In vitro screening of novel treatments for Aphanomyces invadans

Forty‐nine compounds were tested in vitro for fungicidal activity against hyphae of the fish‐pathogenic Oomycete fungus, Aphanomyces invadans. These comprised: (a) chemicals with previous reported activity against Oomycete fungi; (b) chemicals in use in Asia to treat ulcerative disease outbreaks; (c) commercial biocides and fungicides; (d) natural products with potential antimicrobial activity; and (e) surfactants tested separately and in combination with some of the above treatments. No compounds tested proved as effective as malachite green, but some low‐toxicity natural compounds and particular surfactants showed potential for further studies. Some compounds that are currently in use in Asian aquaculture were shown to have no effect on A. invadans hyphae at recommended treatment rates. A range of compounds that demonstrated activity against the mycelium were selected for further testing in a zoospore motility assay. Lower treatment concentrations were required to inhibit zoospore motility than were required to inhibit hyphal growth. Zoospore activity ceased within 1 h of exposure to 2.5 parts per million (p.p.m.) coconut diethanol‐ amide; 1.25 p.p.m. propolis + 0.5 p.p.m. 13/6.5; 5 p.p.m. neem (Azadirachta siamensis) seed extract + 0.01 p.p.m. OP10; 20 p.p.m. tea tree (Melaleuca alternifloria) oil; and 25 p.p.m. D‐limonene + 0.05 p.p.m. E‐Z‐Mulse^TM. The treated spores were shown to be non‐viable in culture medium. Selected compounds were further tested for ability to inhibit zoospore production by A. invadans mycelium over a 72‐h period. In toxicity trials, silver barb, Barbodes gonionotus (Bleeker), exposed to 2.5 p.p.m. coconut diethanolamide; 2.5 p.p.m. OP10; 0.5 p.p.m. E‐Z‐Mulse^TM; 20 p.p.m. neem seed extract + 0.5 p.p.m. OP10; and 5 p.p.m. D‐limonene + 0.5 p.p.m. E‐Z‐Mulse^TM suffered no mortalities and no obvious behavioural changes. Similarly, rainbow trout, Oncorhynchus mykiss (Walbaum), exposed to 25 p.p.m. propolis + 1 p.p.m. OP10; 10 p.p.m. neem seed extract + 0.01 p.p.m. 13/6.5; and 10 p.p.m. D‐limonene + 0.01 p.p.m. OP10 suffered no mortalities and no obvious behavioural changes.     

Molecular characterization of the fish-pathogenic fungus Aphanomyces invadans

Aphanomyces invadans (Saprolegniaceae) is a peronosporomycete fungus associated with the serious fish disease, epizootic ulcerative syndrome (EUS), also known as mycotic granulomatosis. In this study, interspecific relationships were examined between A. invadans isolates and other aquatic animal pathogenic Saprolegniaceae, and saprophytic Saprolegniaceae from EUS-affected areas. Restriction fragment length polymorphisms and sequences of ribosomal DNA confirmed that A. invadans is distinct from all other species studied. A sequence from the internal transcribed spacer region ITS1, unique to A. invadans, was used to design primers for a PCR-based diagnostic test. Intraspecific relationships were also examined by random amplification of polymorphic DNA using 20 isolates of A. invadans from six countries. The isolates showed a high degree of genetic homogeneity using 14 random ten-mer primers. This provides evidence that the fungus has spread across Asia in one relatively rapid episode, which is consistent with reports of outbreaks of EUS. Physiological distinctions between A. invadans and other Aphanomyces species based on a data set of 16 growth parameters showed remarkable taxonomic congruence with the molecular phylogeny.     

Investigation and management of an outbreak of multispecies mycobacteriosis in Australian lungfish (Neoceratodus fosteri) including the use of triple antibiotic treatment

Disease due to non‐tuberculous mycobacteria (NTM) is common in fish. Current recommendations focus on outbreak management by depopulating entire fish stocks and disinfecting tanks. Treatment is not advocated. Treatment may be appropriate, however, where individual, valuable fish are concerned. ZSL London Zoo managed an outbreak of mycobacteriosis in a valuable group of imported F1 captive‐bred Australian lungfish (Neoceratodus fosteri) by depopulation, isolation, extensive testing and daily oral antibiotic treatment. Four species of Mycobacterium (M. marinum, M. fortuitum, M. chelonae and M. peregrinum) were involved in this outbreak, each with unique antibiotic sensitivities. Triple therapy with rifampicin, doxycycline and enrofloxacin for 8 months was the most effective antibiotic combination, resulting in full disease resolution. No side effects were noted and, more than 18 months post‐treatment, no recurrence had occurred. This is the first report of mycobacterial disease in lungfish and the first report of a polymycobacterial outbreak in fish involving these four species of Mycobacterium. This report demonstrates the value of extensive isolation and identification. Also, as therapies currently advised in standard texts did not reflect the antibiotic sensitivity of the NTM found in the fish reported here, we recommend that antibiotic treatment should always be based on sensitivity testing.     

