Case report: Blindness associated with Learedius learedi trematode infection in a green sea turtle, Chelonia mydas, of the northern Red Sea.

Spirorchiid blood flukes are widespread in sea turtles, causing disease and mortality in their populations, with high prevalence in several ocean basins. Besides being leading parasitic causes of sea turtle strandings in several parts of the world, these infectious agents can cause endocarditis, vasculitis, thrombosis, miliary egg granulomas, and aneurysms, which ultimately may compromise the survival of green sea turtles. More severe cases may also result in multifocal granulomatous meningitis or pneumonia, both of which can be fatal. Herein, we report the first case of severe trematode infection, Caused by Learedius learedi, in a green sea turtle in the northern Red Sea; this infection is associated with bilateral blindness. Necropsy revealed multiple granulomas with intralesional trematode eggs in the optic nerve, eyes, spleen, heart, and lungs. The parasite was identified as Learedius learedi through specific primers of the ribosomal genome and COI sequences obtained from GenBank. Altogether, these findings emphasize the importance of recognizing the systemic nature of this particular fluke infection to ultimately protect the lives of these marine animals and ensure the sustainability of these species in the wild.

Low prevalence of Contracaecum third-stage larvae parasitizing Sea of Galilee fisheries: 1-year survey after 57 years of no information.

Freshwater and marine ecosystems are a suitable habitat for parasitic nematodes of the genus Contracaecum (family: Anisakidae) to complete their complex life cycle. Several fish species of the Sea of Galilee (Lake Kinneret) were reported in 1964 as second intermediate/paratenic hosts of Contracaecum spp. larvae. The lack of taxonomically relevant morphological features of these larvae hindered their proper identification. Here we report the results of a 1-year survey conducted in 2021, 57 years after the first (and only) such survey. We analyzed 352 specimens from 10 fish species (native and non-native) of the Sea of Galilee (Israel) ichthyofauna. We compared our results with those of the first parasitological survey conducted by Paperna in 1964; the overall prevalence of nematodes referable to Contracaecum larvae was 16.8% and 0.85% in 1964 and in 2021, respectively. Different from the first survey that identified Contracaecum larvae morphologically, we used both morphological and molecular tools. Two wild native cyprinids-Jordan himri (Carasobarbus canis) and Jordan barbel (Luciobarbus longiceps)-were infected (a single specimen each) with Contracaecum quadripapillatum larvae in their abdominal cavity. A single specimen of blue tilapia (Oreochromis aureus) was infected with two larvae of Contracaecum multipapillatum E, localized in the pericardial cavity. The findings of our study, which is part of a large project focused on Contracaecum spp. infecting both piscivorous birds and fish collected in Israel, advance our knowledge about the distribution and host range of this potentially zoonotic parasite in fishery products of the Sea of Galilee.

Identification of Mycobacterium pseudoshottsii in the Eastern Mediterranean.

Among the numerous pathogenic nontuberculous mycobacteria (NTM), which may cause disease in both poikilothermic and homoeothermic organisms, members of the unique clade Mycobacterium ulcerans/Mycobacterium marinum (MuMC) may cause disease in both fish and humans. Here, we describe the emergence of Mycobacterium pseudoshottsii, one of the four MuMC members, in Israel. For many years, M. marinum was the dominant NTM that was diagnosed in Israel as a fish pathogen. To the best of our knowledge, this is the first isolation and genomic characterization of M. pseudoshottsii infecting edible fish from two different fish species farmed in offshore sea cages in the eastern Mediterranean as well as in a recirculating aquaculture system in Israel. We compared the M. pseudoshottsii whole-genome sequences to all available genomic sequences of MuMC in free, publicly accessible databases. IMPORTANCE Mycobacterium pseudoshottsii was first detected in 1997 in the USA, infecting wild striped bass (Morone saxatilis). Since then, several reports from different countries worldwide have shown its capacity to become established in new regions as well as its pathogenicity to saltwater and euryhaline finfish of different genera. Our phylogenetic analysis revealed that the Mycobacterium ulcerans/Mycobacterium marinum clade (MuMC) is divided into two main branches: one that includes M. marinum and M. pseudoshottsii, and the second, which includes other M. marinum isolates as well as two isolates of M. shottsii. Our results reinforce the proposition that the geographical distribution of M. pseudoshottsii is much more extensive than is commonly believed. The emergence of M. pseudoshottsii in different parts of the world and its pathogenic traits that affect finfish of different genera may be a cause for concern among fish farmers, researchers, and environmental organizations.

