Binding specificities of mono- and polyclonal antibodies to the protozoan oyster pathogen Perkinsus mannus

Abstract

Binding specificities of mono-and polyclonal antibodies to the protozoan oyster pathogenPerkinsus marinusChristopher F. ~u n g a n ' , Bob S. ~o b e r s o n~ ABSTRACT: In an effort to develop direct detection methods for Perkinsus marinus cells in host tissue and environmental samples, the production of both mono-and polyclonal antibodies specific for this apicomplexan oyster pathogen was attempted. A particulate hypnospore immunogen was isolated from infected oyster hemolymph following incubation in Ray's fluid thioglycollate medium. Polyclonal antisera produced in both mice and rabbits following immunization with this preparation exhibited high antibody titers for pathogen cell epitopes, but not for host oyster tissue epitopes. At least some antibodies to hypnospore antigenic determinants recognized common epitopes on trophozoite cells in infected oyster tissues, as well as on zoospores and other proliferative cells produced by incubation of infected oyster hemolymph in sterile seawater Polyclonal antibodies also recognized a diffusable, noncellular substance present in host oyster tissues in areas surrounding focal lesions. Rabbit polyclonal antibodies did not recognize 2 Dermocystldium species infecting salmonid fishes or Haplosporidlum nelsoni infecting eastern oysters Crassostrea virginica. These antibodies did recognize many, but not all. Perkinsus species infecting selected molluscan hosts worldwide. Monoclonal antibodies produced to date apparently recognize epitopes unique to the hypnospore immunogen.

Perkinsus Protozoan Infecton in Short-necked Clam Tapes (=Ruditapes) philippinarum in Japan.

“…marinus (Choi et al, 1992;Dungan and Roberson, 1993). The method, once it is established, is specific and rapid.…”

“…For the diagnosis of Perkinsus species, histological method , culture method using RFTM (Fisher and Oliver, 1996), immunological method using Perkinsus-specific antibodies (Choi et al, 1992;Dungan and Roberson, 1993), and molecular biological method have been proposed. Histological method has been commonly used.…”

Perkinsus marinus (Apicomplexa) as a potential source of oyster Crassostrea virginica mortality in coastal lagoons of Tabasco, Mexico

“…Antibody was generously supplied by C. F. Dungan, Cooperative Oxford Laboratory, Oxford, MD, USA. Production and specificity of the antibody is described by Dungan & Roberson (1993). Biocell goat anti-rabbit IgG gold probe and Blocell light microscopy silver enhancement reagents (Goldmark Biologicals, Phillipsburg, NJ, USA) were utilized to visualize specific binding of the primary antibody.…”

Comparison of diagnostic techniques to detect the clam pathogen Perkinsus olseni

“…Screening methods recommended by the OIE are histopathology and incubation on Ray's fluid thioglycollate medium (RFTM) (OIE 2006). In addition, other methods have been developed, especially for the oyster parasite P. marinus, including mol-ecular methods based on PCR (Robledo et al 1998, Elandalloussi et al 2004, Russell et al 2004) and immunological detection (Choi et al 1991, Dungan & Roberson 1993. Internal transcribed spacers (ITS-1 and ITS-2) are non-coding regions of the RNA gene cluster that are less conserved than the ribosomal RNA coding genes and are suitable to distinguish among proximal species.…”