Isolation and purification of developmental stages ...

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Title Isolation and purification of developmental stages of Perkinsus karlssoni (Apicomplexa: Perkinsea), a parasite affecting bay scallops Argopecten irradians
Author(s) S. K. Whyte, Richard J. Cawthorn, R. J. MacMillan, B. Despres
Journal Diseases of Aquatic Organisms
Date 1993
Volume 15
Issue 3
Start page 199
End page 205
Abstract Isolation and purification of the trophozoite and schizont stages of the protozoan Perkinsus karlssoni , a parasite affecting Argopecten irradians , was achieved by macerating infected host tissue. Prior culture of infected tissue in fluid thioglycollate medium was unnecessary either for detecting Perkinsus karlssoni in the host tissue, or for collecting developmental stages of the parasite for further studies. The maceration technique is rapid and simple to perform, yielding numerous viable trophozoites and schizonts which are relatively free of contaminating host material. Trophozoites and schizonts demonstrated good retention of structural integrity following isolation from host tissue. These stages were further purified by separation from host material using a Percoll gradient and were harvested at the 50 to 60% interface. Development of the successive parasite stages was observed to proceed in vitro when trophozoites and schizonts were transferred to petri dishes containing sterile seawater fortified with antibiotics and incubated at 26 degree C. Light and electron microscopy were used to document the development of zoosporangia in vitro. The terminal stage of development was the release of biflagellate zoospores after 3 d at 26 degree C. All stages of the parasite could be used for further studies including development of specific diagnostic aids.
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