75 Flukes of the Family Microscaphidiidae Looss, 1900 (Trematoda: Digenea) from the Green Sea Turtle, Chelonia mydas L. (Reptilia: Chelonia) in the Philippines with Description of Three New Species Salcedo L. Eduardo1·, DVM, MS, PhD and Jose L. Diaz2, DVM 'Department of Veterinary Paraclinical Sciences, College of VeterinaryMedicine, University of the Philippines Los Banos, Laguna 4031, Philippines: and 'Protected Areas and Wildlife Bureau, Department of Environment and Natural Resources, Quezon City, Philippines ABSTRACT A total of 15 green sea turtles, Chelonia mydas L. were examined at various times for parasites from the coastal areas of Bohol and Zamboanga, Philippines. Among others, trematodes belonging to the family Microscaphidiidae Looss, 1900 were recovered following the concentration sedimentation decantation technique. Specimens were stained in borax carmine, cleared in oil of cloves and mounted in Canada balsam for microscopic examination. Twelve species including 3 new species of trematodes representing 6 genera of the family Microscaphidiidae Looss, 1900 were identified. These are: Microscaphidium reticulare (van Beneden, 1854) Looss, 190L Microscaphidium aberans Looss, 1902, Microscaphidium warui Blair, 1986; Deuterobaris proteus (Brandes, 1891) Looss, 1900; Angiodictyum parallelum(Looss, 1901) Looss, 1902; Angiodictyum glossoides Blair, 1986; Angiodictyum longumBlair, 1986: Angiodictyum undulatumn. sp.; Angiodictyum dejesusi n. sp.; Polyangium manueli n. sp.; Octagium sagitta (Looss, 1899) Looss, 1902 and Polygorgyra choladus Blair, 1986. Angiodictyum undulatum, A. dejesusi and Polyangium manueli were described and illustrated as new species. A. undulatum differs from all species of the genus by its lateral margin thrown into folds giving an undulating appearance of the worm, vitellaria present only in the median area and their absence in lateral fields. A. dejesusi is differentiated from the closely related species," A. Jongum by its vitellaria extending anteriorly to level of anterior testis and not beyond it, the pharyngeal-position of the genital pore and the relatively short esophagus. P manueli differs from P linguatulum, the only valid species of the genus, by its ovoid shape, the limited extent of vitellaria which is behind the testes and the more anterior location of the genital pore just behind the pharynx. This is the first report of these trematodes in the Philippines constituting a new locality record for these species. Keys to separate the genera of the family Microscaphidiidae and species of the genera Microscaphidium and Angiodictyum occurring in marine turtles in the Philippines are provided. Keywords: Angiodictyum dejesusi, Angiodictyum undulatum, Chelonia mydas, Microscaphidiidae, Philippines, Polyangium manueli ------------------ Philipp. J. Vet.Med., 45: 75·85, 2008 INTRODUCTION are: Polystomoides cyclemydis, a monogenetic trematodes from Cyclemys dentata (serrated The chelonian (turtle) fa una of the freshwater turtle) in Palawan (Fischthal & Philippines consist of 8 species, 3 freshwater Kuntz, 1964) and Spironoura duyagi, a and 5 marine (Alcala, 1986; Wildlife nematode from Cuora amboinensis (Malayan Conservation Society of the Philippines, 1997). fresh water turtle) and Cyclemys dentata in As far as the authors are aware, only two Novaliches, Luzon and Palawan (Tubangui & parasites have been reported so far from only Villaamil, 1933; Schmidt & Kuntz, 1972). As two turtles species in the Philippines. These far as can be ascertained no report on the *FOR CORRESPONDENCE parasites of the other species of turtles 76 SL EDUARDO AND JL DIAZ including the sea or marine turtles. The true Physiological salt solution was added and sea turtles, Chelonia mydas (green sea turtle) shaken to relax any worm present. This was and Eretmochelys imbricata (hawksbill sea then processed following the concentration turtle) had a wide distribution locally but sedimentation decantation technique, worldwide these are listed by the International repeatedly done until the supernatant fluid is Union for Conservation of Nature (IUCN) as clear. The resulting sediment was stored in 10% globally endangered (Wildlife Conservation formalin solution and brought to the laboratory Society of the: Philippines, 1997; DENR-UNEP, for further examination. The preserved 1997). Although many species of parasites sediment was examined for parasite in the particularly trematodes from marine turtles in laboratory using a Zeiss Stemi DV4 other parts of the world have been recorded stereomicroscope. Parasites were collected and (Gupta, 196L Rodriguez, 196L Chat­ post-fixed in formalin-alcohol-acetic acid topadhyaya, 1972; Groschaft et al, 1977; Blair, solution (FAA) for at least 48 hours and stored 1983, 1986, 1987; Dyer