Monitoring infection with Piscine myocarditis virus and development of cardiomyopathy syndrome in farmed Atlantic salmon (Salmo salar L.) in Norway

An epidemiological study was carried out in Norway in 2015–2018, investigating the development of infection with Piscine myocarditis virus (PMCV) and development of cardiomyopathy syndrome (CMS) in farmed Atlantic salmon. Cohorts from 12 sites were followed and sampled every month or every other month from sea transfer to slaughter. PMCV was detected at all sites and in all sampled cages, and fish in six sites developed clinical CMS. The initial infection happened between 1 and 7 months post‐sea transfer, and the median time from infection with PMCV until outbreak of CMS was 6.5 months. Generally, fish from sites with CMS had higher viral titre and a higher prevalence of PMCV, compared to sites that did not develop clinical CMS. The virus persisted until the point of slaughter at most (11 out of 12) of the sites. The detection of PMCV in all sites suggests that PMCV is more widespread than previously known. Screening for PMCV as a tool to monitor impending outbreaks of CMS must be supported by observations of the health status of the fish and risk factors for development of disease.     

A natural infection by the red sea bream iridovirus‐type Megalocytivirus in the golden mandarin fish Siniperca scherzeri

An outbreak of a Megalocytivirus infection was found in the golden mandarin fish Siniperca scherzeri during September and October 2016, in Korea. Phylogeny and genetic diversity based on the major capsid protein (MCP) and adenosine triphosphatase (ATPase) genes showed a new strain. Designated as GMIV, this strain derived from the golden mandarin fish was suggested to belong to the red sea bream iridovirus (RSIV)‐subgroup I. Additionally, this train clustered with the ehime‐1 strain from red sea bream Pagrus major in Japan and was distinguished from circulating isolates (RSIV‐type subgroup II and turbot reddish body iridovirus [TRBIV] type) in Korea. The infection level, evaluated by qPCR, ranged from 8.18 × 10² to 7.95 × 10⁶ copies/mg of tissue individually, suggesting that the infected fish were in the disease‐transmitting stage. The diseased fish showed degenerative changes associated with cytomegaly in the spleen as general sign of Megalocytivirus infection. The results confirm that the RSIV‐type Megalocytivirus might have crossed the environmental and species barriers to cause widespread infection in freshwater fish.     

Yersiniosis in Atlantic cod,Gadus morhua (L.), characterization of the infective strain and host reactions

A disease outbreak in farmed Atlantic cod caused by Yersinia ruckeri is reported. Mortality started following vaccination of cod reared in two tanks (A and B). The accumulated mortality reached 1.9% in A and 4.8% in B in the following 30 days when treatment with oxytetracycline was applied. Biochemical and molecular analysis of Y. ruckeri isolates from the cod and other fish species from fresh and marine waters in Iceland revealed a high salinity‐tolerant subgroup of Y. ruckeri serotype O1. Infected fish showed clinical signs comparable with those of Y. ruckeri ‐infected salmonids, with the exception of granuloma formations in infected cod tissues, which is a known response of cod to bacterial infections. Immunohistological examination showed Y. ruckeri antigens in the core of granulomas and the involvement of immune parameters that indicates a strong association between complement and lysozyme killing of bacteria. Experimental infection of cod with a cod isolate induced disease, and the calculated LD₅₀ was 1.7 × 10⁴ CFU per fish. The results suggest that yersiniosis can be spread between populations of freshwater and marine fish. Treatment of infected cod with antibiotic did not eliminate the infection, which can be explained by the immune response of cod producing prolonged granulomatous infection.     

