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Emerging Diseases of Aquatic Organisms Carmelo Iaria

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Emerging Diseases of Aquatic Organisms Carmelo Iaria Emerging diseases of aquatic organisms Carmelo Iaria THESIS FOR THE DEGREE OF PHILOSOPHIAE DOCTOR PHD COURSE IN APPLIED BIOLOGY AND EXPERIMENTAL MEDICINE UNIVERSITY OF MESSINA DEPARTMENT OF CHEMICAL, BIOLOGICAL, PHARMACEUTICAL AND ENVIRONMENTAL SCIENCES NOVEMBER 2019 University of Messina THESIS FOR THE DEGREE OF PHILOSOPHIAE DOCTOR PHD COURSE IN APPLIED BIOLOGY AND EXPERIMENTAL MEDICINE CURRICULUM IN BIOLOGICAL AND ENVIRONMENTAL SCIENCES XXXII PROGRAM SSD VET/03 Emerging diseases of aquatic organisms Carmelo Iaria Supervisor Professor Fabio Marino Doctoral Examination Board Professor Domenico Britti Professor Patrizia Trifilò Professor Vasileios Bakopoulos Doctor Monique Mancuso Coordinator: Professor Maria Assunta Lo Gullo ACADEMIC YEAR 2018-2019 To my family, People who define my world i SOMMARIO Il presente studio è stato effettuato con l'obiettivo primario di valutare agenti patogeni e malattie nei pesci provenienti da allevamenti di acquacoltura, pesci utilizzati come animali da laboratorio e organismi acquatici selvatici, che sono stati identificati durante il dottorato di ricerca, nell’ambito del progetto di ricerca in Biologia Applicata e Medicina Sperimentale presso il Centro di Ittiopatologia Sperimentale della Sicilia (CISS) e durante l’attività diagnostica routinaria dell’Unità di Patologia Veterinaria dell’Università di Messina. Sono stati evidenziati gli effetti negativi delle infezioni subcliniche sui risultati della ricerca, sulla salute degli animali utilizzati a fini scientifici, sulla salute degli animali destinati al consumo umano e sulla salute degli organismi acquatici selvatici. Le malattie dei pesci utilizzati ai fini della ricerca scientifica possono alterare i risultati sperimentali, aumentare la variabilità dei dati e impedire la riproducibilità sperimentale. Le malattie negli organismi di allevamento e selvatici possono causare gravi perdite finanziarie nelle aziende d’acquacoltura e nelle attività di pesca, nonché possono essere indicatori di problematiche ambientali. A tal fine, sono state effettuate analisi istopatologiche su un totale di 3831 organismi acquatici, dei quali 571, appartenenti a sei specie marine (Dicentrarchus labrax, Sparus aurata, Argyrosomus regius, Mugil cephalus, Lithognathus mormyrus, Coelorinchus caelorhincus), otto specie di acqua dolce (Danio rerio, Carassius auratus, Xiphophorus variatus, Poecilia reticulata, Abramis brama, Carassius carassius, Salmo trutta fario e Cobitis bilineata) e due specie di molluschi bivalvi (Pinna nobilis e Pinna rudis), risultavano positivi a differenti patologie. Numerose malattie sono state riscontrate durante questo studio. In particolare, la micobatteriosi e la mixosporidiosi sono state considerate le patologie più significative nei teleostei destinati alla ricerca scientifica; fotobacteriosi, enteromixiosi, criptocarioniasi e leyomioma sono state considerate le patologie più significative nei teleostei provenienti da impianti di acquacoltura; parassitosi dovute a Clinostomum complanatum, Lophoura edwardsi e Cystidicoloides ephemeridarum, tumori quali papilloma, fibroma e schwannoma e micobatteriosi sono state considerate le patologie più significative negli organismi acquatici provenienti da ambienti selvatici. Questi risultati possono essere utili per ampliare le conoscenze riguardo diverse malattie in varie specie ittiche sia allevate che selvatiche. al fine di comprenderne meglio l’eziologia e i meccanismi di patogenesi, nonché allertare gli operatori del settore sulle emergenze sanitarie nuove e ricorrenti. ii ABSTRACT The present research was undertaken with the prime objective of evaluating pathogens and diseases in fish from aquaculture farms, laboratory fish, and wild aquatic organisms, which were identified during the Ph.D. research project in Applied Biology and Experimental Medicine in the Centre for Experimental Fish Pathology of Sicily (CISS) and the Unit of Veterinary Pathology of the University of Messina. Adverse effects of subclinical infections on research results, the importance of animal health regarding welfare, the health of animals intended for human consumption and the health of wild aquatic organisms are likewise highlighted. Diseases in fish used for research can alter experimental outcomes, increase the variability of data, and impede experimental reproducibility. Diseases in farmed and wild organism can cause severe financial losses in aquaculture and fishery companies, as well as being indicators of environmental related distress. For this purpose, 571 diseased fish of different species (out of a total of 3831 fish analysed) that belonged to six marine species (Dicentrarchus labrax, Sparus aurata, Argyrosomus