First report on the serum chemistry and haematology of free-ranging dusky (Carcharhinus obscurus) and sandbar (Carcharhinus plumbeus) sharks in the eastern Mediterranean Sea.

Shark assessments in the Mediterranean Sea are still scarce, and serum chemistry and haematological data have yet to be reported for wild dusky (Carcharhinus obscurus) or sandbar (Carcharhinus plumbeus) shark populations in the Mediterranean Sea. Herein, blood samples were obtained from adult dusky (n = 23) and sandbar (n = 14) sharks from an aggregation site near the Hadera power and desalination plants in Israel in the winters of 2016-20. Several serum chemistry analytes were characterized with relation to stress, body size and environmental conditions. Glucose concentrations were higher, while total cholesterol concentrations were lower in dusky sharks than in sandbar sharks, potentially due to distinct metabolic pathways utilized during the capture-related activity by both species. However, differences in sex and size are noted and should be considered. The blood cell morphology of both species was consistent with previous findings for sandbar sharks. Atypical monocytes were noted in one dusky shark. Preliminary and exploratory reference intervals for female dusky sharks were calculated for glucose, triglycerides, total cholesterol, total protein and creatine kinase. These data must be viewed with caution due to the potential influence of capture-related stress on analyte concentrations and activities and the fact that only females were employed in the calculations. Moreover, the sampling site is adjacent to coastal power and desalination plants, which may significantly affect shark physiology. Although limited, this novel database on dusky and sandbar shark serum chemistry and haematology aspects is essential as a first attempt to obtain data on these species in the eastern Mediterranean Sea and for future conservation and long-term biomonitoring efforts.

First description of a Gammaherpesvirus in a common dolphin (Delphinus delphis) from the Eastern Mediterranean Sea.

In September 2020, a male common dolphin (Delphinus delphis) was found dead on a beach near Bat-Yam, Israel. A small, raised, well circumscribed penile lesion (i.e., mass) was identified and removed for histology and molecular characterizations. By histology, the penile mass presented focal keratinization of the squamous epithelium and a mild ballooning of acanthocytes in lower epithelium levels, as well as features compatible with viral plaques, and tested positive for a gammaherpesvirus through molecular characterization analyses. Tissue samples from the lungs, liver, and spleen, however, tested negative for herpesvirus infection. The gammaherpesvirus detected herein is similar to other isolates found in several areas worldwide in different cetacean species. This is the first reported case of gammaherpesvirus infection in dolphins from the eastern Mediterranean Sea, indicative of the need for long-term assessments to create viral infections databases in cetaceans, especially in a climate change context, which is likely to intensify infectious disease outbreaks in marine mammals in the future.

Preliminary study of shark microbiota at a unique mix-species shark aggregation site, in the Eastern Mediterranean Sea.

Sharks, as apex predators, play an essential ecological role in shaping the marine food web and maintaining healthy and balanced marine ecosystems. Sharks are sensitive to environmental changes and anthropogenic pressure and demonstrate a clear and rapid response. This designates them a "keystone" or "sentinel" group that may describe the structure and function of the ecosystem. As a meta-organism, sharks offer selective niches (organs) for microorganisms that can provide benefits for their hosts. However, changes in the microbiota (due to physiological or environmental changes) can turn the symbiosis into a dysbiosis and may affect the physiology, immunity and ecology of the host. Although the importance of sharks within the ecosystem is well known, relatively few studies have focused on the microbiome aspect, especially with long-term sampling. Our study was conducted at a site of coastal development in Israel where a mixed-species shark aggregation (November-May) is observed. The aggregation includes two shark species, the dusky (Carcharhinus obscurus) and sandbar (Carcharhinus plumbeus) which segregate by sex (females and males, respectively). In order to characterize the bacterial profile and examine the physiological and ecological aspects, microbiome samples were collected from different organs (gills, skin, and cloaca) from both shark species over 3 years (sampling seasons: 2019, 2020, and 2021). The bacterial composition was significantly different between the shark individuals and the surrounding seawater and between the shark species. Additionally, differences were apparent between all the organs and the seawater, and between the skin and gills. The most dominant groups for both shark species were Flavobacteriaceae, Moraxellaceae, and Rhodobacteraceae. However, specific microbial biomarkers were also identified for each shark. An unexpected difference in the microbiome profile and diversity between the 2019-2020 and 2021 sampling seasons, revealed an increase in the potential pathogen Streptococcus. The fluctuations in the relative abundance of Streptococcus between the months of the third sampling season were also reflected in the seawater. Our study provides initial information on shark microbiome in the Eastern Mediterranean Sea. In addition, we demonstrated that these methods were also able to describe environmental episodes and the microbiome is a robust measure for long-term ecological research.