et al., 1991), none have in 80% ethyl alcohol until further study. been reported from these hosts in the Specimens were then stained in aceta alum Philippines. We have had opportunities to carmine overnight, destained in 1% examine at various times marine turtles for hydrochloric acid solution until the desired color parasites from the coastal areas of Bohol and (pale pink) is reached, neutralized in 1% Zamboanga, Philippines. This paper dealing solution of sodium hydroxide to stop further with the species of the family Microscaphidiidae destaining. Specimens were then gradually from the green sea turtle has been written to dehydrated in increasing grades of ethyl record some of our findings contributing to our alcohol, cleared in oil of cloves and mounted knowledge of invertebrate biodiversity in the whole in Canada balsam. In addition, country. Such information on the health aspects representative specimens of thick and muscular is also vital in conservation programs when worms were stained and cleared as above but these animals are raised initially in captivity. were hand sectioned in the sagittal, cross and Furthermore, chelonians (turtles) belong to an longitudinal planes following the procedure of ancient group dating back 200 million years to Eduardo (1982) for amphistomes. These were the Jurassic era. Consequently, their parasites then arranged accordingly in the proper are of great importance from an evolutionary sequence on a glass slide and mounted in perspective due to their presumed ancient Canada balsam. This technique has enabled the origin (Jakes et al.,2001). The other trematode observation of the internal organs of thick families including those recovered from the specimens which otherwise are not readily seen hawksbill sea turtle (Eretmochelys imbricata) in whole mounts. will form the subject of a separate paper. Measurements were made with the aid of a calibrated eyepiece micrometer. Photo· MATERIALS AND METHODS micrographs were taken using an Olympus BX51 research microscope with digital camera A total of 15 green sea turtles (Chelonia attachment. Drawings were made with the aid mydas L.) were examined at various times for of an Olympus drawing apparatus attached to parasites, twelve from Zamboanga City and an Olympus CX31 research microscope. three from Jagna, Bohol. In almost all cases, Additional microscaphidiid materials only the digestive tract was made available for deposited at the Swedish Museum of Natural examination. The digestive tract was tied by History were examined for comparative segment with a thread to separate one section purposes by the senior author during his visit from the other, and to avoid free flowof contents there as follows: Microscaphidium reticulare from one segment to another. The mucosa of (NHR Nos. 1908, 1914, 2649, 2650); M. each digestive section was scraped with a fine adherens (NHR Nos. 1889, 1905, 2648, 2649); knife to dislodge any attached parasite and the Angiodictyum parallelum (NHR Nos. 1890, contents placed in a wide-mouth bottle. 1895, 1908); A. longum (NHR Nos. 867, 2934, FLUKES IN GREEN SEA TURTLES 77 2961); Polyangium linguatulum (NHR Nos. recently have been sufficiently described by 850, 891, 1893, 1932, 2651). Many of these Blair (1986, 1987), these are no longer described specimens were reexamined and identified by here but photographs of them are provided Blair (1986). based on the Philippine materials. The three Taxonomic allocation of the trematodes at new species however are described and the family and generic levels follows that of illustrated by photographs and drawings below, Jones et al. (2005). and other taxonomic data are also provided. Voucher and type (holotype and paratype) specimens were deposited in the Parasite Description of new species: Collection of the College of Veterinary Medicine, UPLB under the accession number indicated Angiodictywn undulatum n. sp. for each species. (Figs. 8, 13, 14) RESULTS (Based on one whole and several fragmented specimens, the latter however were not included Twelve species representing six genera of in the measurements) the family Microscaphidiidae were identified Body is elongate, measures 5.820 mm long in the present collections. Of these, nine are and 2.250 mm wide, linguiform with sides known species but are reported for the firsttime almost parallel, rounded on both ends. Marginal in the Philippines constituting new locality bodies are absent. Lateral margins are thrown record for these species, while three are species into folds giving a characteristic undulating new to science. The known species are listed in appearance of the worm. Edges of folds are the table showing their organ location, marked by fine muscle fibers. accession number of voucher specimens and Pharynx is barrel·shaped,