Beaver dams shift desert fish assemblages toward dominance by non‐native species (Verde River,Arizona, USA)

The reintroduction of beaver (Castor canadensis) into arid and semi‐arid rivers is receiving increasing management and conservation attention in recent years, yet very little is known about native versus non‐native fish occupancy in beaver pond habitats. Streams of the American Southwest support a highly endemic, highly endangered native fish fauna and abundant non‐native fishes, and here we investigated the hypothesis that beaver ponds in this region may lead to fish assemblages dominated by non‐native species that favour slower‐water habitat. We sampled fish assemblages within beaver ponds and within unimpounded lotic stream reaches in the mainstem and in tributaries of the free‐flowing upper Verde River, Arizona, USA. Non‐native fishes consistently outnumbered native species, and this dominance was greater in pond than in lotic assemblages. Few native species were recorded within ponds. Multivariate analysis indicated that fish assemblages in beaver ponds were distinct from those in lotic reaches, in both mainstem and tributary locations. Individual species driving this distinction included abundant non‐native green sunfish (Lepomis cyanellus) and western mosquitofish (Gambusia affinis) in pond sites, and native desert sucker (Catostomus clarkii) in lotic sites. Overall, this study provides the first evidence that, relative to unimpounded lotic habitat, beaver ponds in arid and semi‐arid rivers support abundant non‐native fishes; these ponds could thus serve as important non‐native source areas and negatively impact co‐occurring native fish populations.     

Outbreak of hirame rhabdovirus infection in cultured spotted sea bass Lateolabrax maculatus on the western coast of Korea

In this study, we determined the cause of a disease outbreak in spotted sea bass, Lateolabrax maculatus reared in culture cages on the western coast of Korea in 2013. The major signs in the diseased fish exhibited were haemorrhaging on the membranes of the abdomen, gastrointestinal organs and opercular gills, as well as an enlarged spleen. No external morphological signs of infection were visible, except for a darkening in colour. No parasites or pathological bacteria were isolated from the diseased fish; however, epithelioma papulosum cyprini (EPC) cells inoculated with tissue homogenates from the diseased fish showed cytopathic effects (CPEs). Virus particles in the EPC cells were bullet‐shaped, 185–225 nm long and 70–80 nm wide, characteristic of Rhabdoviridae. Polymerase chain reaction analyses of homogenized tissues from the diseased fish and supernatants of cell cultures with CPEs indicated specific, 553‐bp‐long fragments corresponding to the matrix protein gene of the hirame rhabdovirus (HIRRV). Phylogenetically, the HIRRV phosphoprotein gene of spotted sea bass was more closely related to phosphoproteins from Chinese and Polish HIRRV strains than from other Korean strains. To our knowledge, this is the first report of HIRRV infection in cultured spotted sea bass.     

Sensitivity and specificity of real‐time PCR and bacteriological culture for francisellosis in farm‐raised Nile tilapia (Oreochromis niloticus L.)

Despite the worldwide occurrence of Francisella noatunensis subsp. orientalis (Fno) infection in farmed tilapia, sensitivity and specificity estimates of commonly used diagnostic tests have not been reported. This study aimed to estimate the sensitivity and specificity of bacteriological culture and qPCR to detect Fno infection. We tested 559 fish, sampled from four farms with different epidemiological scenarios: (i) healthy fish in a hatchery free of Fno; (ii) targeted sampling of diseased fish with suggestive external clinical signs of francisellosis during an outbreak; (iii) convenience sampling of diseased and clinically healthy fish during an outbreak; and (iv) sampling of healthy fish in a cage farm without a history of outbreaks, but with francisellosis reported in other farms in the same reservoir. The qPCR had higher median sensitivity (range, 48.8–99.5%) than culture (range, 1.6–74.4%). Culture had a substantially lower median sensitivity (1.6%) than qPCR (48.8%) to detect Fno in carrier tilapia (farm 4). Median specificity estimates for both tests were >99.2%. The qPCR is the superior test for use in surveillance and monitoring programmes for francisellosis in farmed Nile tilapia, but both tests have high sensitivity and specificity which make them fit for use in the diagnosis of Fno outbreaks.     

Aeromonas jandaei and Aeromonas veronii caused disease and mortality in Nile tilapia,Oreochromis niloticus (L.)