Two Decades of Coastal Dolphin Population Surveys in Israel, Eastern Mediterranean.

Along the Mediterranean coast of Israel, two near-shore dolphin species are prevalent; Tursiops truncatus (least concern, IUCN) and Delphinus delphis (endangered, IUCN). Ship-board surveys and sporadic sightings over the last two decades have shown that the two differ in distribution-T. truncatus is found along the entire coast and D. delphis only in the south. The environmental and anthropological factors affecting these species' spatial distribution and determining their habitat preferences in this area are largely unknown. This work is a first attempt at summarizing 20 years of observations and studying habitat preferences for both species, by use of Generalized Additive Models. T. truncatus was found to be present in all areas of the continental shelf where survey effort coverage was sufficient, with a high affinity towards bottom trawlers. Model results showed D. delphis distribution to be associated to (shallow) water depths, though the factors driving their limited latitudinal distribution currently remain unknown. It is evident that T. truncatus and D. delphis are present in segregated areas of the Israeli continental shelf and T. truncatus currently sustains a delicate balance with continuously shifting human activities, while the drivers of D. delphis distribution are more specified, yet still not fully understood.

Photobacterium damselae subspecies damselae Pneumonia in Dead, Stranded Bottlenose Dolphin, Eastern Mediterranean Sea.

Photobacterium damselae subspecies damselae, an abundant, generalist marine pathogen, has been reported in various cetaceans worldwide. We report a bottlenose dolphin in the eastern Mediterranean Sea that was found stranded and dead. The dolphin had a severe case of chronic suppurative pneumonia and splenic lymphoid depletion caused by this pathogen.

Unraveling Metabolite Provisioning to Offspring Through Parental Fluids: A Case Study of the Brazilian Guitarfish, Pseudobatos horkelii.

Elasmobranchs have a very distinct metabolism, and many aspects related to the energetic dynamics of these animals remain poorly investigated. The reproductive period is particularly vulnerable for viviparous species, as part of the energy reserves of the parental biomass is reallocated for gamete production and embryo development. In this context, this study aimed to characterize parental metabolite provisioning to the offspring (both sperm and developing embryos) of the Brazilian Guitarfish, Pseudobatos horkelii, through glucose, ß-hydroxybutyrate, triglycerides, and total cholesterol determinations in the uterine liquid (UL) and serum of pregnant females and in the seminal fluid (SF) and serum of males during the copulation period. No significant difference was observed for the analyzed markers between the UL and SF. Except for triglycerides, higher in female serum samples, all other energy markers were present at similar concentrations in the serum of both females and males. When comparing female UL and serum, significant differences were observed for triglycerides and total cholesterol. No differences were observed between SF and serum in males. The results indicate that all markers are being made available to offspring, possibly complementary to the yolk in the case of maternal liquid, and as an additional source for sperm mobilization required during egg fertilization in the case of the paternal fluid. Correlations between the markers in the parental matrices were also noted, compatible with the metabolic pathways activated during energy mobilization in vertebrates. Moreover, distinct marker predominance patterns were also noted for both UL and SF. Energy mobilization characterization directed to offspring through parental fluids aids in unraveling metabolic dynamics during the reproduction stage while also providing support for stress physiology studies to evaluate the indirect effects of parental allostatic overload in both sperm and developing embryos. Finally, energy mobilization assessments of parental fluids may also help elucidate how internal fertilization and viviparity evolved in this very distinct taxonomic group.

