodies in the egg masses 1996: 41Ð47. of some tropical Opisthobranchia. Journal of Molluscan Studies 49: 232Ð241. Swennen, C., 1998. Two new gastropods, Elysia bangtawaensis and E. siamensis from southern Thailand. Bulletin Zoölogisch Coleman, N., 2008. Nudibranchs encyclopedia. Neville Coleman’s Museum, University of Amsterdam, 16 (6), 1997: 33Ð39. Underwater Geographic Pty Ltd, Australia, 416 pp. Swennen, C., 2001. Two new sacoglossans (Gasteropoda; Händeler, K., Y. P. Grzymbowski, P. J. Krug, H. Wägele, Opisthobranchia) from Thailand. Beaufortia 51 (3): 75Ð81. 2009. Functional chloroplasts in metazoan cells Ð a unique evolutionary strategy in animal life. Frontiers in Zoology, 6: Verrill, A. E., 1901. Additions to the fauna of the Bermudas from 28 (18 pages; doi:10.1186/1742-9994-6-28). the Yale Expedition of 1901 with notes on other species. Transactions of the Connecticut Academy of Sciences 11: Jagtap,T.G., P. S. Shenai-Tirodkar, S. B. Savant, V. A. Kulkarni, 15Ð62. & Z. A. Ansari, 2009. Elysia bangtawaensis Swennen (Nudibranch) from the mangrove habitat of Mandovi estuary (Goa) central westcoast India. Current Science 96: 30Ð33.

37 THE RAFFLES BULLETIN OF ZOOLOGY 2011

THE RAFFLES BULLETIN OF ZOOLOGY 2011 59(1): 39–46 Date of Publication: 28 Feb.2011 © National University of Singapore

A NEW SUBSPECIES OF AMPHIDROMUS (AMPHIDROMUS) ATRICALLOSUS FROM SINGAPORE (MOLLUSCA: GASTROPODA: CAMAENIDAE)

Siong Kiat Tan Rafﬂ es Museum of Biodiversity Research, Department of Biological Sciences, National University of Singapore, Block S6, Science Drive 2, #03-01, Singapore 117546, Republic of Singapore Email: [email protected]

Sow Yan Chan VBox 888313, Singapore 919191. Email: [email protected]

Somsak Panha Animal Systematics Research Unit, Department of Biology, Faculty of Science, Chulalongkorn University, Bangkok, 10330, Thailand Email: [email protected]

ABSTRACT. – The arboreal land snail of the genus Amphidromus Albers, 1850, found in Singapore is often identiﬁ ed in the literature as Amphidromus atricallosus perakensis Fulton, 1901. Studies of A. atricallosus perakensis material from various localities in Peninsular Malaysia showed that the Singapore population is distinct. Results from earlier studies on molecular phylogeny and allozyme analyses also suggest genetic discontinuity from other A. atricallosus subspecies. The Singapore population is herein described as A. atricallosus temasek, new subspecies, and can be differentiated from A. atricallosus perakensis by several morphological characters.

KEY WORDS. – Singapore, Gastropoda, Camaenidae, Amphidromus atricallosus temasek, new subspecies.

INTRODUCTION The recent revision of the subgenus Amphidromus by Sutcharit & Panha (2006) clarified some of the confusion. Four The arboreal land snails of the genus Amphidromus Albers, subspecies of A. (Amphidromus) atricallosus (Gould, 1843) 1850, are well known within Southeast Asia (Fulton, 1896; with distinct shell characters and geographic distribution are Collinge, 1903; Bartsch & Solem, 1971; Benthem-Jutting, currently recognized, the nominate subspecies, A. atricallosus 1949), particularly for their colourful shells and their curious classiarius Sutcharit & Panha, 2006, A. atricallosus dimorphic coiling (Pilsbry, 1900; Berry, 1963; Sutcharit et leucoxanthus (von Martens, 1864), and A. atricallosus al., 2007; Schilthuizen & Looijestijn, 2009). Two distinct perakensis Fulton, 1901. Amphidromus atricallosus subgenera of the genus Amphidromus are currently recognised perakensis, although ﬁ rst described from Perak, Malaysia, has (Sutcharit et al., 2007; Chan & Tan, 2010), and within the been widely documented in Singapore (e.g., Ng & Lim, 1992; respective subgenera, several species groups are known Chou et al., 1994; Ho, 1995 [as A. atricallosus]; Sutcharit and members are characteristically similar in shell form & Panha, 2006; Chou & Tan, 2008). Records suggest that (Pilsbry, 1900; Laidlaw & Solem, 1965). Based on locality this subspecies is rather widespread in Peninsular Malaysia information given in original descriptions and museum (Laidlaw & Solem, 1961; Sutcharit & Panha, 2006), but its collections, many species and subspecies have been reported elusiveness is illustrated by Sutcharit & Panha (2006) who from West Malaysia and Singapore (Pilsbry, 1900; Berry, mentioned the present distribution is conﬁ ned to the southern 1963; Laidlaw & Solem, 1961; Maassen, 2001). However, part of Peninsular Malaysia and Singapore. many of these records remain dubious and the taxonomic status of several species remains uncertain without reliable Recently, we had the opportunity to examine some topotypic locality information (Laidlaw & Solem, 1961). material of A. atricallosus perakensis and comparison with material from Singapore revealed subtle but consistent differences in shell morphology. Additional material from

39 Tan et al. – New Amphidromus from Singapore other localities in Peninsular Malaysia was obtained for National Park Survey, 30 Apr.1992; 1 D SH 45.0 x SD 26.4 (ZRC examination (see Comparative material), and we conclude 1994.4118), outside Nee Soon range, Seletar Reservoir Park (NS that the differences are consistent enough to recognise the 43), coll. H. K. Lua, 1994; 1 D (CUMZ 2133), Nee Soon Nature Amphidromus from Singapore (hitherto referred to as A. Reserve, coll. S. Panha, 26 Jun.1998; 2 D (CUMZ 2067), Nee Soon Nature Reserve, coll. S. Panha & Ms Loua, 19 Feb.2000; atricallosus perakensis) as a distinct and yet undescribed 10 D (CUMZ 2198), Nee Soon Nature Reserve, coll. S. Panha & subspecies. Additionally, results of recent phylogenetic C. Sutcharit, 19 Feb.2001; 11 D (CUMZ 2633), Nee Soon Nature studies imply that the Singapore population is not closely Reserve, coll. S. Panha & P. Prasarnkok, 04 Mar.2004; 2 S (1 juv.) related to the other A. atricallosus subspecies (see Sutcharit SH 43.4 x SD 26.8 (ZRC.MOL.2843), Nee Soon Swamp Forest, et al., 2007; Prasankok et al., 2007). The description of this coll. H. H. Tan & A. F. S. L. Lok, 13 Oct.2008; 1 D (juv.) (ZRC. Amphidromus from Singapore as a new subspecies forms MOL.3059), Nee Soon Swamp Forest, coll. Y. C. Ng, Sep.2010; the basis of this study. 1 D SH 43.5 x SD 27.1 (ZRC.MOL.3057), Nee Soon Swamp Forest, coll. S. K. Tan & S. Y. Chan, 03 Oct.2010; 1 S SH 38.9 x SD 25.1 (ZRC.MOL.3060), Nee Soon Swamp Forest, edge of swamp forest, behind Nee Soon Range, coll. P. X. Ng, B. Y. Q. MATERIALS AND METHODS Ng & C. K. Yeo, 03 Nov.2010.

The types are deposited in the Zoological Reference Collection Non-type material. – 3 D SH 42.1 x SD 24.9–SH 42.9 x SD 26.4 (ZRC) of the Raffl es Museum of Biodiversity Research, (ZRC 1989.1109–1111), no data; 10 D (9 juv.), SH 42.7 x SD 26.5 National University of Singapore, and Chulalongkorn (ZRC 1989.1135–1144), no data. SINGAPORE: 1 D (juv.) (ZRC University, Museum of Zoology (CUMZ), Bangkok, 1990.1528), Nee Soon, on tree along road, coll. CLM, Sep.1969; Thailand. Additional material mentioned in the text is 3 S (juv.) (ZRC 1998.3373-3377), Nee Soon Swamp Forest, coll. identifi ed by the following abbreviations: CBB (Collection C. M. Yang et al., 9 Sep.1988; 1 D SH 44.3 x SD 25.4 (ZRC of B. Boo), CSY (Collection of Chan S.-Y.), FJK (Collection 1990.10741), Nee Soon Swamp Forest, NW sector, in leaf litter on open scrubland, coll. K. Lim, 9 May 1990; 3 S (juv.), 2 D (broken, of Foon J. K.), M.E. (Collection of M. E. Marzuki), TSK not measured) (ZRC.MOL.3061) Nee Soon Swamp Forest, dead on (Collection of Tan S. K.). Other abbreviations mentioned ground, coll. S. K. Tan, 2009; 2 D (juv.) (CSY 409.3.4.30), Pulau are: SH = shell height; SD = shell diameter; D = dextral; Tekong, dead on ground among low shrubs, coll. S. Y. Chan, 25 S = sinistral. Shell height is measured as the apex to the Dec. 1993; 1 S SH 43.7 x SD 26.2 (CSY 409.3.4.31), Dairy Farm, lowest part of the basal side of the peristome parallel to dead on road path beside forest, coll. S. Y. Chan, 25 Oct. 1994; the coiling axis, and shell diameter is the widest part of 1 S SH 39.5 x SD 24.8, Central Catchment area, on artifact, coll, the body whorl (including the lip) perpendicular to the R. Koh, Apr. 1995; 1 D SH 37.4 x SD 25.1 (TSK 11017), Mandai coiling axis. Measurements were taken in mm. In lots with Road Track 7, on fi shtail palm, coll. S. K. Tan, 04 Apr.1999; 2 multiple specimens, sizes shown indicate the dimension of the D (juv.), (ZRC.MOL.3065), Lower Peirce Reservoir, arboreal on leaves, forest edge, coll. A. F. S. L. Lok, 23 Jul.2010. MALAYSIA: smallest and largest specimens examined. Shells that have not 1 D (broken, not measured) (CSY 409.3.4.14), Gunung Ledang developed the expanded and thickened outer lip are deemed (Mount Ophir), Johor, dead, among leaf litter in forest, coll. S. Y. juvenile (juv.), and are not taken into consideration. Chan et al, 13 Oct.1996.

Diagnosis. – Shell relatively large, ovate, chirally dimorphic TAXONOMIC ACCOUNT coiling. Whorls 5½–6¾, slightly convex, surface generally smooth with indistinct axial striations or growth lines. Colour CAMAENIDAE PILSBRY, 1895 uniformly yellow with a thin white subsutural line (shells of living animals yellowish-green due to the dark colouration of Amphidromus ALBERS, 1850 soft body showing through), varix always absent. Columella simple, straight. Periostracum thin, transparent. Peristome Amphidromus (Amphidromus) atricallosus in mature animals white, thickened and expanded except (Gould, 1843) for parietal side, outer lip reﬂ ected but not adnate. Parietal callus usually inconspicuous, very rarely thickened to being Amphidromus (Amphidromus) atricallosus temasek, whitish or translucent. new subspecies (Figs. 1A, 2A, 3A) Animal body very pale greyish white in colour, reticulated, the recessed parts being darker, light brown around the head Amphidromus atricallosus perakensis – Ng & Lim, 1992: 262; Chou to greyish on the body. Mantle edge dark cream coloured to et al., 1994: 73; Sutcharit & Panha, 2006: 21–22, Fig. 4. F–G, brown around the peristome. Upper and lower tentacles dark Fig. 11. A–C, Fig. 12. C; Prasankok et al., 2007; Sutcharit et al, 2007; Chou & Tan, 2008: 57; Lok & Tan, 2008. yellow, central part of the head between the upper tentacles Amphidromus atricallosus – Ho, 1995: 108–109. occasionally pigmented with yellow, foot with longitudinal yellow stripe on either side. Material examined. – SINGAPORE: Holotype: 1 S SH 44 x SD 25.6 (ZRC.MOL.3058), Nee Soon Swamp Forest, Singapore, Distribution. – The distribution of A. atricallosus temasek, coll. S. K. Tan & S. Y. Chan, 03 Oct.2010. Paratypes: 2 D SH new subspecies, is largely confined to Singapore, and 42.7 x SD 26–SH 46.4 x SD 27.7, 2 S SH 40.1x SD 25–SH 40.8 tentatively, Johor, southern Peninsular Malaysia (Sutcharit & x SD 25.1 (ZRC.MOL.2992), Nee Soon, coll. E. Alfred; 1 D SH Panha, 2006 [part]; Prasankok et al., 2007 [part]; this study). 46.9 x SD 28.3 (ZRC.1992.3163), Nee Soon Swamp Forest, coll. Known Singapore localities include: Seletar (Hanitsch, 1908),

40 THE RAFFLES BULLETIN OF ZOOLOGY 2011

Nee Soon Swamp Forest, Bukit Timah Nature Reserve, southern part of Peninsular Malaysia and the Riau islands of Mandai, and Pulau Tekong (Ng & Lim, 1992; Chou & Indonesia. In Singapore, these arboreal snails are apparently Tan, 2008; Lok & Tan, 2008). Although a specimen from restricted to the remnant forests. Preference for particular Johor (CSY 409.3.4.14) is provisionally determined to be tree species has not been observed or reported, and the this subspecies in the material examined, identification animals can be found on a wide variety of plants and on was based on a weathered and broken specimen with only man-made structures. the body whorl intact. Veriﬁ cation with better preserved material would be required. Available evidence suggests Etymology. – The subspecies epithet is derived from A. atricallosus temasek to be endemic, but this has to be Temasek, the historical name of the type locality Singapore. veriﬁ ed with greater sampling effort, particularly around the It is used as a noun in apposition.

Fig. 1. A, Amphidromus atricallosus temasek, new subspecies: Holotype, SH 44 x SD 25.6 (ZRC.MOL.3058), Nee Soon Swamp Forest, Singapore; B–E Amphidromus atricallosus perakensis (Fulton, 1901): B, SH 45 x SD 26.4 (TSK 11041), Bukit Serdam, Pahang, West Malaysia; C, SH 47.1 x SD 25.3 (CBB), Tapah Hill, Perak, West Malaysia; D, SH 47.2 x SD 27.6 (ZRC.MOL.3064), Pulau Tulai, near Pulau Tioman, West Malaysia; E, SH 46.4 x SD 24.2 (ZRC.MOL.3062), Gunung Genting, Perak, West Malaysia; F, Amphidromus atricallosus atricallosus (Gould, 1843): SH 45 x SD 25.7 (CSY 409.4.4.16), Ranong, Thailand; G, Amphidromus atricallosus leucoxanthus (von Martens, 1864): SH 51 x SD 28.5 (CSY 409.3.49.1), Chantaburi, East Thailand.

41 Tan et al. – New Amphidromus from Singapore

Remarks. – The thin white subsutural line and simple and & Panha, 2006, is endemic to Koh Tachai, off Pangnga, straight columella that is never twisted is diagnostic. Shells in the Andaman Sea and is similar to the nominotypical of juveniles can also be separated from A. atricallosus subspecies, but can be separated by its consistently smaller perakensis Fulton, 1901, by lack of a white zone around the size and being invariably sinistral. Amphidromus atricallosus columella (see Fig. 2). Although the convexity of the whorls leucoxanthus (von Martens, 1864), is also similar in shell of A. atricallosus perakensis is evidently variable throughout form to the nominotypical subspecies, but differs by the lack West Malaysia populations, shell whorls of A. atricallosus of a darkened parietal callus (Fig. 1. G) and its colour varies temasek, new subspecies, seem to be generally more convex from yellowish to white shells with brown streaks which in profi le and resulting in the suture appearing more sunken is also called form laidlawi Solem, 1965. And except for in comparison. Living A. atricallosus temasek individuals are A. atricallosus perakensis, one or more varices are usually easily distinguished by the soft body colouration, and a pale present in shells of the other subspecies (e.g., Fig. 1. G). yellowish-green shell with a thin white subsutural line (Fig. For details, see Sutcharit & Panha (2006). 3. A). The SEM images of the radula and genital system of A. atricallosus temasek, new subspecies, are described Fulton (1901) who had numerous specimens (50 D, 48 S) in detail and fi gured in Sutcharit & Panha (2006, as A. (A.) remarked that the columellar character is constant in all atricallosus perakensis [CUMZ 2198; herein designated examined specimens. However, Sutcharit & Panha (2006) paratypes]; radula, pg. 16: Fig. 11A–C; genital system, pg. observed that the columella varies from simple and straight 17: Fig. 12C). to strong and folded, apparently based on insuffi cient material and the assumption that material examined, including lectotype and paralectotype of A. perakensis, and recent DISCUSSION material from Pulau Tulai and Singapore, are conspecifi c. None of the A. atricallosus temasek examined in this study Amphidromus atricallosus temasek, new subspecies, is most has a twisted columella. Very rarely, the columella may similar in shell morphology to A. atricallosus perakensis appear slightly curved, but is never twisted or developed Fulton, 1901, which it has been hitherto confused with. into a plate. Conversely, the columella is rather consistently Amphidromus atricallosus perakensis was fi rst described as twisted in the examined specimens from various Malaysian a distinct species based on the characteristic twisted plait localities (see Comparative material). Fulton (1901) also of the columella (Fulton, 1901; Collinge, 1903; Laidlaw, noted that the extent of this character is variable, from rather 1933; Benthem-Jutting, 1949). Laidlaw & Solem (1961) inconspicuous to forming a plate-like projection or tooth in and Solem (1965) regarded perakensis as a mere form of many, presumably mature, individuals. Nevertheless, even A. atricallosus which was described briefl y in Gould, 1843, in specimens without a signifi cant plait in the columella, the and more fully described and fi gured in Gould, 1844, which slight twist often forms a shallow groove at the parietal side additionally mentioned a specimen without the dark parietal which is not seen in A. atricallosus temasek. Thus even when callus as possibly coming from Singapore. Current consensus the variables are considered, we concur with Fulton (1901) recognises perakensis as a valid subspecies (Maassen, 2001; that this character is generally consistent for A. atricallosus Sutcharit & Panha, 2006; Sutcharit et al., 2007; Prasankok perakensis (see Fig. 1). et al., 2007; Lok & Tan, 2008). Amphidromus atricallosus temasek, new subspecies, is allopatric with the other The thin white subsutural line of A. atricallosus temasek recognised subspecies and with respective distinct shell is distinctive. Shells from Malaysian localities, with the characters, confusion is unlikely. Amphidromus atricallosus exception of Pulau Tulai, have a broader white subsutural atricallosus (Gould, 1843), is characterised by its slightly band that diffuses into the yellow part of the shell. The Pulau fl attened whorls, and a dark brown to black parietal callus Tulai population is problematic because the subsutural band (Fig. 1. F). Amphidromus atricallosus classiarius Sutcharit is barely discernible in most of the examined specimens

Fig. 2. A–C, Shells of juveniles: A, SH 31 x SD 19.7, juvenile shell of Amphidromus atricallosus temasek, new subspecies, showing the white columella; B–C, Amphidromus atricallosus perakensis (Fulton, 1901), juveniles, has a white zone bordering the columella: B, SH 33.9 x SD 22.4, Tapah Hill, Perak, West Malaysia; C, SH 34.7 x SD 25, Pulau Tulai, West Malaysia.

42 THE RAFFLES BULLETIN OF ZOOLOGY 2011

Fig. 3. A–B, Live animal colouration: A, Amphidromus atricallosus temasek, new subspecies, holotype (ZRC.MOL.3058); B, Amphidromus atricallosus perakensis (Fulton, 1901), Pulau Tulai, West Malaysia. (Photographs by Tan Heok Hui)

43 Tan et al. – New Amphidromus from Singapore although it is broad where present and other characters suggest as a member of the A. atricallosus group (Maassen, 2001; affi nity with typical A. atricallosus perakensis. It is possible Sutcharit & Panha, 2006; Prasankok et al., 2007; Lok & that we are dealing with a separate or incipient subspecies Tan, 2008). here. A number of specimens from the more northerly states of Peninsular Malaysia have a brown line at the suture on Comparative material. – Amphidromus (Amphidromus) early whorls (e.g., Fig. 1. E), but this appears to have no atricallosus perakensis Fulton, 1901. 1 D (ZRC 1989.1108), taxonomic bearing as the character is inconstant. The white SH 51.7 x SD 29.5, no data. WEST MALAYSIA. Perlis: subsutural line of A. atricallosus temasek is however, of little 1 D (broken, not measured) (CSY 409.3.4.11), Bukit Keteri use when examining eroded or weathered shells. (Twin Peaks), near a cement factory, Chuping Mukim, dead collected at foot of limestone hill, coll. S. Y. Chan & L. The shells of live A. atricallosus temasek appear green, and Nguang, 27 Apr.1996; Kedah: 1 D SH 53.4 x SD 29.5, it is often known by the common name of green land snail 1 S SH 50.5 x SD 30.9 (ZRC 1990.10272–10273), Pulau or green tree snail in Singapore (Ng & Lim, 1992; Chou Langkawi, coll. 1968; Perak: 1 D SH 48.8 x SD 26.8 (ZRC & Tan, 2008; Lok & Tan, 2008). In comparison, shells of 1975.4.2.10), Sungai Siput, Perak, coll. 1936; 3 D SH 47.2 A. atricallosus perakensis from Pahang, and Pulau Tulai, x SD 23.8–SH 50.7 x SD 28 (ZRC 1975.4.2.14–16), Gapis; except for juveniles, do not appear as greenish while alive. 18 D (all broken, not measured) (CSY 409.3.4.6), Sungei And even though empty shells are invariably yellow, A. Kuala Lipang (Chinese cave temple, ‘Kwang Yin Tong’) atricallosus temasek is by and large a deeper shade of yellow about 9 km NNE of Kampar town, Sungei Siput Selatan, (very pale ochre yellow versus pale sulphur yellow of typical Kinta District, dead collected in limestone rock crevices A. atricallosus perakensis). Differences in colour and tone and forest debris bordering foot of hill, coll. S. Y. Chan et of the shells on living snails and empty shells between A. al, 28 Dec.1995; 1 S SH 52.6 x SD 25.8, 1 D (juv.) (CSY atricallosus temasek and A. atricallosus perakensis are 409.3.4.7), Gunung Terendum (Chinese cave temple, ‘Wan likely due to shells of the former being thinner. The soft Shi Kong’), about 9 km SSE of Ipoh City, road to Kramat body colouration also shows remarkable differences. The from Gopeng Town towards Pahang border, Kinta District, mantle edge of A. atricallosus temasek is cream coloured dead collected at base and slopes of the limestone hill, coll. to brown, whilst it is orange in animals from Bukit Serdam S. Y. Chan et al , 22 Feb.1996; 4 D SH 50.5 x SD 27.9–SH and Pulau Tulai, Pahang. However, the body of the Pulau 51.8 x SD 28.2, 11 S (2 juv.) SH 48.8 x SD 27.5–SH 51.4 x Tulai animals is almost entirely orange, while the Bukit SD 28 (CBB), Tapah Hill, on tree trunks, coll. M. Yeh, 1998; Serdam animals have an orange head and stripe on either 3 D SH 48.5 x SD 26.2, SH 53.0 x SD 28.3, SH 55.3 x SD side of the foot, the other parts being milky white in colour. 29.8, 1 S SH 47.7 x SD 24.7 (CSY 409.3.4.23), same data Photographs of animals from Perak showed a similarly as previous; 8 D SH 47.1 x SD 26.8-SH 55.5 x SD 28.3, 7 patterned animal with much lighter colouration (light yellow S SH 45.5 x SD 24.5-SH 54.9 x SD 28.8 (CSY 409.3.4.32), instead of orange) and with very faint, almost non-existent, Kuala Woh Recreation Park, Perak, on tree trunks, coll. M. yellow longitudinal stripe, but otherwise similar to the Yeh, 1998; 3 S (2 juv.) SH 46 x SD 24, 3 D (fragments, Pahang animals. The implications and full extent of inter- not measured) (FJK CAM 0006), Gua Tempurung, Gunung population variations regarding soft body colouration is not Tempurung, Dead and scattered around base of a tree beside understood, and at this stage inconclusive. Nevertheless, the a limestone cliff, coll. J. K. Foon, 12 Aug.2005; 2 S (1 typical colouration of the animals of A. atricallosus temasek juv.) SH 50 x SD 28, 2 D (1 juv.) SH 47 x 26 (FJK CAM is distinct from those of any Malaysian population that we 0004), Kampung Ulu Geroh (Orang Asli village), Gopeng, have observed thus far. on concrete lamp post early morning after rain, coll. J. K. Foon, Sep.2007; 1 S SH 46.4 x SD 24.2 (ZRC.MOL.3062 Probably due to the lack of prominent or distinctive [ex CUMZ]), Gunung Genting, on tree near road from Ipoh morphological features, the number of accepted subspecies town to Tanjung Rambutan, 27 Sep.2007; 1 S SH 49.7 x in the genus Amphidromus is relatively high and the SD 24.5, 2 D SH 43.9 x SD 23–SH 48.4 x SD 26.5 (both prevailing tendency is to apply subspecies conservatively in with apex broken) (ZRC.MOL.3063 [ex CUMZ]), Gunung spite of consistent morphological differences and allopatric Genting, Ipoh, dead on ground and rock, 27 Sep.2007; distribution. But despite the superfi cial similarities in shell Kelantan: 1 S SH 49.3 x SD 26.4 (ZRC 1975.4.2.8–9), form and colour, A. atricallosus temasek is ostensibly not Batu Tongkat, coll. M. W. F. Tweedie, 5 Sep.1939; 1 D closely affi liated with the other recognised A. atricallosus (broken, not measured) (CSY 409.3.4.29), dead collected, subspecies. Recent studies on molecular phylogeny and coll. G. Davison, 1990; 1 S (apex broken, not measured) (FJK allozyme analyses suggest signifi cant genetic discontinuity CAM 0025), Orang Asli Village, Kampung Jidim, Lojing (see Sutcharit et al., 2007; Prasankok et al., 2007). The Highlands, along an Orang Asli trail, coll. J. K. Foon, 24 same studies also suggest that one or more of the presently Dec.2005; 1 D SD 23.3 (apex broken) (M.E.2009/0205), Gua recognised subspecies, including the population from Musang, dead in cave fl oor, coll. M. E. Marzuki, 19 May Singapore (as A. atricallosus perakensis), might be valid 2009; 1 S SH 50.05 x SD 23.3 (M.E.2010/0149), FELDA full species, and the morphological differences mentioned in Ciku 4, dead on forest fl oor at foot of limestone hill, coll. this study can probably be ascribed to allopatric speciation. M. E. Marzuki, 17 Feb.2010; Pahang: 1 S SH 54.3 x SD Until in-depth phylogenetic revisions of the genus or family 26.1 (ZRC 1975.4.2.12), Bukit Chintamani, coll. 1935; 1 S are conducted, A. atricallosus temasek is here conservatively SH 50.4 x SD 24.8 (ZRC 1975.4.2.11), Sungai Telom, Kuala regarded a subspecies since it has been unanimously regarded Terla, coll. 1935; 1 D SH 51.2 x SD 29.2 (apex broken)

44 THE RAFFLES BULLETIN OF ZOOLOGY 2011

(ZRC), Kuala Tahan, coll. E. R. Alfred; 1 S (broken, not Bartsch, P. F. & A. Solem, 1971. Notes on a collection of non- measured) (CSY 409.3.4.18), Bukit Chintamani, at foot of marine Mollusca from Pulau Aur, an island off the east coast limestone hill, in banana plantation, coll. S. Y. Chan et al, of Malaya. Federation Museums Journal, 16: 91–95. 03 Jan.1997; 2 S SH 38.2 x SD 22.9, SH 35.9 x SD 23.0 Benthem-Jutting, W. S. S. van, 1949. On a collection of non-marine (CSY 409.3.4.28), near Bera, on ground (dead) in the forest; Mollusca from Malaya in the Raffl es Museum, Singapore, with 4 S SH 36.7 x SD 22.5–SH 40.8 x SD 23.9, 4 D SH 36.4 an appendix on cave shells. Bulletin of the Raffl es Museum, x 22.7–SH 41.8 x 22.5, same data as last; 1 S SH 46.9 x 19: 50–77. SD 24.1 (CSY 409.3.4.20), Bukit Serdam, dead on forest Berry, A. J., 1963. An introduction to the non-marine Molluscs of fl oor, coll. S. Y. Chan et al, 03 Jan.1997; 1 S (juv.) (CSY Malaya. Malayan Nature Journal, 17: 1–17. 409.3.4.33), Kota Gelanggi, dead on forest fl oor beside hill, Chan, S. Y. & S. K. Tan, 2010. On two new species of Amphidromus coll. S. Y. Chan et al, 25 May 1997; 8 D SH 48.0 x SD (Gastropoda: Camaenidae) from the Lesser Sunda Islands, 25.8 – SH 51.2 x SD 30.3, 7 S SH 46.7 x SD 27.4 – SH Indonesia. Raffl es Bulletin of Zoology, 58(2): 245–249. 52.7 x SD 27.5 (CSY 409.3.4.22), Bukit Serdam, Raub Chou, L. M., P. K. L. Ng & K. K. P. Lim, 1994. Animalia. In: Wee, District, live collected, on trees next to limestone hill (Bukit Y. C. & P. K. L. Ng (eds.), A First Look at the Biodiversity in Serdam), coll. S. Y. Chan et al, 06 May 2001; 1 D SH 51.8 Singapore. National Council on the Environment, Singapore. x SD 30.7, 1 S SH 45 x SD 26.4 (TSK 11041), same data Pp 70–106. as last; 2 S SH 46.8 x SD 25.0, SH 48.0 x SD 24.7 (CSY Chou, L. M. & K. S. Tan, 2008. Fauna: invertebrates. Corals, 409.3.4.24), Cameron Highlands, coll. H. E. Ng, 28 Jan. 2003; worms and molluscs. In: Davison, G. W. H, P. K. L. Ng & 2 S (broken, not measured) (FKJ CAM 0005), Dusun Eco H. C. Ho (eds.), The Singapore Red Data Book: Threatened Resort (between Bukit Tinggi and Bentong, along the Plants and Animals of Singapore. 2nd Edition. The Nature Karak Highway), Bentong, Pahang, long dead specimens Society (Singapore), Singapore. Pp. 39–61. collected on forest fl oor, coll. J. K. Foon, 14 Dec.2005; 1 S Collinge, W. E., 1903. Report on the non-operculate land Mollusca. SH 48.0 x SD 25.7 (M.E.2009/0030), Gunung Jebak Puyuh, Fasciculi Malayenses, Zoology, 2: 205–218. dead on forest fl oor, coll. M. E. Marzuki, 22 Mar. 2009; 1 Fulton, H., 1896. A list of the species of Amphidromus, Albers, D SH 48.1 x 24.9 (M.E.2010/0287), Bukit Mengabor, dead with critical notes and descriptions of some hitherto undescribed on forest fl oor at foot of limestone hill, coll. M. E. Marzuki, species and varieties. Annals and Magazine of Natural History, 21 Feb.2010; 1 S (broken, not measured) (M.E.2010/0035), Series 6, 17: 66–93, Pls. V–VII. Bukit Kilat, dead on forest fl oor at foot of limestone hill, Fulton, H., 1901. Descriptions of new species of Xesta, Amphidromus coll. M. E. Marzuki, 16 Feb.2010; 1 D SH 50.3 x SD 26.4 and Cyclostoma from Madagascar and Perak. Journal of (M.E.2010/0065), Bukit Keling, dead on forest floor at Malacology, 8: 103–104, Pl. IX. foot of limestone hill, coll. M. E. Marzuki, 16 Feb.2010; 5 Gould, A. A., 1843. [S]hells not long since announced as having S (2 S broken; 1 S juv.) SH 44 x SD 25–SH 46 x SD 26 been received from Rev. Francis Mason, missionary at Tavoy, (FJK CAM 0022), Kampung Salang, Pulau Tioman, long in British Burmah. In: Regular meeting—Dr. C. T Jackson, dead specimens collected in cleared coastal bushland, coll. Vice-President, in the Chair. September 6th, 1843. Proceedings J. K. Foon, 16 Mar.2009; 1 S, SH 47.2 x SD 27.6 (ZRC. of the Boston Society of Natural History, 1: 139–141. MOL.3064), Pulau Tulai, off Pulau Tioman, coll. P. K. L. Gould, A. A., 1844. Descriptions of land shells from the province Ng, 23 Aug.2003; 3 D (1 juv.) SH 44.7 x SD 26–SH 47.5 x of Tavoy, in British Burmah. Proceedings of the Boston Society SD 27.1, 4 S SH 44.5 x SD 27.9–SH 47.2 x SD 27.6 (ZRC. of Natural History, 4: 452–459. MOL.86), Pulau Tulai, off Pulau Tioman, coll. P. K. L. Ng Hanitsch, R., 1908. Guide to the Zoological Collections of the et al., 23 Aug.2003; Negri Sembilan: 1 D SH 49.4 x SD Raffles Museum, Singapore. Straits Times Press, Limited, 28.1 (ZRC 1975.4.3.2.13), Bukit Tangga. Singapore. 21 pls + 112 pp. Ho, W. H., 1995. A Review of the Land-snail Fauna of Singapore. Raffl es Bulletin of Zoology, 43(1): 91–113. ACKNOWLEDGEMENTS Laidlaw, F. F., 1933. A list of the land and freshwater Mollusca of the Malay Peninsula. Part 2 (Land Mollusca, Pulmonata). We would like to thank P. K. L. Ng for advice and important Journal of the Malayan Branch Royal Asiatic Society, 11(2): discussions, and also for suggesting the epithet. Ang W. 211–234. F., B. Boo, B. Dharma, Foon, J. K., A. Lok, Lua H. K., Laidlaw, F. F. & A. Solem, 1961. The land snail Genus M. E. Marzuki, B. Ng, Ng. P. X., Ng H. E., L. Nguang, Amphidromus. A synoptic catalogue. Fieldiana: Zoology, Tan H. H., Teo S., M. Yeh, and Yeo C. K. are gratefully 41(4): 505–677. acknowledged for their generosity and help with information Lok, A. F. S. L. & S. K. Tan, 2008. A review of the Singapore and specimens. We would also like to thank M. Low for status of the green tree snail, Amphidromus atricallosus help with literature, and Tan H. H. for photographs of the perakensis Fulton, 1901 and its biology. Nature in Singapore, live snails. Comments and suggestions from the anonymous 1: 225–230. reviewers improved this article. Maassen, W. J. M., 2001. A preliminary checklist of the non-marine molluscs of West-Malaysia. “A handlist”. De Kreukel, Extra Editie 2001: 1–155. LITERATURE CITED Martens, E. von, 1864. Diagnosen neuer Heliceen aus dem ostasiatischen Archipel. Monatsberichte der Königlichen Albers, J. C., 1850. Die Heliceen, nach natürlicher Verwandtschaft Preussischen Akademien der Wissenschaften zu Berlin, 1861: systematisch geordnet. Verlag Theodor Christian Friedrich 523–529. Enslin, Berlin. 262 pp. 45 Tan et al. – New Amphidromus from Singapore

Ng, P. K. L. & K. K. P. Lim, 1992. The conservation of the Nee Soon Schilthuizen, M. & S. Looijestijn, 2009. The sexology of the chirally freshwater swamp forest of Singapore. Aquatic Conservation: dimorphic snail species Amphidromus inversus (Gastropoda: Marine and Freshwater Ecosystems, 2(3): 255–266. Camaenidae). Malacologia, 51(2): 379–387. Pilsbry, H. A. 1895. Manual of Conchology. Second Series: Solem, A. 1965. Land snails of the Genus Amphidromus from Pulmonata, Volume 9. Academy of Natural Sciences Thailand (Mollusca: Pulmonata: Camaenidae). Proceedings of Philadelphia. i–xlviii + 366 pp, 71 pls. the United States National Museum, 117(3519): 615–627. Pilsbry, H. A. 1900. Manual of Conchology. Second Series: Sutcharit, C. & S. Panha, 2006. Taxonomic review of the tree Pulmonata, Volume 13. Academy of Natural Sciences snail Amphidromus Albers, 1850 (Pulmonata: Camaenidae) in Philadelphia. 253 pp., 72 pls. Thailand and adjacent areas: Subgenus Amphidromus. Journal Prasankok, P., H. Ota, M. Toda & S. Panha, 2007. Allozyme of Molluscan Studies, 72: 1–30. variation in the Camaenid tree snails Amphidromus atricallosus Sutcharit, C., T. Asami & S. Panha, 2007. Evolution of whole-body (Gould, 1843) and A. inversus (Müller, 1774). Zoological enantiomorphy in the tree snail genus Amphidromus. Journal Science, 24: 189–197. of Evolutionary Biology, 20(2): 661–672.

