Evidence of Centrocestus formosanus (Nishigori, 1924) in Zebrafish (Danio rerio)

Diagnosis of Centrocestus formosanus Infection in Zebrafish (Danio rerio) in Italy: A Window to a New Globalization-Derived Invasive Microorganism

“…The present study reports on the occurrence of C. formosanus metacercariae in the gills of zebrafish previously intended for research. To the authors' knowledge, this is the second report of C. formosanus infections in zebrafish imported in Italy [32]. Since the scarce and fragmentary data in the European literature [19,32,40] are probably due to underestimated and underdiagnosed infections, we propose to increase the awareness and ameliorate the diagnostic investigations, to shed light on this zoonosis, using a morphological and molecular approach.…”

“…To the authors' knowledge, this is the second report of C. formosanus infections in zebrafish imported in Italy [32]. Since the scarce and fragmentary data in the European literature [19,32,40] are probably due to underestimated and underdiagnosed infections, we propose to increase the awareness and ameliorate the diagnostic investigations, to shed light on this zoonosis, using a morphological and molecular approach. In particular, we propose for the first time a fast and specific diagnostic method based on a species-specific primer pair to detect the presence of this new invasive species in Italy.…”

“…Extracted DNA samples were stored at −80 • C until the molecular analyses described here. The detection of C. formosanus DNA was performed by PCR targeting the ribosomal internal transcribed sequence 2 (ITS2), using the primer pair 3S (5 -GGTACCGGTGGATCACTCGGCTCGTG-3 ) and BD2 (5 -TATGCTTAAATTCAGCGGGT-3 ), previously described [10,32]. As these primers are not specific for C. formosanus, we designed the species-specific reverse primer ITS2_Centr_R (5 -CGTGCAATGTTTGCATCGGA-3 ) and used it together with the forward primer ITS2_Centr_F (5 -ATGAAGAGCGCAGCCAACT-3 ) to amplify a 393 bp fragment of the ITS2 region of C. formosanus.…”

“…Among the numerous freshwater fish species affected by C. formosanus [23,25], zebrafish (Danio rerio) have been considered susceptible to infection, but only three reports have been described to date [10,21,32].…”

“…Danio rerio is a freshwater fish native to Asia, although it is widely distributed worldwide, probably due both to aquarists' predilection and researchers' interest as an animal model [32][33][34][35][36]. On the one hand, D. rerio is one of the top 30 species in the ornamental fish industry, whose on-going growth (approximately 120 countries and 1.5 billion ornamental fish per year) has led to difficulties in supply, traceability, sustainability, susceptibility to disease, and antibiotic resistance [35,37].…”