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      Invasive crayfish and crayfish plague on the move: first detection of the plague agent Aphanomyces astaci in the Romanian Danube

      , , , , , ,
      Diseases of Aquatic Organisms
      Inter-Research Science Center

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          Abstract

          Native European crayfish, such as Astacus leptodactylus, are threatened, among other factors, by the crayfish plague agent Aphanomyces astaci, dispersed by invasive North American crayfish. Two of these invaders, Pacifastacus leniusculus and Orconectes limosus, have extended their distribution in the River Danube catchment; the latter was detected for the first time in Romania in 2008. We monitored, at monthly intervals for over 2 yr, occurrence of native A. leptodactylus and invasive O. limosus at 6 sites on the Romanian Danube and checked for the invasive species in 4 of its tributaries. Between January 2009 and March 2011, the relative abundances of O. limosus steadily increased with time, while the native A. leptodactylus dramatically decreased in abundance. O. limosus expanded downstream at a rate of ca. 15 km yr-1; in August 2011, it was already present in the upper 105 km of the Romanian Danube. An agent-specific real-time PCR analyses demonstrated the presence of A. astaci DNA in at least 32% of the analysed invasive (n = 71) and 41% of the native (n = 49) crayfish coexisting in the Danube. Furthermore, A. astaci was also detected in A. leptodactylus captured about 70 km downstream of the O. limosus invasion front (at the time of sampling). Assuming a steady rate of expansion, O. limosus may invade the sensitive Danube delta area in the mid-2060s, even without long-distance dispersal. The crayfish plague agent, however, may reach the delta substantially earlier, through dispersal downstream among populations of native crayfish.

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          Most cited references33

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          A review of the ever increasing threat to European crayfish from non-indigenous crayfish species

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            Geographical spread of bacterial and fungal diseases of crustaceans.

            J Alderman (1996)
            The author discusses the way in which bacterial and fungal diseases of crustaceans can be and have been transmitted via commercial movements of stocks. Two classic examples of such diseases are presented, namely gaffkaemia (a bacterial disease of marine lobsters) and crayfish plague (a fungal disease of freshwater crayfish). In both cases, there is ample evidence that much of the disease spread can be linked to the commercial movement of infected animals caught from infected wild stocks and transported over large distances for wholesale and retail. In the case of crayfish plague, much of the relentless spread of this disease through Europe for nearly 140 years can also be liked to the movement of contaminated fishing equipment without disinfection.
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              A quantitative TaqMan MGB real-time polymerase chain reaction based assay for detection of the causative agent of crayfish plague Aphanomyces astaci.

              Here we present the development and first validation of a TaqMan minor groove binder (MGB) real-time polymerase chain reaction (RT-PCR) method for quantitative and highly specific detection of Aphanomyces astaci, the causative agent of crayfish plague. The assay specificity was experimentally assessed by testing against DNA representative of closely related oomycetes, and theoretically assessed by additional sequence similarity analyses comparing the primers and probe sequences to available sequences in EMBL/GenBank. The target of the assay is a 59 bp unique sequence motif of A. astaci found in the internal transcribed spacer 1 of the nuclear ribosomal gene cluster. A standard curve for quantification was established by setting up a four-fold dilution series with genomic A. astaci DNA. The absolute limit of detection (LOD(abs)), defined as the lowest concentration yielding a false negative probability<5% was found to be approximately 5 PCR forming units (PFU
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                Author and article information

                Journal
                Diseases of Aquatic Organisms
                Dis. Aquat. Org.
                Inter-Research Science Center
                0177-5103
                1616-1580
                February 17 2012
                February 17 2012
                : 98
                : 1
                : 85-94
                Article
                10.3354/dao02432
                22422132
                12f06299-bdc7-4ebf-94c3-3bc0781a144a
                © 2012
                History

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