Diseases caused by motile aeromonads in freshwater fish have been generally assumed to be linked with mainly Aeromonas hydrophila while other species were probably overlooked. Here, we identified two isolates of non‐A. hydrophila recovered from Nile tilapia exhibiting disease and mortality after exposed to transport‐induced stress and subsequently confirmed their virulence in artificial infection. The bacterial isolates were identified as Aeromonas jandaei and Aeromonas veronii based on phenotypic features and homology of 16S rDNA. Experimental infection revealed that the high dose of A. jandaei (3.7 × 10⁶ CFU fish^?1) and A. veronii (8.9 × 10⁶ CFU fish^?1) killed 100% of experimental fish within 24 h, while a 10‐fold reduction dose killed 70% and 50% of fish, respectively. When the challenge dose was reduced 100‐fold, mortality of the fish exposed to A. jandaei and A. veronii decreased to 20% and 10%, respectively. The survivors from the latter dose administration were rechallenged with respective bacterial species. Lower mortality of rechallenged fish (0%–12.5%) compared to the control groups receiving a primary infection (37.5%) suggested that the survivors after primary infection were able to resist secondary infection. Fish exposed to either A. jandaei or A. veronii exhibited similar clinical signs and histological manifestation.     

First report of Shewanella sp. and Listonella sp. infection in freshwater cultured loach,Misgurnus anguillicaudatus

A novel disease outbreak occurred among cultured loach, Misgurnus anguillicaudatus in China. The diseased fish displayed lethargy, loss of appetite, black discolouration, anasarca and ventral skin hyperaemia. Clinical signs were observed in most of the fish examined, which consisted of discoloured liver, swollen spleen and petechial haemorrhage in the intestine and peritoneal membranes. The causative agent was found to be two species of bacteria by experimental infection, identified as Shewanella sp. and Listonella sp. The result showed that these were likely to be Shewanella putrefaciens and Listonella anguillarum respectively. Bacterial identification consisted of physiological and biochemical tests as well as 16S rRNA sequence analysis. In addition, antibiotic susceptibility testing was performed, revealing that Shewanella putrefaciens and Listonella anguillarum were susceptible to enrofloxacin, ciprofloxacin, levofloxacin, pipemidicacid and norfloxacin. This is the first report of S. putrefaciens and L. anguillarum from cultured loach.     

Response of the riverine fish community to the construction and operation of a diversion hydropower plant in central Chile

• 1. Chilean rivers have a large potential for hydropower development, and they also contain a unique native fish fauna with a high level of endemism. Several diversion hydropower plants have recently been constructed in Chile; however, the response of fish communities to these new hydropower plant designs is not well known.
• 2. Responses of native and non‐native fish to the construction and operation of a new hydropower plant that diverts water from two rivers were quantified. The Laja River is highly regulated and manipulated with three older (40 yr) dam‐based hydropower plants and irrigation diversions located upstream from the new facility. In contrast, the Rucúe River has no other hydropower facilities and is comparatively undisturbed.
• 3. Prior to construction, the Laja River had a fish community with lower species richness compared with the Rucúe River. The fish community structure in the Laja River was dramatically altered after the new hydropower facility began operation. On the other hand, in the Rucúe River, even though abundance of fish declined, there was less of a change in the total fish community structure. The fish community in the Rucúe River exhibited greater resistance to change compared with the Laja River.
• 4. The species most affected were the introduced salmonids and an endangered native species Percilia irwini.
• 5. Although diversion hydropower designs may have less impact than traditional dam‐based hydropower facilities, results of this study indicate that diversion hydropower structures can cause large changes in the fish community.

Copyright © 2006 John Wiley & Sons, Ltd.     

Antimicrobial susceptibility pattern of Flavobacterium columnare isolates collected worldwide from 17 fish species

Flavobacterium columnare is the causative agent of columnaris disease in diverse fish species worldwide. Although columnaris is an important disease, the antimicrobial susceptibility pattern of F. columnare is not well studied. Thus, the purpose of this study was to test the in vitro antimicrobial susceptibility of 97 F. columnare isolates collected worldwide between 1987 and 2011 from 17 fish species. The broth microdilution technique was utilized for reliable testing of these fastidious organisms. None of the isolates displayed acquired resistance to florfenicol, gentamicin, ormetoprim‐sulfadimethoxine and trimethoprim‐sulfamethoxazole. Acquired resistance to chloramphenicol was detected in 1%, to nitrofuran in 5%, to oxytetracycline in 11% and to enrofloxacin, flumequine and oxolinic acid in 10%, 16% and 16% of the isolates, respectively, as reflected by a bimodal or trimodal distribution of their minimum inhibitory concentrations (MICs). One isolate showed acquired resistance towards several antimicrobial agents including erythromycin. Another isolate revealed acquired resistance towards – amongst others – ampicillin. The isolates displaying acquired resistance originated from ornamental fish species or Vietnamese catfish, except for two isolates coming from wild channel catfish in which acquired resistance was encountered towards oxytetracycline only. Fifty per cent of the resistant isolates from ornamental fish were shown to have acquired resistance against three classes of antimicrobial agents, assigning these isolates as multiple resistant. These data might indicate less prudent use of antimicrobials especially in ornamental fish species.     