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THE RAFFLES BULLETIN OF ZOOLOGY 2011 59(1): 47–51 Date of Publication: 28 Feb.2011 © National University of Singapore

EVIDENCE OF THE EARLIEST FRESHWATER DECAPOD FOSSIL FROM SOUTHEAST ASIA (CRUSTACEA: DECAPODA: BRACHYURA)

Sebastian Klaus Goethe-Universität, Department of Ecology and Evolution, Siesmayerstrasse 70A, 60054 Frankfurt am Main, Germany

Department of Biological Science, National University of Singapore, 14 Science Drive 4, Singapore 117543, Republic of Singapore Email: [email protected]

Madelaine Böhme Senckenberg Center for Human Evolution and Palaeoecology (HEP), Eberhard-Karls-Universität Tübingen, Institute for Geoscience, Sigwartstrasse 10, 72076 Tübingen, Germany Email: [email protected]

Simon Schneider Bayerische Staatssammlung für Paläontologie und Geologie, Richard-Wagner-Strasse 10, 80333 München, Germany Email: [email protected]

Jérôme Prieto Senckenberg Center for Human Evolution and Palaeoecology (HEP), Eberhard-Karls-Universität Tübingen, Institute for Geoscience, Sigwartstrasse 10, 72076 Tübingen, Germany

Ludwig-Maximilians-Universität München, Department of Earth and Environmental Sciences, Palaeontology and Geobiology, Richard-Wagner-Strasse 10, 80333 München, Germany Email: [email protected]

Boungnavong Phetsomphou Viangphoukha Coal Mine Co. LTD, Lao PDR Email: [email protected]

ABSTRACT. – We report the ﬁ rst evidence for Miocene brachyuran freshwater crabs from Southeast Asia, a hotspot of extant freshwater crab biodiversity. This conﬁ rms the presence of potamoid freshwater crabs in Southeast Asia during the Miocene, as suggested by previous molecular clock estimates. The specimen (one claw fragment) originates from the Middle to Late Miocene site of Vieng Phouka, Lao People’s Democratic Republic. Sedimentological and palaeontological data indicate the former existence of alternating swampy and lacustrine environments, inhabited by the crab and a low-diversity gastropod fauna.

KEYWORDS. – Miocene, Laos, Vieng Phouka, Potamidae, Gecarcinucidae.

INTRODUCTION and occur in Southeast Asia as the families Potamidae and Gecarcinucidae, together comprising about 12% of the total The fossil record of freshwater decapod crustaceans is number of currently described brachyuran species (Ng et al., generally very sparse (Glaessner, 1969; Rode & Babcock, 2008; Yeo et al., 2008; Cumberlidge et al., 2009). 2003; Klaus & Gross, 2010), especially in tropical Asia. In contrast, Southeast Asia represents a biodiversity hotspot However, the fossil record of these families is restricted for extant primary freshwater brachyurans (Yeo et al., to the geographic periphery regarding current regions of 2008; Cumberlidge et al., 2009; Klaus et al., 2009). Primary species richness. For example, Potamon sivalense was freshwater crabs can be considered as those brachyuran described by Glaessner (1933) from the Siwalik beds families that lack any marine members (see Yeo et al., 2008), (Middle Miocene to Pliocene) in the northern part of the

47 Klaus et al.: The earliest freshwater decapod fossil from Southeast Asia

Indian Subcontinent, but without information on the exact section of lignites, dark lignitic marls, grey marls, and bodies location and age. Several claw fragments originate from of fi ne sand (Fig. 3). Occasionally, allochthonous carbonized the Tatrot Formation within the Siwalik beds (2.5 Ma) that tree-trunks (>2 m in length, up to 1.2 m in diameter) occur were attributed a closer relationship with the gecarcinucid in the lignite seams. The bases of several marl levels that genus Sartoriana Bott, 1969 (Klaus & Gross, 2010). Late overly lignite seams show root traces and the development Pleistocene subfossil specimens of the potamid freshwater of stump horizons (bed # 2, 5, 7), and often contain fossil crab genus Geothelphusa Stimpson, 1858 are known from resin. The partly fi ne-laminated and pyrite-bearing grey clay sediments on Okinawa Island and Honshu (Karasawa, marls contain small freshwater gastropods (e.g., Bithynia 1997; Naruse et al., 2003, 2006), and Holocene subfossil Leach, 1818; Bithyniidae) in moderate abundance. The freshwater crabs (family Potamidae) were found in northern crab claw fragment is also derived from one of these marl Vietnam (Rabett et al. 2008). horizons (lower part of bed # 5). The fi ne sands, which are locally rich in plant debris, usually show erosional bases, Fossils are of major importance for zoologists. They serve while ripple-marks may occur at their top. Altogether, these as direct evidence for past distribution patterns and thus data indicate a swamp environment driven by a periodically contribute to biogeographic inferences. Although fossils can pending groundwater level. During high stand periods, the only supply minimum ages for taxa, they are a widely used swamp was fl ooded, forming a shallow lake (gastropod and tool for estimating mutation rates and calibrating molecular crab-bearing grey marl) that was supplied with fi ne-grained clocks (for freshwater crabs see Daniels et al., 2006; Klaus et sandy sediments by infl owing rivers. al., 2010; Shih et al., 2009). Here, we report the fi rst evidence of Neogene freshwater brachyurans in tropical Asia. Reliable age indicators are absent from the sediments. However, the Vieng Phouka Basin obviously is structurally related to the N-S to NE-SW striking Neogene rift basins of MATERIAL Northern Thailand (Morley, 2001), as it is positioned in their direct prolongation. Assuming a similar tectonostratigraphic Description. – A single claw fragment, either part of propodus evolution for the basins along these major Southeast-Asian or dactylus, three conical teeth visible, one smaller tooth fault systems, a mid to late Miocene age may be suggested adjoined by two larger teeth to the left and right (see Fig. for the sediments at Vieng Phouka. This estimation already 1). A fourth tooth is only preserved in part. The length implies a potential age gradient between western and eastern of the fragment is 2.3 mm. The specimen is stored in the basins, as previously emphasized (Morley, 2001, Songtham paleontological collection of the University of Tuebingen et al., 2003) or neglected (Chaimanee et al., 2007). (collection number: GPIT/CU/337).

Location. – Vieng Phouka (or Viengpuka, Viengphoukha, DISCUSSION Viangphouka) Coal Mine (coordinates: N 20.5833, E 101.0583), Luang Namtha Province, Lao People’s Democratic The critical question concerning the claw specimen is its Republic (see Fig. 2). taxonomical assignment. There have been efforts to assign chelae fragments of fossil freshwater crabs to extant species Geology, palaeoenvironment and stratigraphy. – The by means of dentition patterns (Pretzmann, 1971; Bachmayer opencast mine of Vieng Phouka exposes an 85 metre thick & Pretzmann, 1971), and by using rigorous morphometric analyses (Ashkenazi et al., 2005). However, for both approaches large sample sizes, entire propodi and dactyli, and especially a priori knowledge about extant relatives for comparison are required. None of these criteria are fulﬁ lled

Fig. 1. Fragment of the brachyuran chela (either propodus or dactylus). Two larger teeth (no. 1 and 3) and one intertooth (no. 2) are completely preserved, with evidence of a fourth tooth. The specimen is stored in the paleontological collection of the University of Tuebingen (collection number: GPIT/CU/337). Fig. 2. Location of Vieng Phouka in northwestern Laos.

48 THE RAFFLES BULLETIN OF ZOOLOGY 2011 with the present fragment. Thus we can only give a broad The sesarmid genera Sesarmoides Serène & Soh, 1970, delimitation of taxonomic identity of the claw fragment, Labuanium Serène & Soh, 1970 and Karstarma Davie & mainly based on the distribution pattern of extant groups Ng, 2007 can be completely freshwater adapted, the latter and their known fossil record. being semi-terrestrial (see Cumberlidge et al., 2009; Davie & Ng, 2007; Ng, 2004). Finally, Geosesarma De Man, 1892, The shape and pattern of the preserved teeth clearly indicate a is found in terrestrial habitats (Ng, 1988; 2004). Also several brachyuran origin of the fragment. Astacid crayﬁ sh comprise species of Gecarcinidae (land crabs), Hymenosomatidae and the oldest known Asian freshwater decapods (Upper Jurassic semiterrestrial Ocypodoidea can be regarded as potential of the Jehol Group in Northern China, see Taylor et al., origin of the claw fragment, as these can disperse far into 1999). However, they do not possess comparably toothed the inland (Ng, 2004; Yeo et al., 2008). However, chelae chelae. Moreover, both extant and fossil astacids of East morphologies differ from the present claw fossil (P. K. L. Asia are restricted to more temperate climates. Ng; T. Naruse, pers. comm.). These brachyuran groups have virtually no fossil record, although fossils of marine The present fossil may be attributed to different taxa within Sesarmidae (Sesarma Say, 1817) and Varunidae (Miosesarma the Brachyura. Many Asian species of the Grapsoidea, for Karasawa, 1989; Cyclograpsus H. Milne Edwards, 1837; example, invade or even wholly inhabit the freshwater Helice De Haan, 1833; but also freshwater adapted, Pliocene habitat (see Yeo et al., 2008). The genera Eriocheir De Eriocheir De Haan, 1835) have been reported from Miocene Haan, 1835 and Varuna H. Milne Edwards, 1830 (Varunidae) and Pliocene sediments of Japan (Karasawa & Narita, 2000; enter estuaries, and are also found further up in rivers. Karasawa & Kato, 2001). Assuming that the transition

Fig. 3. Sedimentologic proﬁ le and fossil content of the Vieng Phouka coal mine.

49 Klaus et al.: The earliest freshwater decapod fossil from Southeast Asia into freshwater and terrestrial environments within these Bott, R., 1969. Die Flußkrabben aus Asien und ihre Klassifi kation. euryhaline, secondary freshwater species may represent a Senckenbergiana biologica, 50: 359–366. rather recent, post-Miocene, phenomenon (Yeo et al., 2008), Chaimanee, Y., Yamee, C., Marandat, B. & Jaege, J.-J., 2007. First the present claw fragment is more likely to be of primary Middle Miocene rodents from the Mae Moh Basin (Thailand): freshwater crab origin. Although the ontogenetic age and biochronological and paleoenvironmental implications. Bulletin fi nal adult size of the fossil crab remains elusive, the small of Carnegie Museum of Natural History, 39: 157–163. size of the claw fragment could indicate that it belongs to Cumberlidge, N., Ng, P. K. L., Yeo, D. C. J., Magalhães, C., a sub-adult specimen that originated not far away from the Campos, M. R., Alvarez, F., Naruse, T., Daniels, S. R., Esser, Vieng Phouka site. In contrast, most secondary freshwater L. J., Attipoe, F. Y. K., Clotilde-Ba, F. L., Darwall, W., McIvor, brachyurans have a marine larval development and would A., Baillie, J. E. M., Collen, B. & Ram, M., 2009. Freshwater reach inland habitats more likely as adults. crabs and the biodiversity crisis: Importance, threats, status, and conservation challenges. Biological Conservation, 142: 1665–1673. Primary freshwater crabs are present in Eurasia as two families, the Potamidae and the Gecarcinucidae, and both Daniels, S. R., Cumberlidge, N., Pérez-Losada, M., Marijnissen, S. A. E. & Crandall, K. A., 2006. Evolution of Afrotropical freshwater have their maximum of species diversity in tropical Asia crab lineages obscured by morphological convergence. (Yeo et al., 2008; Cumberlidge et al., 2009; Klaus et al., Molecular Phylogenetics and Evolution, 40: 227–235. 2009). The preserved rounded crushing teeth resemble the Davie, P. J. F. & Ng, P. K. L., 2007. A new genus for cave- character state found in several Southeast Asian gecarcinucid dwelling crabs previously assigned to Sesarmoides (Crustacea: genera, while potamids tend to have sharper teeth. Also the Decapoda: Brachyura: Sesarmidae). – Raffles Bulletin of palaeoenvironment of the Vieng Phouka site, as inferred from Zoology, Supplement 16: 227–231. the sedimentological and palaeontological data, suggests that Glaessner, M. F., 1933. Tertiary crabs in the collection of the a gecarcinucid affi liation is more likely: extant potamids occur British Museum. The annals and magazine of natural history, predominantly at higher elevations in clear streams, while zoology, botany and geology, 12: 12–13. gecarcinucid freshwater crabs dominate lowland habitats, Glaessner, M. F., 1969. Decapoda. In: Moore, R. C. (ed.) Treatise like fl oodplains or lacustrine environments (Ng, 1988). on Invertebrate Paleontology, Part R. Arthropoda 4, vol. 2, R400–R533; Boulder and Lawrence (Geological Society of Divergence time estimates of potamid (Shih et al., 2009) America and University of Kansas Press). and gecarcinucid (Klaus et al., 2010) phylogenies, based on Haan, H. M. De, 1833–1849. Crustacea. In: von Siebold, P. F. (ed.) molecular clock approaches, do not confl ict with the age of Fauna Japonica, sive Descriptio animalium, quae in itinere the present fossil, as Southeast Asian lineages presumably per Japoniam, jussu et auspiciis superiorum, qui summum in predate the Pliocene. On the contrary, the claw fragment India Batavia imperium tenent, suscepto, annis 1823– 1830 provides independent evidence for the presence of primary collegit, notis, observationibus a adumbrationibus illustravit. freshwater crabs in Southeast Asia during the Miocene. Lugduni Batavorum, fasc. 1–8: I–xxi+vii– xvii+ix–xvi+1–243, Nevertheless, for an unambiguous morphological assignment pls. 1–55, A–Q, circ., pl. 2. to one of these families further material will be necessary, Karasawa, H., 1989. Decapod crustaceans from the Miocene ideally with preserved carapaces. Mizunami Group, central Japan, Part 1. Superfamily Thalassinoidea, Leucosioidea and Grapsidoidea. Bulletin of the Mizunami Fossil Museum, 16: 1–28. ACKNOWLEDGMENTS Karasawa, H., 1997. Geothelphusa dehaani (White, 1847), a fossil land crab from the fi ssure deposits of Yage, Inasa-cho, Shizuoka The authors thank the offi cials of the Viangphoukha Coal Prefecture, Japan. Science report of the Toyohashi Museum of Mine Co. LTD for access to the outcrops, as well as Natural History, 7: 25–26. August Ilg (Duesseldorf) for providing the plot of fi gure 2. Karasawa, H. & Kato, H., 2001. The systematic status of the genus Peter K. L. Ng, Darren C. J. Yeo (Singapore), and Tohru Miosesarma Karasawa, 1989 with a phylogenetic analysis Naruse (Taketomi, Okinawa) gave valuable comments on within the family Grapsidae and a review of fossil records the manuscript. (Crustacea: Decapoda: Brachyura). Paleontological Research, 5: 259–275. Karasawa, H. & Narita K., 2000. A fi rst fossil of Eriocheir japonica LITERATURE CITED (de Haan, 1835) (Crustacea, Decapoda, Brachyura) from the lower Pliocene Okubo Formation in Nagano Prefecture, central Japan. Research Report of the Shinshushinmachi Fossil Ashkenazi, S., Motro, U., Goren-Inbar, N., Biton, R. & Rabinovich, Museum, 3: 1–3 R., 2005. New morphometric parameters for assessment of body size in the fossil freshwater crab assemblage from the Klaus, S., Brandis, D., Ng, P. K. L., Yeo, D. C. J. & Schubart, C. Acheulian site of Gesher Benot Ya’aqov, Israel. Journal of D., 2009. Phylogeny and Biogeography of Asian Freshwater Archaeological Science, 32: 675–689. Crabs of the family Gecarcinucidae (Brachyura: Potamoidea). In: Martin, J. W., Crandall, K. A. & Felder, D. L. (eds.), Crustacean Bachmayer, F. & Pretzmann, G., 1971. Krebsreste aus den Issues 18: Decapod Crustacean Phylogenetics. Boca Raton, altpliozänen Süßwasserablagerungen des Eichkogels bei Florida: Taylor & Francis/CRC Press. pp. 509-531. Mödling, Niederösterreich. Annalen des Naturhistorischen Museums Wien, 75: 283–291. Klaus, S. & Gross, M., 2010. Synopsis of the fossil freshwater crabs of Europe (Brachyura: Potamoidea: Potamidae). Neues Jahrbuch für Geologie und Paläontologie, 256: 39–59.

50 THE RAFFLES BULLETIN OF ZOOLOGY 2011

Klaus, S., Schubart, C. D., Streit, B. & Pfenninger, M., 2010. Pretzmann, G., 1971. Scheren und Scherenbezahnung bei Potamon When Indian crabs where not yet Asian – evidence for Eocene (Crustacea: Decapoda). Annalen des Naturhistorischen Museums proximity of India and Southeast Asia from freshwater crab Wien, 75: 489–493. biogeography. BMC Evolutionary Biology, 10: 287. Rabett, R., Barker, G., Hunt, C. O., Naruse, T., Piper, P., Raddatz, Leach, W. E., 1818. In: Abel, C. (ed.) Narrative of a journey E., Reynolds, T., Nguyêñ, V. S., Stimpson, C., Szabó, K., in the interior of China, and of a voyage to and from that Nguyêñ, C. T. & Wilson, J., 2008. The Tràng An Project: country, in the years 1816 and 1817; containing an account Late-to-Post-Pleistocene Settlement of the Lower Song Hong of the most interesting transactions of Lord Amherst’s embassy Valley, North Vietnam, 2009. Journal of the Royal Asiatic to the court of Pekin, and observations on the countries which Society, Series 3, 19(1): 83–109. it visited. Vol. 1, 1-420 pp, London: Longman, Hurst, Rees, Rode, A. L. & Babcock, L. E., 2003. Phylogeny of fossil and extant Orme & Brown. freshwater crayfi sh and some closely related nephropid lobsters. Man, J.G. De, 1892. Decapoden des indischen Archipels. In: Weber, Journal of Crustacean Biology, 23: 418–435. M. (ed.) Zoologische Ergebnisse einer Reise in Niederländisch Say, T., 1817. An account of the Crustacea of the United States. Ost-Indien. Vol. 2. 265–527 pp, plates 15–29. Journal of the Academy of Natural Sciences of Philadelphia, Milne Edwards, H., 1830. Varune. Varuna. Crust. In: Saint-Vincent, 1 (1-2): 57-63, 65-80, 97-101, 155–160. B. de (ed.) Dictionnaire Classique d’Histoire Naturelle. Vol. Serène, R. & C. L. Soh, 1970. New Indo-Pacifi c genera allied to 16. Paris: Rey et Gravier & Amable Gobin et Cie. 511 pp. Sesarma Say 1877 (Brachyura, Decapoda, Crustacea). Treubia, Milne Edwards, H., 1837. Histoire naturelle des Crustacés 27: 387–416. comprenant l’anatomie, la physiologie et la classifi cation de Shih, H.-T., Yeo, D. C. J. & Ng, P. K. L., 2009. The collision of ces animaux. Paris, Librairie Encyclopédique de Roret. Vol. the Indian plate with Asia: molecular evidence for its impact 2: 1–531. Atlas, 1837: 1–32, pls. 1–42. on the phylogeny of freshwater crabs (Brachyura: Potamidae). Morley, C. K., 2001. Combined escape tectonics and subduction Journal of Biogeography, 36: 703–719. rollback-back arc extension: a model for the evolution of Songtham, W., Raranasthien, B., Mildenhall, D. C., Singharajwarapan, Tertiary rift basins in Thailand, Malaysia and Laos. Journal S. & Kandharosa, W., 2003. Oligocene–Miocene climatic of the Geological Society, 158: 461–474. changes in northern Thailand resulting from extrusion tectonic Naruse, T., Karasawa, H., Shokita, S., Tanaka, T. & Moriguchi, M., of Southeast Asian landmass. ScienceAsia, 29: 221–233. 2003. A fi rst fossil record of the terrestrial crab, Geothelphusa Stimpson W., 1858. Prodromus descriptionis animalium tenuimanus (Miyake & Minei, 1965) (Decapoda, Brachyura, evertebratorum in expeditione ad Oceanum Pacificum Potamidae) from Okinawa Island, central Ryukyus, Japan. Septentrionalem missa, C. Ringgold et Johanne Rodgers Crustaceana, 76: 1211–1218. ducibus, observatorum et descriptorum. Pars V. Crustacea Naruse, T., Shokita, S. & Ng, P. K. L., 2006. A revision of the Ocypodoidea. Proceedings of the Academy of Natural Sciences Geothelphusa levicervix species group (Crustacea: Decapoda: of Philadelphia 10: 93–110. Brachyura: Potamidae), with descriptions of three new species. Taylor, R. S., Schram, F. R. & Shen, Y.–B., 1999. A new crayfi sh Journal of Natural History, 40(13/14): 759–781. family (Decapoda: Astacida) from the Upper Jurassic of Ng, P. K. L., 1988. The Freshwater Crabs of Peninsular Malaysia China, with a reinterpretation of other Chinese crayfi sh taxa. and Singapore. Shinglee, Singapore. p. 1–156. Paleontological Research, 3: 121–136. Ng, P. K. L., 2004. Crustacea: Decapoda, Brachyura. In: Freshwater Yeo, D. C. J., Ng, P. K. L., Cumberlidge, N., Magalhães, C., Invertebrates of the Malaysian Region. C. Yule and H. S. Yong, Daniels, S. R. & Campos, M. R., 2008. Global diversity editors, Malaysian Academy of Sciences, pp. 311–336. of crabs (Crustacea: Decapoda: Brachyura) in freshwater. Ng, P. K. L., Guinot D. & Davie, P. J. F., 2008. Systema Hydrobiologia; 595: 275–286. Brachyurorum. Part I. An annotated checklist of extant brachyuran crabs of the world. Raffl es Bulletin of Zoology, 17: 1–286.

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THE RAFFLES BULLETIN OF ZOOLOGY 2011 59(1): 53–60 Date of Publication: 28 Feb.2011 © National University of Singapore

GUIDE TO THE AQUATIC HETEROPTERA OF SINGAPORE AND PENINSULAR MALAYSIA. 6. MESOVELIIDAE, WITH DESCRIPTION OF A NEW NEREIVELIA SPECIES FROM SINGAPORE

Yang Chang Man c/o Rafﬂ es Museum of Biodiversity Research, National University of Singapore, Kent Ridge, Singapore 117546, Republic of Singapore Email: [email protected]

Dennis H. Murphy c/o Rafﬂ es Museum of Biodiversity Research, National University of Singapore, Kent Ridge, Singapore 117546, Republic of Singapore

ABSTRACT. – This is the sixth part of a series of guide to the aquatic Heteroptera of Singapore and Peninsular Malaysia. A key to three genera and ﬁ ve species of the family Mesoveliidae of Southeast Asia is provided. A new species of Nereivelia is described from Singapore mangroves.

KEY WORDS. – Mesoveliidae, Mesovelia, Nereivelia, new species, Singapore, Peninsular Malaysia, key.

INTRODUCTION and species. The male genital segments are relatively large. Females are larger in size than males, with a well developed The family Mesoveliidae are small semi-aquatic bugs (1.0–3.5 ovipositor and serrated gonapophyses (Fig. 16). mm body length) commonly known as water treaders or pondweed bugs. They usually live on marginal floating The family Mesoveliidae currently contains 12 genera and 46 vegetation amid stagnant water, slow fl owing streams of known species belonging to two subfamilies: Madeoveliinae both freshwater and brackish water, or in moist secretive and Mesoveliinae (Andersen, 1982; Andersen & Polhemus, habitats (e.g. soil or leaf litter of wet forest, under decayed 2003; Andersen & Weir, 2004). The genus Mesovelia is logs on the intertidal mangroves). widely distributed with 27 species, while the other genera are rare, scattered and have restricted ranges of distribution The head is usually distinctly prolonged and defl ected in (Andersen & Polhemus, 1980; Andersen, 1999; Andersen front of the eyes, and the eyes are usually large, with 20 & Polhemus, 2003). There are fi ve mesoveliid species in to 30 ommatidia in each eye. The eyes of Cryptovelia are three genera (Mesovelia, Nereivelia, Cryptovelia) recorded vestigial, with only 3–4 ommatidia each. The antennae are from Southeast Asia (Fernando & Cheng, 1974; Polhemus long and fl agelliform, with segments III and IV longer and & Polhemus, 1989; Murphy, 1990; Yang et al., 1997, 1999; thinner than segments I and II. The head bears 3 pairs of Andersen, 1999). cephalic trichobothria dorsally, and the rostrum is long, usually reaching to the metacoxa. A pair of longitudinal Mesovelia is yellowish green in colour with some brownish ridges is present ventrally on the head of Cryptovelia and patterns. They are very common and widespread in fresh Nereivelia (Fig. 11). The macropterous form is known only water habitats, rarely in estuarine waters. They are frequently in the genus Mesovelia, and has a pair of ocelli on the found running on the fl oating leaves of water plants, such posterior part of the head, three closed cells in the forewing, as Lemna, Nymphea and Pistia. There are two known and the scutellum elevated. There are no ocelli in apterous species, Mesovelia horvathi Lundblad, 1933 and M. vittigera forms. The thoracic segments vary in mid-length among Horváth, 1895 (=orientalis Kirkaldy, 1901) (Figs. 1–3) in different genera and species. The legs are slender, with the Southeast Asia. Mesovelia vittigera is an old world species, hind leg longest, and the tarsi are three segmented with extremely common and very widespread in Europe, Africa, two apical claws. The abdomen has 7 mediotergites and 5 and Asia to Australia. Mesovelia horvathi is common in laterotergites in the male, versus 8 and 6–7 respectively in south Asia, Japan, the Philippines to New Guinea and female. The scent gland orifi ce is distinct and situated in Australia (Polhemus & Polhemus, 2000; Andersen & Weir, different positions on mediotergite IV in different genera 2004). Macropterous and apterous forms are known in both

53 Yang & Murphy: Guide to Mesovelidae, new species of Nereivelia species, but the macropterous form is not common. The 4. Mid-length of pronotum subequal to combined length of macropterous form is larger in size, and often collected by meso- and metanotum. Scent gland orifi ce before middle of light traps. Zettel and Tran (2009) recorded a possible new mediotergite IV. Male posterior margin of abdominal tergite species closely related to M. horvathi from Pulau Langkawi VIII simple (Figs. 6–9) .. N. murphyi Polhemus & Polhemus – Mid-length of pronotum greater than combined length of (northwest coast of Peninsular Malaysia). meso- and metanotum. Scent gland orifi ce behind middle of mediotergite IV. Male posterior margin of abdominal tergite Nereivelia is a group of very small bugs (1.70–2.20 mm), VIII concave (Figs.10–16) ...... N. polhemorum, new species dark brown in colour, fl ightless and living under decayed logs in the intertidal zone of mangrove swamps. Nereivelia murphyi Polhemus & Polhemus, 1989 (Figs. 6–9) is so far FAMILY MESOVELIIDAE Douglas & Scott only recorded from southern Thailand and Singapore. A second species, N. polhemorum, new species from the same Mesovelia Mulsant & Ray habitats in Singapore is described here. Mesovelia horvathi Lundblad A key to the genera and species of Mesoveliidae is provided, (Figs. 1, 2) including the genus Cryptovelia which is so far only recorded from the moist soil-litter of a rainforest in Sarawak, East Mesovelia horvathi Lundblad, 1933: 190 (type locality, Malaysia (Andersen, 1999), but further exploration may Sumatra). prove it to have a wider distribution in the region. Mesovelia japonica Miyamoto, 1964: 199 (Synonymised by Polhemus & Polhemus 2000:208). The material used for this study consists primarily of alcohol- preserved specimens and some dry-mounted specimens in the Material examined. – SINGAPORE: numerous apterus males, Zoological Reference Collection (ZRC), National University females and a few macropterous forms collected from different locations in vegetated pools, ponds, reservoirs, sides of streams, of Singapore. Specimens of Austrovelia queenslandica 1980 – 2002. Malipatil & Monteith, 1983 were loaned from the CSIRO (Australia). All measurements are given in millimetres. W. MALAYSIA (all apterus form unless stated): Johore: 10 males, Measurements of the new species refer to the holotype and 6 females, Pontain, coll. C.M. Yang, 4 Mar. 1992; 1 female, Gunong allotype. The holotype and paratypes are deposited in ZRC, Pulai, 20 May 1993; 4 males, 1 female, Endau Rompin, Sungei with additional paratypes in the Natural History Museum of Vienna, Austria (NHMW) and the Bernice P. Bishop Museum, Honolulu (BPBM).

KEY TO GENERA AND SPECIES OF MESOVELIIDAE

1. Body slender, size 2.0– 3.5 mm in male. Head length distinctly less than thorax length. Pronotum mid-length shorter or subequal to mesonotum in apterous form. Stout spines on antennae and legs. Apterous and macropterous ...... 3 Mesovelia – Body shorter or suboval in shape, size about or less than 2 mm. Head prolonged, subequal length to thorax. Pronotum longer than mesonotum. Spines absent on antennae and legs. Only apterous form known ...... 2

2. Eyes vestigial with 3–4 ommatidia. Mesonotum about 2 times the length of metanotum (Fig. 5); size small, 1.0–1.5 mm .... Cryptovelia (one species, stysi Andersen, recorded from Sarawak). – Eyes large with 24–30 ommatidia. Mesonotum subequal to length of metanotum, size larger, 1.70–2.20 mm ...... 4 Nereivelia

3. Posterior margin of middle femur with only 1–2 black spines distally in both sexes, male abdominal sternum VIII without middle black hairs, anteriorly with a pair of black tuft of hairs (Figs. 1, 2) ...... M. horvathi Lundblad – Posterior margin of middle femur armed with row of black spines (10–17) in both sexes. Male abdominal sternum VIII with a tuft of stout black hairs on the middle and one paired patch of pale hairs laterally (Fig. 3) ..... M. vittigera Horváth Fig. 1–2. Mesovelia horvathi Lundblad. 1. apterous male, dorsal view; 2. apterous male, ventral view.

54 THE RAFFLES BULLETIN OF ZOOLOGY 2011

Pantai Burung, coll. C.M. Yang, 13 Jul. 2001;1 male, Sungei Material examined. – SINGAPORE: numerous apterus males, Temantang, Mawai, coll. H.K. Lua, 24 Feb. 1995; Selangor: 1 females and a few macropterous forms collected from different macropterous male, Templer Park, coll. C.M. Yang, 15 Nov. 1995; locations of Singapore, both freshwater and brackish water habitats, 10 males, 6 females, Ulu Gombak, coll. H.K. Lua, 6 May 1996; 2 1980 – 2002. males, North Selangor peat swamp, Coll. H.K. Lua, 17 Jun. 1991; Pahang: 12 males, 13 females, Pulua Tioman (Sungei Keliling, W. MALAYSIA (all apterus form unless stated): Johore: 5 Sungei Paya, Sungei Dungun), Jun. 1997; 1 macropterous male, macropterous males, 21 males, 27 females, Pontain, coll. K.L. Yeo, Lake Chini, coll. M. Balke, 17 Apr. 1997; 1 male, 4 females, Sungei 14 May 1992; 3 males, 1 macropterous male, 5 females, Layang Belat, coll. B. Tan & G. Sumita, 15 May 1995; Parak: 4 females, Layang, coll. C.M. Yang, 4 Mar. 1991; 1 female, Tg. Sedili, Sungei Kenderong, Grik, coll. H.K. Lua, 21 Feb. 1997; 1 male, Mawai, coll. K.L. Yeo, 15 Jul. 1992; Malacca: 1 macropterous 2 females, Sungei Lebey, coll. H.K. Lua, 17 Feb. 1997; Perlis: 1 female, Alor Gajah, 28 Oct. 1963; Selangor: 2 macropterous males, macropterous male, 4 females, Sintok Padang, coll. H.K. Lua, 19 1 macropterous female, Kuala Selangor, coll. K.L. Yeo, 15 May Feb. 1997; Trengganu: 1 male, 2 females, coll. H.K. Lua, 28 Oct. 1991; 1 female, Ulu Gombok, 22 Nov. 1995; Pahang: 13 males, 1998; 1 male, Sekayu, coll. B.Tan & G. Sumita, 16 May 1995; 2 20 females from Pulua Tioman (Sungei Raya, Sungei Genting, males, Pulua Redang, coll. H.K. Lua, 25 Jun. 1992. Sungei Paya, Sungei Keliling, Sungei Kelepak), Jun. 1997; 1 female, Lake Chini, Coll. K.L.Yeo, 16 Apr. 1997; Kelantan: 4 Remarks. – This species is very common in Singapore and macropterous males, 2 males, 6 macropterous females, 1 female, Malaysia, also collected from brackish water. Widespread Sungei Hulu Besut, coll. H.K. Lua, 20 Mar. 1992; Trengganu: 2 from China, Japan to Australia (Polhemus & Polhemus, males, 9 females, North Rantau, Abang, swamp, coll. H.K. Lua, 2000; Andersen & Weir, 2004). 18 Mar. 1992.

Remarks. – This species is less common then Mesovelia Mesovelia vittigera Horváth horvathi in our collections from Singapore and Malaysia. (Fig. 3) Widespread in warm area in Europe, North Africa, and Asia to Australia (Polhemus & Polhemus, 2000). Both species also Mesovelia vittigera Horváth, 1895: 160 (type locality Egypt). recorded from Pulau Langkawi (Zettel & Tran, 2009). Mesovelia orientalis Kirkaldy, 1901: 808 (described from Sumatra; synonymised by Horváth, 1915).

Fig. 3. Mesovelia vittigera Horváth, apterous male, ventral view.

Fig. 4. Nereivelia polhemorum, new species, apterous female, Fig. 5. Cryptovelia stysi Andersen, apterous male, dorsal view dorsal view. (after Andersen, 1999).

55 Yang & Murphy: Guide to Mesovelidae, new species of Nereivelia

Nereivelia Polhemus & Polhemus Nereivelia polhemorum, new species Nereivelia murphyi Polhemus & Polhemus (Figs. 4, 10–16) (Figs. 6–9) Material examined. – All specimens apterous. Holotype and Nereivelia murphyi Polhemus & Polhemus, 1989: 73-82 (type allotype: Singapore, Mandai mangroves, under wet wood on locality Thailand). intertidal muddy ground at 2.3 m above chart datum. 20 Mar. 1992, coll. D. H. Murphy (ZRC). Material examined. – SINGAPORE: 1 apterus male, Sungei Buloh East, mangrove, coll. D.H. Murphy, 22 Mar.1992; 2 apterus Paratypes: SINGAPORE: 4 males, same data as holotype; 2 males, females, Sungei Buloh East, mangrove, coll. D.H. Murphy, 21 1 female, Sungei Buloh East mangroves, under imbedded timber Jul.1987 and 8 Apr. 1992. on very wet muddy ground 2.4m above marine datum (above mean HWN) in air ﬁ lled cavities, along with abundant Collembola Remarks. – This species is very rare and only known from (Axelsonia and Dicranocentrus), 22 Mar. 1992, coll. D.H. Murphy; mangroves of Thailand and Singapore. Living in cavities of 1 female, Sungei Buloh East mangroves, on lobster mound surface, 2.6m above marine datum, 1 Mar. 1991, coll. D. H. Murphy; 1 decayed logs in intertidal mangrove swamps. female, Sungei Buloh mangroves, on lobster mound surface, 22

Fig. 6–9. Nereivelia murphyi Polhemus & Polhemus (after Polhemus & Polhemus, 1989). 6. apterous male, dorsal view; 7. apterous female, dorsal view; 8–9. male abdominal terminalia; 8. lateral view; 9. dorsal view.

56 THE RAFFLES BULLETIN OF ZOOLOGY 2011

Mar. 1992, coll. D. H. Murphy; 1 male, 1 female, Sungei Buloh extending beyond hind coxa, length of segments III:IV, mangroves, 12 Apr. 1999, coll. D. H. Murphy (ZRC, NHMW, 0.67:0.18. Antenna long, slender and fl agelliform, slightly BPBM). shorter than body length (1.71:1.80). Segments I and II thick, without spines, III and IV much longer, fl agelliform with Size. – males 1.70–1.85 (holotype 1.80, width 0.69); females long setae; segment II shortest. Length of antenna segments 1.90–2.09 (allotype 2.03, width 0.81). I–IV, 0.32:0.18:0.58:0.63. Description of holotype. – Body dark brown, lighter brown Thorax length along midline subequal to head length on legs and antenna, darker on thorax. Body elongate oval, (0.41:0.39), matte, lateral margins rounded. Dorsal margins covered with fi ne and coarse semi-erect hairs, denser on with groups of small shining disc-like plates (Figs. 12, 13). abdomen (Fig. 10). Pronotum longest on midline, slightly longer than combined length of meso- and metanotum which are almost subequal Head moderately sub-quadrate; matte, broadly rounded in length (0.22:0.10:0.09). Posterior margins of pro- and anterior and strongly defl ected. Head almost as long as mesonotum straight, that of metanotum curved in middle. width across eyes (0.39:0.38). Interocular space 0.21. Three Greatest width of thorax (metanotum) slightly narrower than equally spaced trichobothria present. A longitudinal row of greatest abdomen width (0.65:0.69). Legs slender, covered pits present on vertex, diverging dorsally and meeting at with setae, without spines. Hind leg longest. Fore femur posterior margin of head; some small shining disc-like plates slightly more incrassate than other femora. Tibiae slender, present near interocular space. Eye normal, large, spherical equal or subequal length to fore- and mid-femur. Hind with about 24 ommatidia, and two long and curved ocular tibia longest. Tarsi with three segments, tarsal III almost setae. Ocelli absent. Venter of head with a broad, raised, 2 times the length of tarsal I; all tarsi bearing two equal ridged plate, widening posteriorly, demarcated by a very terminal claws. low longitudinal carina on each side (Fig. 11). Rostrum

Fig. 10–13. Nereivelia polhemorum, new species. 10. apterous male, dorsal view; 11. lateroventral view of head showing the longitudinal carina and the raised ridged plate; 12. dorsal view of abdomen and thorax, showing disk-like plates (D) and the scent oriﬁ ce (S); 13. surface of thorax showing disc-like plates.

