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The effect of sunlight and salinities (10, 20, 39, and 60 psu) on the survival of Vibrio parahaemolyticus strains carrying either (thermostable direct hemolysin) tdh, the (thermostable related hemolysin) trh, and both or none of them were studied in water microcosms stabilized at 20°C using plate count agar and acridine orange direct viable count. All V. parahaemolyticus strains exposed to sunlight rapidly lose their culturability and evolve into a viable but non-culturable state (VBNC). However, the tdh positive strains remain more culturable than the non-virulent or trh positive strain but statically insignificant. At tested salinities, the survival time was higher at 10, 20, and 60 psu compared with that observed in seawater (39 psu). In seawater under dark condition, Vibrio strains remain culturable for up to 200 days with a significant difference between strains (p 0.05). Furthermore, the non-pathogenic strain survives longer than the virulent ones. At different salinities, a better adaptation is observed at 10 and 20 psu compared with 39 and 60 psu. Resuscitations essays performed on VBNC bacteria in a nutrient broth at 20°C and 37°C does not show any revivification. PRACTITIONER POINTS: Effect of sunlight and salinities on the survival of V. parahaemolyticus in the marine environment. Resuscitation essay performed on viable but no cultivable bacteria. Microscope motility examines show that all strains exposed to sunlight remain motile after the loss of cultivability.

MedAID (Mediterranean Aquaculture Integrated Development) is a European H2020 Research and Innovation Project that started in 2017. The main objective was to increase the overall competitiveness and sustainability of the whole value chain of the Mediterranean marine aquaculture sector by improving its technical and business performance and by shifting to a sustainable market-oriented approach with a more positive social and consumer perception. In the field of disease control and welfare MedAID aims to provide essential components for a better health and welfare management system for the Mediterranean marine aquaculture industry overall and at company level. One of the gaps addressed by MedAID was the lack of reference methodologies for disease diagnostics for European seabass and Gilthead seabream, the main species produced in Mediterranean marine fish farming. The present Diagnostic Manual was conceived in the form of up-to-date guidelines providing current methodologies for a harmonised approach to the health challenges due to viral and bacterial pathogens in the farming of seabass and seabream.

Aims: To detect the possible coexistence of striped jack nervous necrosis virus (SJNNV) and red-spotted grouper nervous necrosis virus (RGNNV) genotypes in a single fish, a methodology based on the combination of PCR amplification and blot hybridization has been developed and applied in this study. Methods and Results: The degenerate primers designed for the PCR procedure target the T4 region within the capsid gene, resulting in the amplification of both genotypes. The subsequent hybridization of these amplification products with two different specific digoxigenin-labelled probes resulted in the identification of both genotypes separately. The application of the RT-PCR protocol to analyse blood samples from asymptomatic wild meagre (Argyrosomus regius) specimens has shown a 46·87% of viral nervous necrosis virus carriers. The combination of RT-PCR and blot hybridization increases the detection rate up to 90·62%, and, in addition, it has shown the coexistence of both genotypes in 18 out of the 32 specimens analysed (56·25%). Conclusions: This study reports the coexistence of betanodaviruses belonging to two different genotypes (SJNNV and RGNNV) in wild fish specimens. Significance and Impact of the Study: This is the first report demonstrating the presence of SJNNV and RGNNV genotypes in the same specimen. This study also demonstrates a carrier state in this fish species for the first time.