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

Abstract

Centrocestus formosanus is a digenetic trematode with a complex life cycle, involving invertebrate and vertebrate hosts, humans included. In particular, it causes gill lesions and mortality in freshwater fish species, and gastrointestinal symptoms in infected humans. Here, we describe the occurrence of C. formosanus infection in zebrafish imported in Italy and propose a newly designed species-specific primer pair to ameliorate the diagnostic investigations for C. formosanus. Gill arches of 30 zebrafish were examined for the presence of encysted metacercariae under a stereomicroscope and processed through molecular analyses targeting the ribosomal internal transcribed sequence 2 (ITS2). Although C. formosanus distribution was originally restricted to Asia, it has been subsequently reported in new countries, revealing itself as an invasive species and raising important concerns for biodiversity, economy, scientific research, as well as animal and public health. Given the crucial role played by the ornamental fish industry in spreading this parasite, there is an urgent need for control measures to prevent the introduction and establishment of C. formosanus in non-endemic areas, including Europe. We also suggest developing new strategies in microbiology and epidemiology to better explore this new globalization-derived invasive species.

DNA Methylation Profiles of Tph1A and BDNF in Gut and Brain of L. Rhamnosus-Treated Zebrafish

“…Fishes were fed twice daily with sterilized commercial food (Sera Vipagran, Heiensberg, Germany). The room, water temperatures and illumination were maintained according to the standards of Zebrafish care [ 19 , 27 ]. As reported by our previous studies [ 19 , 27 ], all fish were treated in accordance with the Directive of the European Parliament and of the Council on the Protection of Animals Used for Scientific Purposes (Directive 2010/63/EU), and in agreement with the Bioethical Committee of the University Federico II of Naples (authorization protocol number 47339-2013).…”

“…The room, water temperatures and illumination were maintained according to the standards of Zebrafish care [ 19 , 27 ]. As reported by our previous studies [ 19 , 27 ], all fish were treated in accordance with the Directive of the European Parliament and of the Council on the Protection of Animals Used for Scientific Purposes (Directive 2010/63/EU), and in agreement with the Bioethical Committee of the University Federico II of Naples (authorization protocol number 47339-2013). Following behavioral testing, the animals were euthanized by immersion in overdose 500 mg/L −1 of 3-aminobenzoic acid ethyl ester (MS-222) buffered with sodium bicarbonate (1:2 ratio solution), to pH 7.4 (Sigma–Aldrich, St Louis, MO, USA), and brain and gut tissues were taken and stored at −80 °C.…”

Good practices in the rearing and maintenance of zebrafish (Danio rerio) in Brazilian laboratories

“…In 2008, the WHO, the World Organization for Animal Health (OIE), and the Food and Agriculture Organization of the United Nations (FAO) created the "One World, One Health" initiative, in which the term "One Health" expresses the inseparability of human health, animal health, and ecosystem health (or environmental health) (123,124) . Parasitic diseases of zebrafish have relevance to one health (89,125) . From this point of view, the health of a zebrafish colony can impact the health of researchers and bioterists, as well as the people they live with and the environment; therefore, some recommendations are essential.…”

“…Furthermore, transgenic or wild zebrafish are potential reservoirs of pathogens that can contaminate the natural environment, consequently infecting wild fish and other animals in general, including humans (Table 2). Indeed, the widely distributed occurrence of zebrafish worldwide is already causing concern about introducing diseases that were not reported in the past in specific geographic regions (89,125) . To avoid contamination of the environment and the risk of infection, it is recommended that zebrafish aquatic facilities coordinators be aware of the provisions of current legislation.…”

Freshwater as a Sustainable Resource and Generator of Secondary Resources in the 21st Century: Stressors, Threats, Risks, Management and Protection Strategies, and Conservation Approaches

“…It lives in intermediate hosts such as aquatic gastropods, fish, and amphibians, and in definitive hosts such as aquatic birds, mammals, and humans. The parasite affects gill health and causes respiratory problems for host fish [ 209 ]. Francis-Floyd et al [ 210 ] estimate the tropical fish losses infested with this parasite at USD 3 million per year (cited by Mitchell et al [ 211 ]).…”