57 Yang & Murphy: Guide to Mesovelidae, new species of Nereivelia

Measurements of legs of holotype middle on tergite IV (Fig. 12). Laterotergites broad and slightly raised. Laterotergite IV broadest (0.13). Femur Tibia Tarsal I Tarsal II Tarsal III Fore leg 0.52 0.52 0.048 0.048 0.085 Genital segments relatively large and distinctly protruding Middle leg 0.52 0.54 0.048 0.054 0.097 from the pregenital abdomen. Longer than width (0.30:0.27). Hind leg 0.69 0.78 0.061 0.073 0.115 Posterior margin of segment VIII concave. Pygophore subovate, proctiger rectangular. Paramere small and Abdomen broad, shining, covered with dense and long symmetrical curved dorsally with pointed apex (Fig. 14). semi-erect setae. Mediotergites tapering in width towards the abdominal apex. Tergite I more shining and with less Description of allotype. – Similar in structure to male. Head setae. Tergites II–IV same length, tergite V shortest, tergite length, 0.46; width across eyes, 0.43; Length of antenna VII longest. Length of tergites I–VII, 0.07:0.09:0.09:0.09:0 segments I–IV, 0.30:0.18:0.60:0.64. Thorax shorter in length .06:0.08:0.23. Scent oriﬁ ce distinct, located slightly behind to head (0.38:0.46). Pronotum length greater than combined length of meso- and metanotum, 0.21:0.090:0.083. Body width gradually increase from thorax (max. width: 0.72) to abdomen tergite V (width 0.81) and tapering toward the tip of abdomen (Fig. 4).

Measurements of legs of allotype

Femur Tibia Tarsal I Tarsal II Tarsal III Fore leg 0.55 0.55 0.048 0.048 0.097 Middle leg 0.56 0.56 0.048 0.050 0.097 Hind leg 0.73 0.78 0.061 0.073 0.109

Abdomen broad, length 1.18, tergite I shortest, tergite II equal in length to tergite VI, tergites III–V same length, tergite VIII longest. Length of tergites I–VIII, 0.09:0.12:0 .10:0.10:0.10:0.12:0.15:0.19. Laterotergites broad, slightly raised. Genital segments relatively large. First gonocoxa large, gonoplac and proctiger small (Fig. 15). Ovipositor large, gonapophyses elongate with strongly serrated teeth (Fig. 16).

Macropterous form unknown.

Distribution. – Currently known only from the mangroves in Sungei Buloh and Mandai in Singapore.

Etymology. – The speciﬁ c epithet dedicates this species jointly to father and son, John T. and Dan A. Polhemus whose generous guidance have been invaluable in our work with the aquatic Heteroptera.

Ecology. – Nereivelia polhemorum was found in the same habitat as the N. murphyi Polhemus & Polhemus, mostly under decayed logs on the intertidal zone of mangroves. They shelter in the air ﬁ lled cavities of the dead wood during high tide and emerge during low tide to feed on small arthropods (Murphy pers. obs.). Nereivelia is now very rare in Singapore; several efforts have been made to obtain more material but failed. This is because of the “International Coastal Cleanup Program” of clearing “unsightly” timber and polyethylene bags from the mangroves since 1997 (Ng & Sivasothi, 1999), which effectively removed the air ﬁ lled cavities which are Fig. 14–16. Nereivelia polhemorum, new species. 14. male inhabited by many enigmatic arthropods. abdominal terminalia, lateral view showing paramere (P), proctiger (Pr), pygophore (Py); 15. Female abdominal terminalia, lateral view showing gonocoxa 1 (Gx1), gonocoxa 2 (Gx2), gonoplac Remarks. – Nereivelia polhemorum shares the same (Gp), proctiger (Pr) and gonapophyses (Go); 16. Ovipositor. Scale characters with Austrovelia queenslandica Malipatil & bars: 0.1 mm. Monteith, by having the scent orifice opening behind

58 THE RAFFLES BULLETIN OF ZOOLOGY 2011

Table1. Differences between Austrovelia queenslandica, Nereivelia murphyi and N. polhemorum, new species.

Austrovelia queenslandica Nereivelia murphyi Nereivelia polhemorum Size (mm.); Shape 0.95–1.50; broad oval 1.70–2.20; elongate oval 1.70–2.09; elongate oval Ventral Head plate-like raised with distinct plate-like raised, weak ridges; plate-like raised, weak ridges, ridges; strongly carinate weakly carinate weakly carinate Eyes normal or reduced, ca 20 normal, ca 30 ommatidia normal, ca 24 ommatidia ommatidia Thorax Surface normal surface with low tubercles surface with disc-like plates on dorsal margins mid-length of segments I:II:III TI = TII + TIII TI = TII + TIII TI > TII+ TIII (0.10 : 0.05 : 0.05) (0.15 : 0.07 : 0.07) (0.22 : 0.10 : 0.09) Posterior margin of metanotum curved straight or slightly curved slightly curved Abdomen swollen, dorsal strongly convex normal normal Mediotergite I : II MI < 1/2 as long as MII MI subequal length to MII MI subequal length to MII Scent orifi ce of mediotergite IV behind middle before middle slightly behind middle Lateral abdominal pleura simple with longitudinal sulcus simple Laterotergites narrow broad and slightly raised broad and slightly raised Posterior margin of male concave simple concave abdomen 8 the middle of mediotergite IV, and by the absence of a Andersen, N. M., 1999. Cryptovelia stysi sp.n. from Borneo with a longitudinal sulcus on the abdominal pleura (see tables by reanalysis of the phylogeny of the Mesoveliidae (Heteroptera: Polhemus & Polhemus, 1989 and Andersen & Polhemus, Gerromorpha). Acta Societatis Zoologicae Bohemoslovacae, 2003). However, Austrovelia differs from the former in the 63: 5–18. distinctive dorsally swollen abdomen and the body size. We Andersen, N. M. & J. T. Polhemus, 1980. Four new genera of have examined paratypes of A. queenslandica and listed their Mesoveliidae (Hemiptera, Gerromorpha) and the phylogeny characters for separating the two genera in Table 1. Our new and classifi cation of the family. Entomogica Scandinavia, 11: species of Nereivelia resembles the type species, N. murphyi, 369–392. by its body size and shape. It can be distinguished by the Anderson, N. M. & D. A. Polhemus, 2003. A new genus of thorax having lesser and shorter setae and shining disc-like Terrestrial Mesoveliidae from the Seychelles (Hemiptera: plates, by the scent gland orifi ce situated slightly behind Gerromorpha). Journal of the New York Entomological Society, 111(1): 12–21. middle of mediotergite IV, and by the concave posterior margin of male abdominal segment VIII. Other differences Andersen N. M. & T. Weir, 2004. Mesoveliidae, Hebridae, between the two species are summarised in Table 1. and Hydrometridae of Australia (Hemiptera: Heteroptera: Gerromorpha), with a reanalysis of the phylogeny of semiaquatic bugs. Invertebrate Systematics, 18: 467–522 ACKNOWLEDGEMENTS Fernando, H. & L. Cheng, 1974. A preliminary checklist of the semiaquatic Hemiptera in Malaya and Singapore. Federation Museums Journal, 19: 21–44. We thank Herbert Zettel (Vienna) for his helpful discussion, to the late Nils Andersen (Copenhagen) for his valuable Malipatil, M. B. & G. B. Monteith, 1983. One new genus and comments and discussions on this secretive mesoveliid, four new species of Terrestrial Mesoveliidae (Hemiptera: Gerromorpha) from Australia and New Caledonia. Australian to Tom Weir (CSIRO, Canberra) for loan of paratypes of Journal Zoology, 31: 943–955. Austraovelia queenslandica, to Lua Hui Kheng, Esther Clews, Yuchen Ang, Tan Swee Hee (NUS) and Tran Anh Duc Murphy, D. H., 1990. Walkers on water – An account of the pleuston of Singapore. In: Chou L. M. & P. K. L. Ng (eds), Essays in (Hanoi University of Science) for their help in SEM, digital Zoology, Papers Commemorating the 40th Anniversary of the images and other assistance, to Peter K. L. Ng (RMBR) for Department of Zoology, National University of Singapore. kind support to make this work possible. Singapore, 153–168. Ng, P. K. L. & N. Sivasothi, 1999 (eds). A Guide to the Mangroves of Singapore 1. The Ecosystem and Plant Diversity. Singapore LITERATURE CITED Science Centre. Singapore, 51–53. Polhemus, J. T. & D. A. Polhemus, 1989. A new mesoveliid genus Andersen, N. M., 1982. The Semiaquatic bugs (Hemiptera, and two new species of Hebrus (Heteroptera: Mesoveliidae, Gerromorpha). Phylogeny, adaptations, biogeography, and Hebridae) from intertidal habitats in Southeast Asian mangrove classifi cation. Entomonograph, 3: 455 pp. swamp. Raffl es Bulletin of Zoology, 37(1&2): 73–82.

59 Yang & Murphy: Guide to Mesovelidae, new species of Nereivelia

Polhemus, J. T. & D. A. Polhemus, 2000. The genus Mesovelia Yang, C. M., T. C. M. Wong, H. K. Lua & L. M. Koh, 1999. A Mulsant & Rey in New Guinea Heteroptera : Mesoveliidae. checklist of aquatic and semi-aquatic bugs (Insecta: Heteroptera) Journal of the New York Entomological Society, 108(3–4): from Pulau Tioman, Peninsular Malaysia. Rafﬂ es Bulletin of 205–230. Zoology, Supplement 6: 277–288 Yang, C. M., H. K. Lua & K. L. Yeo, 1997. Semi-aquatic bug Zettel, H. & A. D. Tran, 2009. First inventory of the water bugs (Heteroptera: Gerromorpha) fauna in the nature reserves of (Heteroptera: Nepomorpha, Gerromorpha) of Langkawi Singapore. Garden’s Bulletin, Singapore, 49(2): 313–320. Island, Kedah, Malaysia. Rafﬂ es Bulletin of Zoology, 57(2): 279–295.

60 THE RAFFLES BULLETIN OF ZOOLOGY 2011

THE RAFFLES BULLETIN OF ZOOLOGY 2011 59(1): 61–67 Date of Publication: 28 Feb.2011 © National University of Singapore

NEW MYRMOTERAS ANTS (HYMENOPTERA: FORMICIDAE) FROM THE SOUTHEASTERN PHILIPPINES

Herbert Zettel Entomological Department, Natural History Museum Vienna, Burgring 7, 1010 Vienna, Austria Email: [email protected]

Daniela Magdalena Sorger Entomological Department, Natural History Museum Vienna, Burgring 7, 1010 Vienna, Austria & Department of Biology, North Carolina State University, Raleigh, NC 27695, USA E-mail: [email protected]

ABSTRACT. – A key to Philippine species of the formicine ant genus Myrmoteras Forel, 1893 is presented and two new species are described: Myrmoteras (Myagroteras) glabrum, new species from the island of Camiguin and Myrmoteras (Myagroteras) mcarthuri, new species from the island of Leyte.

KEY WORDS. – Formicidae, Myrmoteras, Myagroteras, new species, Philippines, Leyte, Camiguin.

INTRODUCTION MATERIAL AND METHODS

Species of the genus Myrmoteras Forel, 1893 are among Specimens. – Specimens are dry mounted on card squares or the most bizarre ant forms and unique among Formicinae triangles. Examination of specimens was carried out with a by having mandibles that form into a specialized trap-jaw LEICA Wild M10 binocular microscope; measurements were mechanism – a character that, however, has convergently taken at magnifi cations of up to 128×. Digital photographs evolved in other subfamilies (Odontomachus and Anochetus were taken with a Leica DFC camera attached to a Leica in Ponerinae; Dacetini in Myrmicinae). The small Myrmoteras MZ16 binocular microscope with the help of Image Manager ants with cryptic living habits in leaf litter have been rarely IM50 and stacked and processed with Helicon 5.0 and Adobe collected in the past, although more specialized collecting Photoshop 7.0 programmes, respectively. methods have yielded a relatively high species diversity, especially on Borneo and Sulawesi (Agosti 1992). Holotypes are deposited in the Entomological Collection of the University of San Carlos, Cebu City, paratypes in Myrmoteras is the only genus of the tribus Myrmoteratini the Natural History Museum Vienna and in the authors’ Emery, 1895 (Bolton 2003) and has a chiefly Oriental reference collections. distribution from India to the Philippines, Sulawesi, and Lombok of the Lesser Sunda Islands; limited to the east Terminology. – Terminology and method of description by Weber’s Line (Agosti 1992; Tab. 1). Both subgenera, chiefl y follow Moffett (1985). Characters of diagnoses are Myrmoteras s.str. and Myagroteras Moffett, 1985, are not fully repeated in the descriptions. probably monophyletic entities (Moffett 1985). Only Myagroteras is known east of Huxley’s Line (Philippines, Measurements and indices. All measurements are in Sulawesi, Lombok), and two Philippine-endemic species millimetres. have been described in the past: Myrmoteras (Myagroteras) TL Total length. Length of outstretched ant measured williamsi Wheeler, 1919 and Myrmoteras (Myagroteras) from apex of closed mandibles to apex of gaster. insulcatum Moffett, 1985. In this study we add two further HW Head width. Maximum width of head, in full-face species. However, the knowledge on Myrmoteras in the view behind eyes (excluding eyes). Philippines is extremely fragmentary (see Fig. 13), and HL Head length, in full-face view, excluding mandibles, much more specifi c fi eld work with leaf litter sampling is measured at full-face view along midline, from required in order to provide a more complete picture than anterior clypeal margin to posterior margin of occipital we can present now. lobe.

61 Zettel & Sorger: Myrmoteras ants from the Philippines

EL Eye length. Maximum diameter of eye. Diagnosis of worker. – Colour mainly dark brown. Dorsum ML Mandibular length. Length of closed mandible of head smooth. Median frontal sulcus prominent. Gap measured in straight line from mandible apex to between clypeus and frons narrow. Labrum not strongly lateral clypeus tooth on the same side as mandible projecting, without long trigger hairs. Mandible with apical being measured, when basal length of mandible shaft part not bent ventrad, with two minute preapical denticles. perpendicular to viewing angle. Disk of pronotum smooth. Dorsum of propodeum strongly MI Mandibular index. ML/HL × 100. convex. Middle tibia strongly dilated, approximately 4.0–4.5 SL Scape length. Length of antennal scape, measured in times longer than wide. straight line, excluding basal condyle. SI Scape index. SL/HW × 100. Description of worker. – Measurements of holotype worker: PW Pronotal width. Maximum width of pronotum in TL 4.48, HW 0.87, HL 0.88, EL 0.57, ML 1.21, MI 137, dorsal view. SL 0.95, SI 109, PW 0.54, WL 1.28. WL Mesosoma length. Diagonal length of alitrunk in lateral view, from frontal-most point of declivitous Measurements of paratype workers: Range of 19 workers: area of pronotum to posterior-most point of apex of TL 4.30–4.72, HW 0.83–0.90, MI 135–141, WL 1.18–1.29. metapleural lobe. Measurements of worker with smallest HW: TL 4.30, HL 0.85, EL 0.53, ML 1.20, MI 141, SL 0.88, SI 106, PW 0.52, WL 1.18. Measurements of worker with largest HW: TAXONOMY OF PHILIPPINE SPECIES TL 4.65, HL 0.91, EL 0.59, ML 1.27, MI 140, SL 0.99, SI 110, PW 0.56, WL 1.27. Key to Philippine species of Myrmoteras (workers and gynes) Colour: Dark brown, posterior half of mesosoma and petiole almost black. (One callow worker with soft integument pale, 1 Head (Fig. 1) and pronotum (Fig. 3) smooth and shiny .... 2 brown and yellow.) Mandibles and palpi yellow. Antennae – Head (Figs. 7, 11) and pronotum (Fig. 9) granulate and dull. brownish yellow. Legs dark brown except bases of tibiae ...... 3 light brown and tarsi pale yellowish. 2 Median frontal sulcus distinct (Fig. 1). Body dark brown ..... Structures: Head (Fig. 1) with sparse pilosity, longest hairs ca...... M. glabrum, new species – Median frontal sulcus absent. Body dark orange red ...... 0.08 mm long. Frons, temples and vertex smooth and shiny...... M. insulcatum Frons with few faint rugulae anteriorly, median frontal sulcus narrow, neither reaching anterior frons margin nor anterior 3 Posterior face of temple (behind ridge) finely granulate. ocellus. Antennal scrobes indistinct, but anteriomedially Mandible Index 1.45 (Fig. 7). Petiolar node wide (lateral limited by short, curved rugae. Temple with blunt ridge aspect, Fig. 8). Antennomeres 2 and 3 of subequal length (Fig. separating a ﬂ at anterior from slightly convex posterior face. 7) ...... M. mcarthuri, new species Clypeus with broad posteromedial elevation, with some – Posterior face of temple (behind ridge) smooth, without any faint rugae, mostly near margins; anterior margin deeply sculpture. Mandible Index 1.30 (Fig. 11). Petiolar node narrow concave. Mandibles long, smooth, dentition see Figure 1. (lateral aspect). Antennomere 2 approximately 1.5 times as long as antennomere 3 (Fig. 11) ...... M. williamsi Maxillary palp with 6, labial palp with 4 segments. Antenna with moderately slender funiculus, each segment less than twice as long as broad (except long ultimate), antennomere 3 SPECIES ACCOUNTS slightly shorter than antennomere 2 (in holotype, measured at leading edge, 0.82 times). Mesosoma, node and gaster (Figs Myrmoteras (Myagroteras) glabrum, new species 2, 3) smooth and sparsely pilose, longest hairs ca. 0.12 mm (Figs. 1–6, 13) long: Pronotum low and evenly convex, summit virtually level with anterior of mesonotum. Mesonotum with low, Etymology. – The Latin adjective glaber means smooth, and the but distinct rugae, irregularly arranged, individually varying epithet refers to the smooth surface of the whole body. from predominately longitudinal to predominately transverse; with pair of distinct ridges extending from metanotal tubercle Material examined. – Holotype (worker) from the Philippines, forward. Metanotal groove visible as feebly impressed notch Camiguin Island, municipality of Mambajao, barangay Tupsan, in lateral aspect. Meso- and metapleura smooth. Propodeum Macaw Cold Spring, ca. 10 m a.s.l., 09°11'N, 124°46'E, 17 March dorsally smooth, in some individuals a few faint rugae 2010, leg. H. Zettel & C. V. Pangantihon (# 517) (label see Fig. indicated. Middle and hind tibiae strongly dilated, middle 4). tibia of holotype 4.3 times as long as wide. Foretibia slender. Paratypes: 17 workers and 1 alate gyne from the same nest as the Node of petiole slender in lateral aspect. Gaster smooth. holotype. 1 worker from the Philippines, Camiguin Island, west of Mambajao proper, Katibawasan spring area, ca. 350 m a.s.l., 15 Description of gyne. – Measurements of paratype gyne: TL March 2010, leg. H.Zettel & C. V. Pangantihon (# 515). 1 worker 4.62, HW 0.91, HL 0.91, EL 0.58, ML 1.20, MI 131, SL 0.95, from the Philippines, Camiguin Island, municipality of Mambajao, SI 104, PW 0.61, WL 1.30. Length of forewing 2.88. barangay Dagoocan, spring area, c. 350 m a.s.l., 14 February 2007, leg. H. Zettel (# 472).

62 THE RAFFLES BULLETIN OF ZOOLOGY 2011

Colour: Similar to worker, but head lighter than mesosoma, gynes (Fig. 6). Pronotum laterally with faint granulation. more orange brown. Wings pale, veins yellow. Mesoscutum posteromedially with some faint rugae. Meso- and metapleura with few longitudinal rugae. Dorsum of Structures: Very similar to worker. MI hardly smaller (Fig. propodeum with low but rather distinct rugae. Wing venation 5). Mesosoma differently structured as typical for winged as typical for the genus.

Figs. 1–6. Myrmoteras (Myagroteras) glabrum, new species. 1–4, paratype worker (Natural History Museum Vienna HYM # 01-0000069, Antbase.net # 00991; HW = 0.85 mm, TL = 4.35 mm), 5–6, paratype gyne (coll. H. Zettel, Vienna; HW = 0.91 mm, TL = 4.62 mm): (1, 5) Head, full face view. (2, 6) Habitus, lateral view. (3) Habitus, dorsal view. (4) Labels. © (1–4) NHMW Image Database & www. antbase.net, (5–6) www.antbase.net, published with permission.

63 Zettel & Sorger: Myrmoteras ants from the Philippines

Comparative notes. – Revisions by Moffett (1985) and the two combinations at couplet 12; but when ignoring dark Agosti (1992) treat all species of Myrmoteras except one colour and following 12–, workers of M. glabrum again key later described species, M. cuneonodum Xu, 1998 from with M. bakeri. The differences between M. bakeri and M. Yunnan, southwestern China (Xu 1998). This Myagroteras glabrum are the following: While M. bakeri is an orange species (erroneously listed as a species of the nominate red species, M. glabrum is dark brown with yellowish subgenus by Bolton et al. 2007) differs from all Philippine mandibles, antennae and tarsi. The dorsum of the propodeum species by head sculpture (smooth, but centre rugose). Both is almost fl at in M. bakeri, but strongly convex in M. glabrum revisions contain identifi cation keys. When using Moffett’s Furthermore, in M. glabrum the middle tibia is strongly (1985) key, Myrmoteras (Myagroteras) glabrum keys with dilated and ca. 4.0–4.5 times as long as broad. This equals M. bakeri Wheeler, 1919, a species recorded from Borneo a “Tibia Width Index” (= middle tibia width / tibia length × and Peninsular Malaysia. When using Agosti’s (1992) key, 100) of ca. 22–25, but Moffett (1985) describes this index the character combination of M. glabrum “body dark brown for M. bakeri differently in key (16–20), diagnosis (> 23) + coxae and femora dark brown + mesosoma length < 1.5 and description (26–27). Differences between M. glabrum mm + propodeum smooth and shining” does not fi t any of and other Philippine species are described in the key.

Figs. 7–12. 7–10, Myrmoteras (Myagroteras) mcarthuri, new species paratype worker (Natural History Museum Vienna HYM # 01-0000068, Antbase.net # 00953; HW = 0.91 mm, TL = 4.75 mm). 11–12, Myrmoteras (Myagroteras) williamsi, gyne from the type locality (coll. H. Zettel, Vienna; HW = 0.96 mm, TL = 4.60 mm): (7, 11) Head, full face view. (8) Habitus, lateral view. (9) Habitus, dorsal view. (10, 12) Labels. © (7–10) NHMW Image Database & www.antbase.net, (11–12) www.antbase.net, published with permission.

64 THE RAFFLES BULLETIN OF ZOOLOGY 2011

Myrmoteras (Myagroteras) mcarthuri, new species Leyte State University, ca. 50 m a.s.l., 19 March 2005, leg. H. (Figs. 7–10, 13) Zettel & C. V. Pangantihon (# 421).

Etymology. – We name this species in honour of our friend and Diagnosis of worker. – Colour mainly medium brown. most enthusiastic myrmecologist Archie McArthur from the South Dorsum of head densely granulate and dull. Median frontal Australia Museum. Coincidally, the type locality is on the island sulcus anteriorly obliterate, posteriorly deep. Gap between where General Douglas MacArthur (1880–1964) landed in 1944 clypeus and frons narrow. Posterior face of temples (behind to end the Japanese occupation of the Philippines. ridge) ﬁ nely granulate. Labrum not strongly projecting, without long trigger hairs. Mandible with apical part not bent Material examined. – Holotype (worker) from the Philippines, Leyte Island, Baybay, at lower slopes of Mt. Pangasugan, on ground ventrad, with two minute preapical denticles; Mandible Index under and between stones at banks of Calbiga-a River near Leyte 145–154. Funiculus of antenna slender, antennomere 3 ca. State University, ca. 50–100 m a.s.l., 20–21 March 2005, leg. H. twice as long as broad and hardly shorter than antennomere Zettel & C. V. Pangantihon (# 422) (label see Fig. 10). 2. Disk of pronotum densely granulate and dull. Mesonotum strongly depressed. Dorsum of propodeum strongly convex. Paratypes: 7 workers from the same nest as the holotype. 3 workers Middle tibia weakly dilated, more than 5 times as long as from the Philippines, Leyte Island, Baybay, Lago-Lago River near wide. Petiolar node wide in lateral aspect.

Description of worker. – Measurements of holotype worker: TL 5.21, HW 0.97, HL 0.96, EL 0.58, ML 1.49, MI 154, SL 1.12, SI 115, PW 0.64, WL 1.46.

Measurements of paratype workers: Range of 10 workers: TL 4.93–5.27, HW 0.92–1.01, MI 145–152, WL 1.38–1.52. Measurements of worker with smallest HW: TL 4.94, HL 0.95, EL 0.56, ML 1.40, MI 147, SL 1.06, SI 115, PW 0.60, WL 1.38. Measurements of worker with largest HW: TL 5.27, HL 1.00, EL 0.60, ML 1.48, MI 148, SL 1.17, SI 116, PW 0.65, WL 1.50.

Colour: Medium brown, gaster slightly lighter than head and mesosoma, propodeum weakly infuscated in some specimens. Mandibles and antennae brownish yellow. Palpi yellow. Legs medium brown except tarsi brownish yellow.

Structures: Head (Fig. 7) with moderately dense pilosity, longest hairs ca. 0.13 mm long. Frons, temples densely granulate and dull (but granulation obsolete at ventro- posterior part of temples). At anterior of frons granulation more or less confl uent to longitudinal rugae; median frontal sulcus narrow, posteriorly deeply impressed and reaching anterior ocellus, anteriorly not reaching frons margin. Antennal scrobes small, shiny, anteromedially limited by short, curved rugae. Temple with sharp ridge separating a concave anterior from fl at posterior. Clypeus with slender medial elevation, with some faint longitudinal rugae, mostly near margins; anterior margin deeply concave. Mandibles very long, smooth, dentition see Figure 7. Maxillary palp with 6, labial palp with 4 segments. Antenna with slender funiculus, each segment about twice as long as broad (except long ultimate), antennomere 3 hardly shorter than antennomere 2 (in holotype, measured at leading edge, 0.96 times). Fig. 13. Map of the Philippines with distributional data of Myrmoteras taken from literature and new fi ndings: green – Los Mesosoma, node and gaster (Figs. 8, 9) with relatively Baños, type locality of M. williamsi and M. insulcatum; white dense pilosity, longest hairs ca. 0.13 mm long: Pronotum – Baguio, additional record of M. williamsi by Moffett (1985); low and evenly convex, summit at level with anterior of yellow – Dumaguete, record of “M. williamsi” by Creighton mesonotum. Mesonotum and propodeum with granulation (1930) and Moffett (1985), possibly based on an undescribed overlaid by some irregular rugae. Pair of distinct ridges species; red – M. glabrum, new species (three close localities near extending from metanotal tubercle forward are present. M mcarthuri, Manbajao); blue – . new species (two close localities Metanotal groove visible as impressed notch in lateral near Baybay).

65 Zettel & Sorger: Myrmoteras ants from the Philippines

Tab. 1: List of Myrmoteras species including information on known morphs and distribution. Data compiled from Moffett (1985), Agosti (1992), Xu (1998), and this paper. Type localities are marked with *. Gyne Male Genus Subgenus Species Worker Distribution Myrmoteras Myrmoteras barbouri Creighton, 1930 x x Indonesia: Java*. Malaysia: Peninsular, Sabah, Sarawak. Singapore. Myrmoteras Myrmoteras baslerorum Agosti, 1992 x Indonesia: Sumatra*. Myrmoteras Myrmoteras binghami Forel, 1893 x Burma*. Thailand. Myrmoteras Myrmoteras brachygnathum Moffett, 1985 x x India*. Myrmoteras Myrmoteras ceylonicum Gregg, 1956 x Sri Lanka*. Myrmoteras Myrmoteras iriodum Moffett, 1985 x x Indonesia: Kalimantan*. Malaysia: Sarawak, Peninsular. Myrmoteras Myrmoteas mjoebergi Wheeler (in Creighton, 1930) x Malaysia: Sarawak*. Myrmoteras Myrmoteras scabrum Moffett, 1985 x India*. Myrmoteras Myagroteras arcoelinae Agosti, 1992 x Malaysia: Sabah*. Myrmoteras Myagroteras bakeri Wheeler, 1919 x x x Malaysia: Sabah*, Peninsular. Myrmotears Myagroteras brigitteae Agosti, 1992 x x Indonesia: Bali, Lombok*. Myrmoteras Myagroteras chondrogastrum Moffett, 1985 x x Malaysia: Sarawak*. Myrmoteras Myagroteras cuneonodum Xu, 1998 x China*. Myrmoteras Myagroteras danieli Agosti, 1992 x x x Malaysia: Sabah*. Myrmoteras Myagroteras diastematum Moffett, 1985 x x Malaysia: Sarawak*, Sabah. Myrmoteras Myagroteras donisthorpei Wheeler, 1916 x x x Indonesia: Kalimantan. Malaysia: Sarawak*, Sabah. Myrmoteras Myagroteras elfeorum Agosti, 1992 x Indonesia: Sulawesi*. Myrmoteras Myagroteras estrudae Agosti, 1992 x x Indonesia: Sumatra*. Myrmoteras Myagroteras glabrum new species x x Philippines: Camiguin*. Myrmoteras Myagroteras indicum Moffett, 1985 x x x India*. Myrmoteras Myagroteras insulcatum Moffett, 1985 x Philippines: Luzon*. Myrmoteras Myagroteras ivani Agosti, 1992 x x Indonesia: Sulawesi*. Myrmoteras Myagroteras jacquelineae Agosti, 1992 x Indonesia: Sulawesi*. Myrmoteras Myagroteras karnyi Gregg, 1954 x Indonesia: Mentawai Archipelago*. Myrmoteras Myagroteras marianneae Agosti, 1992 x Indonesia: Sulawesi*. Myrmoteras Myagroteras maudeae Agosti, 1992 x Indonesia: Sulawesi*. Myrmoteras Myagroteras mcarthuri new species x Philippines: Leyte*. Myrmoteras Myagroteras morowali Moffett, 1985 x x Indonesia: Sulawesi*. Myrmoteras Myagroteras nicoletteae Agosti, 1992 x x Indonesia: Sulawesi*. Myrmoteras Myagroteras susanneae Agosti, 1992 x x Indonesia: Sulawesi*. Myrmoteras Myagroteras tonboli Agosti, 1992 x Malaysia: Sabah*. Myrmoteras Myagroteras toro Moffett, 1985 x x Indonesia: Sulawesi*. Myrmoteras Myagroteras williamsi Wheeler, 1919 x x x Philippines: Luzon*. Myrmoteras Myagroteras wolasi Moffett, 1985 x Indonesia: Sulawesi*. aspect. Meso- and metapleura with longitudinal striation, based on sexuals (two gynes and one male) from Los Baños, partly some granulation visible in addition. Middle and hind Laguna, Luzon Island (Wheeler 1919; Fig. 13). Moffett tibiae moderately dilated, middle tibia of holotype 5.4 times (1985) redescribed gynes (types and additional specimens) as long as wide. Foretibia slender. Node of petiole thickened from Luzon and Negros and workers from Dumaguete, in lateral aspect. Gaster smooth. Negros Island. Although he noted important differences between the material from the two islands, he considered Comparative notes. – Myrmoteras mcarthuri is similar to them as conspecifi c, following Creighton (1930). M. williamsi and has been initially mistaken for this species by the fi rst author. However, close examination of material Myrmoteras williamsi gynes (Fig. 11) collected at the from Leyte yielded the discovery that in fact it represents type locality and deposited in the fi rst author’s collection a distinct new species. Myrmoteras williamsi was described agree perfectly with the original description of this

66 THE RAFFLES BULLETIN OF ZOOLOGY 2011 taxon. Myrmoteras mcarthuri differs in some important Myrmoteras (Myagroteras) insulcatum Moffett, 1985 characteristics from M. williamsi: In M. mcarthuri the posterior face of the temples, behind a blunt ridge, is Notes. – This taxon is only known from the holotype gyne granulate, where it is smooth and shiny in M. williamsi. The originating from Mt. Makiling, Laguna, Luzon Island. The Mandible Index is larger in M. mcarthuri (145–154) than in specimen was illustrated in detail by Moffett (1985) and is M. williamsi (124–130; measurements partly from Moffett available in colour at the Discover Life website by Alpert 1985) (comp. Figs. 7 and 11). In lateral aspect, the petiolar et al. (2010). The specimen is unique in the genus by the node is wide in M. mcarthuri (Fig. 8), but narrow in M. combination of an absent median frontal sulcus and smooth williamsi. Another very obvious difference is found in the and shiny surfaces of head and pronotum. antenna: The funiculus of M. williamsi is stouter than that of M. mcarthuri, especially the basal joints (except antennomere 2) are not much longer than broad in M. williamsi, while ACKNOWLEDGEMENTS their length is almost twice their width in M. mcarthuri. The length of antennomere 3 is approximately 0.7 times Clister V. Pangantihon (University of San Carlos, Cebu the length of antennomere 2 in M. williamsi, but almost City) is sincerely thanked for his help in ﬁ eld work, Archie 1.0 times in M. mcarthuri. In addition, the length of setae, McArthur (South Australia Museum) for a linguistic review, especially on pronotum, is slightly longer in M. mcarthuri and two anonymous reviewers for their useful comments. than in M. williamsi.

Although this compares a gyne with a worker, all characters LITERATURE CITED mentioned do not seem to be morph-related, at least there are no differences in the differential diagnoses of worker Agosti, D., 1992. Revision of the ant genus Myrmoteras of the and gynes of other species (see Moffett 1985). Malay Archipelago (Hymenoptera: Formicidae). Revue Suisse de Zoologie, 99(2): 405–429. Myrmoteras mcarthuri differs from Moffett’s (1985) Alpert, G. D., General D. M. & V. Samarita, 2010. Ants of description of “M. williamsi” workers from Negros, as the Philippines. , retrieved on 20 June scapes, and mandibles” which is absent in M. mcarthuri. 2010. We suspect that this Negros material will turn out being Bolton, B., 2003. Synopsis and classifi cation of Formicidae. Memoirs another undescribed species. of the American Entomological Institute, 71: 1–370. Bolton, B., Alpert, G., Ward P. S. & P. Naskrecki, 2007. Bolton’s catalogue of ants of the world: 1758-2005. Harvard University Myrmoteras (Myagroteras) williamsi Wheeler, 1919 Press, Cambridge, MA, CD-ROM. (Figs. 11–12) Creighton, W. S., 1930. A review of the genus Myrmoteras (Hymenoptera, Formicidae). Journal of the New York Material examined. – 2 gynes and 1 male from Luzon, Laguna, Entomological Society, 38: 177–192, pl. 11. Mt. Makiling, 13–18 November 1992 (label see Fig. 12) (in the Moffett, M. W., 1985. Revision of the genus Myrmoteras fi rst author’s collection). (Hymenoptera: Formicidae). Bulletin of the Museum of Comparative Zoology, 151(1): 1–53. Notes. – Although Moffett (1985) describes workers from Negros, only sexuals (gynes and males) from Central Luzon belong to M. Wheeler, W. M., 1919. The ants of Borneo. Bulletin of the Museum williamsi without doubt. At the Discover Life website, Alpert et of Comparative Zoology, 63: 43–147. al. (2010) present another record from Camarines Sur in southern Xu, Z., 1998. Two new record genera and three new species of Luzon; however, this record should be reconfi rmed, since the Formicidae (Hymenoptera) from China. Entomologica Sinica, fi rst author studied males from the same province which did not 5: 121–127. fully agree with the M. williamsi-male from the type locality. See comparative notes of M. mcarthuri.

67 THE RAFFLES BULLETIN OF ZOOLOGY 2011

THE RAFFLES BULLETIN OF ZOOLOGY 2011 59(1): 69–75 Date of Publication: 28 Feb.2011 © National University of Singapore

FOUR NEW SPECIES OF TRURLIA JAŁOSZY SKI FROM SUMATRA AND BORNEO (COLEOPTERA: STAPHYLINIDAE: SCYDMAENINAE)

P. Jałoszy ski Museum of Natural History, Wrocław University, Sienkiewicza 21, 50-335 Wrocław, Poland Email: [email protected]

ABSTRACT. – Four new species are described in the hitherto monospeciﬁ c genus Trurlia Jałoszy ski (Coleoptera: Staphylinidae: Scydmaeninae): T. acutangula, new species and T. biarcuata, new species from Sumatra (Indonesia); T. constricta, new species from Sabah (East Malaysia); and T. semengohensis new species from Sarawak (East Malaysia). Habitus and important diagnostic characters of the new taxa, including aedeagi, are illustrated.

KEY WORDS. – Coleoptera, Staphylinoidea, Cephenniini, Trurlia, new species, Oriental.

