Koi herpesvirus (KHV) is the causative agent of a koi herpesvirus disease (KHVD) inducing high mortality rates in common carp and koi (Cyprinus carpio). No widespread effective vaccination strategy has been implemented yet, which is partly due to side effects of the immunized fish. In this study, we present an evaluation of the purification of infectious KHV from host cell protein and DNA, using the steric exclusion chromatography. The method is related to conventional polyethylene glycol (PEG) precipitation implemented in a chromatographic set-up and has been applied for infectious virus particle purification with high recoveries and impurity removal. Here, we achieved a yield of up to 55% of infectious KHV by using 12% PEG (molecular weight of 6 kDa) at pH 7.0. The recoveries were higher when using chromatographic cellulose membranes with 3-5 μm pores in diameter instead of 1 μm. The losses were assumed to originate from dense KHV precipitates retained on the membranes. Additionally, the use of >0.6 M NaCl was shown to inactivate infectious KHV. In summary, we propose a first step towards a purification procedure for infectious KHV with a possible implementation in fish vaccine manufacturing.

This research aimed to investigate the present status of disease prevalence and usage of aqua drugs for various aquaculture operations in the Narsingdi region of Bangladesh. Data were collected through the market survey, preset questionnaire interview, personal contact, and participatory rural appraisal tools. Amongst the respondents, the maximum percentages were found practicing mixed cultures of carp, tilapia, and pangas. The respondents suggested that epizootic ulcerative syndrome, saprolegniasis, streptococcosis, tail and fin rot and bacillary necrosis are common fish diseases in the area. About 140 drugs of different companies used in aquaculture for different purposes such as disease treatment, growth enhancement, water quality improvement, toxic gas removal, improvement of feed conversion ratio. Zeolite, rotenone, disinfectant, oxygen precursors, ammonia reducers, and probiotics were applied for pond preparation, water, and soil quality maintenance, while 30 different antibiotics were used for the purpose of treatment. Among the available antibiotics, oxytetracycline, ciprofloxacin, enrofloxacin, erythromycin, sulphadiazine, and trimethoprim were found extensively used by the fish farmers. Four enzymes and eighteen growth promoters were identified as being utilized to enhance digestion and boost up the production. This study elicited various issues connected with application and administration of such aqua chemicals, including farmers\' ignorance about their usage, proper doses, application methods, withdrawal period, and the human health concerns associated with their irresponsible use. However, the consequences of these chemical products to the environment, animal health, and human health required further study for the betterment of mankind.

This study describes the patterns of mortality and investigates the sources of variation in mortality during the marine phase of commercial salmon farming. The study included daily mortality records from stocking to harvest of 21 million salmon from ten hatcheries in 136 fish-groups (fish in the same cage from the same hatchery). The fish was stocked in 2017-2018 at 21 marine farms within two Norwegian companies. The sources of variation in mortality were investigated using multilevel linear regression models with \'fish-group\' nested within \'farm\' as a random effect, cross-classified with \'hatchery\'. In the final model, \'fish-group\' was the source of most variation (70%). Furthermore, the mortality categories \'smolt-related mortality\', \'infectious diseases\' and \'handling and treatment\' were responsible for 10%, 17% and 29% of the total number of dead fish respectively. Overall, the study shows that smolt-related mortality is one of the major causes of death in the first part of the production, while handling and treatment was the dominating cause of mortality in total. Mortality varied by fish-group to a large extent. This means that targeted preventive strategies to decrease mortality for individual fish-groups might be more effective than overall measures at farm or hatchery level.

The purpose of this study was to determine the phylogenetic relationships among the primary betanodavirus strains circulating in Tunisian coastal waters. A survey was conducted to investigate nodavirus infections at 15 European sea bass Dicentrarchus labrax and gilthead sea bream Sparus aurata farming sites located along the northern and eastern coasts of Tunisia. The primary objective of the study was to create epidemiological awareness of these infections by determining phylogenetic relationships between the main betanodavirus strains circulating during the period 2012-2019, using RNA1 and/or RNA2 genome segments. Approximately 40% (118 of 294) tissue pools tested were positive for betanodavirus. Positive pools were distributed across all of the sampling sites. While fish mortalities were always correlated with the presence of virus in sea bass, a severe outbreak was also identified in sea bream larvae in 2019. Phylogenetic analysis revealed that almost all Tunisian strains from both sea bass and sea bream irrespective of outbreaks clustered within the RGNNV genotype. It is noteworthy that samples collected during the 2019 outbreak from sea bream contained both RNA1 and RNA2 fragments belonging to the RGNNV and SJNNV genotype, respectively, an indication of viral genome reassortment. To our knowledge, this is the first report of reassortant betanodavirus in Tunisia.

In 2014 the contribution of aquaculture to supply food for human consumption overtook wild-caught fish for the first time. Despite improvements in the aquaculture industry, it has been estimated that as much as 10% of all cultured aquatic animals are lost because of infectious diseases, amounting to >10 billion USD in losses annually on a global scale. Vaccination to prevent disease is used routinely in finfish aquaculture, especially for Atlantic salmon (Salmo salar), while in a limited capacity (or not at all) in many other fish species due to lack of vaccines, poor performance or cost. There has, nevertheless, been impressive progress in fish vaccine development over the last 4 decades with 24 licenced fish vaccines now commercially available for use in a variety of fish species. These comprise whole killed, peptide subunit, recombinant protein, DNA and live attenuated vaccines. Challenges do, however, still exist as the majority of commercial vaccines are killed whole cell pathogen preparations administered by intraperitoneal injection. This may not be the optimal route to deliver some vaccines, but lack of effective adjuvants and basic knowledge on immune response has hindered progress in the development of mucosal vaccines. The cost of injecting fish may also be prohibitive in some countries leading to disease treatment (e.g. with antibiotics) rather than using preventative measures. It is important that these issues are addressed as the industry continues to grow globally. Exciting opportunities exist for rapid development of fish vaccines in the future, with continued reduction in cost of technologies (e.g. of whole genome sequencing), regulations changing (e.g. DNA vaccines can now authorised in Europe), the introduction of novel antigen expression and delivery systems (such as virus-like particles, VLPs), development of novel adjuvants and advancements in the elucidation of basic mechanisms of mucosal immunity. Development of effective mucosal vaccines and optimisation of their delivery will facilitate novel vaccine development, and enable the aquaculture industries in LMIC to use vaccination routinely in the future. In addition, effective use of emergency (autogenous) vaccines will assist in tackling emerging disease challenges.