Publication Date:
2022-05-25
Description:
Author Posting. © National Shellfisheries Association, 2005. This article is posted here by permission of National Shellfisheries Association for personal use, not for redistribution. The definitive version was published in Journal of Shellfish Research 24 (2005): 719-731, doi:10.2983/0730-8000(2005)24[719:NSRRGC]2.0.CO;2.
Description:
Western Long Island Sound (LIS) lobsters collected by trawl surveys, lobstermen and coastal residents during 2000 to 2002 were identified histologically as infected with a parasome-containing amoeba. Primers to conserved SSU rRNA sequences of parasome-containing amoebae and their nonparasome-containing relatives were used to amplify overlapping SSU rRNA fragments of the presumptive parasite from gill, antenna, antennal gland and ventral nerve cord of infected lobsters. The consensus sequence constructed from these fragments had 98% or greater nucleotide sequence identity with SSU rRNA gene sequences of strains of Neoparamoeba pemaquidensis and associated with high confidence in distance- and parsimony-based phylogenetic analyses with strains of Neoparamoeba pemaquidensis and not members of the family Paramoebidae, e.g., Paramoeba eilhardi. Primers designed to SSU rRNA sequences of the lobster amoeba and other paramoebid/vexilliferid amoebae were used in a nested polymerase chain reaction (PCR) protocol to test DNA extracted from formalin-fixed paraffin-embedded tissues of lobsters collected during the 1999 die-off, when this amoeba initially was identified by light and electron microscopy and reported to be a paramoeba of the genera Paramoeba or Neoparamoeba (Mullen et al. 2004). All sequences amplified from 1999 lobsters, with the exception of one, had 98% to 99% identity to each other, and the 1999 PCR product consensus had 98% identity to Neoparamoeba pemaquidensis strains CCAP 1560/4 (AF371969.1) and 1560/5 (AF371970.1). Molecular characterization of the amoeba from western LIS lobsters by direct amplification circumvents a collective inability to culture the organism in vitro, provides insight into the molecular epidemiology of neoparamoebiasis in American lobster, and allows for PCR-based detection of infected lobsters for future research and diagnostics.
Description:
Funding for this work was provided by the
Connecticut Department of Environmental Protection under Long
Island Sound Research Fund Grant No. CWF 333-R to S. Frasca;
and by the Connecticut Sea Grant College Program, Grants No.
LR/LR-4 to R. Gast and No. LR/LR-5 to P. Gillevet and C.
O’Kelly, through the US Department of Commerce, National
Oceanic and Atmospheric Administration (NOAA), Award
NA16RG1364.
Keywords:
Homarus americanus
;
Lobster
;
Molecular phylogeny
;
Neoparamoeba pemaquidensis
;
Paramoebiasis
;
PCR
;
Small-subunit rRNA
Repository Name:
Woods Hole Open Access Server
Type:
Article
Format:
application/pdf
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