INTRODUCTION and mounted in Canada balsam on pins under card with specimens. Illustrations of aedeagi were prepared by free- The recently discovered genus Trurlia Jałoszy ski is one of hand drawing. Habitus photos were taken as sets of multiple the most unusual taxa of the Scydmaeninae, with the antennae shots at different depths of focus with Olympus C-750 camera composed not of eleven, but only ten antennomeres and Raynox macro lenses, fi nal pictures were assembled with (Jałoszy ski, 2009). As discussed in the original description, CombineZ freeware. The measurement convention follows the two terminal antennomeres are entirely fused and show that of Jałoszy ski (2009). The elytral index is the length of no trace of septum; only a shallow impression indicates the elytra divided by their combined width. In order to provide site of fusion. Moreover, the resulting compound antennomere unambiguous data to identify type specimens, full contents forms a large, oval club, most similar to that known in some of locality and identifi cation labels are cited in the original Histeridae. Apomorphies of Trurlia include also a pair of form and style (i.e., as it appears on the labels), and the lateromedian foveae on the pronotum, which are trichome- symbol “\” is used to indicate the end of line. like structures and most likely represent openings of glands. Repositories of the studied materials: This remarkable genus shares a number of characters with MHNG – Muséum d’Histoire Naturelle, Geneva, Cephennomicrus Reitter, Cephennula Jałoszy ski and some Switzerland other, yet undescribed genera that seem to form a distinct PCPJ – Private collection of Paweł Jałoszy ski, Pozna , lineage within the Cephenniini (Jałoszy ski, submitted). Poland Trurlia has been known so far from a single holotype of T. insana Jałoszy ski, 2009 from Western Sumatra (Bukittinggi, Lembah Anai) and a few females representing undescribed TAXONOMY species from West Malaysia. In the present paper four new species are described from Sumatra and Borneo. The small Trurlia acutangula, new species number of specimens found among nearly 20 thousand dry- (Figs. 1a, 2a, 3a, b) mounted Oriental Scydmaeninae surveyed by the author in various museums confi rms that species of Trurlia are either Material examined. – Holotype. Male, two labels: “SUMATRA: very rare or lead a cryptic life in habitats requiring special Aceh # 25a \ Mt Leuser NP, 300- \ 500 m, Ketambe, 23- \ methods of collecting. All specimens seen so far have been 30.XI.1989, Löbl \ Agosti, Burckhardt” [white, printed], “TRURLIA collected by sifting leaf litter in moist forest fl oor. \ acutangula m. \ P. JAŁOSZY SKI, ‘2010 \ HOLOTYPUS” [red, printed] (MHNG). Paratypes. 1 male, 9 females, same data as holotype (MHNG, PCPJ). MATERIALS AND METHODS Diagnosis. – Anterior part of frons and clypeus with fi ne Dry-mounted specimens were relaxed in warm water and transverse wrinkles; basic vestiture of pronotal disc well dissected; aedeagi were dehydrated in isopropanol and xylene visible under magnifi cation 40×; front angles of pronotum in

69 Jałoszy ski – New Trurlia from Sumatra and Borneo males strongly and in females slightly but distinctly projecting located just posterior to middle, each with frontomesal fringe anterad; pronotum in posterior half barely noticeably of long setae. Surface of disc densely covered with large, narrowing toward base; ante-basal pronotal groove arcuate; elongate, sharply marked and deep punctures separated by humeral carinae shorter than half length of elytra; aedeagus spaces as long as half puncture diameters; basic setae very in ventral view with blunt, subtriangular apex and simple short, sparse and recumbent but well visible, additionally (i.e., not narrowed) apices of parameres. pronotum with several pairs of very long, strongly curved and suberect macrosetae (Fig. 2a). Male. – Body (Figs. 1a, 2a) strongly convex, oval, with distinct but shallow constriction between pronotum and Elytra oval and very short, broadest distinctly anterior to elytra; pigmentation chestnut-brown with slightly lighter middle, length 0.64–0.68 mm, width 0.63–0.65 mm, elytral antennae, palps and legs, vestiture light brown. Body length index 1.02–1.04; humeral carinae slightly divergent caudad, 1.14–1.20 mm. as long as 0.48× of elytral width; surface of elytra covered with punctures much smaller, shallower and sparser than Head moderately large in relation to pronotum, length 0.13 those on pronotum, separated by spaces 2–3× as long as mm, width 0.30 mm, vertex and posterior part of frons puncture diameters; each elytron with about 14 long, strongly confl uent and convex, evenly and densely covered with large, curved and suberect macrosetae (Fig. 2a). Hind wings fully elongate, sharply marked and deep punctures separated by developed. spaces half as long as puncture diameters, anterior part of frons and clypeus with smaller punctures and fi ne transverse Aedeagus (Figs. 3a, b) 0.26 mm in length, in ventral view and oblique wrinkles; supraantennal tubercles small and strongly narrowing toward blunt, subtriangular apex; internal weakly raised; eyes large and strongly convex, coarsely armature simple and lightly sclerotized, composed of faceted; antennal club large and oval, length of antennae median tubular structure located in distal half of aedeagus; 0.35 mm. parameres not reaching apex of median lobe, with simple apices, each with three setae: long apical and short median Pronotum broadest near middle, barely noticeably narrowing subapical seta inserted close to each other and long proximal toward base, in strictly dorsal view trapezoidal with front subapical seta. angles strongly protruding anterad and their frontomesal surfaces flattened and expanded; length 0.38-0.40 mm, Female. – Slightly larger and stouter than male, with less width 0.55-0.56 mm; lateral margins microserrate, slightly protruding front angles of pronotum and lateral margins concave just behind front angles, rounded in middle and behind front angles rounded; body length 1.18–1.28 mm nearly straight in posterior fi fth; hind angles nearly right and (mean 1.24 mm); length of head 0.13–0.16 mm (mean blunt; posterior margin with three shallow emarginations; 0.15 mm), width 0.31–0.34 mm (mean 0.32 mm); length internal ante-basal foveae small but distinct, connected by of antennae 0.35–0.39 mm (mean 0.38 mm); length of sharply marked, arcuate groove; oval lateromedian foveae pronotum 0.38–0.43 mm (mean 0.41 mm), width 0.60–0.63

Fig. 1. Dorsal habitus of holotype males. Trurlia acutangula new species (a), Trurlia biarcuata new species (b), Trurlia constricta new species (c), Trurlia semengohensis new species (d). Scale bars = 0.2 mm.

70 THE RAFFLES BULLETIN OF ZOOLOGY 2011 mm (mean 0.61 mm); length of elytra 0.65–0.70 mm (mean printed] (MHNG). Paratypes: 2 females, same data as holotype, 0.68 mm), width 0.65–0.68 mm (mean 0.67 mm), elytral except one specimen from altitude of 250 m (MHNG, PCPJ). index 1.00–1.04. Diagnosis. – Anterior part of frons and clypeus with fi ne Etymology. – The Latin adjective acutangulus (“sharp- transverse wrinkles; basic vestiture of pronotal disc not angled”) refers to the projected front angles of the pronotum visible under magnifi cation 40×; front angles of pronotum in in this species. strictly dorsal view barely visible, not projecting; pronotum in posterior half slightly but distinctly narrowing toward Distribution. – Sumatra (Indonesia: province Aceh). base; ante-basal pronotal groove distinctly biarcuate; humeral carinae slightly longer than half length of elytra; aedeagus Remarks. – The projected anteriorly and broadened in ventral view with subrectangular apex and parameres frontomesally front angles of the pronotum, remotely with simple (i.e., not narrowed) apices, each bearing three resembling a similar modifi cation common in Cryptophagidae, long setae. are very unusual character in the Cephenniini. In most taxa belonging in this tribe the front angles of the pronotum Male. – Body (Figs. 1b, 2b) strongly convex, oval, with are blunt and not visible or barely visible in dorsal view. distinct but shallow constriction between pronotum and A similar character state can be seen in T. semengohensis elytra; pigmentation chestnut-brown with slightly lighter described below, but in males of the latter species the apices antennae, palps and legs, vestiture light brown. Body length of front angles are distinctly less projected and broadened 1.15 mm. than those in females of T. acutangula. Head moderately large in relation to pronotum, length 0.13 mm, width 0.30 mm, vertex and frons confl uent and convex, Trurlia biarcuata, new species evenly and densely covered with large, elongate, sharply (Figs. 1b, 2b, 3c, d) marked and deep punctures separated by spaces half as long as puncture diameters, clypeus with very fi ne transverse Material examined. – Holotype. Male, two labels: “SUMATRA: wrinkles; supraantennal tubercles small and weakly raised; W Sum. #19 \ Anai V. Nat. Res. \ 10 km W Padangpanjan \ 200 m, eyes large and strongly convex, coarsely faceted; antennal 17,XI.1989, Löbl \ Agosti, Burckhardt” [white, printed], “TRURLIA club large and oval, length of antennae 0.39 mm. \ biarcuata m. \ P. JAŁOSZY SKI, ‘2010 \ HOLOTYPUS” [red,

Fig. 2. Simpliﬁ ed outline of left body half in dorsal view and distribution of macrosetae. Trurlia acutangula new species (a), Trurlia biarcuata new species (b), Trurlia constricta new species (c), Trurlia semengohensis new species (d). Scale bars = 0.2 mm.

71 Jałoszy ski – New Trurlia from Sumatra and Borneo

Pronotum broadest between middle and anterior third, Aedeagus (Figs. 3c, d) 0.33 mm in length, in ventral view weakly narrowing toward base, in strictly dorsal view distinctly narrowing toward separated, subrectangular apex; trapezoidal with front angles weakly protruding anterad internal armature simple and lightly sclerotized, composed of and barely visible; length 0.35 mm, width 0.55 mm; lateral median tubular structure in apical half of aedeagus; parameres margins microserrate, in anterior half rounded, in posterior not reaching apex of median lobe, with simple apices, each half nearly straight; hind angles slightly obtuse and blunt; bearing three equally long setae: apical and median subapical posterior margin with three shallow emarginations; internal setae inserted very close to each other, proximal subapical ante-basal foveae small but distinct, connected by sharply seta remarkably distant from apex. marked, biarcuate groove; oval lateromedian foveae located in middle, each with frontomesal fringe of long setae. Female. – Externally indistinguishable from male, only Surface of disc densely and unevenly covered with large, slightly larger; body length 1.18–1.19 mm; length of head mostly nearly circular or slightly elongate, sharply marked 0.13–0.15 mm, width 0.30 mm; length of antennae 0.38–0.39 and deep punctures separated by spaces 0.5-1.5× as long as mm; length of pronotum 0.38–0.40 mm, width 0.55–0.58 puncture diameters; basic setae extremely short, sparse and mm; length of elytra 0.65–0.66 mm, width 0.63–0.64 mm, recumbent, visible under magnifi cations ≥80×, additionally elytral index 1.04. pronotum with several pairs of very long, strongly curved and suberect macrosetae (Fig. 2b). Etymology. – The Latin adjective biarcuatus refers to the shape of the ante-basal pronotal groove in this species. Elytra oval and very short, broadest distinctly anterior to middle, length 0.68 mm, width 0.65 mm, elytral index 1.04; Distribution. – Sumatra (Indonesia: province West humeral carinae nearly parallel to each other, as long as 0.56× Sumatra). of elytral width; surface of elytra covered with punctures much smaller, shallower and sparser than those on pronotum, Remarks. – Trurlia biarcuata has a relatively unremarkable separated by spaces 1.5–2× as long as puncture diameters; external morphology and identifi cations must be confi rmed each elytron with ca. 20 long, strongly curved and suberect by examination of the unique aedeagus. macrosetae (Fig. 2b). Hind wings not studied.

Fig. 3. Aedeagus in ventral (a, c) and lateral (b, d) views. Trurlia acutangula new species (a, b), Trurlia biarcuata new species (c, d). Scale bar = 0.1 mm.

72 THE RAFFLES BULLETIN OF ZOOLOGY 2011

Trurlia constricta, new species covered with large, sharply marked and deep punctures (Figs. 1c, 2c, 4a, b) separated by spaces comparable to puncture diameters; frons slightly impressed, with punctures slightly smaller than those Material examined. – Holotype. Male, two labels: “SABAH: on vertex but equally distinct and dense; supraantennal Poring Hot \ Springs, 500 m \ 6.V.1987 \ Burckhardt - Löbl” [white, tubercles small but very distinct; eyes large and strongly printed], “TRURLIA \ constricta m. \ P. JAŁOSZY SKI, ‘2010 convex, coarsely faceted; antennal club large and oval, length \ HOLOTYPUS” [red, printed] (MHNG). Paratypes: 5 females, of antennae 0.38 mm. same data as holotype (MHNG, PCPJ). Pronotum broadest near middle, strongly narrowing toward Diagnosis. – Anterior part of frons and clypeus with sparse base, in strictly dorsal view nearly semicircular with barely punctures, without wrinkles; basic vestiture of pronotal disc visible front angles; length 0.38 mm, width 0.55 mm; barely noticeable under magniﬁ cation 40×; front angles of lateral margins microserrate, strongly rounded in anterior pronotum in both sexes weakly projecting anterad, barely half and weakly but distinctly rounded in posterior half; visible in strictly dorsal view; pronotum in posterior half hind angles slightly obtuse and blunt; posterior margin strongly narrowing toward base; humeral carinae longer shallowly biemarginate with barely marked and very short than half length of elytra; aedeagus in ventral view slightly median emargination; internal ante-basal foveae small but constricted laterally in subapical region, with oval, rounded distinct, connected by sharply marked, arcuate groove; oval apex, and apex of each paramere rapidly narrowed at median lateromedian foveae located distinctly posterior to middle, subapical seta. each with frontomesal fringe of long setae. Surface of disc densely covered with moderately large, mostly circular Male. – Body (Figs. 1c, 2c) strongly convex, oval, with or slightly elongate, sharply marked and deep punctures deep constriction between pronotum and elytra; pigmentation separated by spaces as long as 0.5-1 puncture diameter; moderately dark brown with slightly lighter antennae, palps basic setae very short, sparse and recumbent, discernible and legs, vestiture light brown. Body length 1.18 mm. under magniﬁ cation 40×, additionally pronotum with several pairs of very long, strongly curved and suberect macrosetae Head moderately large in relation to pronotum, length 0.15 (Fig. 2c). mm, width 0.30 mm, vertex convex, evenly and densely

Fig. 4. Aedeagus in ventral (a, c) and lateral (b, d) views. Trurlia constricta new species (a, b), Trurlia semengohensis new species (c, d). Scale bar = 0.1 mm.

73 Jałoszy ski – New Trurlia from Sumatra and Borneo

Elytra oval and very short, broadest between middle and Male. – Body (Figs. 1d, 2d) strongly convex, oval, anterior third, length 0.65 mm, width 0.63 mm, elytral index with shallow constriction between pronotum and elytra; 1.04; humeral carinae feebly divergent caudad, as long as pigmentation moderately dark reddish-brown with slightly 0.6× elytral width; surface of elytra covered with punctures lighter antennae, palps and legs, vestiture light brown. Body much smaller and much shallower but only slightly sparser length 1.10 mm. than those on pronotum, separated by spaces 1–1.5× as long as puncture diameters; each elytron with ca. 15 long, Head moderately large in relation to pronotum, length 0.10 strongly curved and suberect macrosetae (Fig. 2c). Hind mm, width 0.30 mm, vertex convex, unevenly and densely wings not studied. covered with moderately large, slightly elongate, sharply marked and deep punctures separated by spaces 0.5-1.5× Aedeagus (Figs. 4a, b) 0.25 mm in length, in ventral view as long as puncture diameters; frons slightly fl attened, with moderately narrowing toward apex and slightly constricted similar punctures as those on vertex; supraantennal tubercles in subapical region; apex separated, oval and rounded; small and weakly raised; eyes large and strongly convex, internal armature very lightly sclerotized, barely discernible, coarsely faceted; antennal club large and oval, length of containing median bell-shaped structure located near middle antennae 0.43 mm. of aedeagus; parameres reaching apex of median lobe, with modifi ed apices and three unequal setae: moderately long Pronotum equally broad between hind angles and middle, in apical seta inserted on long and narrow “projection”; short strictly dorsal view trapezoidal with front angles distinctly median and very long proximal subapical setae approximate protruding anterad and their frontomesal surfaces slightly to each other and distant from apical seta. fl attened and expanded; length 0.35 mm, width 0.51 mm; lateral margins microserrate, straight just behind front angles, Female. – Externally indistinguishable from male, on average rounded near middle and straight in posterior half; hind slightly larger and stouter; body length 1.20–1.24 mm (mean angles nearly right and blunt; posterior margin with three 1.22 mm); length of head 0.15 mm, width 0.30–0.33 mm shallow emarginations; internal ante-basal foveae small but (mean 0.31 mm); length of antennae 0.35–0.40 mm (mean distinct, connected by sharply marked, arcuate groove; oval 0.38 mm); length of pronotum 0.40–0.41 mm (mean 0.41 lateromedian foveae located just posterior to middle, each mm), width 0.58–0.60 mm (mean 0.59 mm); length of elytra with frontomesal fringe of long setae. Surface of disc very 0.65–0.68 mm (mean 0.66 mm), width 0.63–0.65 mm (mean densely covered with moderately large, slightly elongate, 0.64 mm), elytral index 1.00–1.04. sharply marked and deep punctures separated by spaces approximately equal to half puncture diameter; basic setae Etymology. – The Latin adjective constrictus (fem. constricta) extremely short, sparse and recumbent, barely noticeable refers to the distinct constriction between the pronotum and under magnifi cation 80×, additionally pronotum with several elytra in this species. pairs of very long, strongly curved and suberect macrosetae (Fig. 2d). Distribution. – Borneo (Eastern Malaysia: Sabah). Elytra oval and relatively elongate, broadest distinctly anterior Remarks. – This is another unremarkable species; certain to middle, length 0.65 mm, width 0.59 mm, elytral index identifi cations can be made only on the basis of the unique 1.11; humeral carinae nearly parallel, as long as 0.58× of aedeagus. Females can be determined by direct comparison elytral width; surface of elytra covered with punctures much with males. smaller, and shallower but only slightly sparser than those on pronotum, separated by spaces 1–1.5× as long as puncture diameters; each elytron with ca. 15 long, strongly curved Trurlia semengohensis, new species and suberect setae (Fig. 2d). Hind wings not studied. (Figs.1d, 2d, 4c, d) Aedeagus (Figs. 4c, d) 0.40 mm in length, in ventral view Material examined. – Holotype. Male, two labels: “SARAWAK distinctly narrowing toward tapered, trapezoidal apex; internal \ Semengoh For. \ 11miSW Kuching \ R.Taylor, 5.68” [white, armature relatively darkly sclerotized and complex, in lateral printed], “TRURLIA \ semengohensis m. \ P. JAŁOSZY SKI, view showing very large and elongate conical structure ‘2010 \ HOLOTYPUS” [red, printed] (MHNG). occupying most of apical part of aedeagus; parameres not reaching apex of median lobe, with modifi ed apices and Diagnosis. – Anterior part of frons and clypeus with small three unequal setae: very long apical seta inserted on long punctures, without wrinkles; basic vestiture of pronotal disc and narrow “projection”, very short median and very long extremely short, not visible under magnifi cation 40×; front proximal subapical setae approximate to each other and angles of pronotum in males distinctly projecting anterad; distant from apical seta. pronotum in posterior half not narrowing toward base; humeral carinae longer than half length of elytra; aedeagus Female. – Unknown. in ventral view with abruptly tapered, trapezoidal apex, and apex of each paramere rapidly narrowed at median Etymology. – Named after the Semengoh Forest in Sarawak, subapical seta. Borneo.

74 THE RAFFLES BULLETIN OF ZOOLOGY 2011

Distribution. – Borneo (Eastern Malaysia: Sarawak). LITERATURE CITED

Remarks. – In addition to the diagnostic characters Jałoszy ski, P., 2008. Cephennula gen. nov. (Coleoptera, listed above, Trurlia semengohensis is distinct and easily Scydmaenidae), with four species from Malaysia and Indonesia. distinguishable from all congeners on the basis of its very Bulletin of National Science Museum Tokyo, Ser. A, 34(4): small body reaching only 1.10 mm (all other species have 197–208. 1.14–1.28 mm in length). Trurlia insana, though much larger Jałoszy ski, P., 2009. Trurlia, a new Oriental genus of the tribe (1.28 mm), is most similar to T. semengohensis in general Cephenniini (Coleoptera: Scydmaenidae). European Journal body shape, which is more elongate than that of any other of Entomology, 106: 261–274. species described in this paper; the elytral indexes of these Reitter, E., 1884. Beitrag zur Pselaphiden- und Scydmaeniden- two species are 1.12 and 1.11, respectively, while the elytra Fauna von Java und Borneo. II. Verhandlungen der Zoologisch- of T. acutangula, T. biarcuata and T. constricta are very Botanischen Gesellschaft in Wien, 33: 387–428. stout and their indexes range from 1.00 to 1.04.

ACKNOWLEDGMENTS

I express my thanks to Dr. Giulio Cuccodoro and Dr. Ivan Löbl for hosting me during my visit to the MHNG; Dr. Cuccodoro handled the loan of materials used in the present study.

75 THE RAFFLES BULLETIN OF ZOOLOGY 2011

THE RAFFLES BULLETIN OF ZOOLOGY 2011 59(1): 77–82 Date of Publication: 28 Feb.2011 © National University of Singapore

BREVIBORA CHEEYA, A NEW SPECIES OF CYPRINID FISH FROM MALAY PENINSULA AND SUMATRA

Te-Yu Liao Department of Vertebrate Zoology, Swedish Museum of Natural History, SE-104 05 Stockholm, Sweden. E-mail: [email protected] Department of Zoology, Stockholm University, SE-106 91 Stockholm, Sweden

Heok Hui Tan* Rafﬂ es Museum of Biodiversity Research, Department of Biological Sciences, National University of Singapore, Kent Ridge, Singapore 119260, Republic of Singapore. *: corresponding author; e-mail: [email protected]

ABSTRACT. – Brevibora cheeya is a new species of cyprinid ﬁ sh from the Malay Peninsula and Sumatra. It is distinguished from its only congener, B. dorsiocellata, by the complete lateral line, more scales along the lateral row and larger size.

KEY WORDS. – Taxonomy, Brevibora cheeya, Brevibora dorsiocellata, Southeast Asia, Biodiversity.

INTRODUCTION Brittan (1954). According to his defi nition, the lateral line is complete when it extends beyond the insertion of last Brevibora is a monotypic genus diagnosed by the large, anal-fi n ray; if incomplete, the lateral line may end at any black dorsal-fi n blotch and less than nine predorsal vertebrae point before the insertion of last anal-fi n ray. Predorsal (Liao et al., 2010). Brevibora dorsiocellata (Fig. 1) is scales were counted along the dorsum midline from the fi rst characterized amongst others, by having an incomplete lateral scale behind the head to the scale in front of the dorsal-fi n line (Duncker, 1904). However, variation of the lateral line insertion. Transverse line scales were counted on a projected length has been reported on by subsequent workers (Brittan, straight line from the origins of the dorsal and pelvic fi ns 1972; Roberts, 1989). Examination of various populations of (F-J in Fig. 2). Circumpeduncular scale row was the number B. dorsiocellata reveals that some populations have complete of longitudinal scale rows around the caudal peduncle. All lateral line while other populations have incomplete lateral measurements and counts were conducted on the left side line. This character is consistent within each population. of the body whenever possible. The purpose of this paper is to describe a new species Measurements on specimens were taken point-to-point to of Brevibora with complete lateral line from the Malay the nearest 0.1 mm with a pair of digital calipers. Points Peninsula and Sumatra. used in measurements are given in Fig. 2. Numbers in parentheses used in description refer to number of specimens examined. MATERIAL AND METHODS Cleared-and-stained specimens were prepared according to Counts were based on radiographs and cleared-and-stained Taylor and Van Dyke (1985) with slight modifi cations. (C&S) specimens. Caudal vertebrae referred to those vertebrae bearing a haemal-spine, including the compound Terminologies of color pattern followed Brittan (1954) unless centrum, and the remaining vertebrae were counted as noted. Vernacular group names follow Fang et al. (2009), abdominal vertebrae, including the vertebrae forming the viz. rasborins for the genus Rasbora and related genera. Weberian apparatus. The last two rays of the dorsal fi n articulating on the same pterygiophore are counted as one. Specimens examined are from the Natural History Museum, All perforated scales along the lateral line were counted as London (BMNH), the Swedish Museum of Natural History lateral line scales. Scales of lateral row were counted along (NRM) and the Zoological Reference Collection, Raffl es the row of the lateral line, including scales on the caudal Museum of Biodiversity Research, National University of fi n. Defi nition of lateral line completeness follows that of Singapore (ZRC).

77 Liao & Tan – Brevibora cheeya, new cyprinid ﬁ sh from Malaysia

TAXONOMY et al., 14 Sep.1991. - ZRC 42852, 4, 20.0-21.5 mm SL; Malaysia: Johor, Sungei Kayu; H.H. Tan et al., 12 Mar.1998. Brevibora cheeya, new species (Figs. 3, 4) Diagnosis. – Brevibora cheeya is distinguished from B. dorsiocellata by the complete lateral line (25-30 perforated Material examined. – .Holotype – ZRC 51965, 26.6 mm SL; scales vs. 4-9), more scales along the lateral row (29-32 vs. Malaysia: Terengganu, Rantau Abang, 56 km to Kuala Terengganu; 25-27), larger size (up to 39.0 mm SL vs. 23.0 mm SL); P.K.L. Ng et al, 16 May.1995. and larger head and prepectoral length (head length 28.5- Paratypes - ZRC 40196, 10, 20.4-27.3 mm SL; same data as 30.0% SL, vs. 24.4-27.9; prepectoral length 29.6-32.9% holotype. - CMK 8156, 20, 12.9-22.8 mm SL, ZRC 24127, 17, SL, vs. 25.8-28.8). 13.3-22.3 mm SL; same locality as holotype; M. Kottelat et al., 18 Mar.1992. - ZRC 51966, 46, BMNH 2010.1.11.1-70, 70, 14.1-25.5 mm SL; Malaysia: Terengganu, Rantau Abang, 56 km to Kuala Description. – Refer to Fig. 3 for overall body shape and to Terengganu; L. Rüber et al., 28 Feb.2009. Table 1 for morphometric data. Body laterally compressed, rather elongated. Head short. Snout pointed. Mouth terminal, Additional non-type material. – ZRC 39158, 11 (1 C&S), 11.4-28.0 reaching vertical of anterior orbit. Dorsal-fi n origin at highest mm SL; Indonesia: Sumatra: Jambi: Berbak Nature Reserve, Sungai point of body outline. Predorsal contour almost straight, with Air Hitam Dalam; H.H. Ng & S.H. Tan, 16-17 Jun.1995. - ZRC a slight depression on snout. Ventral outline slightly curved 43116, 1, 19.6 mm SL; Indonesia: Sumatra: Riau: Indragiri, Sungai from tip of snout to the middle of caudal peduncle. Barbel, Jakil; H.H.Tan et al., 25 Nov.1996. - ZRC 30911, 2 ex., 30.4-30.9 keel and tubercles absent. mm SL; Sumatra: Banka island, Kampung Jelit; M. Kottelat et al., 2 Mar.1993. - ZRC 3206, 406, 10.9-38.8 mm SL; Malaysia: Pahang, Tasek Bera; C.C. Lindsey, 29 Mar.1963. - ZRC 18826, 7, Dorsal-fi n origin opposite or slightly ahead to pelvic-fi n 25.9-31.9 mm SL; Malaysia: Selangor, Tanjung Malim; P.K.L. Ng origin, tip of last branched ray opposite to anal-fi n origin, pointed, posterior outline rather straight. D. ii.6 (11). Longest pectoral-fi n ray (1st or 2nd branched ray) almost reaching pelvic-fi n origin. P. ii.11 (2), ii.12 (8). Pelvic fi n triangular, longest ray almost reaching anal-fi n origin. V. ii.8 (10). Anal-fi n origin closely behind vent, posterior outline concave, longest ray not reaching caudal fi n A. iii.5 (11). Caudal fi n deeply forked.

Lateral line complete with 25-30 perforated scales [25 (1), Fig. 1: Brevibora dorsiocellata. ZRC 42313, 22.7 mm SL; Indonesia: Sumatra: Jambi: Sungai Alai: 19.5 km to Muara Tebo 26 (2), 27 (5), 28 (1), 29 (1), 30 (1)], last two to six scales from Muara Bungo. not perforated. Lateral row with 29-32 scales [29 (3), 30 (3), 31 (3), 32 (2)], including two scales on base of caudal ﬁ n, lateral row descending from 1st to 4th scale and rather horizontal between 5th and 7th, then descending from 8th scale onwards to caudal-ﬁ n base (scale number based on holotype). Predorsal scales 10-11 [10 (9), 11 (2)]. Transverse

Fig. 2: Points used in measurements of rasborins species. 1, Standard length (SL): A-N, from tip of upper jaw to end of hypural plate. 2, Total length (SL): A-H, from tip of upper jaw to tip of Fig. 3: Brevibora cheeya. ZRC 51965, holotype, 26.6 mm upper caudal-fi n lobe. 3, Head length: A-E, from tip of upper jaw SL; Malaysia: Terengganu, Rantau Abang, 56 km to Kuala to posterior edge of opercle. 4, Head depth: at level of posterior Terengganu. margin of orbit (D). 5, Orbital diameter: B-D, between horizontal margins of osseous orbit. 6, Snout length: A-B, from tip of upper jaw to anterior margin of osseous orbit. 7, Interorbital width: distance between upper margins (C) of each osseous orbit. 8, Predorsal length: A-F, from tip of upper jaw to dorsal-fi n origin. 9, Preanal length: A-K: from tip of upper jaw to anal-fi n origin. 10, Prepectoral length: A-I, from tip of upper jaw to pectoral-fi n origin. 11, Prepelvic length: A-J, from tip of upper jaw to pelvic- fi n origin. 12, Dorsal hypural length: F-N, from anterior edge of dorsal-fi n insertion to end of hypural plate. 13, Body depth: at level of dorsal fi n origin (F). 14, Caudal peduncle length: L-N, from end of anal-fi n base to end of hypural plate. 15, Caudal peduncle Fig. 4: Brevibora cheeya. ZRC 39158, 11.4 mm SL; Indonesia: Jambi depth: narrowest part of caudal peduncle (G-M). Province: Berbak Nature reserve, Sungai Air Hitam Dalam.

78 THE RAFFLES BULLETIN OF ZOOLOGY 2011 line scales ½ 4/1/3 ½ (11). One scale between lateral-line anterior middle dorsal fi n, intensely pigmented with a defi ned scale row and the pelvic-fi n origin. Circumpeduncular scales outline, oval in shape, covering two unbranched and fi rst four ½ 3/1/1 ½ (11). Pectoral axillary scale rudimentary. Pelvic branched rays. Other fi ns hyaline, with some melanophores axillary scale well developed, corresponding to 1/3 of pelvic scattered on fi n rays. Melanophores most intense on distal fi n in length. half of anal fi n rays, similar to a blotch in overall shape. Inter-radial membrane of anal fi n transparent. It is not sure Predorsal vertebrae 8-9 [8 (10), 9 (1)], abdominal vertebrae whether the blotch on anal fi n due to stronger pigmentation 16 (11), caudal vertebrae 15-16 [15 (10), 16 (1)], total caused by environmental factors. In the smallest specimen of vertebrae 31-32 [31 (10), 32 (1)]; pharyngeal teeth in three ZRC 39158 (Fig. 4) the distal half of its pelvic fi n rays with rows, 5, 4, 2; 2, 4, 5 (1), rasborin process present. highly scattered melanophores and pigmentation on its anal fi n rays more intense than larger specimens. It is postulated Coloration in preservative. – Dorsum dark brownish, ventral that this pigmentation fades out upon maturity. gradually lighter. Abdomen yellowish. Dorsum of head dark brownish. Tip of snout more densely pigmented than adjacent Distribution and field notes. – Brevibora cheeya is area, exhibiting an appearance of dusky lips. Suborbital distributed both in the westerly and easterly fl owing river yellowish. Gill cover rather transparent with brownish basins of Peninsular Malaysia (western: Selangor; eastern: pigment. Iris whitish, black dorsally and ventrally. Dorsal Terengganu, Pahang, Johor), Central Sumatra (Jambi, Riau) stripe prominent, half of dorsal-scale row in width. Reticulate and Banka Island (Fig. 5). Brevibora dorsiocellata appears pattern on sides distinct. Melanophores densely scattered to be restricted to the Muar river basin in Malaysia, but along distal margin of scales in three or four rows, more this could be due to insuffi cient sampling (based on ZRC prominent in dorsal half of body. An axial streak running from material and second author pers. obs.); but occurs in Central two scales in front of caudal-fi n base to the upper edge of Sumatra, where B. cheeya is also present, but seemingly in gill opening, fading out anteriorly. Dark lateral stripe absent. different sub-basins. The Brevibora dorsiocellata group is Supra-anal pigment present as a streak and connected with also present in Borneo, and will be covered in more detail subpeduncle streak, both blackish. A distinct black blotch on in future works.

The type locality in Malaysia is in a coastal heath forest (Fig. 6). This habitat consists mainly of Melaleuca (Myrtaceae) stands on sandy and peaty substrate. The waters are tannin stained, slow ﬂ owing, and acidic (pH 4.5, as measured in February 2009). Syntopic species recorded from the type locality include: Boraras maculatus, Cyclocheilichthys apogon, Osteochilus spilurus, Parachela maculicauda, P. oxygastroides, Rasbora einthovenii, R. trilineata, Systomus johorensis, Trigonopoma gracile, T. pauciperforatum (Cyprinidae), Lepidocephalichthys furcatus, Pangio alcoides, P. semicincta (Cobitidae), Kryptopterus macrocephalus, Ompok leiacanthus, Wallago leerii (Siluridae), Hemibagrus nemurus, Pseudomystus leiacanthus (Bagridae), Clarias batrachus, C. meladerma (Clariidae), Parakysis verrucosa (Akysidae), Aplocheilus panchax (Aplocheilidae), Hemirhamphodon pogonognathus (Hemiramphidae), Monopterus albus (Synbranchidae), Nandus

Fig. 5: Map of Sumatra and Malay Peninsula showing the distribution of Brevibora cheeya (square; hollow symbol denotes the type locality) and B. dorsiocellata from comparative material Fig. 6: Type locality of Brevibora cheeya at Rantau Abang, (triangle; hollow symbol denotes the type locality). Terengganu, Malaysia (February 2009).

79 Liao & Tan – Brevibora cheeya, new cyprinid fi sh from Malaysia are from 12 specimens including two of paralectotypes BMNH 1913.5.24.13-14 and ten B. dorsiocellata . SD = standard deviation. Measurements of Brevibora cheeya Brevibora n 25.8 26.2-29.3 27.8 1.0 n 16.5 16.5-18.8 17.7 (n=7) 0.9 length of anal fi range mean SD range mean SD n=12 mean range (n=10) SD mean Paratype range of ZRC 42313. Differences in range variation are indicated bold. Holotype length length Standard length (SL; in mm) deapth In % of SL total 0.8 (n=10) (n=10) 2.6 head length body length caudal peduncular depth 26.6 28.5 28.5-30.0 29.3 137.3 137.1-143.0 139.2 0.5 caudal peduncular length 141 1.7 26.3 24.4-27.9 26.5 24.4-26.7 25.6 27.0 length 1.0 0.7 137.2-144.7 predorsal 25.8-28.4 length preanal 72.3 70.2-73.9 1.1 20.4-27.3 length prepectoral 73.6 70.6-76.8 1.3 51.2 51.2-55.2 53.5 prepelvic 1.1 53.5 48.8-56.0 dorsal-hypural 50.8 49.3-52.2 50.7 1.3 53.8 52.7-54.6 53.7 1.1 11.9 depth 0.6 17.0-22.1 length of dorsal fi 52.7 51.0-55.4 54.4 51.8-57.9 18.5 1.4 length 1.3 diameter In % of head length (HL) width 11.7-13.0 head 16.6-18.5 snout 29.6 orbital 63.5 59.5-63.9 61.8 interorbital cheeya 1.3 Brevibora 32.4 28.3-32.8 30.5 35.1 33.3-40.0 37.2 12.3 60.3-65.5 1.6 62 2.2 35.1 33.8-41.0 37.7 28.3 23.4-30.9 34.5-37.7 17.3 1.1 2.1 36 1.7 37.3 29.8-43.4 29.6-32.9 1.1 2.3 0.5 0.6 31 10.5-13.6 17.3-20.7 0.8 dorsiocellate Brevibora 11.8 18.5 25.8-28.8 0.5 0.8 27.7 0.8 Table 1: Morphometry of

80 THE RAFFLES BULLETIN OF ZOOLOGY 2011 nebulosus (Nandidae), Pristolepis grooti (Pristolepididae), cheeya. In some rasborins with complete lateral line, the last Belontia hasseltii, Betta imbellis, B. waseri, Luciocephalus few scales are usually not perforated, commonly observed pulcher, Parosphromenus paludicola, Trichopodus leerii, T. in the Rasbora daniconius and R. trifasciata species groups trichopterus, Trichopsis vittata (Osphronemidae), Channa (Brittan, 1954), and this is probably the reason why Brittan bankanensis, C. lucius and C. striata (Channidae) (Kottelat (1954) made a unique deﬁ nition of lateral line completeness et al., 1992; unpublished data). (see MATERIAL AND METHODS).

