The family Mesocoeliidae Dollfus, 1929 (Digenea) is composed by parasites of the digestive organs of a number of amphibian and reptile species including R. marina. This family has two genera: Mesocoelium Odhner, 1910 and Pintneria Poche, 1907(Pojmanska 2008. This paper describes the morphological and molecular characteristics of a parasite of the genus Mesocoelium, which appear to be unique and have thus led us to propose a new species. The digestive tube of each toad was isolated in phosphate buffered solution pH 7.4 and analyzed for trematodes. For light microscopy analysis, 10 parasites were fixed with 2% glacial acetic acid, 3% formaldehyde, 95% ethanol 70º, dehydrated in increasing ethanol concentrations, stained in alcoholic chloridric carmine, bleached in methyl salicylate and mounted in Entellan TM . Illustrations were drawn with camera lucida attached to an Olympus BX41 microscope. Measurements are presented in millimetres, unless otherwise indicated, as means and standard deviations, with the range of values in parentheses.
MATERIALS AND METHODS
FourteenFor scanning electron microscopy (SEM), 20 specimens were fixed with 2.5% glutaraldehyde, post-fixed in 1% OsO 4 0.8% K 2 Fe (CN) 6 , gold sputtered and observed in a Jeol JSM-5310 scanning electron microscope.Molecular characterization of three parasites was based on complete SSU rDNA gene, which after DNA extraction through the ChargeSwitch gDNA Mini Tissue Kit (Invitrogen Life Technologies), was amplified in two overlapping fragments using the primers 18S-E/18S-A27 and 18S-8/cestode-6 according to Olson and Caira (1999). The polymerase chain reactions (PCRs) were carried out in 25 µL final volume, containing 5-10 ng of DNA, 50 mM KCl, 2 mM MgCl 2 , 10 mM Tris-HCl, 50 µM of each DNTP, 0.5 µM of each oligonucleotide and one unit of Taq DNA polymerase (Invitrogen). Reactions using the 18S-E/18S-A27 primers were denatured for 5 min at 95ºC followed by 35 cycles, 1 min thermal denaturing cycles at 94ºC, 1 min at 68ºC for annealing and 1 min extension at 72ºC, with a final 7 min extension at 72ºC. Reactions using the 18S-8/cestode-6 primers were amplified following the same procedure, except for annealing temperature of 64ºC. Amplicons were electrophoresed in a 1% agarose gel, excised and puri-