Age dependency of nervous necrosis virus infection in barramundi Lates calcarifer (Bloch)

Age‐dependent susceptibility to nervous necrosis virus (NNV) was demonstrated for barramundi (Lates calcarifer). The experiment used juvenile barramundi produced from a single spawning that were challenged consecutively by immersion with a redspotted grouper nervous necrosis virus (RGNNV) isolate. The dose and environmental conditions (35 ppt salinity and 30 °C) were constant. Fish and water were sampled longitudinally for histopathology and RT‐qPCR analysis to examine the evolution of the disease, virus replication, immune response and release of virus into water. Viral nervous necrosis (VNN) disease occurred in barramundi challenged at 3 and 4 weeks of age while fish challenged at 5, 7 and 9 weeks of age developed subclinical infection. Replication of NNV occurred faster and the concentration of virus reached higher concentrations in the younger fish with clinical disease. Virus isolation and qPCR tests indicated that infectious NNV was released from carcasses into water when fish were affected with clinical disease but not when NNV infection was subclinical. Based on these observations, we consider that carcasses from clinically infected fish have a potentially important role in the horizontal transmission of NNV, and barramundi juveniles should be protected from exposure to NNV until they are 5 weeks of age and reach the disease resistance threshold.     

Identification and pathogenicity study of emerging fish pathogens Acinetobacter junii and Acinetobacter pittii recovered from a disease outbreak in Labeo catla (Hamilton, 1822) and Hypophthalmichthys molitrix (Valenciennes, 1844) of freshwater wetland in West Bengal,India

The disease outbreaks in aquaculture system of wetlands are the major cause of fish mortality. Among various bacterial septicaemic diseases, fish mortality caused by Acinetobacter spp. is recently reported in different fish species. Fish disease outbreak was investigated in a wetland of West Bengal, India to identify the aetiological factors involved. The moribund fish were examined and subjected to bacterial isolation. Two bacterial causative agents were identified as Acinetobacter junii and Acinetobacter pittii by biochemical characterization and 16S rRNA gene amplification. Both the isolates were oxidase‐negative, nitrate‐negative, catalase‐positive and indole‐negative. The molecular identification using 16S rRNA gene sequencing and phylogenetic tree analysis further confirmed the two Acinetobacter spp. with 97%–99% similarity. The antibiotic resistance patterns of these two bacteria revealed that both of them were resistant to β‐lactam, cefalexin, cephalothin, amoxyclav, cefuroxime, cefadroxil, clindamycin, vancomycin and penicillin. In addition, A. pittii was also resistant to other antibiotics of cephams group such as ceftazidime and cefotaxime. In the challenge experiment, both A. junii and A. pittii were found to be pathogenic with LD₅₀ of 1.24 × 10⁵ and 1.88 × 10⁷ cfu/fish respectively. Histopathological examination of gill, liver and kidney revealed prominent changes supporting bacterial septicaemia. The investigation reports for the first time on concurrent infection by A. junii and multidrug‐resistant (MDR)‐A. pittii as emerging fish pathogens to cause severe mortality in Labeo catla and Hypophthalmichthys molitrix in a freshwater wetland.     

Impact of farm management on expression of early mortality syndrome/acute hepatopancreatic necrosis disease (EMS/AHPND) on penaeid shrimp farms in Thailand

Asian shrimp farming industry has experienced massive production losses due to a disease caused by toxins of Vibrio bacteria, known as early mortality syndrome/acute hepatopancreatic necrosis disease (EMS/AHPND) for the last 5 years. The disease can cause up to 100% cumulative pond mortality within a week. The objective of this study was to identify factors associated with AHPND occurrence on shrimp farms. A case–control study was carried out on shrimp farms in four provinces of Thailand. Factors related to farm characteristics, farm management, pond and water preparation, feed management, post‐larvae (PL) shrimp and stock management were evaluated. Multivariable logistic regression analysis identified factors affecting AHPND occurrence at the pond level. Chlorine treatment, reservoir availability, use of predator fish in the water preparation, culture of multiple shrimp species in one farm and increased PL stocking density contributed to an increased risk of AHPND infection, while delayed first day of feeding, polyculture and water ageing were likely to promote outbreak protection. Additionally, the source of PL was found to be associated with AHPND occurrence in shrimp ponds, which requires further study at the hatchery level. Identification of these factors will facilitate the development of effective control strategies for AHPND on shrimp farms.     

