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ABSTRACTAmyloodiniosis and vibriosis are serious diseases in European seabass (Dicentrarchus labrax) hatcheries with noticeable high mortality. This study was conducted on tank-cultured D. labrax frys at a private marine hatchery near Mariout Lake (Alexandria, Egypt). Frys showed a high mortality rate (70%), lethargy, darkening, asphyxia, ascites, and velvety skin appearance. Both infectious agents were presumptively identified in all investigated frys. The identities of the two recovered agents were confirmed by molecular assay and phylogenetic analysis. On the tissue level, histopathological examination of skin, splenic, and renal tissue indicated severe alterations due to the direct impacts of both infections. On the cellular level, scanning electron micrographs showed both protozoal and bacterial pathogens on/in gill epithelial cells in solitary and colonial forms. Vibrio alginolyticus showed variable results for tested antibiotics, with a higher sensitivity to florfenicol. A successful control strategy was strictly adopted to overcome infections and stop mortalities. Copper sulphate and hydrogen peroxide were efficiently applied to tank water to overcome A. ocellatum infections. Further, florfenicol was effectively used to overcome systemic V. alginolyticus infections. The efficacy of treatments was confirmed by the absence of infectious agents in randomly collected fish samples. To the best of the authors’ knowledge, this study is one of the earliest Egyptian studies that dealt with the dilemma of mass kills associated with external parasitic/systemic bacterial infections among hatchery-reared European seabass.

The amyloodiniosis outbreak was documented with high mortality of percula clownfish (Amphiprion percula) and subsequently, various therapeutics were evaluated to control the infection. The affected fish exhibited symptoms such as discoloration of the skin and jerky movement with severe respiratory stress. Microscopic examination of the gill and body surface of the infested moribund fish showed the presence of brown coloured spherical shaped trophonts with the size range of 150–300 µM. Copper sulphate (10 ppm) and hydroxychloroquine phosphate (10 ppm) treatment with 14 days of continuous bath showed 76 ± 5.3 and 66 ± 4.3% survival rates. However, formalin (10 ppm) and malachite green (10 ppm) treated groups showed a survival of 41 ± 1.7 and 32.7 ± 1.2% respectively. The present results suggest that, use of copper sulphate to treat amyloodiniosis in clownfish will relatively more effective than other treatment options. The findings will be helpful to mitigate amyloodiniosis in marine finfish aquaculture, particularly marine ornamentals. [ABSTRACT FROM AUTHOR]

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The ectoparasite Amyloodinium ocellatum is a dinoflagellate that causes severe morbidity and mortality in both brackish and marine warmwater aquaculture fishes worldwide. A. ocellatum has a triphasic life cycle based on a free-living flagellate (the dinospore), a parasitic stage (the trophont) and a resting and reproductive cyst (the tomont). Current chemical treatments have shown some efficacy in eliminating dinospores but fail to inactivate the tomonts. Here we evaluated the efficacy of alternative treatments in vitro through sporulation tests and the quantification of dinospore production and motility. Hydrogen peroxide and peracetic acid efficiently decreased dinospore production at low concentrations, but total inactivation of tomonts was only achieved with higher dosages. Tomont inactivation was also observed with disinfectants such as sodium hypochlorite and Virkon S. This work provides insights into effective and environmentally friendly alternatives for the elimination of resistant forms of the marine parasite A. ocellatum.

Abstract: Amyloodinium ocellatum is a highly pathogenic dinoflagellate parasite with global distribution that causes high mortalities in the culture of tropical and sub-tropical marine and estuarine fishes. Diagnosis typically occurs through gross examination following the onset of morbidity, at which point treatment is of limited benefit. In the present study, a new molecular diagnostic tool for the rapid detection of A. ocellatum (AO) was developed using the loop-mediated isothermal amplification method (LAMP). The AO-LAMP assay designed is highly specific using a set of four primers – two outer and two inner primers targeting six different regions on the 5′ end of the Small Subunit rDNA region (SSU rDNA) of A. ocellatum. The AO-LAMP assay, optimized for 25–30min at 62°C, amplified the DNA from A. ocellatum extracted from both water and gill tissue samples and did not amplify DNA from four closely related dinoflagellate species. The detection limit of the AO-LAMP assay was 10fg, exceptionally higher than the conventional PCR (1pg). In addition, the standardized AO-LAMP assay was capable of detecting single tomonts and trophonts; the assay was not affected by the presence of possible inhibitory substances present in environmental water samples or gill samples. The AO-LAMP assay developed in the present study provides a novel useful tool for the simple, rapid and sensitive detection of A. ocellatum in water and gill tissue samples, which could assist in the early detection and improved control of A. ocellatum infections in aquaculture systems. [Copyright &y& Elsevier]

Background and Objective: Amyloodinium ocellatum infects the gills and skin of both marine and brackish water fishes. The aim of the present study was to examine pathogenesis, prevalence, trials for treatment and histopathological alterations of Amyloodinosis in naturally infested Asian Seabass Barramundi Lates calcarifer and Hamour Epinephelus polyphekadion in Ismailia Governorate, Egypt., Materials and Methods: A total number of 1447 Red Sea cultured Seabass (Lates calcarifer) broadstock and a total number of 53 Red Sea cultured Hamour, Epinephelus polyphekadion broadstock were collected and subjected for the study. Fishes showed symptoms of sudden death and respiratory distress besides Amyloodiniosis on gills and skin. All fishes were treated with various treatment protocols while gills of naturally infected fishes were examined histopathologically., Results: The clinical signs of infested fishes were flashing, surfacing, off food and respiratory distress. The intensity of infestation of Amyloodiniosis was more sever in Asian Seabass than Epinephelus polyphekadion while treatment of choice was copper sulphate (prolonged bath), freshwater bath and formalin consequently., Conclusion: Treatment of choice for Amyloodinium ocellatum infestation in Asian Seabass was copper sulphate (prolonged bath) followed by freshwater bath then formalin.

Amyloodinium ocellatum, the causative agent of amyloodiniosis (marine velvet, velvet disease), affects marine and brackish fish in various warm and temperate habitats. We recorded disease outbreaks with high morbidity and mortality rates in marine-cultured European seabass Dicentrarchus labrax fry at 2 locations in northwest Egypt. The sudden outbreak, high morbidity and mortality rates, and skin lesions with a velvety appearance in affected fish all indicated A. ocellatum infection. This was further confirmed by microscopic findings of the parasitic stage (trophonts) in skin and gill smears. While ecological factors including water temperature and salinity were all amenable to parasite establishment and propagation, mortality rates differed between the 2 farms, with rates of mortality well correlated with prevalence and intensity of A. ocellatum infections. Characterization by PCR targeting rDNA gene fragments and subsequent DNA sequencing and phylogenetic analysis further confirmed the molecular identity of the A. ocellatum isolate, which was genetically similar to isolates from other geographical locations. Finally, an improved treatment method using dual hyposalination and copper sulfate exposure to increase the efficiency and decrease the toxicity of copper sulfate was tested. The gradual reduction in water salinity coupled with copper sulfate treatment was more efficient at controlling the disease than only applying copper sulfate. To our knowledge, this is the first parasitological and molecular characterization of A. ocellatum in marine cultures in Egypt. The high molecular identity and close phylogenetic relationship further confirmed the monophyletic nature of A. ocellatum isolates.