regius, Mugil cephalus, Lithognathus mormyrus, Coelorinchus caelorhincus), to eight fresh water species (Danio rerio, Carassius auratus, Xiphophorus variatus, Poecilia reticulata, Abramis brama, Carassius carassius, Salmo trutta fario and Cobitis bilineata) and two species of bivalve molluscs (Pinna nobilis and Pinna rudis) were examined for histopathological health monitoring. Several diseases have been found during this study. In particular, mycobacteriosis and myxosporidiosis were seen to be the most significant diseases in our research aquatic organisms; photobacteriosis, enteromyxiosis, cryptocarioniasis and leyomioma were seen to be the most significant diseases in our aquaculture aquatic organisms; Clinostomum complanatum, Lophoura edwardsi and Cystidicoloides ephemeridarum parasitosis, papilloma, fibroma and schwannoma tumours and mycobacteriosis were seen to be the most significant diseases in our wild aquatic organisms. These results may be useful in gathering wider knowledge on the occurrence of several diseases in various species and the environment, as well as in alerting industry operators to new and recurring health emergencies. In this way, the pathogenic mechanisms of aetiological agents may be better understood. iii ACKNOWLEDGEMENTS The accomplishment of this thesis work was made possible due to the participation of several co-workers, and different research teams and laboratories. First of all, I would like to thank my supervisor, Professor Fabio Marino, master of life, science and method, for all the teachings, support and his driving me every day on this path. My sincere thanks also to the PhD Course Coordinator, Professor Maria Assunta Lo Gullo, for help and guidance. To Professor Nunziacarla Spanò, for having been my go-to person during all my student career. Professor Giovanni Lanteri, master of techniques and laughter mate. Professors Battesimo and Francesco Macrì, as well as Professor Giuseppe Mazzullo for all support given during the last few years. I further acknowledge the grant obtained in 2016 from the University of Messina to support my PhD project. The support and contribution of research teams of veterinary pathology and marine ecology of the University of Messina, in particular Dr. Fabiano Capparucci, Dr. Giovanni De Benedetto, Dr. Sabrina Natale, Dr. Serena Savoca, Dr. Gioele Capillo and Dr. Giuseppe Panarello, in all kinds of daily laboratory activities and sampling procedures, was essential to the success of this thesis. I am really grateful to all the staff of Keldur, Institute for Experimental Pathology, University of Iceland, Reykjavik, for letting me have such a great, precious time during my PhD abroad internship, in learning skills relating to fish pathology and immunology. A warm thanks to my internship supervisor, Dr. Birkir Þór Bragason, master in fish immunology and molecular biology, for all his teachings, kindness and helpfulness, to Dr. Árni Kristmundsson and Dr. Sigríður Guðmundsdóttir for having kindly pre-reviewed my thesis. I would also like to thank all co-authors of papers involved in this study, and in particular research teams from the University of Messina in the persons of Professors Gabriella Gaglio, Giuseppe Piccione, Francesco Fazio, Antonio Ieni and Domenico Trombetta; the University of Naples, in the persons of Professor Francesca Carella; the University of Catania, in the person of Professor Maria Violetta Brundo; the Experimental Zooprophylactic Institute of Sicily, in the person of Dr. Daniele Macrì and the Biological Station Anton Dohrn of Messina in the persons of Dr. Teresa Bottari for all support and collaboration in research activities and all analysis provided during the last few years. I sincerely thank the Reviewers, Professor Giovanni Di Guardo, Professor Gionata De Vico, Professor Jorge Manuel de Oliveira Fernandes and Doctor Sepand Rastegar and the Doctoral Examination Board, Professor Domenico Britti, Professor Patrizia Trifilò, Professor Vasileios iv Bakopoulos, and Doctor Monique Mancuso for their interest, expertise contribution and constructive criticism. A proper thanks is due to journal editorial boards for allowing permission to use the scientific papers to support this thesis, in particular: to the Laboratory Animals SAGE journal, to the Zebrafish journal and Mary Ann Liebert, Inc. publisher, to the Journal of Aquatic Animal Health and The American Fisheries Society, to the Cahiers De Biologie Marine and Station Biologique de Roscoff and to the International Journal of Environmental Research and Springer Nature publisher. Finally, and not least, my parents for their patience and support during the doctoral years. Also, I wish to express my gratitude to other family members and friends for their encouragement. DECLARATION OF CONTRIBUTION I was responsible for the conception and management of the thesis project.
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