Host,agent and environment interactions affecting Nervous necrosis virus infection in Australian bass Macquaria novemaculeata

Australian bass Macquaria novemaculeata were challenged by immersion with nervous necrosis virus (NNV) at different ages and under controlled conditions to investigate factors affecting disease expression. Fish challenged at 3 weeks of age with 10³ TCID₅₀/ml and higher doses developed clinical disease; a lower dose of 10² TCID₅₀/ml resulted in incidence below 100% and 10¹ TCID₅₀/ml was insufficient to cause infection. Additionally, fish were challenged at 5, 6 and 13 weeks of age at 17 and 21°C to assess the role of the age of the host and water temperature on disease expression. Although Australian bass challenged at all ages had evidence of replication of NNV, only those challenged at 3 weeks of age (20 and 24 days post?hatch [dph]) developed clinical disease. Higher water temperature had an additive effect on disease expression in larvae challenged at 24 dph, but it did not affect the disease outcome in older fish. Finally, isolates of NNV derived from fish with clinical or subclinical disease presentations caused similar cumulative mortality and clinical signs when larvae at 24 dph were challenged, suggesting that agent variation was not responsible for variation in clinical presentation in these field outbreaks of NNV infection.     

Infection experiments with Aphanomyces invadans in four species of estuarine fish

Along the eastern seaboard of the US, Atlantic menhaden, Brevoortia tyrannus, develop characteristic ulcerative lesions, a condition termed ulcerative mycosis. These lesions are identical to those seen across Asia in fish affected by epizootic ulcerative syndrome, a condition caused by the fungus-like oomycete Aphanomyces invadans. Young-of-the-year menhaden inhabiting estuarine environments are the primary species affected in the USA and little is known about the factors involved in the initiation of the lesions, or why menhaden are predominantly infected. Atlantic menhaden, hogchoker, Trinectus maculatus, striped killifish, Fundulus majalis, and mummichog, Fundulus heteroclitus, were inoculated with A. invadans (80 zoospores per fish) to explore species differences in infection and lesion development. All four species developed lesions. Killifish developed frank lesions similar to those observed in menhaden but the gross lesions occurred later, approximately 5-10 days after those on menhaden. Hogchoker and mummichog did not develop gross skin ulcers; rather, their lesions appeared as reddened areas under the epidermis. Mummichogs also showed evidence of significant healing with a well-developed granuloma and significant myocyte regeneration. These experiments show that species barriers as well as ecological barriers can explain some of the factors involved in the development of lesions in, and specificity of the water mould for, menhaden.     

Characterization of a megalocytivirus from cultured rock bream,Oplegnathus fasciatus (Temminck & Schlege), in China

Megalocytivirus belongs to the Iridoviridae family and is known to affect finfish. Megalocytivirus epizootics have been reported to occur in several cultured fish species in China; however, no megalocytivirus associated with rock bream, Oplegnathus fasciatus (Temminck & Schlege), has been documented. In this study, we characterized for the first time a megalocytivirus, rock bream iridovirus (RBIV)‐C1, detected in cultured rock bream in a fish farm in China that had been inflicted with a high‐mortality disease outbreak. Sequence analysis of three conserved genes showed that RBIV‐C1 shares over 90% overall identities with a number of known megalocytiviruses. Electron microscopic examination revealed RBIV‐C1 as hexagonal particles similar to those reported for megalocytiviruses. In vivo infection study indicated that, following inoculation into rock bream, RBIV‐C1 induced 100% mortality and upregulated the expression of Mx, IL‐1β and IL‐8. The infected fish exhibited pathological signs similar to those observed in naturally diseased fish. Furthermore, studies in a turbot (Scophthalmus maximus L.) model indicated that RBIV‐C1 induced acute infection in turbot that led to 100% mortality. These results indicate that RBIV‐C1 is highly virulent to rock bream as well as turbot and that RBIV‐C1 is closely related to a number of previously reported megalocytivirus and likely a genetic variant of the latter.