Description of Perkinsus andrewsi n. sp. Isolated from the Baltic Clam (Macoma balthica) by Characterization of the Ribosomal RNA Locus, and Development of a Species‐Specific PCR‐Based Diagnostic Assay

Journal of Eukaryotic Microbiology - Tập 48 Số 1 - Trang 52-61 - 2001
Cathleen A. Coss1, José A. Fernández Robledo1, Gregory M. Ruiz2, Gerardo R. Vasta1
1Center of Marine Biotechnology, University of Maryland Biotechnology Institute, Baltimore, Maryland 21202, USA.
2Smithsonian Environmental Research Center; Edgewater Maryland 21037 USA

Tóm tắt

ABSTRACT. A Perkinsus species was isolated from the baltic clam Macoma balthica and an in vitro culture established under conditions described for P. marinus. As reported previously, morphological features remarkable enough to clearly indicate that this isolate is a distinct Perkinsus species were lacking. In this study, regions of the rRNA locus (NTS, 18S. ITS1, 5.8S. and 1TS2) of this isolate were cloned, sequenccd, and compared by alignment with those available for other Perkinsus species and isolates. Sequence data from the rRNA locus and species‐specific PCR assays indicated not only that Perkinsus sp. from M. balthica was not P. marinus, but it was different from P. atlanlicus and P. olseni. The degree of difference was comparable to or greater than differences between accepted Perkinsus species. In particular, NTS sequence and length were dramatically different from that of P. marinus and P. atlanlicus. Therefore, we formally propose to designate the Perkinsus sp. from M. balthica as a separate species, P. andrewsi n. sp. Primers based on P. andrewsi NTS sequence were used to develop a PCR‐based diagnostic assay that was validated for species‐specificity and sensitivity. PCR‐based assays specific for cither P. andrewsi or P. marinus were used to test for their presence in bivalve species sympatric to M. balthica. Although isolated from M. balthica, P. andrewsi was also detected in the oyster Crassostrea virginica and clams Macoma mitchelli and Mercenaria mercenaria, and could coexist with P. marinus in all four bivalve species tested.

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Journal of Eukaryotic Microbiology

Tập 48 Số 1

52-61

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