Etymology. – Cheeya and Beiya, are two Chinese deities Kottelat & Vidthayanon (1993) provided a list of 47 miniature who hunt ghosts for Yama; “chee” and “bei” mean seven fi sh species from South and Southeast Asia according to the and eight, respectively, and “ya” is an honorable title for a criterion proposed by Weitzman & Vari (1988), being less respected person. Cheeya is tall and Beiya is short; in allusion than 20 mm SL at female maturity or unknown maturity to its relatively larger size as compared to B. dorsiocellata. but never surpassing 26 mm SL in the wild. Brevibora A noun in apposition. dorsiocellata (sensu lato) was not included in their list, and none of the subsequent authors considered this species as a miniature fi sh. However, after this present description of DISCUSSION B. cheeya, B. dorsiocellata (less than 23 mm SL) meets the criterion of a miniature fi sh. Brevibora dorsiocellata was described by Duncker (1904) as a member of Rasbora, based on specimens from Muar A black blotch in the middle of dorsal fi n is a conspicuous (north-western corner of the southern state of Johor in character distinguishing Brevibora from the other rasborins. Malaysia). This species is a popular aquarium fi sh well-known Among species of rasborins, Rasbora atridorsalis and R. for the conspicuous black blotch on its dorsal fi n. Duncker dorsinotata also possess a black blotch on the dorsal fi n (1904) mentioned that B. dorsiocellata has an incomplete (Kottelat & Chu, 1987), but the blotch is at the tip rather lateral line with only eight perforated scales. However, in than in the middle of the fi n. his drawing (Duncker, 1904), the lateral line is complete. Weber & Beaufort (1916) noticed this discrepancy and after Comparative material. – Brevibora dorsiocellata: examination of the types sent by Duncker, they stated that BMNH1913.5.24.13-14, 2 paralectotypes, 17.0 & 19.8 the drawing is erroneous. In addition, the drawing did not mm SL; Malaysia: Johore. - ZRC 2315, 3 paralectotypes, show the unique lateral line pattern shared by Brevibora and 14.4-15.7 mm SL; Malaysia: Negeri Sembilan, Kuala Kottelatia. Meinken (1951) described a subspecies of Rasbora Jelai. - BMNH 1994.12.16.237, 1, 21.6 mm SL; Indonesia: dorsiocellata as R. dorsiocellata macrophthalma, also with a Kalimantan Tengah: Sungai Sebangau: 1 km downstream black blotch on the dorsal fi n and an incomplete lateral line, from Keram Benkari (10km South of Palangka Raya). - which was subsequently synonymized with B. dorsiocellata ZRC 42313, 11 (out of 67), up to 23.0 mm SL; Indonesia: by Brittan (1954). The holotype of B. macrophthalma is Sumatra: Jambi: Sungai Alai: 19.5 km to Muara Tebo from missing (Eschmeyer & Fricke, 2009), so no direct comparison Muara Bungo. ZRC 38494, 2, 9.6-17.8 mm SL; Indonesia: can be made. Moreover, the locality is vague and listed Sumatra: Jambi: Danau Pinang, Sungai Pijoan. - ZRC 38580, only as Malay Archipelago, as it had been obtained from 21, 13.5-19.9 mm SL; Indonesia: Sumatra: Jambi: Danau the aquarium trade. Since B. macrophthalma also possesses Kamining. - ZRC 38563, 33, 10.8-20.9 mm SL; Indonesia: an incomplete lateral line, it would not affect the validity Sumatra: Jambi: Sungai Alai. NRM 57237, 1 (C&S), 19.8 of B. cheeya. We follow Brittan’s opinion in treating B. mm SL; Aquarium. macrophthalma as a synonym of B. dorsiocellata.

Brittan (1954) considered variation in length of lateral line ACKNOWLEDGEMENTS as an intra-species variation in B. dorsiocellata. However, this conclusion was based on only fi ve specimens from three The fi rst author is grateful to Erik Åhlander for translating the localities. Apparently the sample size is insuffi cient and, German literature; Kelvin K. P. Lim for his assistance during indeed, no variation was observed within any population his stay in the Raffl es Museum of Biodiversity Research; by Brittan (1954). Roberts (1989) also noted the difference two anonymous reviewers for their constructive suggestions. in lateral line length, but he adopted the interpretation of Part of the research was conducted in the Raffl es Museum Brittan without additional comments. In material of B. of Biodiversity Research, supported by a grant to the fi rst dorsiocellata from Malay Peninsula and Sumatra, we found author from the Riksmusei Vänner. The second author that lateral line length (complete or incomplete) is a stable thanks Lukas Rüber, Sebastien Lavoué, Norsham Yaacob character within a population and locality, and hence the and Dennis Yong, for assistance and logistic support during lateral line completeness is a good character for species the Malaysian peat swamp survey in 2009. Material collected diagnosis within this genus. Brittan (1954) reported that two and used for research by the second author is funded by specimens of B. dorsiocellata (s. l.) from Pahang (SU 31196) various research grants including RMBR, R-154-000-318- possess incomplete lateral line of 30 and 33 mm SL, one 112, R-264-001-004-272; and the National Geographic grant with 20 pores on the left and 28 on the right, the other with 8509-08 and Natural Environment Research Council grant 24 pores on both sides. Although we did not examine these NE/F003749/1 to Lukas Rüber. two specimens, it is likely that these two specimens are B.

81 Liao & Tan – Brevibora cheeya, new cyprinid ﬁ sh from Malaysia

LITERATURE CITED Kottelat, M., P. K. L. Ng & K. K. P. Lim, 1992. Recent collections of freshwater fish from Terengganu, Peninsular Malaysia. Brittan, M.R., 1954. A revision of the Indo-Malayan fresh-water Malayan Naturalist, 46: 7–12. fi sh genus Rasbora. Monographs of the Institute of Science Liao, T.Y., S.O. Kullander & F. Fang, 2010. A phylogenetic analysis and Technology, Manila. 224 pp. of the cyprinid fi sh genus Rasbora (Teleostei: Cyprinidae). Duncker, G., 1904. Die Fische der malayischen Halbinsel. Zoologica Scripta, 39: 155–176. Mitteilungen aus dem Naturhistorischen Museum in Hamburg, Meinken, H., 1951. Mitteilungen der Fischbestimmungsstelle des 21: 133–207. VDA. Aquarien und Terrarien Zeitschrift, 4: 119–120. Eschmeyer, W. N. & R. Fricke (eds.), 2009. Catalog of Fishes Roberts, T. R., 1989. The freshwater fi shes of western Borneo electronic version (version of 9 September 2009). Available (Kalimantan Barat, Indonesia). Memoirs of the California via http://research.calacadcmy.org/ichthyology/catalog/ Academy of Sciences, 14: 1–10. fi shcatsearch.html. Taylor, W.R. & G.C. Van Dyke, 1985. Revised procedures for Fang, F., M. Noren, T.Y. Liao, M. Källersjö & S.O. Kullander, 2009. staining and clearing small fi shes and other vertebrates for Molecular phylogenetic interrelationships of the south Asian bone and cartilage study. Cybium, 9: 107–119. cyprinid genera Danio, Devario, and Microrasbora (Teleostei, Cyprinidae, Danioninae). Zoologica Scripta, 38: 237–256. Weber, M. & L.F. de Beaufort, 1916. The fishes of the Indo- Australian Archipelago. III. Ostariophysi: II Cyprinoidea, Kottelat, M. & X.L. Chu, 1987. Two new species of Rasbora Apodes, Synbranchi. Brill, Leiden. 455 pp. Bleeker, 1860 from southern Yunnan and northern Thailand. Spixiana, 10: 313–318. Weitzman, S.H. & R.P. Vari, 1988. Miniaturization in South American freshwater fishes; an overview and discussion. Kottelat, M. & C. Vidthayanon, 1993. Boraras micros, a new genus Proceedings of the Biological Society of Washington, 101: and species of minute freshwater fi sh from Thailand (Teleostei: 444–465. Cyprinidae). Ichthyological Exploration of Freshwaters, 4: 161–176.

82 THE RAFFLES BULLETIN OF ZOOLOGY 2011

INTRODUCTION OF AN INDOCHINESE FRESHWATER CRAB SAYAMIA GERMAINI (CRUSTACEA: BRACHYURA: GECARCINUCIDAE) TO TAIWAN: MORPHOLOGICAL AND MOLECULAR EVIDENCE

Hsi-Te Shih Department of Life Science, National Chung Hsing University, 250, Kuo Kuang Road, Taichung 402, Taiwan, ROC Email: [email protected]

Jhy-Yun Shy Department of Aquaculture, National Penghu University, 300 Liu-Ho Road, Magong City, Penghu 880, Taiwan, ROC Email: [email protected] (Corresponding author)

Tohru Naruse Transdisciplinary Research Organization for Subtropical and Island Studies, University of the Ryukyus, 870 Uehara, Taketomi, Okinawa 907-1541, Japan

Ho-Tian Hung Zoology Department, National Museum of Natural Science, 1 Guancian Road, Taichung 404, Taiwan, ROC

Darren C. J. Yeo Department of Biological Sciences, National University of Singapore, 14 Science Drive 4, Singapore 117543, Singapore

Peter K. L. Ng Department of Biological Sciences, National University of Singapore, 14 Science Drive 4, Singapore 117543, Singapore

ABSTRACT. – Freshwater crabs of the genus Sayamia (family Gecarcinucidae) are native to Indochina and northern Peninsular Malaysia. Recently, individuals of Sayamia were collected from the coastal areas of Kaohsiung, southwestern Taiwan. The species is identiﬁ ed as S. germaini (Rathbun, 1902) [from southern Vietnam and Cambodia] by morphological characters as well as 16S rRNA and cytochrome oxidase I DNA sequences. Individuals shared identical haplotypes, which suggest that they are derived from a small founder population. The possible source and pathways of introduction of this alien species and its potential impacts on Taiwanese aquatic biodiversity, human health, and agriculture are discussed.

KEY WORDS. – Sayamia germaini, freshwater crab, alien species, Vietnam, Cambodia, Taiwan, 16S rRNA, cytochrome oxidase I.

INTRODUCTION distributions. The richness of freshwater crabs on Taiwan island (and islets) is the highest among the East Asian islands True (or primary) freshwater crabs are land-locked organisms (Taiwan, the Ryukyus and mainland Japan), with 42 species with direct development of fertilized eggs and maternal belonging to four genera: Geothelphusa, Candidiopotamon, care (Ng, 1988; Cumberlidge & Ng, 2009). Owing to their Nanhaipotamon (family Potamidae) and Somanniathelphusa relatively poor dispersal abilities, they are easily isolated (family Gecarcinucidae) (Shy et al., 1994; Ng et al., 2008; by geographical barriers such as mountains, seas and Shih et al., 2008, 2010, in press; Shih & Shy, 2009; Shih even rivers, with their biogeography often reﬂ ecting past & Ng, in press). While Geothelphusa and Candidiopotamon geological events (see Shih et al., 2009). Not surprisingly, are endemic to the East Asian islands, most species of many species of freshwater crabs have very restricted Nanhaipotamon and Somanniathelphusa are more widely

83 Shih et al. – Introduction of Indochinese freshwater crab to Taiwan distributed on the Asian mainland (Ng & Naiyanetr, 1993; Swofford, 2003) with 2000 bootstrap reiterations of a simple Dai, 1999; Shy & Yu, 1999; Ng et al., 2001; Shih & Ng, heuristic search, tree bisection-reconnection (TBR) branch- in press; Shih et al., in press). swapping, and 100 random addition sequence replicates. All characters were equally weighted. Gaps in the MP tree Recently, specimens of Sayamia Naiyanetr, 1994 (family construction were treated as missing data. The relationships Gecarcinucidae) were collected from two canal systems in of the combined 16S and COI haplotypes in S. germaini Kaohsiung, southwestern Taiwan. Sayamia species are native and its closest congener, S. sexpunctata, were examined by to southern Indochina (Thailand, Cambodia and southern using the program TCS (vers. 1.20, Clement et al., 2000) Vietnam) and northern Peninsular Malaysia (Rathbun, 1902, with gaps treated as a missing data. 1905; Ng, 1988, 1997; Ng & Naiyanetr, 1993; Naiyanetr, 1994; Yeo & Ng, 1999; Ng et al., 2008), and have never been reported from Taiwan, despite the island being extensively RESULTS surveyed and the fauna very well documented (Shy et al., 1994; Shy & Yu, 1999; Ng et al., 2001). Thus, the presence Taxonomy of this genus in Taiwan is certainly the result of anthropogenic introduction. In this paper, we report the presence of the Five adult male specimens (NTOU F10301, 10302, 10303, Indochinese species, Sayamia germaini Rathbun, 1902, in 10306, 10307; CW 46.12–54.58 mm) and two adult females Taiwan. The species is identiﬁ ed using morphological and (NTOU F10304, CW 48.14 mm, ovigerous; NTOU F10305, molecular (16S rRNA and cytochrome oxidase I, COI) 48.69 mm) were collected from a canal near the coast in characters. Possible pathways of introduction into Taiwan Siaogang, Kaohsiung City, southwestern Taiwan in October and potential impacts of the species on native fauna, human and November of 2005; and two females (NCHUZOOL health and agriculture are discussed. 13386, 13387; CW 50.72, 42.41 mm) from a depression

MATERIALS AND METHODS A

Fresh specimens of Sayamia from the introduced areas of Kaohsiung in southwestern Taiwan, and markets of southern Vietnam and Cambodia, within the native range of the genus, were preserved in 70% to 95% ethanol after collection, illustrated with the help of a drawing tube attached to a stereomicroscope, and deposited in the Department of Environmental Biology and Fisheries Science, National Taiwan Ocean University, Taiwan (NTOU) and the Zoological Collections of the Department of Life Science, National Chung Hsing University, Taiwan (NCHUZOOL). The following abbreviations are used: G1 for the male ﬁ rst pleopod and CW for the carapace width. Comparative specimens are deposited in the Muséum national d’Histoire naturelle, Paris (MNHN), and Zoological Reference Collection of the Rafﬂ es Museum of Biodiversity Research, National University of Singapore (ZRC) (Table 1).

Sequences of 16S and COI from the above specimens (n = 24), with Somanniathelphusa taiwanensis Bott, and Siamthelphusa improvisa (Lanchester) used as outgroups (Table 1), were obtained following the method described by Shih et al. (2007) and analysed. The best-ﬁ tting model for sequence evolution of the combined 16S and COI dataset was determined by MrModeltest (vers. 2.2, Nylander, 2005), and selected by Akaike Information Criterion (AIC), with subsequent Bayesian Inference (BI) performed with MrBayes (vers. 3.1.1, Ronquist & Huelsenbeck, 2003). The search was run with four chains for 10 million generations and four independent runs, with trees sampled every 1000 BC generations and the ﬁ rst 500 trees were discarded as the burnin (determined by the average standard deviation of split Fig. 1. Sayamia germaini (Rathbun, 1902). A, dorsal view of a male frequency values below the recommended 0.01; Ronquist (NTOU F10307, CW 46.1 mm) from the coastal region of Kaohsiung, et al., 2005). A consensus maximum parsimony (MP) tree southwestern Taiwan; B, C, G1s of holotype (MNHN-B5162, CW was constructed using the program PAUP* (vers. 4.0b10, 44.2 mm) and NTOU F10301 (CW 50.4 mm)

84 THE RAFFLES BULLETIN OF ZOOLOGY 2011 beside a canal in Renwu, Kaohsiung County (Table 1). The best model selected by MrModeltest was the GTR+I+G The external morphology (Fig. 1A) and the structure of model (Rodríguez et al., 1990) for the combined 16S and COI the G1 (Fig. 1B) of these specimens agree very well with segment of 1170 bp. For MP, a single tree was recovered Sayamia germaini (Rathbun, 1902) collected from southern with a tree length of 301 steps, a consistency index of 0.89, Vietnam (Table 1), including the holotype of the species and a retention index of 0.87. (MNHN-B5162, Fig. 1C), and we have little doubt they are conspecifi c. Sayamia germaini is naturally distributed The phylogenetic tree constructed by BI and MP methods in southern Vietnam (Rathbun, 1902, 1905) and Cambodia (Fig. 2) agrees with the morphological results, with the (this study) (Table 1). haplotypes from Vietnam, Cambodia and Taiwan forming a highly-supported clade of Sayamia germaini, with the closely related S. sexpunctata from southern Thailand and northern DNA analysis Peninsular Malaysia forming a sister clade. Another clade is composed of S. bangkokensis and S. cf. maehongsonensis A ~550 basepair (bp) segment (excluding the primer regions) from Thailand. The bp differences and nucleotide divergence of the 16S from all 24 ingroup specimens was amplifi ed and with the Kimura (1980) 2-parameter distance (K2P distance) aligned; 29 positions were variable and 18 parsimoniously between those haplotypes of COI is shown in Table 2. informative. Among the total number of sequences, 10 For S. germaini, the bp difference is ≤ 7 (1.14%) and the different haplotypes were distinguished (Table 1). The studied nucleotide divergence is ≤ 1.15%; for S. sexpunctata, the segment of 16S sequences was AT rich (69.9%) (T, 33.9%; bp difference is ≤ 4 (0.65%) and the nucleotide divergence A, 36.0%; G, 20.1%; and C, 10.0%). For the COI gene, a 616 is ≤ 0.65%. The minimum interspecifi c bp differences and bp segment was compared, resulting in 9 different haplotypes divergence is 18 (2.92%) and 3.00% (between S. germaini of the ingroups (Table 1). The studied segment of the COI and S. sexpunctata), respectively. Thus, the minimum COI sequence was also AT rich (63.5%) (T, 36.4%; A, 27.1%; divergence is 2.6 times higher between species than within. G, 14.9%; and C, 21.5%). In this gene, 72 positions were Only a single haplotype of 16S and COI was sequenced from variable and 61 parsimoniously informative. the seven S. germaini specimens from Taiwan, compared

Fig. 2. A Bayesian inference (BI) tree of the Sayamia species from Taiwan, Vietnam, Cambodia, Malaysia and Thailand based on the combined 16S rRNA and cytochrome oxidase I genes. Probability values at the nodes represent support values for BI and maximum parsimony (MP). For haplotype abbreviations and detailed localities see Table 1.

85 Shih et al. – Introduction of Indochinese freshwater crab to Taiwan of Environmental Biology and Fisheries Science, National es Museum of Biodiversity Research, National University Singapore, Singapore). species, with the outgroups, used in this study. (CBM = Natural History Museum and Institute, Chiba, Japan; Sayamia elds along Ulu Melaka, Langkawi ZRC 1989.2026-38 1 SYs-1 AB601845 SYs-C1 AB601856 TAIWAN Shihguei, Dounan, Yunlin NCHUZOOL 13001b 1 AB265228 AB265240 TAIWAN Yanhai 2nd Road, Siaogang, Kaohsiung City NTOU F10301, 10302, 10304, 5 SYg-1 AB601842 SYg-C1a AB601853 THAILAND Phitsanulok (in lower northern Thailand) ZRC 1997.108 Thani THAILAND Surat 1 SYm-1 AB601850 SYm-C 1999.0024 ZRC AB601860 1 AB601852 AB601861 THAILAND Bangkok 1989.2159-2165 ZRC 1 SYb AB601849 SYb-C AB601859 MALAYSIA Paddy ﬁ MALAYSIA Paddy maehongsonensis cf. OUTGROUPS Somanniathelphusa taiwanensis Siamthelphusa improvisa Mae Taeng (Taeng River) (in northern Thailand) CBM 1 SYm-2 AB601851 SYm-C AB601860 Bangkok ZRC 1989.2151-2155 1 SYb AB601849 — AB601849 SYb 1 1989.2151-2155 Island (neotype) Bangkok ZRC S. bangkokensis S. THAILAND Khanom (in northern Peninsular Malaysia) Chuping, Perlis Northern Peninsular Malaysia ZRC1985.1956 Malaysia (aquarium purchase in Singapore) ZRC 1 ZRC 2001.1101 SYs-4 ZRC 1984.7352-9, 7689-94 AB601848 1 1 1 — SYs-1 SYs-2 SYs-3 AB601845 AB601846 AB601847 SYs-C1 SYs-C2 SYs-C3 AB601856 AB601857 AB601858 16S no. COI no. COI no. 120°21'29.19"E) 16S NCHUZOOL = Zoological Collections of the Department Life Science, National Chung Hsing University, Taiwan; NTOU (22°41'29.58"N, Taiwan Ocean University, Taiwan; ZRC = Zoological Reference Collection of the Rafﬂ Species S. germaini Takeo Countries Localities S. sexpunctata VIETNAM market in Di Linh, Lam Dong (22°33'25.96"N, 120°22'7.24"E) Renwu, Kaohsiung County CAMBODIA Trapeang Thom Khang Cheung, Tramkak, Voucher number market in Di Linh, Lam Dong CBM 10306, 10307 NZHUZOOL 13319 market in Duong Minh Chau, Tay Ninh NZHUZOOL 13386, 13387 NZHUZOOL 13320 N 2 2 Haplotypes of NZHUZOOL 13319 DDBJ Acc. Haplotypes of SYg-1 SYg-1 DDBJ Acc. 1 1 AB601842 AB601842 4 SYg-2 SYg-C1a SYg-3 SY-C1b SYg-2 AB601843 AB601853 AB601844 AB601854 AB601843 SY-C2 SYg-C1a SY-C2 AB601853 AB601855 AB601855 Table 1. Haplotypes of 16S rRNA and cytochrome c oxidase I (COI) genes

86 THE RAFFLES BULLETIN OF ZOOLOGY 2011

Table 2. Matrix of percentage pairwise nucleotide divergences of K2P distance (lower left) and mean number of differences (including gaps) (upper right) based on 616 bp of cytochrome c oxidase I (COI) between Sayamia species from Taiwan, Vietnam, Cambodia, Malaysia and Thailand. For haplotype abbreviations and detailed localities see Table 1.

S. germaini S. sexpunctata S. bangkokensis S. cf. maehongsonensis SYg-C1a SYg-C1b SYg-C2 SYs-C1 SYs-C2 SYs-C3 SYb-C SYm-C SYg-C1a 1 6 22 21 20 51 49 SYg-C1b 0.16 7 23 22 21 52 50 SYg-C2 0.98 1.15 20 19 18 49 51 SYs-C1 3.69 3.86 3.35 1 4 54 53 SYs-C2 3.51 3.69 3.17 0.16 3 53 52 SYs-C3 3.34 3.51 3.00 0.65 0.49 53 52 SYb-C 8.94 9.13 8.57 9.50 9.31 9.31 20 SYm-C 8.56 8.75 8.95 9.31 9.12 9.12 3.36 with the higher diversity of haplotypes from Vietnam and edge that year (C.-H. Ho, pers. comm.), which indicate Cambodia (Fig. 2, Tables 1, 2). that the species was already established in Taiwan by 2005. Specimens were subsequently also observed in 2008 (J.-F. The haplotype network constructed to further depict the Huang, pers. comm.). Most recently, we also collected relationships among the combined 16S and COI haplotypes specimens from a second site about 15 km north of the fi rst of clades of S. germaini and S. sexpunctata are shown in Fig. observed site at the end of 2010 (Table 1). 3. The combined haplotype SYg-1+SYg-C1a from Taiwan is more central relative to other haplotypes and is therefore The source(s) of the introduced Taiwanese populations of assumed to represent the ancestral haplotype (cf. Clement S. germaini could not be confi rmed. Interestingly though, et al., 2000) (but see Discussion). Kaohsiung City also has a large population of Vietnamese migrant workers, the country from which S. germaini is naturally found. We do not believe this is a coincidence. DISCUSSION Freshwater crabs, especially, gecarcinucid ricefi eld crabs like Sayamia, are commonly eaten in Vietnam (Ng, 1988; Ng & Possible pathways and sources of introduction Kosuge, 1995; Yeo & Nguyen, 1999; Yeo et al., 2008) and it is possible Sayamia may have been introduced to Taiwan Sayamia germaini was fi rst observed in Kaohsiung City, for this purpose. Grocery shops in the area of Kaohsiung Taiwan, in 2005, although the species may have arrived City also specialise in importing fresh Vietnamese cuisine earlier. One ovigerous female was obtained (NTOU F10304) to cater for the large Vietnamese labour force in Kaohsiung. in 2005, and juveniles were also observed at the water’s While it is illegal to do so, it is not inconceivable that businesses may have smuggled in live Sayamia for sale. In markets in Vietnam and Cambodia, Sayamia is regularly sold live for food (e.g., Table 1). The anthropogenic spread of live gecarcinucid crabs for food has precedence. Ng & Naiyanetr (1993) reported that S. bangkokensis, a species native to central Thailand, was introduced to Chiangmai in the northwestern part of the country by migrant workers as a protein source. Sayamia as well as allied genera like Somanniathelphusa and Esanthelphusa are always sold live to keep them fresh (Ng, 1988; Ng & Naiyanetr, 1993). Yet another possibility may be that the crab was brought in by Vietnamese restaurants in Kaohsiung City to serve to the migrant community there. This has happened elsewhere, for example, in the United States, where individuals of the Japanese potamid, Geothelphusa dehaani, were found in a lake at Las Vegas, Nevada, which almost certainly escaped from an adjacent hotel sushi bar (Carlton, 2001). In recent Fig. 3. Genealogical network for the combined 16S rRNA and COI years, freshwater crabs have also become popular aquarium haplotypes observed within the clades of Sayamia germaini (collected subjects, and we have observed specimens of Sayamia from Taiwan, Vietnam and Cambodia) and S. sexpunctata (Malaysia). sexpunctata and S. bangkokensis on sale in Singapore (e.g., The ancestral haplotype, or root of the network, is indicated by a Table 1), and some have even been exported to Europe square. Unlabelled nodes indicate inferred haplotypes not found in and Japan (unpubl. data). As aquarium fi sh and plants are the sampled populations regularly exported from Singapore to Taiwan, the specimens

87 Shih et al. – Introduction of Indochinese freshwater crab to Taiwan in Kaohsiung could also have arrived there via this pathway. species native to the island (Shih et al., 2007). Over the past Regardless of how S. germaini entered southern Taiwan, two decades, So. taiwanensis has become increasingly rare the original introduced populations were fortunate to have due to habitat destruction and the extensive use of insecticides man-modifi ed, earth-lined canals in Kaohsiung that were and herbicides (Shih et al., 2007), and is currently regarded ecologically similar to their natural habitats in Indochina (Ng, as an endangered species internationally (Cumberlidge et 1988; Ng & Naiyanetr, 1993). The climate in Kaohsiung is al., 2009). A serious concern would arise if the range of also tropical and even in winter, is relatively warm, allowing the alien S. germaini population expands northwards to this tropical species to survive. overlap with that of So. taiwanensis, which would result in competition between the two species. As adult S. germaini Based on the molecular data, individuals of the populations grow to twice the size of So. taiwanensis and probably of S. germaini in Kaohsiung City and Kaohsiung County share similar habitat and food preferences, the survival of share identical haplotypes of 16S rRNA and COI (Fig. 1, the latter species would be even more threatened. Not only Tables 1, 2), suggesting a founder effect (Mayr, 1963). could S. germaini directly interfere with and outcompete This indicates that the current two Kaohsiung populations So. taiwanensis for common limited resources such food probably originated from a single introduction event, where and habitat, but it could also have the added advantage of only a few individuals (perhaps even a single ovigerous relative size refuge from predation. female) were originally released (or escaped) into the wild. However, it is not clear which of the two localities, which The presence of S. germaini also raises potential human health are 15 km apart and likely to be connected by waterways, concerns with regards to lung fl uke disease or paragonimiasis. might be the original site of introduction. Although the Fortunately, the lung fl uke (Paragonimus spp.) infection rates Kaohsiung haplotypes appear to be more ancient compared of Taiwanese and Chinese gecarcinucid species is relatively to other haplotypes obtained from southern Vietnam and low compared to that of potamids (see Chiu, 1964; Chung et Cambodia (Fig. 3), there is only 1 bp difference (without gap) al., 1975; Li & Lin, 1994); with So. taiwanensis from Taiwan between the Kaohsiung haplotype and the nearest haplotype reported to be infected with Paragonimus westermanii only in SYg-1+SYg-C1b from Lam Dong, southern Vietnam (Fig. 1915 (locality questioned, Nakagawa, 1917) and So. sinensis 3), a very minor difference that suggests these haplotypes sensu lato from Fujian, China, infected with P. fukienensis originate from one population. Firmer conclusions can in 1936 (Li et al., 1999). The high level of public hygiene therefore only be made after more extensive surveys of the in Taiwan also helps to keep the infection rates low (Li & range of species. Huang, 2002). However, Indochinese Sayamia species have been reported as secondary intermediate hosts of lung fl ukes, e.g., Paragonimus siamensis in “S. germaini” from Thailand Potential impacts and management (Kawashima et al., 1989; Sugiyama et al., 2006; Binchai et al., 2007). Hence, the proliferation of Sayamia and their It is important to note that known Sayamia species have possible associated alien parasites may present problems in a tropical distribution ranging from southern Indochina to controlling this disease in Taiwan. northern Peninsular Malaysia (Rathbun, 1902, 1905; Ng, 1988, 1997; Ng & Naiyanetr, 1993; Yeo & Ng, 1999; Ng et Sayamia germaini may also present economic problems. al., 2008). That the alien population of S. germaini in Taiwan Crabs of the genus Somanniathelphusa and Sayamia inhabit has been established for at least fi ve years in the tropical lentic water bodies and are especially common in ricefi elds climate of southern Taiwan is interesting in itself. How long in China, Taiwan and Indochina. They dig deep burrows into the species will persist in southern Taiwan and whether it the bunds of the ricefi elds, and these have been known to is able to spread north and survive in the cooler latitudes of compromise the structural integrity of the bunds (Ng, 1988; central and northern Taiwan is not clear. In any case, if S. Dai, 1999; Shy & Yu, 1999). In Taiwan, ricefi eld bunds germaini was to spread to other parts of Taiwan, potential are usually relatively narrow (ca. 30 cm in width) and will ecological, human health and agricultural problems can be probably not be able to cope with extensive excavations by anticipated. The present ranges of Taiwanese S. germaini the much larger Sayamia. Any damage to ricefi eld bunds are restricted to two canals located in the highly urbanized by Sayamia burrowing activities may in turn translate into industrial park of Kaohsiung City and the suburban Kaohsiung economic damages in the form of reduction or losses in County, respectively. No other native freshwater crab species rice production and increased repair and maintenance of has been recorded in these areas. The nearest location where these structures. a native freshwater species is known in Chaishan, a coastal hill in Kaohsiung City, where the potamid Geothelphusa Although S. germaini in Taiwan has yet to cause any makatao Shih & Shy, 2009, occurs. However, as the biology obvious or detectable impacts, if the alien population was to and habitat of potamids and gecarcinucids are very different expand, it may affect not only the survival of the endemic (Ng, 1988), there is unlikely to be signifi cant competition and endangered So. taiwanensis, but is also likely to raise between them. For the endemic Taiwanese gecarcinucid, public health issues, and cause serious problems for rice Somanniathelphusa taiwanensis, however, the presence of cultivators through their burrowing activities. If this alien S. germaini presents a different problem. Somanniathelphusa species persists, an attempt at eradication to remove or control taiwanensis is distributed in lowland habitats from central- the species (e.g., see Myers et al., 2000; Lodge et al., 2006; western to southwestern Taiwan and is the only gecarcinucid Roche et al., 2009) should be seriously considered. Although

88 THE RAFFLES BULLETIN OF ZOOLOGY 2011 complete eradication is often diffi cult or impossible (Lodge Chung, H. L., L. Y. Ho, W. C. Ts’ao & C. P. Hsu, 1975. On the et al., 2006), there have been successful instances, mostly metacercariae of some Paragonimus species and other trematodes attributed to early detection, or to limited and localized found in Chinese freshwater crabs. Acta Zoologica Sinica, 21: distribution of the invading species (Myers et al., 2000; 155–165, pl. 2. (in Chinese) Roche et al., 2009), of which, the latter scenario appears to Clement, M., D. Posada & K. A. Crandall, 2000. TCS: a computer be the case with the S. germaini invasion in Taiwan. Recent program to estimate gene genealogiees. Molecular Ecology, 9: surveys suggest that the species seems to be slightly more 1657–1659. widely distributed now than when fi rst detected. There is Cumberlidge, N. & P. K. L. Ng, 2009. Systematics, evolution, nevertheless an opportunity to eradicate it by removing and biogeography of freshwater crabs. Crustacean Issues, 18: reproductively mature individuals or at least lowering the 491–508. population to a non-viable level (see Myers et al., 2000). Such Cumberlidge, N., P. K. L. Ng, D. C. J. Yeo, C. Magalhães, M. R. a program should therefore be done as soon as is practical, Campos, F. Alvarez, T. Naruse, S. R. Daniels, L. J. Esser, F. and could include, among other methods, biological control Y. K. Attipoe, F.-L. Clotilde-Ba, W. Darwall, A. Mcivor, B. by fi sh predation through stocking of predatory fi shes native Collen & M. Ram, 2009. Freshwater crabs and the biodiversity crisis: importance, threats, status, and conservation challenges. to the area (targeted at reducing the population size) and Biological Conservation, 142: 1665–1673. trapping (targeted at reducing the population growth rate) (Hein et al., 2006). These could be supplemented by physical Dai, A. Y., 1999. Fauna Sinica. Arthropoda: Crustacea: Malacostraca: Decapoda: Parathelphusidae, Potamidae. Science Press, Beijing, removal by hand and net collection. These methods must China. (in Chinese) be accompanied by monitoring of the invaded habitat and surrounding unaffected areas to evaluate the effectiveness of Hein, C. L., B. M. Roth, A. R. Ives & M. J. Vander Zanden, 2006. the program. To further complement eradication efforts, the Fish predation and trapping for rusty crayfi sh (Orconectes rusticus) control: a whole-lake experiment. Canadian Journal invaded stretches of the canals should be isolated to slow of Fisheries and Aquatic Sciences, 63: 383–393. any spread of the species (Myers et al., 2000; Lodge et al., 2006), while education, prevention and enforcement measures Kawashima, K., H. Sugiyama & P. Ketudat, 1989. IV-3. Paragonimus infection in crabs in Thailand. In: Kawashima, K. (ed) directed at likely or potential sources of introduction (see Paragonimus in Asia – Biology, Genetic Variation and Speciation. earlier) probably need to be carried out at the same time. Paragonimus Research Report 2. Kyushu University of Health Sciences, Fukuoka, Japan. Pp 75–79. Kimura, M., 1980. A simple method for estimating evolutionary rates ACKNOWLEDGEMENTS of base substitutions through comparative studies of nucleotide sequences. Journal of Molecular Evolution, 16: 111–120. This study was supported by grants from the National Li, Y. S. & Y. M. Huang, 2002. Observation on the paragonimiasis Science Council, Executive Yuan, Taiwan to HTS (NSC96- in Taiwan. Journal of Tropical Medicine 2(2): 105–107. (in 2621-B-005-005-MY3; 98-2621-B-005-001-MY3) and Chinese) JYS (NSC96-2313-B-346-001-MY3), and Council of Li, Y. S. & J. X. Lin, 1994. Investigation on etiology of paragonimaisis Agriculture, Executive Yuan, Taiwan to HTS (98-AGRSCI- in Minhou County, Fujian Province. Wuyi Science Journal, 11: 8.6.1-GNRAFR-e1). Thanks are also due to Chuan-Hsin 162–166. (in Chinese) Ho and Guo-Chen Jiang for the collection of specimens Li, Y. S., J. X. Lin, Y. Z. Chen & C. X. Lin, 1999. Study on from Kaohsiung, Chin-Ko Shih for collection in Vietnam, reasons and changes of infection rate in 10 epidemic areas of Tomoyuki Komai for providing the specimen tissue collected paragonimiasis in Fujian Province. Chinese Journal of Parasitic from Cambodia and Thailand, I-Chu Chien for helping the Disease Control, 12(4): 275–277. (in Chinese) molecular work, and Jung-Hsiang Lee, Shen-Horn Yen and Lodge, D. M., S. Williams, H. J. MacIsaac, K. R. Hayes, B. Jung-Fu Huang for the crab information from Kaohsiung. Leung, S. Reichard, R. N. Mack, P. B. Moyle, M. Smith, D. We acknowledge two anonymous referees who improved A. Andow, J. T. Carlton & A. McMichael, 2006. Biological this manuscript. invasions: recommendations for U.S. policy and management. Ecological Applications, 16: 2035–2054. Mayr, E., 1963. Animal Species and Evolution. Havard University LITERATURE CITED Press, Cambridge. Myers, J. H., D. Simberloff, A. M. Kuris & J. R. Carey, 2000. Binchai, S., A. Rangsiruji, P. Ketudat, Y. Morishima & H. Sugiyama, Eradication revisited: dealing with exotic species. Trends in 2007. Molecular systematics of a new form of Paragonimus Ecology and Evolution, 15: 316–320. westermani discovered in Thailand. Southeast Asian Journal of Tropical Medicine and Public Health, 38 (supplement 1): Naiyanetr, P., 1994. On three new genera of Thai ricefi eld crabs 92–96. allied to Somanniathelphusa Bott, 1968 (Crustacea: Decapoda: Brachyura: Parathelphusidae). The Raffl es Bulletin of Zoology, Carlton, J. T., 2001. Introduced Species in U.S. Coastal Waters: 42: 695–700. Environmental Impacts and Management Priorities. Pew Oceans Commission, Arlington, Virginia. Nakagawa, K., 1917. Human pulmonary distomiasis caused by Paragonimus westermanni. Journal of Experimental Medicine, Chiu, J. K., 1964. Prevalence of Paragonimus infection in crab 26: 297–323. hosts in Taiwan. Bulletin of the Institute of Zoology, Academia Sinica, 3: 63–73. Ng, P. K. L., 1988. The Freshwater Crabs of Peninsular Malaysia and Singapore. Shing Lee Publishers LTD., Singapore.

