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In Vitro Propagation of Two Perkinsus spp. Parasites from Japanese Manila Clams Venerupis philippinarum and Description of Perkinsus honshuensis n. sp.
Author(s) -
DUNGAN CHRISTOPHER F.,
REECE KIMBERLY S.
Publication year - 2006
Publication title -
journal of eukaryotic microbiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.067
H-Index - 77
eISSN - 1550-7408
pISSN - 1066-5234
DOI - 10.1111/j.1550-7408.2006.00120.x
Subject(s) - biology , parasite hosting , internal transcribed spacer , ribosomal rna , protozoa , bay , bivalvia , microbiology and biotechnology , zoology , mollusca , gene , genetics , world wide web , computer science , civil engineering , engineering
.Perkinsus species are destructive parasites of commercial Manila clams, Venerupis philippinarum , in Japan, Korea, and Spain. However, in vitro parasite cultures from this important host clam are not available. Tissues of Manila clams collected during April 2002 in Gokasho Bay, Japan harbored Perkinsus sp. parasites at a 97% prevalence (28/29) of moderate‐ and high‐intensity infections. Perkinsus sp. cells in tissue samples were enlarged in alternative Ray's fluid thioglycollate medium, before propagation in DME:Ham's F‐12 Perkinsus sp. culture medium. Enlarged parasite hypnospores zoosporulated at high frequencies to release motile zoospores, which gave rise to continuous schizogonic cell lines that also zoosporulated continuously at low frequencies. Four Perkinsus sp. in vitro isolates comprising two distinct morphotypes were cryopreserved, cloned, and archived for public distribution. For three isolates of one morphotype, nucleotide sequences of the ribosomal DNA internal transcribed spacer region, of the large subunit rRNA gene, and of actin genes, were consistent with those reported for P. olseni . Similar sequences from one morphologically unique isolate differed from those of all described Perkinsus species. These results show that at least two Perkinsus spp. infect Japanese Manila clams, and that one represents a new species, Perkinsus honshuensis n. sp.