The alveolate protozoan parasite Perkinsus olseni infects a range of marine bivalves inhabiting tidal flats and shallow subtidal zones, causing considerable damage to shellfish industries. Infection by P. olseni is typically assessed using Ray’s fluid thioglycollate medium (RFTM) assay, with gill tissue often employed as a diagnostic proxy for whole-body infection. However, the diagnostic reliability of gill-based assays across diverse ecological settings—particularly under low-infection conditions—remains uncertain. In this study, we investigated tissue-specific distribution and the diagnostic performance of the RFTM assay in detecting P. olseni in Manila clams (Ruditapes philippinarum) collected from 6 tidal flats along Korea’s west and south coast. The assay was applied to 6 different tissues, including gills and visceral mass. Infection prevalence reached 100% at most sites, except at Padori (90%). Whole-body infection intensity ranged from 0.1×104 to 3.7×106 cells per gram of tissue. The visceral mass consistently harbored the largest proportion of parasites (27.8%–49.0%), followed by the mantle (17.4%–30.6%) and gills (19.4%–25.2%). Gill infection levels correlated strongly with whole-body infection intensity (r²=0.6–0.95), supporting their diagnostic value in high-infection areas. However, at Padori—where infection levels were lowest—the efficacy of the gill assay dropped to 56%, resulting in a 44% false negative rate. These results underscore the limitations of relying solely on gill tissue in low-infection environments and highlight the need for a context-dependent diagnostic approach. A dual-tissue strategy incorporating both gill and whole-body samples is recommended to improve diagnostic accuracy in P. olseni surveillance of Manila clam populations.
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Life cycle stages, including daughter sporocysts, cercariae, and metacercariae, of Parvatrema duboisi (Dollfus, 1923) Bartoli, 1974 (Digenea: Gymnophallidae) have been found in the Manila clam Ruditapes philippinarum from Aphaedo (Island), Shinan-gun, Jeollanam-do, Korea. The daughter sporocysts were elongated sac-like and 307-570 (av. 395) μm long and 101-213 (av. 157) μm wide. Most of the daughter sporocysts contained 15-20 furcocercous cercariae each. The cercariae measured 112-146 (av. 134) μm in total length and 35-46 (av. 40) μm in width, with 69-92 (av. 85) μm long body and 39-54 (av. 49) μm long tail. The metacercariae were 210-250 (av. 231) μm in length and 170-195 (av. 185) μm in width, and characterized by having a large oral sucker, genital pore some distance anterior to the ventral sucker, no ventral pit, and 1 compact or slightly lobed vitellarium, strongly suggesting P. duboisi. The metacercariae were experimentally infected to ICR mice, and adults were recovered at day 7 post-infection. The adult flukes were morphologically similar to the metacercariae except in the presence of up to 20 eggs in the uterus. The daughter sporocysts and metacercariae were molecularly (ITS1-5.8S rDNA-ITS2) analyzed to confirm the species, and the results showed 99.8-99.9% identity with P. duboisi reported from Kyushu, Japan and Gochang, Korea. These results confirmed the presence of various life cycle stages of P. duboisi in the Manila clam, R. philippinarum, playing the role of the first as well as the second intermediate host, on Aphae-do (Island), Shinan-gun, Korea.
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