89 Shih et al. – Introduction of Indochinese freshwater crab to Taiwan

Ng, P. K. L., 1997. On two new species of freshwater crabs Shih, H.-T. & P. K. L. Ng, in press. Diversity and biogeography of the genera Sayamia and Heterothelphusa (Brachyura, of freshwater crabs (Crustacea: Brachyura: Potamidae, Parathelphusidae) from southern Thailand. Crustaceana, 70: Gecarcinucidae) from East Asia. Systematics and Biodiversity. 710–719. Shih, H.-T. & J.-Y. Shy, 2009. Geothelphusa makatao sp. nov. Ng, P. K. L., D. Guinot & P. J. F. Davie, 2008. Systema (Crustacea: Brachyura: Potamidae), a new freshwater crab from Brachyurorum: Part I. An annotated checklist of extant brachyuran an uplifted Pleistocene reef in Taiwan. Zootaxa, 2106: 51–60. crabs of the world. The Rafﬂ es Bulletin of Zoology, Supplement, Shih, H.-T., J.-Y. Shy & J.-H. Lee, 2010. A new freshwater crab of the 17: 1–296. genus Geothelphusa (Brachyura, Potamidae) from southwestern Ng, P. K. L. & T. Kosuge, 1995. On a new Somanniathelphusa Taiwan. Crustaceana Monographs, 14: 661-675. Bott, 1968, from Vietnam (Crustacea: Decapoda: Brachyura: Shih, H.-T., D. C. J. Yeo & P. K. L. Ng, 2009. The collision of the Parathelphusidae). Proceedings of the Biological Society of Indian plate with Asia: molecular evidence for its impact on the Washington, 108: 61–67. phylogeny of freshwater crabs (Brachyura: Potamidae). Journal Ng, P. K. L. & P. Naiyanetr, 1993. New and recently described of Biogeography, 36: 703–719. freshwater crabs (Crustacea: Decapoda: Brachyura: Potamidae, Shih, H.-T., X.-M. Zhou, G.-X. Chen, I-C. Chien & P. K. L. Ng, Gecarcinucidae and Parathelphusidae) from Thailand. Zoologische in press. Recent vicariant and dispersal events affecting the Verhandelingen, Leiden, 284: 1–117. phylogeny and biogeography of East Asian freshwater crab genus Ng, P. K. L., C.-H. Wang, P.-H. Ho & H.-T. Shih, 2001. An Nanhaipotamon (Decapoda: Potamidae). Molecular Phylogenetics annotated checklist of brachyuran crabs from Taiwan (Crustacea: and Evolution (doi:10.1016/j.ympev.2010.11.013). Decapoda). National Taiwan Museum Special Publication Series, Shy, J. Y., P. K. L. Ng & H. P. Yu, 1994. Crabs of the genus 11: 1–86. Geothelphusa Stimpson, 1858 (Crustacea: Decapoda: Brachyura: Nylander, J. A. A., 2005. MrModeltest, vers. 2.2. Program distributed Potamidae) from Taiwan, with descriptions of 25 new species. by the author. Evolutionary Biology Centre, Uppsala University, The Rafﬂ es Bulletin of Zoology, 42: 781–846. Uppsala, Sweden. Shy, J. Y. & H. P. Yu, 1999. Freshwater Crabs of Taiwan. National Rathbun, M. J., 1902. Description des nouvelles espèces de Museum of Marine Biology/Aquarium, Pingtung, Taiwan. (in Parathelphusa appartenant au Muséum de Paris. Bulletin Chinese) du Muséum National d’Histoire Naturelle, Paris, 1902(3): Sugiyama, H., Y. Morishima, A. Rangsiruji, S. Binchai, P. Ketudat 184–187. & M. Kawanaka, 2006. Application of multiplex PCR for species Rathbun, M. J., 1905. Les crabes d’eau douce. Nouvelles Archives discrimination using individual metacercariae of Paragonimus du Muséum d’Histoire Naturelle, Paris, (4)7: 159–323, pls. occurring in Thailand. Southeast Asian Journal of Tropical 13–22. Medicine and Public Health, 37(supplement 3): 48–52. Roche, D. G., M. E. Torchin, B. Leung & S. Binning, 2009. Swofford, D. L., 2003. PAUP*: Phylogenetic Analysis Using Localized invasion of the North American Harris mud crab, Parsimony (*and Other Methods), vers. 4. Sinauer Associates, Rhithropanopeus harrisii, in the Panama Canal: implications for Sunderland, Massachusetts. eradication and spread. Biological Invasions, 11: 983–993. Yeo, D. C. J. & P. K. L. Ng, 1999. The state of freshwater crab Rodríguez, F., J. L. Oliver, A. Marín & J. R. Medina, 1990. The taxonomy in Indochina (Decapoda, Brachyura). In: Schram, F. R. general stochastic model of nucleotide substitution. Journal of & J. C. von Vaupel Klein (eds.), Crustaceans and the Biodiversity Theoretical Biology, 142: 485–501. Crisis, Proceeding of the 4th International Crustacean Congress, Ronquist, F. & J. P. Huelsenbeck, 2003. MRBAYES 3: Bayesian 1998, vol. I. Brill, Leiden, The Netherlands. Pp 637–646. phylogenetic inference under mixed models. Bioinformatics, Yeo, D. C. J., P. K. L. Ng, N. Cumberlidge, C. Magalhaes, S. 19: 1572–1574. R. Daniels & M. R. Campos, 2008. Global diversity of crabs Ronquist, F., J. P. Huelsenbeck & P. van der Mark, 2005. MrBayes 3.1 (Crustacea: Decapoda: Brachyura) in freshwater. Hydrobiologia, Manual. Available via http://mrbayes.csit.fsu.edu/manual.php. 595: 275–286. Shih, H.-T., S.-H. Fang & P. K. L. Ng, 2007. Phylogeny of the Yeo, D. C. J. & X. Q. Nguyen, 1999. Description of a new species freshwater crabs genus Somanniathelphusa Bott (Decapoda: of Somanniathelphusa (Decapoda, Brachyura, Parathelphusidae) Parathelphusidae) from Taiwan and the coastal regions of China, from Vietnam. Crustaceana, 72: 339–349. with notes on their biogeography. Invertebrate Systematics, 21: 29–37. Shih, H.-T., T. Naruse & D. C. J. Yeo, 2008. A new species of Geothelphusa Stimpson, 1857, from Taiwan (Crustacea: Brachyura: Potamidae) based on morphological and molecular evidence, with notes on species from western Taiwan. Zootaxa, 1877: 37–48.

90 THE RAFFLES BULLETIN OF ZOOLOGY 2011

DISTRIBUTION OF SMALL CETACEANS IN THE NEARSHORE WATERS OF SARAWAK, EAST MALAYSIA

Gianna Minton Cindy Peter Andrew Alek Tuen Institute of Biodiversity and Environmental Conservation, Universiti Malaysia Sarawak, 94300 Kota Samarahan, Sarawak, Malaysia Email: [email protected] (GM- corresponding author), [email protected] (CP), [email protected] (AAT)

ABSTRACT. – Between June 2008 and September 2009, 56 days of small boat surveys were conducted off the coast of Sarawak, Malaysia with the aim of recording cetacean distribution. These surveys, which focused on the Miri, Bintulu-Similajau and Kuching regions, comprised 173 hours of survey effort and covered 2851 km of pre-determined systematic tracks. Surveys were clustered into three sets of seasonal snapshots: June-July, September-October and March-April. A total of 115 cetacean sightings were made, of which 65 were on-effort and used in analyses of cetacean encounter rates in relation to habitat characteristics. Species observed included (in order of frequency) Irrawaddy dolphins (Orcaella brevirostris), fi nless porpoises (Neophocaena phocaenoides), Indo-Pacifi c bottlenose dolphins (Tursiops aduncus) and Indo-Pacifi c humpback dolphins (Sousa chinensis).

One hundred and ten of 115 sightings were made in less than 10 m water depth, highlighting the importance of nearshore coastal habitats for these species. Despite an apparent overlap in habitat, Irrawaddy dolphins showed a statistically signifi cant affi liation with areas of shallower depth and closer proximity to shore and river mouths than fi nless porpoises or bottlenose dolphins. This preference for nearshore areas renders the species vulnerable to threats such as fi sheries by-catch and habitat degradation from coastal development. Irrawaddy dolphins were more frequently encountered in Kuching, while the highest encounter rate for fi nless porpoises was in the Bintulu-Similajau region. Depictions of encounter rates in these regions in relation to survey effort in 2 km × 2 km grid-cells give an indication of the preferred habitats of Irrawaddy dolphins, and show that the highest encounter rates in both Kuching and Similajau occurred in areas that are destined for major coastal developments. The information presented here should help researchers and managers design effective future research and conservation strategies.

KEYWORDS. – Malaysia, Sarawak, South China Sea, distribution, Irrawaddy dolphin, ﬁ nless porpoise.

INTRODUCTION aduncus, Ehrenberg, 1833) and Indo-Pacific humpback dolphin (Sousa chinensis, Osbeck, 1765). However, these Prior to 2008, formal research on the marine mammals sources do not provide detailed information on the species’ of Sarawak was limited. Various incidental and historical fi ne-scale distribution or habitat preferences. records were reported by naturalists in the region (e.g. Lydekker, 1901; Gibson-Hill, 1949). Beasley and Jefferson Throughout their range, the documented preference of all (1997) also conducted some preliminary surveys, while aerial four of these species for nearshore habitats exposes them to and boat surveys conducted jointly by the Sarawak Forestry a number of threats, including by-catch in fi sheries (Dolar Corporation (SFC), the University Malaysia Sabah (UMS) et al., 2002; Read et al., 2006), habitat loss and degradation and the Sabah Wildlife Department provided further reports (e.g. Jefferson et al., 2009), decreased fi tness from pollution/ and overviews of the species present and their distribution contaminants (e.g. Reeves et al., 2003; Adams et al., 2008), (e.g. Jaaman, 2006; Bali et al., 2008). These collective sources high levels of vessel traffi c, underwater noise and dolphin of information on cetacean populations in Sarawak indicated watch tourism (Lusseau, 2003; Constantine et al., 2004; that the species most commonly encountered in nearshore Beijder et al., 2006; Lusseau et al., 2007). The IUCN Red waters were the Irrawaddy dolphin (Orcaella brevirostris, List of Endangered Species (IUCN, 2008) classifi es both Owen in Gray, 1866), finless porpoise (Neophocaena Irrawaddy dolphins and fi nless porpoises as “Vulnerable”, phocaenoides, Cuvier, 1829), bottlenose dolphin (Tursiops while humpback dolphins are considered “Near-Threatened”.

91 Minton et al. Nearshore cetaceans in Sarawak

Bottlenose dolphins, most likely due to taxonomic uncertainty allowed 74% of survey time to be spent on effort, with a about different populations around the globe, are considered coverage probability of 34.1%. “Data-defi cient”. Five freshwater populations of Irrawaddy dolphins in Southeast Asia are already listed as “Critically Survey vessels were open-decked and fi breglass-hulled, Endangered” (IUCN, 2008). approximately seven to 10 m in length, and used either single or double outboard engines ranging from 90 to 200 The threats that affect these nearshore species throughout hp. A minimum of three observers stood on decks or benches their range are also known to be present in Borneo (Dolar resulting in an eye height approximately three meters above et al., 1997; Kreb & Budiono, 2005) and in Sarawak, sea level. All three observers searched with the naked eye, Malaysia, specifi cally (Jaaman, 2006; Jaaman et al., 2009). with one observer at the bow scanning a cone of about 30° Detailed scientifi c surveys are of great importance in order and two observers on either side of the boat scanning arcs to assess more accurately each species conservation status of approximately 90° to port and starboard (e.g. Parra et al., and conservation requirements. As an important fi rst step in 2006). A minimum of two experienced cetacean observers this process, a series of small boat surveys was conducted were always on board, while other observers undergoing in three locations along Sarawak’s coastline with the aims training had varying degrees of experience. Due to the high of: 1) assessing more accurately the species composition, turnover of observers undergoing training, and the need for distribution and habitat requirements of coastal cetaceans all observers to be standing together at deck level rather than in the region; 2) collecting baseline data on dolphin separated by elevated observation platforms, the team opted distribution and habitat use; and 3) facilitating management to implement methods appropriate for calculating relative and conservation efforts. The results of these surveys as rather than absolute abundance. presented here should enable managers and researchers to identify areas of key importance for future conservation and Transects were navigated at a steady speed of 10 knots (18.5 research efforts and lay the foundation for more detailed km/h) and observers rotated through different positions on and targeted study both within Sarawak and in other parts the boat at the end of each transect line (roughly half-hour of the species’ range. intervals) to avoid fatigue. When cetaceans were detected, search effort was suspended to collect data on the group composition and behaviour following standardized data MATERIAL AND METHODS collection methods (e.g. Smith & Reeves, 2000). Positional data for both survey tracks and sightings were collected Data Collection. – Small boat surveys were conducted along using a handheld GPS unit. the Sarawak coast of the Kuching, Bintulu-Similajau, and Miri regions (Fig. 1). These areas were chosen for reasons Effort was recorded to the nearest minute throughout each of logistical practicality, as well as their contrasting habitat survey day in order to distinguish between time spent in composition. The Miri and Similajau coastlines both run optimal survey conditions (from here on referred to as from approximately north to south and are interspersed on-effort), transiting to or from the start and end points of with rivers and streams of varying sizes at fairly regular transect lines, working with cetacean groups, or simply off intervals. The Kuching coastline runs from east to west survey effort. Only sightings made on-effort were used in and comprises a complex and interconnected series of wide the generation of encounter rates, which were calculated rivers and mangrove channels, which empty into three major both in relation to hours of survey effort and the distance estuaries/bays. The Kuching study area includes some of the covered during on-effort survey time. Weather conditions waterways encompassed in the Kuching Wetland National that could affect sighting probability were recorded on an Park, while the Bintulu-Similajau study area includes the hourly basis, or upon a rapid condition change, and search entire coastline bordering the Similajau National Park. A further reason for choosing the Kuching and Similajau areas was the large-scale coastal developments planned in both places.

Transects extended up to 15 km offshore and were systematically orientated at 45° angles to the primary coastline to ensure they were independent of habitat features and environmental gradients, and to allow for detection of cetacean density gradients alongshore as well as onshore/ offshore (e.g. Dawson et al., 2008). The survey design function in DISTANCE (Thomas et al., 2009) was used to compare the coverage probability and proportion of survey time spent in transit between transects when transects were separated by 2, 3, 4, 5 and 6 km. A transect separation of 4 km was selected as this allowed a compromise between intense survey coverage and a realistic amount of survey effort. For the Kuching area, for example, the 4 km spacing Fig. 1. Three main study areas in Sarawak.

92 THE RAFFLES BULLETIN OF ZOOLOGY 2011 effort was suspended during heavy rain and/or Beaufort These interpolated values were used to assign depth values Sea-states of four or higher. to dolphin sightings where no echo-sounder was available on the research vessel. Group size was estimated after approach, and behaviour classifi ed into standardized categories of traveling, feeding Sightings made while on effort were also imported into (direct feeding observed), probable feeding (dive patterns Google Earth in order to measure the distances to the nearest consistent with feeding activity or fi sh observed at the surface land and nearest river mouth. Only on-effort sightings were near dolphins), resting/milling, socializing, or undetermined used to avoid the bias that might be introduced by including (e.g. Parra, 2006; Lusseau, 2006). Some of the survey vessels sightings made while speeding close to shore to the start and used were equipped with a depth sounder, in which case end-points of transect lines. Distance to land was measured depth was recorded at the point of sighting. When a depth to the nearest 100 m. Distance to the nearest river mouth sounder was not available, depths at locations of sightings was measured by creating a fixed mid-point in the line were interpolated during data processing. The presence of connecting the “corners” of the river mouth and measuring fi shing effort in proximity to cetacean groups was noted, the distance from the sightings to this midpoint. Sighting and the number of boats or set gillnets within an estimated locations inland/upriver of the midpoint were assigned a 50 m of the group recorded at each sighting. value of “0”. The resulting distributions of sighting location characteristics for each species were tested for normality Photo-identifi cation. – Whenever a group of dolphins or using the Anderson–Darling test. The tests showed a non- porpoises was sighted, active survey effort was suspended, normal distribution for all three characteristics for both the boat departed from the transect line and efforts were Irrawaddy dolphins and fi nless porpoises, and therefore, made to approach the group as close as possible without the non-parametric Kruskal-Wallis as well Wilcoxon ranked disturbing their natural behaviour. Digital SLR cameras sum tests with correction for continuity were used to test with either 200 mm or 300 mm zoom lenses were used to the signifi cance of the differences between each species’ take photos of the left and/or right sides of dorsal fi ns of as sighting locations with respect to the depth, distance to land many individuals in the group as possible. and distance to river mouths. Bottlenose dolphins were not included in these analyses due to the small sample size of For Irrawaddy and Indo-Pacific humpback dolphins, only three on-effort sightings. photographs showing left or right sides of dorsal fi ns in suffi cient detail were cropped and digitally enhanced and On-effort tracks in the Kuching and Similajau areas were entered into a custom-designed MS Access® database which analysed in ArcGIS 9.3 and overlayed with a 2 km × 2 km allowed for storage of sighting information as well as on- grid. The number of on-effort Irrawaddy dolphin sightings screen comparison of photographs. Left and right dorsal fi n in each grid cell was divided by the sum of on-effort survey photos were treated as two separate data sets. Photographs tracks within each grid cell to generate an encounter rate were assigned scores of 1-4 for both overall quality and for each cell. Cells with varying encounter rates were distinctiveness following the protocols described by Friday then colour-shaded to provide a graphic indication of high et al. (2000) with a score of 4 indicating excellent quality density areas. or a high level of distinctiveness, and a score of 1 indicating very poor quality or lack of distinguishing features on the dorsal fi n area. Matching of photographs was conducted on- RESULTS screen, and included photographs of every score in both the quality and distinctiveness categories. Unique ID numbers Sightings and encounter rates. – Surveys completed as were assigned to each individual dolphin after matching to of September 2009 are detailed in Table 1. The completed all previously collected photographs. tracks and sightings made during the surveys are depicted in Figures 2 a-c. During 56 days (over 389 hours) a total of Data analysis. – Survey tracks and precise sighting locations 5,555 km was covered, of which 2851 km and 173 hours recorded with the handheld GPS were downloaded at the end were spent on-effort. A total of 115 cetacean sightings were of each day and saved in different formats for later processing logged, of which 65 were on-effort and used in encounter in Google Earth® and/or ArcMap®. Sighting details were rate and habitat analyses. entered into a custom-designed MS Access database, and both tracks and sightings were imported into Google Earth The number of dolphin or porpoise groups sighted per total and ESRI’s ArcMap for viewing, mapping and analysis. number of hours spent on the boat across all areas was 0.30 groups per hour, while the on-effort encounter rate was 0.38 Depth values were obtained from Arc-GIS compatible rasters groups of cetaceans per hour. Irrawaddy dolphins were the of British Admiralty charts (issued by Seazone®). Digitized most frequently sighted species, with an encounter rate of bathymetry points were generated by manually assigning 0.20 sightings per hour of search effort and the highest depth values to each depth point on the chart, and various encounter rate in Kuching (0.30 sightings per hour). Finless functions were tested to determine the “best fi t” model for porpoises were sighted at a rate of 0.13 sightings per hour interpolating depths in our study area between the known of effort across all regions, with the highest encounter rate depth values of the chart. The function chosen was Inverse in Similajau (0.22 groups per hour). Distance Weighted (IDW) with 0.5 nautical mile grid size.

93 Minton et al. Nearshore cetaceans in Sarawak

Table 1: Survey dates, distance covered, hours spent surveying and number of dolphin sightings per survey period in each region.

Dates Surveyed Total Hours on Total Irrawaddy Total Finless Other Distance effort Irrawaddy encounter Finless porpoise species covered (hrs:mins) dolphin per hour porpoise encounter (km) sightings of effort sightings per hour of effort Kuching 15-17 July ‘08 319.57 9:00 5 0.333 2 0.222 1 (unid) a 16-18 Sept ‘08 299.33 10:56 6 0.366 1 0.091 1 (Tursiops) 14-16 Oct ‘08 254.34 9:04 7 0.551 1 0.000 8, 13 Nov and 11 Dec ‘08 114.82 3:52 4 0.000 0 0.000 4-6 Mar ‘09 279.403 10:19 3 0.194 2 0.097 20-21 May, 6 June 235.01 7:13 6 0.139 2 0.277 21-23 Jul ‘09 320.48 11:47 5 0.255 1 0.000 19-21 Aug ‘09 235.34 7:02 3 0.427 0 0.000 7-10 Sep ‘09 326.10 9:14 5 0.325 1 0.108 1 (unid) a Kuching total 2384.40 78.27 44 0.288 11 0.088 3 Similajau 11-13 June ‘08 272 6:16 7 0.479 6 0.319 1 (Sousa) 24-26 Sept ‘08 249.37 9:26 1 0.106 3 0.318 18-20 Feb ‘09 334.78 7:35 4 0.132 5 0.132 1 (Sousa) 25-27 Mar ‘09 339.03 9:41 5 0.103 5 0.310 8-10 Jul ‘09 305.18 8:47 2 0.228 2 0.000 5-6 Oct ‘09b 18.34 0 0 0 0 0 Similajau total 1518.7 41:45 19 0.192 20 0.216 2 Miri 24 June, 7-9 Jul ‘08 351.05 13:29 1 0.074 3 0.148 9 Sept and 4 Nov ‘08 199.8 6:22 0 0.000 1 0.157 1 (Tursiops) 16-18 Mar ‘09 258.195 10:15 0 0.000 0 0.000 8-10 Jun ‘09 359.565 11:17 0 0.000 2 0.089 1 (Tursiops) 14-16-Sep ‘09 353.37 11:23 2 0.088 3 0.176 1 (unid) a Miri total 1617.18 52:46 3 0.038 10 0.133 3 Total for all areas 5554.62 172.58 66 0.197 41 0.133 8 a Not identiﬁ ed to species level – sighting too brief and/or distant. bPoor weather prevented surveys at sea – only one short river survey – no cetacean sightings.

Encounter rates were almost certainly affected by the extremely unpredictable in their surfacing patterns. Low “sightability” of the species involved. Bottlenose dolphins light, haze and rain/drizzle contributed to the diffi culty of were sighted from up to an estimated 1 km from the survey obtaining good quality photographs of the left and right sides vessel, with a mean estimated sighting distance of 550 m of individual dolphins’ dorsal fi ns. Following selection and (SD 427). With a mean estimated group size of 20.3 (SD cropping for entry in the database, a total of 102 individual 8.4), these larger dolphins displayed more conspicuous Irrawaddy dolphins were identifi ed from the right side of the surfacing behaviour, including occasional leaps and splashes dorsal fi n, and 101 by their left side. Only 27 individuals in from tail slaps and fast turns. The mean estimated sighting each group were represented by a photograph of a quality distance for Irrawaddy dolphins, on the other hand, was 3 or higher, a quality considered suitable for inclusion in much lower at 126 m from the boat (SD 108 m). Group mark-recapture analysis (Table 2). sizes for this species were smaller, with a mean estimated group size of 4.3 (SD 3.1). Finless porpoises were the least For Irrawaddy dolphins, only nine defi nite matches were conspicuous species, with a mean sighting distance of only made in the Kuching area, and one in the Similajau area. 100 m from the boat (SD 97) and mean estimated group All but one of the matches were based on poor quality size of 2.3 (SD 1.6). photos of distinctively marked individuals. There was a relatively high number of “possible matches”, where dorsal Photo-Identification. – While Irrawaddy dolphins were fi n shapes looked to be the same, but photographs were not encountered relatively frequently, they were elusive and sharp enough and the fi ns were not distinctive enough to

94 THE RAFFLES BULLETIN OF ZOOLOGY 2011

Table 2: Results of Photo Identiﬁ cation of Irrawaddy dolphins after processing in a tailor-made MS Access database.

Left dorsal fi n Right dorsal fi n Year Total no. No. of individuals Total no. No. of individuals of identifi ed represented by of identifi ed represented by individuals Photo quality individuals Photo quality 3 or higher 3 or higher 2007 0 0 6 3 2008 40 7 40 9 2009 62 20 56 15 Totala 101 27 102 27

Confi rmed matches between surveys 5 1 4 0 Possible matches between surveys 12 18 a Note the total is not the sum of all identifi cations as individuals re-sighted from one year to the next are only counted once. show more subtle scarring or markings that would allow and the Bako-Buntal bay on the east. Re-sights included a confi rmation of the match. This indicates that our data individuals photographed between survey years as well as may include a high number of “false negatives” – cases from one month to the next, confi rming likely year-round where the same individual is not being recognized from residence in the survey area. Movement between the two one sighting to the next. Mark-recapture analyses using main bays of the study area indicates that the home range various fi ltering mechanisms for the sighting histories (e.g. of the population under study includes both bays, with selecting only photos with distinctiveness scores of 3 or the furthest distance between sightings being 17 km, but higher) in the sample yielded confl icting results with very separated by a landmass which would take 25 to 30 km to wide confi dence intervals, and were not considered suitably navigate around (Fig. 2c). It is not clear whether movement robust for publication. between the two bays occurs through the river network or around the tip of the Santubong peninsula. The confirmed matches do, however, yield insight into movement of the dolphin population within our study Matches made in the Similajau survey area include one areas. The matches indicate movement between the Salak- Irrawaddy dolphin and one Indo-Pacifi c humpback dolphin. Santubong estuary on the west side of the study area, In both cases, the re-sights indicate that the identified dolphin’s range extends along coastline between the area encompassed by the Similajau National Park and Tanjung Similajau, an area which is being considered for large-scale industrial development.

Sighting distribution and habitat preferences. – Fig. 3 depicts the characteristics of dolphin and porpoise sighting locations with respect to depth, distance from shore and distance from river mouths. Bottlenose dolphins were sighted in depths ranging from 3.8 to 24.3 m, with a mean of 9.9 m (SD 10.3). Finless porpoises were sighted in depths ranging from 1.3 to 16.8 m with a mean of 9.6 m (SD 11.8). Irrawaddy dolphins were encountered in the shallowest depths, ranging from 2.0 to 5.4 m with a mean of 3.3 (SD 1.4). Similarly, Irrawaddy dolphins were found closest to shore (mean of 1.38 km, SD 1.90), and closest to river mouths (mean 3.53 km, SD 3.58). The box and whisker plots demonstrate little or no overlap in the median quartile values of all three habitat variables for all three species. Kruskal-Wallis tests and Wilcoxon ranked sum tests with correction for continuity confi rmed that the differences between Irrawaddy dolphins and fi nless porpoises were statistically signifi cant for all three variables of depth, distance to shore and distance to river mouths, with p-values lower than 0.01 (Table 3). The results indicate that Irrawaddy dolphins were found at locations that were signifi cantly shallower (sample estimate difference of 2.6 m), closer to shore (sample estimate difference of 1.6 Fig. 2. Survey tracks (total track, including on- and off-effort portions) and sightings (on- and off-effort) in the three main survey km), and most signifi cantly closer to river mouths (sample areas of (a) Miri, (b) Similajau, and (c) Kuching. estimate difference of 4.2 km) than fi nless porpoises.

95 Minton et al. Nearshore cetaceans in Sarawak

Irrawaddy dolphin encounter rates (number of sightings Figs. 2 a-c and 4a-b demonstrate the importance of shallow divided by the total number of kilometers searched) per grid coastal waters for all four species sighted during surveys, cell in the Kuching and Similajau study areas are depicted with 110 of 115 sightings taking place in depths of 10 m in Figures 4a and b. In the Kuching area, the cells with or less. While ﬁ gures 4a and b indicate speciﬁ c grid cells the highest encounter rates (darkest shading) are located where Irrawaddy dolphin encounter rates were high during near the four major river mouths in the study area. In the our surveys, readers should realise that these are not the Similajau study area, the two cells with the highest encounter only areas of dolphin distribution, but rather an indication rates are located on either side of the Similajau River and of which types of habitats might be the preferred feeding, Tanjung Similajau – the headland in the northern portion resting or breeding areas for this species. of the study area. With the exception of one sighting made roughly 8 km upriver in the Kuching area, Irrawaddy dolphins were exclusively DISCUSSION encountered in coastal or estuarine areas. While this species may occur in freshwater habitats in other river systems in Species composition and distribution. – This series of Sarawak, they do not appear to frequent the inland freshwater three-day “snapshot surveys” has offered an important portions of the Miri, Similajau, Santubong or Salak rivers, initial insight into the species composition and distribution which were occasionally surveyed during our study. However, of cetaceans in three nearshore areas of Sarawak’s coastline. more consistent survey effort is required in these river systems to determine whether or not this is the case.

The locations of our sightings indicate a high degree of overlap between the distribution of all four species, particularly that of Irrawaddy dolphins and ﬁ nless porpoises in the Kuching and Similajau areas. Comparison of on-effort sightings indicates that Irrawaddy dolphins show a preference for shallower waters closer to shore and river mouths than ﬁ nless porpoises or bottlenose dolphins. Current and ongoing research efforts involve the measurement of water parameters,

Fig. 3. Box and whisker plots showing the minimum and maximum Fig. 4. (a) Kuching and (b) Similajau survey areas with 2k m × values, median and upper and lower quartiles of the values for 2 km grid cells shaded according to encounter rate (number of depth (a), distance to shore (b), and distance to nearest river mouth sightings/distance searched in each grid cell). Only grid cells with (c) for on-effort sightings of Irrawaddy dolphins (n=35), ﬁ nless dolphin sightings are shaded – others are left blank to better display porpoises (n=24) and bottlenose dolphins (n=3). underlying survey effort.

96 THE RAFFLES BULLETIN OF ZOOLOGY 2011

Table 3: Results of Kruskal-Wallis and Wilcoxon rank sum (also known as Mann-Whitney U-) testing for the statistical signifi cance of the differences in depth, distance to shore and distance to nearest river mouth for on-effort sightings of Irrawaddy dolphins (n= 35) and fi nless porpoises (n=24). Bottlenose dolphin sightings (n=3) were not included in the analyses. All resulting p values were considered signifi cant at < 0.01.

Kruskal-Wallis fi nless porpoise vs Irrawaddy dolphin χ2 P-Value df Depth 10.3123 0.001321 1 Distance to shore 12.7501 0.000356 1 Distance to river mouth 16.405 5.115e-05 1 95% confi dence Wilcoxon rank sum test Sample estimate interval of fi nless porpoise vs Irrawaddy of difference in differences in dolphin W P-value location medians location medians Depth 633 0.001019 2.60 1.1- 5.8 Distance to shore 650 0.0003976 1.6 0.8-2.6 Distance to river mouth 674.5 8.618e-05 4.2 2.0-6.0 such as salinity, turbidity and temperature at sighting the Indo-Pacifi c region under the species Sousa chinensis. locations, and may provide a more detailed understanding However, Frère et al. (2008) concluded that humpback of the subtle ecological differences that allow these species dolphins sampled in Australia differ signifi cantly from those to share habitat (e.g. Parra, 2006; Parra et al., 2006). sampled in China and may comprise a new species. Further genetic sampling and morphological studies are required from The external morphology of the bottlenose dolphins that Borneo to determine how signifi cantly humpback dolphins in were observed in the Kuching and Miri study sites was this region may differ from other parts of the species’ range, consistent with T. aduncus. Body size was generally smaller and whether it might also merit a new species nomenclature, and more slender than that of T. truncatus, with a more Sousa borneensis (drawing on the original nomenclature of elongated rostrum a visible dark grey dorsal “cape” as well Sotalia borneensis (Lydekker, 1901)). as speckling on more mature individuals ranging from the ventral surface up toward the fl anks (Wang et al., 2000; Species habitat use and ecology. – Confi rmed observations Wang & Yang, 2009). Their occurrence in relatively shallow of small calves during sightings of Irrawaddy dolphins and nearshore waters is consistent with the habitat described for fi nless porpoises indicate that the nearshore areas of Similajau this species (Wang & Yang, 2009). However, our infrequent and Kuching are used as breeding and nursing grounds for encounters with the bottlenose dolphins indicate that this these species. On 20 August 2009, a sighting was made species may have a wider range which extends further of an unusually large aggregation of Irrawaddy dolphins offshore and/or along the shore than the Irrawaddy dolphins in the western part of our Kuching study area. The group and fi nless porpoises which are observed more consistently of at least 20 individuals, three of which were recognized within our survey areas. from previous surveys in the Kuching region, was tightly packed and displayed frequent body contact consistent with External morphology of the fi nless porpoise observed on a descriptions of gregarious mating or social aggregations by-catch specimen in October 2008 and on photographs of for this species in Trat Province, Thailand (Ponnampalam free-ranging animals is consistent with that described for et al. 2009), and with published descriptions of multi-pod Neophocaena phocaenoides phocaenoides – the subspecies aggregations of killer whales in the Kamchatka peninsula characterized by a wide area of tubercles on the dorsal (Filatova et al., 2009). Anecdotal reports from dolphin watch surface (Jefferson & Hung, 2004; Wang et al., 2008). Our tour operators indicate that these aggregations occur in the observed distributions with respect to habitat type are also Kuching area more frequently in summer months than at consistent with those observed in other areas of the species’ other times of the year when smaller groups of two to six range (Jefferson et al., 2002; Jefferson & Hung, 2004). individuals are more commonly observed. Calves were also observed during sightings of humpback dolphins in the Indo-Pacifi c humpback dolphins were only observed twice off Similajau area, and bottlenose dolphin groups in both the effort. Morphologically, they resemble Sousa chinensis (Parra Kuching and Miri study areas. & Ross, 2009). However, animals appear more robust and more mottled in colouration than those in Hong Kong, with One observation of bottlenose dolphins in the Miri area only a very small percentage of animals (e.g. 1 per group of included sub-groups tightly circling and tail slapping around 15-20 animals) appearing fully “white” or pink (pers. obs.). schools of fi sh visible at the surface, indicating that feeding An analysis of geographic variation in skull morphology takes place in this area approximately 14 km offshore and led Jefferson and Van Waerebeek (2004) to conservatively within 5 km of a number of coral outcrops. Both Irrawaddy label specimens obtained from several locations throughout dolphins and fi nless porpoises were observed on several

97 Minton et al. Nearshore cetaceans in Sarawak occasions chasing fi sh at the surface, both in Similajau and for a hydroelectric power station were considered to be the Kuching. While other priorities may influence cetacean most likely contributing factor to a sudden drop in bottlenose distribution under particular circumstances (e.g. security dolphin calf survival rates (Currey et al. 2009). Although from predation), prey availability is expected to be the we would expect Irrawaddy dolphins to be more tolerant of primary factor in determining habitat choice (e.g. Hastie et freshwater input, signifi cant and rapid unnatural changes to al., 2004). Our observations indicate that both Irrawaddy salinity and turbidity resulting from the fl ood channel may dolphins and fi nless porpoises feed in shallow nearshore prove harmful to this population. Dredging of large volumes areas, and are thus likely specialised in hunting techniques of sand for construction materials is also common in the and dietary preferences that restrict them to the nearshore Salak River estuary (e.g. Naeg & Ten, 2010). Shirakihara habitats. As such, any coastal development that introduces et al. (2007) concluded that high levels of dredging were signifi cant changes to the salinity, turbidity, substrate or responsible for a lower fi nless porpoise density in a portion bathymetry, or introduces waterborne contaminants or toxins of their study area, and attributed the lower numbers to the into the environment may seriously impact the dolphins’ fact that benthic prey may decline following removal of the prey sources, feeding opportunities and overall health (e.g. sandy substrate. Clark et al., 2002; Reeves et al., 2003; Hansen et al., 2004; e.g. Adams et al., 2008). The data presented here provide a baseline for comparison against future surveys which may take place during or after Conservation signifi cance. – Fifty percent of all on-effort planned coastal developments in Sarawak. Surveys should Irrawaddy dolphin sightings made in the Kuching area continue on a monthly or quarterly basis, and should include were in close association (within an estimated 50 m) with new areas not yet covered in this study. Applying the same fi shermen setting gill nets of varying mesh sizes from small methods used for this study will allow direct comparison (6-10 m) fi breglass boats. On many occasions, dolphins were of encounter rates to monitor possible changes in relative observed approaching the boats as nets were being hauled abundance between seasons or between years. These surveys in, and fi shermen were seen throwing cast-off fi sh to the should also incorporate an element of water parameter dolphins. Even though this does not involve the same level sampling to understand more fully habitat preference in of cooperation as that described for the Ayeyarwady River relation to environmental factors such as temperature, salinity in Myanmar (Smith et al., 2009), anecdotal information and turbidity. from fi shermen during our surveys indicate that they feel positively toward the dolphins. At the same time, Jaaman While relative abundance and photo–identifi cation methods et al. (2009) report high levels of by-catch in Sarawak and should be continued, it is also important that future research further research is required to determine the severity of the efforts work toward the generation of absolute abundance threat currently posed by fi shing in our study areas. estimates. Photographs collected through September 2009 did not yield suffi cient data for the application of mark- Currently the Irrawaddy dolphin population in the Kuching recapture analysis to generate absolute abundance estimates region supports a local dolphin-watching industry involving for the Irrawaddy dolphins in the study areas. As such, six to eight separate tour operators and generating an future studies should also make use of a vessel with a higher income of an estimated $308,000 per year (O’Connor et viewing platform, a more consistent and trained team of al. 2009). This industry represents the only established observers, and methods suitable for the use of DISTANCE dolphin-watching tourism in Malaysia. Threats to the dolphin software (Buckland et al., 2001; Thomas et al., 2009) to population in the area may undermine efforts to expand the calculate absolute abundance estimates in those areas shown industry in other parts of the region, and thus provide more to be signifi cant habitats for Irrawaddy dolphins and fi nless economic incentive to protect dolphin populations. porpoises. Given the precarious conservation status of Irrawaddy dolphins and fi nless porpoises throughout their Without longer term data and more detailed water parameter range, obtaining population estimates in this part of the analysis, it is diffi cult to determine precisely why encounter species’ range is of particular global conservation value. rates with Irrawaddy dolphins were so much lower in the Miri region than the Bintulu-Similajau and Kuching regions. The results offer the potential to enact some immediate The sections of Miri coastline covered by the surveys are conservation measures, in the form of improved more heavily developed than those in the other two survey communication, cooperation and information sharing between areas, with a major port at the Baram River to the north, a scientists, government planning bodies, and other relevant number of offshore oil rigs located within the survey area, stakeholders in coastal management and conservation. Those and high levels of pollution recorded in the Baram and Miri bodies responsible for approving new developments (e.g. rivers (NREB, 2005). Planned coastal developments in the planning departments and environment boards responsible Similajau study area include extensive dredging for ports and for reviewing and approving Environmental Impact wharves and the construction of a large industrial park near Assessments), should be urged to include consideration for Tanjung Similajau. In Kuching, the construction of a fl ood cetaceans and the integrity of cetacean habitats into their mitigation channel will transport large volumes of silt laden approval criteria. This is particularly true for Irrawaddy fresh water into the Salak River estuary. In the Doubtful dolphins and fi nless porpoises, whose global conservation Sound of New Zealand, large increases in freshwater input status is precarious. Dolphin watch and other coastal eco- resulting from the construction of a second tailrace tunnel tour operators should also be encouraged to contribute to

98 THE RAFFLES BULLETIN OF ZOOLOGY 2011 conservation and research efforts by systematically collecting Currey, R., S. Dawson, E. Slooten, K. Schneider, D. Lusseau, O. sightings data and reporting them to research teams, as well Boisseau, P. Haase, and J. Williams, 2009. Survival rates for a as by adhering to approach guidelines as published, but declining population of bottlenose dolphins in Doubtful Sound, currently not enforced by government bodies. New Zealand: an information theoretic approach to assessing the role of human impacts. Aquatic Conservation: Marine and Freshwater Ecosystems, 19, 658–670. Although the results include only 56 days of survey effort in three study areas, the data presented here represent some of Dawson, S. M., P. R. Wade, E. Slooten, and J. Barlow, 2008. Design the only fi ne-scale habitat analysis of coastal (as opposed to and fi eld methods for sighting surveys of cetaceans in coastal and riverine habitats. Mammal Review, 38: 19–49. fresh water) populations of Irrawaddy dolphins in Southeast Asia. These results can be used to predict the likely species Dolar, M. L. L., W. Perrin, A.A.S.P. Yaptinchay, S.A.B.H. composition and distribution of coastal cetaceans in similar Jaaman, M.D. Santos, M.N. Alava, and M.S.B. Suliansa, 1997. Preliminary investigation of marine mammal distribution, habitats throughout Sarawak’s coastline and to guide abundance and interactions with humans in the southern Sulu managers in future research and conservation efforts. Sea. Asian Marine Biology 14: 61–81. Dolar, M. L. L., W. Perrin, J. P. Gaudiano, A. A. S. P. Yaptinchay, and J. M. L. Tan, 2002: Preliminary report on a small estuarine ACKNOWLEDGMENTS population of Irrawaddy dolphins (Orcaella brevirostris) in the Philippines. The Raffl es Bulletin of Zoology, Supplement The Sarawak Dolphin Project surveys conducted between No. 10: 155–160. June 2008 and December 2009 were made possible through Filatova, O. A., I. D. Fedutin, T. V. Ivkovich, M. M. Nagaylik, funding from Shell Malaysia, Sarawak Aluminium Company A. M. Burdin, and E. Hoyt, 2009. The function of multi- (SALCO), and the Ocean Park Conservation Foundation pod aggregations of fi sh-eating killer whales (Orcinus orca) Hong Kong. We would like to thank the Sarawak Forestry in Kamchatka, Far East Russia. Journal of Ethology, 27: Corporation for the valuable contribution of staff and 333–341. logistical support to the surveys, and the Sarawak Forestry Frère, C. H., P. T. Hale, L. Porter, V. G. Cockcroft, and M. Department for the relevant permissions to conduct dolphin Dalebout, 2008. Phylogenetic analysis of mtDNA sequences surveys in Sarawak waters. We would also like to thank John suggests revision of humpback dolphin (Sousa spp.) taxonomy Braisby and two anonymous reviewers for their valuable is needed. Marine and Freshwater Research, 59: 259–268. comments and suggestions which led to the considerable Friday, N., T. D. Smith, P. Stevick, and J. Allen, 2000. Measurement improvement of this manuscript. of photographic quality and individual distinctiveness for the photographic identifi cation of humpback whales, Megaptera novaeangliae. Marine Mammal Science, 16: 355–374. LITERATURE CITED Gibson-Hill, C. A., 1949. The whales, porpoises and dolphins known in Malayan waters. Malayan Nature Journal, 4: 44–61. Adams, J., M. Houde, D. Muir, T. Speakman, G. Bossart, and P. Hansen, L. J., L. H. Schwacke, G. B. Mitchum, A. A. Hohn, R. Fair, 2008. Land use and the spatial distribution of perfl uoroalkyl Wells, E. S. Zolman, and P. A. Fair, 2004. Geographic Variation compounds as measured in the plasma of bottlenose dolphins in Polychorinated Biphenyl and Organochlorine Pesticide (Tursiops truncatus). Marine Environmental Research, 66: Concentrations in the Blubber of Bottlenose Dolphins from 430–437. the U.S. Atlantic Coast. Science of the Total Environment, Bali, J., S. A. Jaaman, O. B. Tisen, W. S. Landong, M. K. Zaini, 319: 147–172. C. W. Yee, K. Bakir, and S. Saimin, 2008. Aerial sighting Hastie, G. D., B. Wilson, L. J. Wilson, K. P. Parsons, and P. M. rate of marine life in Sarawak waters. 7th International Thompson, 2004. Functional mechanisms underlying cetacean Scientifi c Symposium of the Intergovernmental Oceanographic Commission -Western Pacifi c (IOC/WESPACT), 21-24 May distribution patterns: hotspots for bottlenose dolphins are linked 2008, Magellan Sutera Hotel, Sutera Harbour, Kota Kinabalu, to foraging. Marine Biology, 144: 403. Sabah, 1–13. IUCN, 2008. Cetacean update of the 2008 IUCN Red List of Beijder, L., A. Samuels, H. Whitehead, N. Gales, J. Mann, R. C. Threatened Species. IUCN Redlist of Endangered Species, http:// Connor, M. Heithaus, J. Watson-Capps, T. Flaherty, and M. cmsdata.iucn.org/downloads/cetacean_table_for_website.pdf. Kreutzen, 2006. Decline in relative abundance of bottlenose Jaaman, S. A. 2006. Marine Mammal Distribution and interactions dolphins exposed to long-term disturbance. Conservation with fisheries in East Malaysia: PhD thesis. University of Biology, 20: 1791–1798. Aberdeen, UK. 1–277 p. Buckland, S. T., D. R. Anderson, K. P. Burnham, J. L. Laake, D. Jaaman, S. A., Y. A. Lah-Anyi, and G. J. Pierce, 2009. The L. Borchers, and L. Thomas, 2001. Introduction to Distance magnitude and sustainability of marine mammal by-catch in Sampling: Estimating Abundance of Biological Populations. fi sheries in East Malaysia. Journal of the Marine Biological Oxford University Press. Association of the United Kingdom, 89: 907–920. Clark, E. D., C. S. Perry, and W. J. Elliott, 2002. Examples of Jefferson, T., S. K. Hung, L. Law, M. Torey, and N. Tregenza, organic and heavy metal contaminants in cetaceans from 2002. Distribution and abundance of fi nless porpoises in Hong coastal waters. Document presented to the 54th meeting of the Kong and adjacent waters of China. The Raffl es Bulletin of International Whaling Commission.1–3 p. Zoology, Supplement No. 10: 43–55. Constantine, R., D. H. Brunton, and T. Dennis, 2004. Dolphin- Jefferson, T. and S. K. Hung, 2004. Neophocaena phocaenoides. watching tour boats change bottlenose dolphin (Tursiops Mammalian Species, 746: 1–12. truncatus) behaviour. Biological Conservation, 117: 299– 307.

99 Minton et al. Nearshore cetaceans in Sarawak

Jefferson, T. A., S. K. Hung, and B. Würsig, 2009. Protecting Ponnampalam, L., E. Hines, S. Mananunsap, A. Ilangakoon, K. small cetaceans from coastal development: Impact assessment Adulyanukosol and L. Morse.2009. The behavior of Irrawaddy and mitigation experience in Hong Kong. Marine Policy, 33: dolphins (Orcaella brevirostris) in Trat Province, Eastern Gulf 305–311. of Thailand, with special reference to observed herd mating Jefferson, T.A. and K. Van Waerebeek. 2004. Geographic behavior. . Poster Presentation at the XVIII Biennial Conference variation in skull morphology of humpback dolphins (Sousa on the Biology of Marine Mammals, Quebec, Canada. October, spp.). Aquatic Mammals, 30: 3–17. 2009. Juried. Kreb, D. and Budiono, 2005. Conservation management of small Read, A., P. Drinker, and S. P. Northridge, 2006. Bycatch of core areas: key to survival of a Critically Endangered population Marine Mammals in U.S. and Global Fisheries. Conservation of Irrawaddy river dolphins Orcaella brevirostris in Indonesia. Biology, 20: 163–169. Oryx, 39: 178–188. Reeves, R. R., B. D. Smith, E. A. Crespo, and G. Notarbartolo Lusseau, D. L., 2003. The effects of tour boats on the behavior di Sciara, 2003. Dolphins, Whales and Porpoises: 2002-2010 of bottlenose dolphins: Using Markov chains to model Conservation Action Plan for the World’s Cetaceans. IUCN, anthropogenic impacts. Conservation Biology, 17: 1785– 1–139 pp. 1793. Shirakihara, K., M. Shirakihara, and Y. Yamamoto, 2007. Lusseau, D. L. 2006. Why do dolphins jump? Interpreting the Distribution and abundance of fi nless porpoise in the Inland behavioural repertoire of bottlenose dolphins (Tursiops sp.) Sea of Japan. Marine Biology, 150: 1025–1032. in DoubtfulSound, New Zealand. Behavioural Processes, 73: Smith, B. D. and R. R. Reeves, 2000. Methods for Studying 257–265. Freshwater Cetaceans: Survey Methods for Population Lusseau, D. L., E. Slooten, and R. J. C. Currey, 2007. Unsustainable Assessment of Asian River Dolphins, in Biology and dolphin-watching tourism in Fjordland, New Zealand. Tourism Conservation of Freshwater Cetaceans in Asia, R. R. Reeves, in Marine Environments, 3: 173–178. B. D. Smith, and T. Kasuya, Eds., IUCN, 97–115. Lydekker, R. 1901. Notice of an apparently new estuarine dolphin Smith, B. D., T. T. Mya, M. C. Aung, W. Hang, and M. Thida, from Borneo. Proceedings of the Zoological Society of London, 2009. Catch composition and conservation management of a 1: 88–91. human-dolphin cooperative cast-net fi shery in the Ayeyarwady River, Myanmar. Biological Conservation, 142: 1042–1049. Naeg, D. and M. Ten, 2010. Wetlands RAMSAR listing safe for now. The Borneo Post. Thomas, L., S. T. Buckland, E. A. Rextad, J. L. Laake, S. Strindberg, C. Hedley, J. R. B. Bishop, T. A. Marques, and NREB (Natural Resources and Environment Board Sarawak). 2005. K. P. Burnham, 2010: Distance software: design and analysis Sarawak Annual Water Quality Report 2005 of distance sampling surveys for estimating population size. O’Connor, S., R. Campbell, H. Cortez and T. Knowles, 2009. Journal of Applied Ecology, 47: 5–14. Whale Watching Worldwide: tourism numbers, expenditures and Wang, J. Y., L.S. Chou and B.N. White. 2000. Differences in expanding economic benefi ts. Yarmouth MA, USA: International external morphology of two sympatric species of bottlenose Fund for Animal Welfare. 1–295 p. dolphins (genus Tursiops) in the waters of China. Journal of Parra, G. J. 2006. Resource partitioning in sympatric delphinids: Mammology 81(4):1157–1165. space use and habitat preferences of Australian snubfi n and Wang, J. Y., T.R. Frasier, S.C. Yang, and B.N. White. 2008. Indo-Pacifi c humpback dolphins. Journal of Animal Ecology, Detecting recent speciation events: the case of the finless 75: 862–874. porpoise (genus Neophocaena). Heredity 101: 145–155. Parra, G. J., R. Schick, and P. J. Corkeron, 2006. Spatial distribution Wang, J. Y. and S. C. Yang, 2009: Indo-Pacifi c bottlenose dolphin, and environmental correlates of Australian snubfi n and Indo- Tursiops aduncus. Encyclopedia of Marine Mammals, 2nd ed. Pacifi c humpback dolphins. Ecography, 29: 1–11. W. Perrin, B. Wursig, and J. G. M. Thewissen, Eds., Elsevier, Parra, G. J. and G. J. B. Ross, 2009. Humpback dolphins, S. 602–608. chinensis and S. Teuszii. Encyclopedia of Marine Mammal, 2nd ed. W. Perrin, B. Wursig, and J. G. M. Thewissen, Eds., Elsevier, 576–581.

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PRELIMINARY REPORT ON MITOCHONDRIAL DNA VARIATION IN MACACA FASCICULARIS FROM SINGAPORE

Michael A. Schillaci Department of Social Sciences, University of Toronto Scarborough, 1265 Military Trail, Toronto, Ontario, M1C 1A4, Canada Email: [email protected] (corresponding author)

Sandy Saravia Department of Cell and Systems Biology, University of Toronto 25 Harbord Street, Toronto, Ontario, M5S 3G5, Canada Email: [email protected]

Benjamin P. Y.-H. Lee Parks Division, National Parks Board Headquarters, Singapore Botanic Gardens, 1 Cluny Road, Singapore 259569, Republic of Singapore Email: [email protected]

Carney Matheson Department of Anthropology, Lakehead University, Thunder Bay, Ontario, Canada Email: [email protected]

ABSTRACT. – Few studies have examined the molecular relationships of a single natural population of macaque species. This preliminary study sought to conduct a phylogenetic analysis of mitochondrial DNA (mtDNA) variation in Singapore’s macaque population and to describe the geographic structure of that variation. Hair samples were obtained from wild Macaca fascicularis fascicularis (n=54) trapped at multiple locations in the Bukit Timah Nature Reserve (BTNR) and the Central Catchment Nature Reserve (CCNR). The mtDNA sequence data obtained from the hair samples were used to generate phylogenetic trees using the neighbor joining and maximum parsimony methods. Based on mtDNA fragments, two groupings were apparent within Singapore, one from BTNR and the other from CCNR. Neighbor joining bootstrap values between 99% (COII) and 82% (rRNA 12S) supported this fi nding. With the exception of the rRNA 12S fragment, similarly strong bootstrap support was observed for the maximum parsimony analysis. The apparent differentiation within Singapore might be due to some form of random drift, such as founder effect, associated with habitat fragmentation stemming from artifi cial modifi cation of the landscape which has occurred since the founding of Singapore in 1819. Fragmentation of the tropical lowland rainforest, therefore, seems to have resulted in two primary sub-populations of macaques on the island.

KEY WORDS. – Long-tailed macaque, phylogeny, mitochondrial DNA, Singapore.

INTRODUCTION and Coke, 2007). Few studies, however, have focused on a single endemic population of macaques (see Hoelzer et A number of studies have examined the molecular al., 1994 ). In this report we examine mitochondrial DNA phylogenetic relationships among macaque species or (mtDNA) variation in an insular population of long-tailed populations across relatively large geographic areas of macaques from Singapore, a relatively small and highly Southeast Asia (e.g., Blancher et al., 2008; Chakraborty et al., urbanized island nation. 2007; Chu et al., 2007; Evans et al., 1999, 2001; Hayasaka et al., 1996; Li and Zhang, 2005; Morales and Melnick, 1998; The island of Singapore would have been covered largely with Smith et al., 2007; Tosi et al., 2000, 2003), or have used lowland evergreen rain forest when it was founded in 1819 molecular phylogenies to estimate the geographic origins (Corlett 1992). Since then, the natural landscape has been of non-endemic human-introduced macaque populations largely modiﬁ ed through various land use decisions in both (Kawamoto et al., 2007; Stevison and Kohn, 2008; Tosi historical and modern times. The initial clearance of primary

101 Schillaci et al. – Mitochondrial DNA variation in Singapore Macaca fascicularis forest on the island after its founding was for the planting MATERIALS AND METHODS of gambier (Uncaria gambir), a cash crop with medicinal properties, and also used in the leather industry (Corlett DNA extraction, PCR amplifi cation and DNA sequencing. – & Turner, 1997). Most of the existing forest fragments in In this study, hair samples were collected from 54 individuals Singapore have been isolated for some 150 years (Corlett of M. fascicularis trapped at six different locations within & Turner, 1997). Much of the deforestation occurred under the Central Catchment Nature Reserve (CCNR) and two British rule from 1819 to 1900 (Corlett & Turner, 1997), locations within the Bukit Timah Nature Reserve (BTNR) in a period which saw rapid economic development and Singapore (Fig. 1). Each location corresponds to a separate population growth. From 1884, the majority of the larger social group or troop. For the purposes of this paper we forest patches were incorporated into forest reserves (Corlett consider all macaque troops on the island to comprise a 1992). All but the central part of Singapore’s forests (totaling single potential breeding population. Hairs were plucked some 1600 ha) was eventually abandoned, and the current using gloved hands, placed into sterile tubes and stored rain forest nature reserves retain most of the remaining in a cool, dark place. Approximately 5-6 hairs from each primary forest patches on the island (Corlett & Turner, 1997). individual were cut with sterile scissors directly into a 1.5 ml However, the construction of roads, reservoirs, golf courses microfuge tube; each section of cut hair was approximately and military facilities further fragmented the forests of the 4 mm in length and did not include the root. Total genomic nature reserves, thereby creating barriers to primate dispersal. DNA was extracted from each sample with the following Currently, the two largest forested nature reserves (BTNR buffer: 1M Tris-HCl (pH 8.0), 0.5 M NaCl, 0.5 M EDTA, and CCNR) are separated by the Bukit Timah Expressway, 0.2% SDS, 0.039 M DTTand 0.1 mg/mL proteinase K a large 6 lane road built in 1985 which experiences a high (Promega). The reactions were incubated overnight at 56oC volume of vehicular traffi c. with rigorous shaking (1000 rpm). Following the addition of 0.10% sodium acetate, the DNA was precipitated with 2.5 Despite its size and urbanicity, Singapore is home to three volumes of chilled 100% ethanol and a 30 minute incubation genera of nonhuman primates (Macaca sp., Presbytis sp. and period on ice. All the DNA samples were furthered purifi ed Nycticebus sp.), and a number of other members of the grand using the HiYield Gel/PCR DNA Extraction Kit (Real Biotech order Archonta, including tree shrews (Order Scandentia, Corp.) according to manufacturer’s instructions. Tupaiidae: Tupaia glis), colugos (Order Dermoptera, Cynocephalidae: Galeopterus variegatus), and 24 extant Using the DNA samples, sections of fi ve mitochondrial species of bat (Order Chiroptera). genes were amplifi ed via the polymerase chain reaction (PCR). PCR was performed using primers for 12s rRNA, Among the nonhuman primate taxa, the long-tailed macaque tRNAGlu- cytochrome b and cytochrome oxidase I, II and [Macaca fascicularis (Raffl es, 1821)], is by far the most III, as previously described by Li and Zhang (2005). The numerous. A census conducted in 2007 (Sha, 2009a) target sequence of each primer pair and its location relative estimated there were between 1,218 and 1,454 macaques to the reference mitochondrial gene is shown in Table 1. on the island (also see Agoramoorthy & Hsu, 2006). Singapore offers a unique and highly urban ecology for its macaques which consists of a patchwork of small primary and secondary forest fragments, disturbed forests, urban parks and even very small forested patches within and adjacent to residential neighborhoods. About 70% of the island’s macaque population resides in Bukit Timah and Central Catchment Nature Reserves which comprise a series of forests and reservoirs located near the center of the island (Sha et al., 2009a, 2009b). As recently suggested by Sha et al. (2009), the majority of the island’s macaque population is confi ned to these two nature reserves, likely as a result of roads and highways.

Recent research on Singapore’s long-tailed macaques has focused on morphological variation (Schillaci et al., 2007), bidirectional disease transmission (Jones-Engel et al., 2006), and human-macaque interactions (Fuentes et al., 2008; Sha et al., 2009a, 2009b). Macaques from Singapore have also been included in the phylogenetic analysis of simian foamy virus (Jones-Engel et al., 2008). To date, however, there has not been an analysis of the molecular genetic variation of Singapore’s macaques. The primary goal of the present research is to conduct a phylogenetic analysis of mitochondrial DNA variation of Singapore’s macaque population and to examine the geographic structure of that Fig. 1. Map of sample locations within the Bukit Timah (BTNR) variation. and Central Catchment (CCNR) Nature Reserves.

102 THE RAFFLES BULLETIN OF ZOOLOGY 2011

Table 1. Size of the ampliﬁ ed sequences and their location relative to the mitochondrial gene sequence.

Mitochondrial gene base pairs (bp) Ampliﬁ ed sequence location Amplicon size (bp) 12s rRNA 948 257–677 421 cytochrome oxidase I (COI) 1539 698–1113 416 cytochrome oxidase II (COII) 684 39–533 495 cytochrome oxidase III (COIII) 784 169–660 492 tRNAGlu- cytochrome b (cytb) 1210 52–471 420

Table 2. Genbank DNA sequences included in phylogenetic analyses.

Taxon Origin GenBank accession number Macaca fascicularis Vietnam AY685714–AY685717, AY685774, AY685775, AY685777, AY685796–AY685799, AY685837–AY685840, AY685878–AY685881 M. fascicularis Not speciﬁ ed M58008 M. mulatta India AY685718, AY685769, AY685800, AY685841 M. mulatta Fujian, China AY685720–AY685722, AY685802–AY685804, AY685843–AY685845, AY685884–AY685886 M. mulatta Hainan, China AY685787 M. mulatta Myanmar AY685783–AY685785, AY685809, AY685727, AY685891, AY685850 M. sylvanus Not speciﬁ ed NC_002764

Amplifi cation of some genes was unsuccessful for some history, we were not able to combine the fi ve gene fragments individuals. into a single analysis (see Li & Zhang 2005: 379) because we were constrained by the availability of comparative Each PCR reaction had a total volume of 25 µl and sequences from the GenBank. We used the sequence data to contained 1x PCR Buffer (Invitrogen), 200 mM of each generate phylogenetic trees using the neighbor joining (NJ) dNTP (Invitrogen), 2 mM MgCl2, 2 U Platinum Taq DNA and Maximum Parsimony (MP) methods (Saitou & Nei, Polymerase (Invitrogen), 0.2 µM of each forward and reverse 1987). Following Chu et al. (2007: 420), when constructing primer and sterile water. Approximately 1µg of DNA sample our NJ phylogenetic trees we used a simple p-distance model was used per reaction. The PCR cycling parameters were because genetic distances estimated using more sophisticated an initial denaturation at 940C for 2 minutes then 45 cycles models often generate large variances which reduce the of 940C for 30 seconds, 600C for 1 minute and 720C for 2 resolution of bootstrapped tree topologies based on short minutes. The PCR products were purifi ed with an ethanol- nucleotide sequences (see Nei & Kumar, 2000). Bootstrap sodium acetate procedure similar to the one described support for nodes was estimated using 2000 replications above. (Felsenstein, 1985). For all trees M. sylvanus was used as the outgroup. All positions containing gaps and missing DNA sequencing was performed using the Big Dye data were excluded. For the MP phylogenetic analysis each Terminator Cycle Sequencing (3.1) kit (Applied Biosystems) three represented a most parsimonious tree from the Close- along with the same primers used in PCR. Each 10 µl Neighbor-Interchange algorithm (Nei & Kumar, 2000), with sequencing reaction consisted of 3 µl of Big Dye Terminator a search level of 2. The initial trees were obtained with Cycle Sequencing Ready Reaction Mix, 0.3 µM of either the random addition of sequences with 100 replicates. The forward or reverse primer and approximately 1 µg of purifi ed branch lengths were calculated using the average pathway DNA. The sequencing cycling parameters were the same to method. Bootstrap values refl ect the percentage of replicate those stated for PCR. The sequencing reactions were purifi ed trees in which the associated taxa clustered together after the same way the PCR reactions were prior to sequencing. 100 replicates (Felsenstein, 1985). The sequences were analyzed by the ABI 3100 Genetic Nucleotide diversity ( ) and average within-group Analyzer. The sequences were edited and aligned using π evolutionary divergence (d ) were calculated as the number BioEdit Sequence Alignment Editor. Along with the 41 M. x of base differences per site from averaging over all sequence fascicularis samples described here, M. sylvanus, M. mulatta pairs (Tamura et al., 2007). Standard errors for within-group and additional M. fascicularis sequences from the GenBank divergence estimates were obtained by a bootstrapping were also included in downstream analyses (Table 2). procedure using 500 replications. The neutral mutation model for each gene fragment for the pooled M. fascicularis sample Data analysis. – Although the fi ve mtDNA sequences are was examined using Tajima’s test of neutrality (Tajima, inherited as effectively one locus without recombination and 1989). All analyses were conducted using the MEGA4 can be assumed to have experienced the same branching computer software package (Tamura et al., 2007).

103 Schillaci et al. – Mitochondrial DNA variation in Singapore Macaca fascicularis

Table 3. Results from Tajima’s neutrality tests for Macaca fascicularis from Singapore and Vietnam. M: number of sites; S: number of segregating sites; ps = S/M; θ = ps/a1; π: nucleotide diversity; D: Tajima’s test statistic (Tajima, 1989).

M S ps θ π D Cytb 52 14 0.03302 0.00731 0.00665 -0.26942 12s rRNA 39 11 0.02619 0.00619 0.00356 -1.30001 COI 54 14 0.03365 0.00739 0.00504 -0.95020 COII1 55 11 0.02222 0.00485 0.00317 -0.99637 COIII 46 26 0.05295 0.01205 0.00673 -1.46814

1The M58008 sequence of unknown provenance was excluded.

RESULTS AND DISCUSSION Singapore formed a monophyletic grouping relative to the small comparative sample of M. fascicularis from Vietnam. The results from Tajima’s neutrality tests indicated the Bootstrap support values for the nodes forming the Singapore neutral mutation hypothesis can explain the observed monophyletic groupings varied from 100–98% (cytb, COI, mtDNA polymorphisms for the combined grouping of M. COII, COIII) to 68% (12s rRNA) for the NJ phylogenies, fascicularis for all ﬁ ve gene fragments (Table 3). The NJ and from 89–99% (cytb, COI, COII, COIII) to 40% (12s and MP phylogenetic analyses yielded trees with almost rRNA) for the MP phylogenies. Four of ﬁ ve bootstrap values identical topologies (cf. Figs. 2 & 3). For each of the therefore strongly support the inference of monophyly, with five mitochondrial genes amplified, the macaques from values ≥ 89%. Consistency and retention indexes of greater

Fig. 2. Neighbor-joining trees for each of ﬁ ve mtDNA gene fragments including: A, cytochrome b; B, 12s rRNA; C, COI; D, COII; E, COIII. Bootstrap support values are presented at each node.

104 THE RAFFLES BULLETIN OF ZOOLOGY 2011

Table 4. Statistics for the most parsimonious tree used in this MP analysis based on the parsimony-informative sites.

Number of trees Consistency Retention Composite Number of Parsimony Informative evaluated Index Index Index Positions Positions Cytb 6258 0.672131 0.874214 0.587586 409 40 12s rRNA 2473 0.750000 0.895522 0.671642 414 19 COI 9773 0.661538 0.862500 0.570577 410 42 COII 7719 0.636364 0.862069 0.548589 493 62 COIII 7829 0.716216 0.905405 0.648466 484 51 than 0.5 for the MP analysis indicate that the phylogenetic number of comparative sequences available in the GeneBank, reconstructions are largely unaffected by recurrent mutation precluding important comparisons with macaques from the and parallel evolution (Feris, 1989) (Table 4). It is important Malay Peninsula and the Indonesian archipelago. to state, however, that the present study is limited by the

A B

50 80 CCNR CCNR BTNR + M18 99 BTNR 44 M. fascicularis AY685837- AY685839 M. fascicularis AY685878, AY685879 98 59 72 M. mulatta AY685886 M. fascicularis AY685840 M. fascicularis AY685880 M. mulatta AY685841 M. fascicularis AY685881 95 73 M. mulatta AY685845 M. mulatta AY685885 55 M. mulatta AY685850 99 64 M. mulatta AY685891 M. mulatta AY685844 81 M. mulatta AY685884 M. mulatta AY685843 M. sylvanus NC002764 M. sylvanus NC002764

5 2

C D

93 CCNR 36 CCNR 90 BTNR

89 BTNR 99 M. fascicularis AY685768, AY685774, AY685775 M. fascicularis AY685714 - AY685716 80 M. fascicularis AY685777 99 M. fascicularis AY685717 M. fascicularis M58008 M. mulatta AY685718 M. mulatta AY685769 M. mulatta AY685727 50 M. mulatta AY685784 85 97 M. mulatta AY685783 83 M. mulatta AY685722 31 34 M. mulatta AY685721 M. mulatta AY685787 85 M. mulatta AY685720 88 M. mulatta AY685785 M. sylvanus NC002764 M. sylvanus NC 002764

5 5

97 CCNR + S31

99 BTNR

80 M. fascicularis AY685799 M. fascicularis AY685796 - AY685798 M. mulatta AY685800

88 48 M. mulatta AY685804 93 M. mulatta AY685809 79 M. mulatta AY685803 M. mulatta AY685802 M. sylvanus NC002764

Fig. 3. Maximum parsimony trees for each of ﬁ ve mtDNA gene fragments including: A, cytochrome b; B, 12s rRNA; C, COI; D, COII; E, COIII. Bootstrap support values are presented at each node.

105 Schillaci et al. – Mitochondrial DNA variation in Singapore Macaca fascicularis

Table 5. Average evolutionary divergence (dx) within macaque groupings for 5 mtDNA gene fragments.

Macaca. fascicularis Macaca mulatta Singapore Vietnam

dx SE dx SE dx SE Cytb 0.00397 0.00165 0.00354 0.00195 0.03538 0.00556 12s rRNA 0.00114 0.00097 0.00716 0.00275 0.02546 0.00528 COI 0.00154 0.00095 0.00120 0.00115 0.03750 0.00646 COII1 0.00178 0.00113 0.00202 0.00135 0.03475 0.00530 COIII 0.00390 0.00127 0.00407 0.00189 0.02846 0.00473

1The M58008 M. fascicularis sequence of unknown provenance was excluded.

With the exception of the 12s rRNA gene fragment, the M. have resulted from any number of changes to the landscape fascicularis grouping exhibited similar levels of within-group associated with the historical development of Singapore. evolutionary divergence as the Vietnam grouping (Table 5). The observed within-group divergence estimates for both M. fascicularis groupings were much less than that CONCLUSION observed for M. mulatta. For all mtDNA gene fragments, two groupings were apparent within Singapore, one from In conclusion, though subject to important limitations BTNR and the other from CCNR. With the exception of one stemming from a dearth of comparative sequences, the results individual for each of two gene fragments (12s rRNA, COIII) of our preliminary study seem to suggest the possibility of these groupings were monophyletic (Appendix). Statistical monophyly for the long-tailed macaques from Singapore support for these groupings varied with NJ bootstrap values relative to other Southeast Asian populations for five between 99% (COII) and 82% (12s rRNA). Lower bootstrap different mitochondrial gene fragments. Additional analyses support values for these groupings were observed for the with a larger comparative sample, however, are needed MP trees. to more adequately assess the possibility of monophyly. Our phylogenetic analysis also indicated there are two Macaque social organization is characterized by strict female largely monophyletic sub-populations within Singapore philopatry with obligatory male migration at adulthood. corresponding to two forested nature reserves.The observed Very large troops may eventually split into separate troops mtDNA differentiation may be attributable to a subset (i.e., according to matrilines, occupying separate geographic home BTNR) of the larger CCNR sub-population becoming isolated by the building of the Bukit Timah Expressway, or some ranges. Because mtDNA is maternally inherited, macaque other artifi cial modifi cation of the landscape. This isolation, social organization produces geographically restricted in conjunction with random genetic drift is likely responsible mitochondrial DNA variation with marked intergroup for the observed geographically mediated population structure heterogeneity (Melnick & Hoelzer, 1992). As discussed by within Singapore. Hoelzer et al. (1994: 451), mitochondrial genome divergence begins when a single mutation differentiates a copy of the Future phylogenetic analyses of mtDNA variation in genome from the ancestral sequence. This differentiated copy Singapore’s macaques should include a larger set of is a non-recombinant clone which will accumulate mutations comparative sequences. Specifi cally, sequences from M. over evolutionary time (Hoelzer et al., 1994). Hoelzer et al. fascicularis populations in Indonesia, Malaysia and Thailand, (1994) point out that this accumulation of mutations can particularly from populations on the Malay Peninsula south produce substantial divergence within a species, even within of the Isthmus of Kra, are needed to more robustly determine populations, without reproductive isolation. For the present the phylogenetic position of the Singapore population. study, it seems unlikely that the divergence observed between the two Singapore sub-populations for each of 5 mtDNA gene fragments would correspond to a geographic barrier merely ACKNOWLEDGMENTS by chance. It seems possible, therefore, that the smaller BTNR group ( n≈125; see Appendix in Sha et al. 2009a), is This research was funded in part by the Caunaught Fund a subset of the larger original CCNR sub-population (n≈902; of the University of Toronto. Trapping and data collection see Appendix in Sha et al. 2009a), that may have become protocols used for this study were approved by the cut off by forest fragmentation associated with the artifi cial institutional animal care and use committee of the University modifi cation or remodifi cation of the landscape surrounding of Toronto (Protocol no. 20005356). The authors are grateful the Bukit Timah area. For example, the construction of to L. Jones-Engel of the University of Washington and S. the Bukit Timah Expressway may have resulted in some Chan of NParks Singapore for facilitating this research. We form of random drift such as founder effect for the BTNR thank S. Oon, W. Ng, and J. Castillo Garcia for their help group. Under this scenario, random drift, in conjunction with with fi eld work and data collection, and D. Nikitovic for isolation from any possible migrating females, resulted in her help with graphics. We also thank the Associate Editor two differentiated mtDNA sub-populations. It is important and two anonymous reviewers for their helpful comments to note, however, that isolation and random drift could and criticisms of earlier drafts.

106 THE RAFFLES BULLETIN OF ZOOLOGY 2011

LITERATURE CITED on the island of Mauritius: an assessment of nuclear and mitochondrial DNA polymorphisms. Journal of Medical Agoramoorthy, G. & Hsu M. J., 2006. Population status of long- Primatology, 37(1): 45–54. tailed macaques (Macaca fascicularis) in Singapore. Mammalia, Li, Q.-Q. & Y.-P. Zhang, 2005. Phylogenetic relationships of 70(3/4): 300–302. the macaques (Cercopithecidae: Macaca), inferred from Blancher, A., M., Bonhomme, B., Crouau-Roy, K. Terao, T. Kitano mitochondrial DNA sequences. Biochemical Genetics, 43(7–8): & N. Saitou, 2008. Mitochondrial DNA sequence phylogeny 375–386. of 4 populations of widely distributed cynomolgus macaque Melnick, D. J. & G. A. Hoelzer, 1992. Differences in male and (Macaca fascicularis fascicularis). Journal of Heredity, 99(3): female macaque dispersal lead to contrasting distributions of 254–264. nuclear and mitochondrial DNA variation. International Journal Chakraborty, D., U. Ramakrishnan, J. Panor, M. Charudutt & A. of Primatology, 13(4): 379–393. Sinha, 2007. Phylogenetic relationships and morphometric Morales, J. C. & D. J. Melnick, 1998. Phylogenetic relationships of afﬁ nities of the Arunachal macaque Macaca munzala, a newly the macaques (Cercopithecidae: Macaca), as revealed by high described primate from Arunachal Pradesh, northeastern India. resolution restriction site mapping of mitochondrial ribosomal Molecular Phylogenetics and Evolution 44 , (2): 838–849. genes. Journal of Human Evolution, 34(1): 1–23. Chu, J.-H., Y.-S. Lin & H.-Y. Wu, 2007. Evolution and dispersal Nei, M. & S. Kumar. 2000. Molecular evolution and phylogenetics. of three closely related macaque species, Macaca mulatta, Oxford University Press, New York. M. cyclopis, and M. fuscata, in eastern Asia. Molecular Phylogenetics and Evolution, 43(2): 418–429. Saitou, N. & M. Nei, 1987. The neighbor-joining method; a new method for reconstructing phylogenetic trees. Molecular Biology Corlett, R.T. 1992. The ecological transformation of Singapore. and Evolution, 4(4): 406–425. Journal of Biogeography, 19(4): 411–420. Schillaci, M. A., L. Jones-Engel, B. P. Y.-H. Lee, A. Fuentes, N. Corlett, R.T. & I.M. Turner, 1997. Long-term survival in tropical Aggimarangsee, G. A. Engel & T. Sutthipat, 2007. Morphology forest remnants in Singapore and Hong Kong. In: Laurance W. Macaca F. & R. O. Bierregaard, Jr. (eds), Tropical Forest Remnants: and somatometric growth of long-tailed macaques ( Ecology, Management and Conservation of Fragmented fascicularis fascicularis) in Singapore. Biological Journal of Communities: 333–345. Chicago University Press. the Linnean Society, 92(4): 675–694. Evans, B. J., J. Supriatna & D. J. Melnick, 2001. Hybridization Sha, J. C. M., M. D. Gumert, B. P. Y.-H. Lee, A. Fuentes, S. and population genetics of two macaque species in Sulawesi, Rajathurai, S. Chan & L. Jones-Engel, 2009a. Status of the Indonesia. Evolution, 55(8):1686–1702. long-tailed macaque Macaca fascicularis in Singapore and implications for management. Biodiversity Conservation Evans, B. J., J. C. Morales, J. Supriatna & D. J. Melnick, 1999. published online March 19, 2009 doi. 10.1007/s10531-009- Origin of the Sulawesi macaques (Cercopithecidae, Macaca) 9616-4 as inferred from mitochondrial DNA phylogeny. Biological Journal of the Linnean Society, 66(4): 539–560. Sha, J. C. M., M. D. Gumert, B. P. Y.-H. Lee, L. Jones-Engel, S. Chan & A. Fuentes, 2009b. Macaque-human interactions and Felsenstein, J., 1985. Conﬁ dence limits on phylogenies: An approach the societal perceptions of macaques in Singapore. American using the bootstrap. Evolution, 39:783–791. Journal of Primatology, 71(10): 825–839. Faris, J. F., 1989. The retention index and the rescaled consistency Smith, D. G., J. W. McDonough & D. A. George, 2007. index. Cladistics, 5:417–419. Mitochondrial DNA variation within and among regional Fuentes, A., S. Kalchik, L. Getter, A. Kwiatt, M. Konecki & L. populations of longtail macaques (Macaca fascicularis) in Jones-Engel, 2008. Characterizing human macaque interactions relation to other species of the Fascicularis group of macaques. in Singapore. American Journal of Primatology, 70(9): American Journal of Primatology, 69(2): 182–198. 879–883. Stevison, L. S. & M. H. Kohn, 2008. Determining genetic Hayasaka, K., K. Fujii & S. Horai, 1996. Molecular phylogeny of background in captive stocks of cynomolgus macaques (Macaca macaques: implications of nucleotide sequences from 896-base fascicularis). Journal Medical Primatology, 37(6): 311–317. pair region of mitochondrial DNA. Molecular Biology and Evolution, 13(7): 1044–1053. Tajima, F., 1989 Statistical methods to test for nucleotide mutation hypothesis by DNA polymorphism. Genetics, 123(3): Hoelzer, G. A., W. P. J. Dittus, M. V. Ashley & D. J. Melnick, 1994. 585–595. The local distribution of highly divergent mitochondrial DNA haplotypes in toque macaques Macaca sinica at Polonnaruwa, Tamura, K., J. Dudley, M. Nei & S. Kumar, 2007. MEGA4: Sri Lanka. Molecular Ecology, 3(5): 451–458. Molecular Evolutionary Genetics Analysis (MEGA) software version 4.0. Molecular Biology and Evolution, 24(8): 1596– Jones-Engel, L., G. A. Engel, M. A. Schillaci, B. Lee, J. Heidrich, 1599 M. Chalise & R. C. Kyes, 2006. Considering human-primate transmission of measles virus through the prism of risk analysis Tosi, A. J. & C. S. Coke, 2007. Comparative phylogenetics American Journal of Primatology, 68(9): 868–879. offer new insights into the biogeographic history of Macaca fascicularis and the origin of the Mauritian macaques. Molecular Jones-Engel, L., C. C. May, G. A. Engel, K. A. Steinkraus, Phylogenetics and Evolutioni, 42(2): 498–504. M. A. Schillaci, A. Fuentes, A. Rompis, M. K. Chalise, N. Aggimarangsee, M. M. Feeroz, R. Grant, J. S. Allan, A. Putra, Tosi, A. J., J. C. Morales & D. J. Melnick, 2000. Comparison of Y- I. N. Wandia, R. Watanabe, L. Kuller, S. Thongsawal, R. chromosome and mtDNA phylogenies leads to unique inferences Chaiwarith, R. C. Keys & M. L. Linial, 2008. Diverse contexts of macaque evolutionary history. Molecular Phylogenetics and of zoonotic transmission of simian foamy viruses in Asia. Evolution, 17(2):133–144. Emerging Infectious Diseases, 14(8):1200–1208. Tosi, A. J., J. C. Morales & D. J. Melnick, 2003. Paternal, maternal, Kawamoto, Y., S. Kawamoto, K. Matsubayashi, K. Nozawa, T. and biparental molecular markers provide unique windows Watanabe, M.-A. Stanley & D. Perwitasari-Farajallah, 2007. onto the evolutionary history of macaque monkeys. Evolution, Genetic diversity of longtail macaque (Macaca fascicularis) 57(6): 1419–1435.

107 Schillaci et al. – Mitochondrial DNA variation in Singapore Macaca fascicularis

Appendix. Sample information and NJ and MP phylogenetic group afﬁ liation (CC=CCNR, BT=BTNR).

ID# Sex Age Location (Troop) Cytb rRNA COI COII COIII S18 F Juvenile Bukit Timah (VC group) BT BT BT S21 F Juvenile Riffl e Range Road (Mid-way) CC CC S23 M Juvenile Riffl e Range Road (Mid-way) CC CC CC S24 F Adult Riffl e Range Road (Mid-way) CC CC CC S25 M Adult Riffl e Range Road (Mid-way) CC S26 M Adult Riffl e Range Road (End) CC CC CC S27 M' Juvenile Riffl e Range Road (End) CC CC CC S28 M Juvenile Riffl e Range Road (End) CC CC CC CC S29 M Adult Riffl e Range Road (End) CC CC CC CC CC S30 F Juvenile Riffl e Range Road (End) CC CC CC CC S31 F Adult Bukit Timah (group B) BT BT BT BT CC S32 F Adult Bukit Timah (group B) BT BT BT S33 F Adult Bukit Timah (group B) BT S34 F' Juvenile Bukit Timah (group B) BT BT BT BT BT S35 M Juvenile Bukit Timah (group B) BT BT BT BT BT S37' F Juvenile Bukit Timah (group B) BT BT BT BT S38 M Subadult Bukit Timah (group B) BT BT BT S39 Bukit Timah (group B) BT BT BT BT BT M1 M' Juvenile Bukit Timah (VC group) BT BT BT BT BT M2 M Juvenile Bukit Timah (VC group) BT BT BT BT M3 F Juvenile Bukit Timah (VC group) BT BT BT BT BT M4 M Juvenile Bukit Timah (VC group) BT M5 M Juvenile Bukit Timah (VC group) BT BT BT BT BT M6 F Adult' Riffl e Range Road (End) CC CC CC CC CC M7 F Adult Riffl e Range Road (End) CC CC CC CC M8 F Juvenile Riffl e Range Road (End) CC CC CC CC CC M9 M Juvenile Riffl e Range Road (End) CC CC CC CC M10 F Adult Riffl e Range Road (End) CC CC CC CC M11 M Adult Upper Peirce Reservoir (Road) CC CC CC CC CC M12 M Subdult Upper Peirce Reservoir (Road) CC CC CC CC CC M13 M Juvenile Upper Peirce Reservoir (Road) CC CC CC CC M14 M Adult Lower Peirce Reservoir CC CC CC CC CC M16 F Subdult Lower Peirce Reservoir CC CC CC CC CC M17 M Infant Lower Peirce Reservoir CC CC CC CC CC M18 M Juvenile Lower Peirce Reservoir CC BT CC CC CC M19 M Adult Bukit Kalang Service Reservoir CC CC CC CC M20 F Adult Bukit Kalang Service Reservoir CC CC CC CC CC M21 M Juvenile Bukit Kalang Service Reservoir CC CC CC M22 F Juvenile Bukit Kalang Service Reservoir CC CC CC CC CC M23 M Adult Upper Seletar CC CC CC CC CC M24 F Juvenile Upper Seletar CC CC CC CC CC M25 M Juvenile Upper Seletar CC CC CC CC CC M26 M Adult Upper Seletar CC CC CC CC CC M27 M Juvenile Upper Seletar CC CC CC CC M28 M Juvenile Upper Seletar CC CC CC CC CC M29 M Juvenile Upper Seletar CC CC CC CC CC M30 F Juvenile Upper Seletar CC CC CC CC CC M31 F Juvenile Upper Seletar CC CC CC CC M32 M Juvenile Upper Seletar CC CC CC CC CC M33 M Adult Lower Peirce Reservoir CC CC CC CC M34 F Subdult Lower Peirce Reservoir CC CC CC CC CC M35 F Juvenile Lower Peirce Reservoir CC CC CC CC CC M36 M Juvenile Lower Peirce Reservoir CC CC CC CC CC M37 M Juvenile Lower Peirce Reservoir CC CC CC CC

108 THE RAFFLES BULLETIN OF ZOOLOGY 2011

NATURAL-LICKS USE BY ORANGUTANS AND CONSERVATION OF THEIR HABITATS IN BORNEAN TROPICAL PRODUCTION FOREST

Hisashi Matsubayashi and Abdul Hamid Ahmad Institute for Tropical Biology and Conservation, Universiti Malaysia Sabah, LB 2073, 88999, Kota Kinabalu, Sabah, Malaysia Email: [email protected] (Corresponding author)

Nobuhiko Wakamatsu, Etsuko Nakazono, and Masaaki Takyu Department of Forest Science, Tokyo University of Agriculture, Sakuragaoka 1-1-1, Setagaya, Tokyo, Japan

Noreen Majalap and Peter Lagan Sabah Forestry Department, LB 68, 90000, Sandakan, Sabah, Malaysia

Jum Raﬁ ah Abd. Sukor Sabah Wildlife Department, 5th Floor, B Block, Wisma MUIS, 88100, Kota Kinabalu, Sabah, Malaysia

ABSTRACT. – The use of natural-licks by orangutans (Pongo pygmaeus) was investigated with camera traps in the Deramakot production forest, Sabah, Malaysian Borneo. The results showed that 1) Although orangutans were in the top three species at all the natural-licks, visitation frequency differed at the natural- licks depending on the surrounding environment; 2) Natural-licks use by orangutans was impacted more by human activity than concentration of the minerals; and 3) Visiting proportion of each orangutan class: fl anged male, female with infant, and others, showed that fl anged male accounted for 31%; female with infant, 17%; and others, 52%; although we had anticipated a bias toward the fl anged male. These results suggest that the natural-licks are key habitats for all classes of orangutans and suitable sites for ground monitoring. Therefore, it is strongly recommended that other production forests adopt protection of natural- licks and managed as a monitoring site for orangutan habitat conservation. Furthermore, as orangutans are vulnerable to human activity, it is necessary to manage natural-licks as protected area with buffer zones of reduced human activity.

KEY WORDS. – Camera trap, monitoring site, Pongo pygmaeus, sustainable forest management.

INTRODUCTION some areas with high biodiversity and frequently used by endangered species must be protected as high conservation The state of Sabah, Malaysian Borneo, has the highest value forests (Forest Stewardship Council, 1996). biodiversity on the island (Hardiono & Alfred, 2005), and the forested area covers about 50% of the state’s total land area. Natural-licks are mineral-rich sources and are benefi cial to However, protected forests represent less than 10%, while large herbivores (Kreulen, 1985). Aggregated distributions production forests (commercial forest reserves) represent of mammals around natural-licks are well known in Africa over 70% of the forested area at present (Sabah Forestry and Neotropical zones (Weir, 1972; McNaughton, 1988; Department, 2009). Montenegro, 2004; Blake et al., 2010; Bravo et al., 2010). In our previous study in the Deramakot Forest Reserve, camera- The Deramakot Forest Reserve is one of the production trap records demonstrated that natural-licks contributed forests located on the upper Kinabatangan River in central minerals for several mammals, and the fi ndings suggest Sabah (Fig.1). The Forest Stewardship Council (FSC) the importance of natural-licks for sambar deer (Rusa certifi ed the Forest Reserve as a well-managed forest in unicolor), which are representative terrestrial herbivores not 1997. The Forest Reserve is divided into 135 compartments only for daily sustenance but also for reproductive support of various sizes with the use of existing roads and other (Matsubayashi et al., 2007a; 2007b). Since 2008, the Sabah natural features (rivers, streams, ridges, and foothills) as Forestry Department has enacted a rule that prohibits any boundaries (Lagan et al., 2007). The forest management is disturbance within a radius of 100 m of a natural-lick. controlled by principles and criteria of the FSC. For instance, This regulation applies to high conservation value forests

109 Matsubayashi et al. - Natural-licks use by orangutans in Sabah in the management plan of Deramakot. However, the use licks by mineral analysis and have used camera traps since of natural-licks by arboreal animals has not been studied 2003 to record mammals at the sites (Matsubayashi et al., much in Borneo. Therefore, it will be necessary for forest 2007a; 2007b). Eleven natural-licks have been identifi ed. management to study the habitats of arboreal animals Four natural-licks each with a different environment, with because these animals are negatively infl uenced by logging similar sizes, D1 (05°19'N 117°28'E), D2 (05°21'N 117°29'E), (Meijaard, 2005). D3 (05°21'N 117°31'E), and D4 (05°19'N 117°34'E), were selected for this study. The natural-licks were approximately To investigate natural-licks use by arboreal animals in 1 m2, and small amounts of water seeped from the soil and detail, we focused on the orangutan (Pongo pygmaeus), a rocks. The seepage water supply is stable all year round. representative arboreal species that is affected by intensity of logging (Felton et al., 2003; Knop et al., 2004; Caldecott Mineral analysis of the natural-licks. – A shallow stagnant & Miles, 2005; Ancrenaz et al., 2008; Husson et al., 2009). water pool was formed at each natural-lick. To determine More than 60 % of orangutans living in Sabah are distributed the background concentration of minerals, we collected 15 in production forests (Ancrenaz et al., 2005). In addition, samples from the natural-licks, and three samples from a pond the orangutan is a high-frequency visitor of the natural-licks that was not affected by natural-licks but was less than 10 m in Deramakot (Matsubayashi et al., 2007a). from them. During each sampling time, water samples were drawn through a 10 ml syringe from more than fi ve points In this study, we anticipated that, since the orangutan is at each water column. After thorough mixing, about 50 ml negatively affected by human activities, their use of natural- of each bulked sample was fi ltered through a syringe fi lter licks might be affected. Therefore, we selected four natural- with 0.2 µm pore size (Whatman, New Jersey, USA). The licks, each with a different environment, and investigated concentrations of dissolved calcium, magnesium, potassium, the preferences of the high-frequency visitors with camera- and sodium in the fi ltered water samples were determined trapping. We also anticipated that more fl anged males might on an atomic absorption spectrometer (GBC Scientific visit natural-licks because they generally spend more time on Equipment Pty Ltd, Victoria, Australia). All analyses were the forest fl oor than females and younger individuals (Cant, conducted at the Chemistry Section of the Forest Research 1987); in addition, we assumed that females with infants Centre, Sabah Forestry Department. might be rare because of their wariness of ground predators (Galdikas, 1988). Therefore, we divided the orangutan into Environmental characteristics of each natural-lick. – The three classes (fl anged male, female with infant, and others environment around each natural-lick was characterised such as unflanged male or female without infant) and as human infl uence, 1) distance from the main roads with investigated natural-licks use by orangutans in detail. heavy traffi c and, 2) distance from the villages, and natural infl uences, 3) distance from the main rivers, 4) altitude and, 5) above-ground biomass of forest around the natural-licks. MATERIALS AND METHODS We digitized geographical information maps (boundary of forest compartments, roads, rivers, and villages) and drew Study site. – This study was conducted from March 2008 a contour map using three-seconds-mesh data of (about 90 to March 2010 in the Deramakot Forest Reserve. The m) called SRTM-3 (Shuttle Radar Topography Mission-3). Deramakot Forest Reserve (05°14'–05°28'N 117°20'–38'E) The distributions of main roads and rivers were described is located on the upper Kinabatangan River in central on the basis of the map of the Sabah Forestry Department Sabah (Fig. 1). The reserve covers 55,083 ha including and the SPOT images. The distributions of four villages 4,000 ha of conservation area, and it ranges from 20 m to in and around the forest reserve were mapped on the basis 350 m above sea level. The climate is humid equatorial with a mean annual temperature of about 26°C. The mean annual precipitation is approximately 3,500 mm (Kleine & Heuveldop, 1993). Monthly rainfall is somewhat seasonal because the air circulation is infl uenced by the northeast monsoon from November to February and by the southwest monsoon from May to August. The Deramakot Forest Reserve is mainly composed of lowland mixed dipterocarp forests dominated by the family Dipterocarpaceae (Dipterocarpus spp., Parashorea spp., and Shorea spp.). Monitoring of the orangutan population has been conducted by aerial line census using a helicopter since 1999 by the Sabah Forestry Department.

The locations of natural-licks in the Deramakot Forest Reserve are better known than those in other production forests because the condition of each compartment was thoroughly surveyed before the Sabah Forest Department Fig. 1. Location maps of Deramakot Forest Reserve in Sabah, made a forest management plan. We have identiﬁ ed natural- Malaysian Borneo (D1 to D4: natural-licks).

110 THE RAFFLES BULLETIN OF ZOOLOGY 2011

of a ﬁ eld survey. Distances from the nearest objects to the natural-licks and altitude were calculated on the basis of the digitized geographical information maps and the contour map. The above-ground biomass of the forest was estimated by the canopy water-contents calculated by Landsat ETM band 4 (near-infrared 0.75–0.90 µm) and band 5 (shortwave IR 1.50–1.75 µm) rectifying from data of ground observation.

Three mesh size, 30, 150 and 210 m including natural-licks were selected for this calculation (210 m was the maximum available data because of the cloud inﬂ uence).

Camera-trap monitoring at the natural-licks. – Camera traps with an infrared triggering system (Capture, Cuddeback, CA, USA) were set up at a total of ﬁ ve camera stations, which

included four natural-licks (D1 to D4) and one small pond Altitude (m) No. of photos Camera days around 10 m near D1 as a control. A camera was tied by belt on the trunk 50 cm above the ground at 2 to3 m apart from a natural-lick. As animals tend to stay at natural-licks longer than at other places, such as on an animal trail, the time delay was set to 15 minutes to reduce ﬂ ash impact on animals and to extend battery life. Maintenance involved Above-ground replacement of the cameras every two months. Moreover, to biomass of forest conﬁ rm whether orangutans consume the water or the soil, video traps (NoFlash, Cuddeback, CA, USA) were set up with a 15-minute time delay, and each recording was 30 or 60 seconds long at D1 and D4.

For comparing the frequency of visitation among species, we counted the numbers of individuals photographed (O’Brien, et al. 2003). When there were many photographs of the same species within one hour, only one was counted. In orangutans,

we set three apparently distinguishable classes, fl anged male, Main road River Village female with infant, and others, such as unfl anged male or female without infant, and investigated the frequency of (m) Distance visitation of each class. To analyse the visiting proportion and pH the visitation time of each orangutan class, when the same individual was continuously photographed more than 1 hour, only the fi rst photo was counted. In this study, we follow the nomenclature of International Union for the Conservation of Nature and Natural Resources (IUCN; 2010).

Statistical analyses. – Comparison of the visitation frequency among the natural-licks was made with a Kruskal-Wallis test. Fisher’s exact test was used to compare the length of stay at natural-licks of the three orangutan classes. A comparison of the visitation frequency of orangutans between the morning (0400 to 1100 hrs) and afternoon (1200 to 1800 hrs) was made with a Mann-Whitney U test. Statistical signiﬁ cance for all tests was set at p<0.05 or p<0.01. Data are presented as the mean ± standard deviation. Ca Mg K Ca Mg Na RESULTS

Chemical analysis and environmental characteristics of the natural-licks. – Table 1 shows the result of chemical

analysis and environmental characteristics of the natural (µg/ml) Minerals licks. The results of the chemical analysis showed that the concentration of minerals of the four natural-licks were signiﬁ cantly higher than those of the control (p<0.05). D4 D1 (4) D2 (3) D3 (4) D4 (4) Control (3) 45.8±4.5 28.2±2.0 9.5±0.9 67.3±7.7 5.9±1.5 125.6±51.2 20.1±6.1 10.7±7.7 24.2±5.9 30.8±10.7 28.8±14.4 154.3±4.6 10.4±1.6 2.5±0.9 21.5±9.0 106.7±9.2 2622.7±1177.2 1.3±0.4 8.4 46.8±12.1 7.7 7.5 8.1 1,300 17.6±12.3 500 2,400 7.6 200 900 500 100 6,600 1300 6,200 4,400 700 388.4±2.7 10,100 247.6±112.5 900 323.8±43.3 6,600 322.3±2.1 217 185 144 388.4±2.7 212 656 670 1839 217 242 494 432 363 14 366 212 (No. of Samples) Table1. Concentrations of the minerals and environmental characteristics natural-licks. Locations

111 Matsubayashi et al. - Natural-licks use by orangutans in Sabah

Table 2. Camera trapped medium to large mammals at the natural-licks.

Order Family Common name (Scientifi c name) Eulipotyphla Erinaceidae Moonrat (Echinosorex gymnura) Primates Hominidae Bornean orangutan (Pongo pygmaeus) Cercopithecidae Pig-tailed macaque (Macaca nemestrina) Rodentia Hystricidae Malayan porcupine (Hystrix brachyura) Long-tailed porcupine (Trichys fasciculata) Carnivora Ursidae Sun bear (Helarctos malayanus) Mustelidae Yellow-throated marten (Martes fl avigula) Oriental small-clawed otter (Aonyx cinerea) Mephitidae Malay badger (Mydaus javanensis) Viverridae Malay civet (Viverra tangalunga) Otter-civet (Cynogale bennettii) Common palm civet (Paradoxurus hermaphroditus) Herpestidae Collared mongoose (Herpestes semitorquatus) Short-tailed mongoose (Herpestes brachyurus) Felidae Sunda clouded leopard (Neofelis diardi) Leopard cat (Prionailurus bengalensis) Proboscidea Elephantidae Asian elephant (Elephas maximus) Cetartiodactyla Suidae Bearded pig (Sus barbatus) Tragulidae Lesser mouse-deer (Tragulus kanchil) Greater mouse-deer (Tragulus napu) Cervidae Bornean yellow muntjac (Muntiacus atherodes) Sambar deer (Rusa unicolor) Bovidae Banteng (Bos javanicus) had the highest concentration of calcium and sodium among natural-lick. The visitation frequencies of three species were the natural-licks. Natural-licks closer to human activity, signifi cantly different at the four natural-licks (p<0.01 each). such as heavy traffi c and village were D3 (200 m from For orangutans, the highest frequency was recorded at D1. the road) and D4 (500 m from the road and 4,400 m from In contrast, sambar deer and bearded pigs had the highest the village), while those with less human activity were D1 frequency at D4. (1,300 m from the road and 6,600 m from the village) and D2 (2,400 m from the road and 6,200 m from the village). Above ground biomass around the natural-licks was not signifi cantly different (p>0.05), and each altitude was similar. D4 was the closest to the river which is recognisable by the SPOT image.

Mammal fauna at the natural-licks and visitation frequency of the top three species. – In total, 23 species of medium to large mammals were recorded at four targeted natural- licks (3,407 photos; 1,655 camera-days; 205.9 photos per 100 camera-days; Table 2). The sambar deer (2,468 photos; 149.1 photos per 100 camera-days), the orangutan (483 photos; 29.2 photos per 100 camera-days; Fig. 2), and the bearded pig (Sus barbatus; 274 photos; 16.6 photos per 100 camera-days), were the most commonly recorded species and represented 94.7% in terms of the frequency of appearance in all photographs. At the control site, only seven species were photographed (14 photos; 212 camera-days; 6.6 photos per 100 camera-days), and orangutans were not recorded.

In all the natural-licks, the top three species were the same; sambar deer, orangutans, and bearded pigs. Fig. 3 shows Fig. 2. Two pairs of females with infants drink the seepage water proportion of visitation frequency by category at each at a natural-lick.

112 THE RAFFLES BULLETIN OF ZOOLOGY 2011

NATURAL-LICKS USE BY ORANGUTANS staying more than 15 minutes was signifi cantly higher than that of females with infants and others (p<0.05). Orangutan behaviour at the natural-licks. – From video traps, 51 movies of orangutans were recorded at D1 (49 Visitation time of orangutans at the natural licks. – Fig. 6 movies per 73 days) and D4 (two movies per 22 days). The shows visitation time of each orangutan class to the natural- video trap results show that orangutans drank water directly licks. Females with infants and others tend to visit more in rather than obtaining it by eating soil at the natural-licks and the afternoon (1200 to 1800 hrs) than in the morning (0400 they sometimes removed stones at a site to obtain access to1100 hrs) (p=0.05 and p<0.05 each), while fl anged males to water. showed no such tendency (p=0.15). In fl anged males, the earliest visit was 0452 hrs; the latest, 1734 hrs; and no Visiting proportion of each orangutan class at the natural- peak visitation was established. In females with infants, licks. – Fig. 4 shows visiting proportion of each orangutan the earliest visit was 0719 hrs; the latest, 1734 hrs; and class: fl anged male, female with infant, and others. The the peak was at 1500 hrs. In others, the earliest visit was fl anged male accounted for 31%; female with infant, 17%; 0541 hrs; the latest, 1805 hrs; and the peaks were at 0600 and the others, 52%. Occasionally, all three classes were hrs and 1600 hrs. sometimes recorded within one day, and different individuals were recorded in the same frame or within short proximity, i.e., 15 minutes (Fig. 2). DISCUSSION

Duration of orangutan visits at the natural-licks. – Fig. 5 Differences in the use of natural-licks among the top three shows duration of the visits by each orangutan class. More species. – This study shows that orangutans were high- than 70% of orangutans stayed less than 15 minutes at the frequency visitors to natural-licks and among the top three natural-licks. 77.2% of females with infants stayed less than species of all natural-licks, which supports the results of a 15 minutes, and none of them stayed more than one hour. previous study (Matsubayashi, et al. 2007a). High-frequency In others, 82.9% stayed less than 15 minutes, and a stay in visitation of orangutans was also confi rmed in the Malua excess of one hour was recorded only once. On the other hand, 55.2% of fl anged males stayed less than 15 minutes, and 9.1% of them stayed more than one hour, and the longest stay was 2 hrs 27 minutes. The percentage of fl anged males

Fig. 5. Duration of the visit of each orangutan class to the natural- licks.

Fig. 3. Proportion of visitation frequency of top 3 species among the natural-licks.

Fig. 4. Visiting proportion of each orangutan class: ﬂ anged male, Fig. 6. Visitation time of each orangutan class to the natural- female with infant, and others. licks.

113 Matsubayashi et al. - Natural-licks use by orangutans in Sabah

Forest Reserve, which is a production forest located on until now. This may be because females with infants are opposite side of the Deramakot; the two are divided by the more vulnerable to ground predators (Galdikas, 1988) and Kinabatangan River (Matsubayashi et al. unpublished data). the risk of descending to the forest fl oor is too high in the Therefore, frequent use of natural-licks by orangutans is a darkness of the early morning. common behaviour in Sabah. Natural-licks as monitoring sites in production forests. The frequency of visits by orangutans differed among the – This study shows the importance of natural-licks for all natural-licks, and D1 had the highest frequency. D1 was orangutans regardless of their class. In addition, natural-licks relatively far from the main roads and did not show apparently are very useful as monitoring sites of orangutans, especially higher concentrations of minerals than other natural-licks of females with infants, who have a high degree of wariness. (Table 1). D1 was also farthest from the river which is More than 60% of orangutans living in Sabah occur in recognisable by a SPOT image, but there were some streams. production forests (Ancrenaz et al., 2005). Therefore, it is Therefore, the area has some other water sources in addition strongly recommended that other production forests protect to D1. On the other hand, D4 with the highest concentration their natural-licks and manage them as monitoring sites for of sodium and close to the main roads showed fewer visits orangutan habitat conservation. This study also shows that by orangutans, and the highest frequency was noted by the decisions of orangutans to visit natural-licks are affected sambar deer and bearded pigs (Fig. 3). The results indicate more by human activity than by the concentration of minerals that the close proximity of a main road leads to a decline at the site. Therefore, for more effi cient conservation of in orangutan visits; however, sambar deer and bearded pigs orangutans, it is necessary to manage nature-licks as protected are more likely to be infl uenced by sodium concentrations. area with buffer zones where human activity is restricted The preference for higher sodium concentrations by sambar (for example reducing number of logged tree or setting skid deer and bearded pigs supports the fi ndings of our previous trails far from the zone). study (Matsubayashi et al. 2007a). These results seem to be dependent on species characteristics such as a high mineral demand and/or degree of wariness. ACKNOWLEDGEMENTS Natural-licks use by orangutans. – The video traps show We express our gratitude to Mr. Rawinder-Ajon (Kampung that the animals drank water directly rather than obtaining Bundu Tuhan) for supporting our fi eldwork. We are grateful it by eating soil at the natural-licks. However, it is still diffi cult to distinguish if they consumed the soil in very small to Datuk S. Mannan, Mr. A. Ahmad, Mr. A. Bagoi, Mr. quantities because MacKinnon (1974) reported geophagy of I. Sunjoto, Mr. S. Intang, Mr. J. Torimo (Sabah Forestry orangutans in Sabah. Department), Dr. T. Kanamori, Dr. H. Samejima, Dr. K. Kitayama (Kyoto University) for their encouragements We had anticipated that the fl anged male would be more likely to our work. This study was supported by the Global to visit the natural-licks. However, fl anged males accounted Environmental Research Aid (F-071) of the Japanese Ministry for only 31% of the total visits, and females with infants, of Environment, and was also supported by a Grant-in-Aid who were assumed to have a higher degree of wariness, for Young Scientists B (No. 20710182) of the Ministry accounted for at least 17% of the visits. This result suggests of Education, Culture, Sports, Science and Technology that natural-licks are important for all orangutan classes as (MEXT). a mineral source. Furthermore, several individuals used natural-licks simultaneously. Evidences in this study include four individuals recorded in the same frame (two pairs of LITERATURE CITED females with infants in Fig. 2), and all orangutan classes were recorded on the same day. In addition, recently, a female Ancrenaz, M., O. Gimenez, L. Ambu, K. Ancrenaz, P. Andau, with an infant was recorded coupling with an adult male at B. Goossens, J. Payne, A. Sawang, A. Tuuga & I. Lackman- Ancrenaz, 2005. Aerial surveys give new estimates for D2. These results suggest that environs of the natural-licks orangutans in Sabah, Malaysia. Plos Biology, 3: 1–8. have secondary function as a communication site. Ancrenaz, M., A. Marshall, B. Goossens, C. van Schaik, J. Pongo pygmaeus Although the proportion of fl anged male visits was lower Sugardjito, M. Gumal & S. Wich, 2008. . In: IUCN 2010. IUCN Red List of Threatened Species. IUCN, than anticipated, fl anged males did tend to stay longer than Gland, Switzerland. Http://www.iucnredlist.org [accessed 22 the members of the other classes. This result suggests that the Apr.2010]. physiological requirement for minerals of the fl anged male Blake, J. G., J. Guerra, D. Mosquera, R. T. Bette, A. Loiselle is much higher and/or that he is at less risk while making & D. Romo, 2010. Use of mineral licks by white-bellied the visit than the members of the other classes. spider monkeys (Ateles belzebuth) and red howler monkeys (Alouatta seniculus) in Eastern Ecuador. International Journal Females with infants and others tended to visit the natural- of Primatology, 31: 471–483. licks in the afternoon rather than in the morning. This Bravo, A., K. E. Harms & L. H. Emmons, 2010. Puddles created suggests that they tended to visit after foraging, although by geophagous mammals are potential mineral sources for the reason remains unclear. Early morning visitation (before frugivorous bats (Stenodermatinae) in the Peruvian Amazon. 0719 hrs) of females with infants had not been recorded Journal of Tropical Ecology, 26: 173–184.

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Caldecott, J. & L. Miles, (eds.) 2005. World Atlas of Great Knop, E., P. I. Ward & S. A. Wich., 2004. A comparison of Apes and their Conservation. Prepared at the UNDP World orangutan density in a logged and unlogged forest on Sumatra. Conservation Monitoring Centre. University of California Biological Conservation, 120: 183–188. Press, Berkeley, USA. Lagan, P., S. Mannan & H. Matsubayashi., 2007. Sustainable use Cant, J. G. H., 1987. Positional behavior of female Bornean of tropical forest by reduced-impact logging in Deramakot orangutans (Pongo pygmaeus). American Journal of Forest Reserve, Sabah, Malaysia. Ecological Research, 22: Primatology, 12: 71–90. 414–421. Felton, A. M., L. M. Engstöm, A. Felton, A., & C. D. Knott, MacKinnon, J., 1974. The behavior and ecology of wild orangutans 2003. Orangutan population density, forest structure and fruit (Pongo pygmaeus) Animal Behavior, 22: 3–74. availability in hand-logged and unlogged peat swamp forests Matsubayashi, H., P. Lagan, N. Majalap, J. Tangah, J. R. A. Sukor in west Kalimantan, Indonesia. Biological Conservation, 114: & K. Kitayama, 2007a. Importance of natural licks for the 91–101. mammals in Bornean inland tropical rain forests. Ecological Forest Stewardship Council, 1996. FSC international standard, FSC Research, 22: 742–748. principles and criteria for forest stewardship. Forest Stewardship Matsubayashi, H., P. Lagan, J. R. A. Sukor & K. Kitayama, Council A. C., Bonn. 2007b. Seasonal and daily use of natural licks by sambar deer Galdikas, B. M. F., 1988. Orang utan diets, range and activity (Cervus unicolor) in a Bornean tropical rain forest. Tropics, at Tanjung Putting, Central Borneo. International Journal of 17: 81–86. Primatology, 9: 1–31. McNaughton, S. J., 1988. Mineral nutrition and spatial concentrations Hardiono, M. & J. R. Alfred, 2005. Borneo: Treasure Island at of African ungulates. Nature, 334: 343–345. Risk. WWF Germany Meijaard, E., D. Sheil, R. Nasi, D. Augeri, B. Rosenbaum, D. Husson, S. J., S. A. Wich, A. J. Marshall, R. D. Dennis, M. Iskandar, T. Setyawati, M. Lammertink, I. Rachmatika, A. Ancrenaz, R. Brassey, M. Gumal, A. J. Hearn, E. Meijaard, Wong, T. Soehartono, S. Stanley & T. O’Brien, 2005. Life after T. Simorangkir & I. Singleton, 2009. Orangutan distribution, logging. Center for International Forestry Research. density, abundance and impacts of disturbance. In: Wich, Montenegro, O. L. 2004., Natural licks as keystone resources for S.A., S. S. U. Atmoko, T. M. Setia & C. P. van Schaik. (eds), wildlife and people in Amazonia. PhD dissertation, University Orangutans. Pp. 77–96. of Florida, Gainesville, Florida. IUCN, 2010. 2010 IUCN Red List of Threatened species. www. O’Brien, T. G., M. F. Kinnaird & H. T. Wibisono, 2003. Crouching iucnredlist.org. tigers, hidden prey: Sumatran tiger and prey populations in a Kleine, M. & J. Heuveldop, 1993. A management planning concept tropical forest landscape. Animal Conservation, 6: 131–139. for sustained yield of tropical forests in Sabah, Malaysia. Forest Sabah Forestry Department, 2009. Annual report 2008. Ecology and Management, 61: 277–297. Sandakan. Kreulen, D. A., 1985. Lick use by large herbivores: a review of Weir, J. S., 1972. Spatial distribution of elephants in an African beneﬁ ts and banes of soil consumption. Mammal Review, 15: National Park in relation to environmental sodium. Oikos, 107–123. 23: 1–13.

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