Morphological description and molecular characterization of Contracaecum larvae (Nematoda: Anisakidae) parasitizing market-size hybrid tilapia (Oreochromis aureus x Oreochromis niloticus) and red drum (Sciaenops ocellatus) farmed in Israel.

Nematodes belonging to the genus Contracaecum (family: Anisakidae) are heteroxenous parasites with a complex life cycle. Contracaecum larvae infecting farmed fish and fishery products are economically important causing market rejection in massive infection and may have zoonotic potential. In Israel, Contracaecum larvae have been described morphologically in several fish species; however, none of these descriptions were supported by molecular tools. In 2019-2020, hybrid tilapia (Oreochromis aureus x Oreochromis niloticus) and red drum (Sciaenops ocellatus), farmed in polyculture were found to be heavily infected with nematodes referable to Contracaecum larvae. Prevalence of infection in hybrid tilapia and red drum was 53.8% and 40.9%, respectively. A combined (morphological and molecular) approach revealed that both infected fish species were parasitized by the same species of Contracaecum, although larvae in hybrid tilapia were localized in the pericardial cavity whereas in red drum, they were observed in the abdominal cavity. Genetic analysis of internal transcribed spacer rDNA and cox2 mtDNA showed high similarity to unidentified Contracaecum larvae detected in several fish species in Ethiopia, Egypt and Kenya. In this study, molecular and morphological analyses place the possible new species in the C. multipapillatum complex and was provisionally named C. multipapillatum E. Further analyses combining morphological and molecular approaches are required on adult specimens collected from piscivorous birds living in the same area to support the identification of a potentially new species.

A survey of arsenic, mercury, cadmium, and lead residues in seafood (fish, crustaceans, and cephalopods) from the south-eastern Mediterranean Sea.

Seafood is capable of bioaccumulating heavy metals (HM), making it a potentially major dietary source of HM for humans. Presently, little data exists on seafood from the eastern-most boundary of the Mediterranean Sea. This study aims to provide exposure insight of the Israeli population to HM through the consumption of locally caught seafood by assessing the levels of arsenic, mercury, cadmium, and lead in raw tissues of seafood. A wide survey of local fisheries was conducted providing 296 samples from 11 different species, including seven fish, two crustacean, and two cephalopod species. Total arsenic, cadmium, and lead were analyzed by graphite-furnace atomic absorption. Total mercury was measured by cold-vapor mercury analyzer. Arsenic speciation was performed by anion chromatography-inductively coupled plasma sector field mass spectrometry. Results suggested that the total arsenic concentrations were significantly higher in crustaceans and cephalopods than fish. Arsenic speciation revealed two samples that exceed 1 mg/kg of inorganic arsenic, whereas methylated arsenic was below the detection limit. Elevated mercury levels were detected in the commercial benthic species Mullus barbatus (red mullet), cadmium was detected in one-third of the samples, and lead detected in eight samples. Comparing the results to health guidelines, 99.4% of seafood tested in this study abide with acceptable levels of heavy metals in seafood, as defined by both Israeli and European Union guidelines.

Physiological Impairment as a Result of Bile Accumulation in an Apex Predator, the Tiger Shark (Galeocerdo cuvier Péron & Lesueur, 1822).

Physiological adaptations have evolved to help sharks face rapid periods of feast. Tiger sharks are generalist apex predators that rely on a high-lipid/protein diet. To achieve a satisfactory nutritional condition, proper lipid absorption and hepatic storage are needed. Bile secretion in sharks is low and sporadic but increases during short periods of fasting. The present study describes a physiological impairment caused by bile accumulation in juvenile tiger sharks, possibly due to prolonged fasting. These evidences suggest that, even though sharks have adaptations that prevent them from dying from starvation, alarming physiological alterations might occur. Future studies are needed to assess how such a condition can affect wild populations, as well as possible sublethal consequences that could impact their long-term survival.

Multilocus sequence analysis of diverse Streptococcus iniae isolates indicates an underlying genetic basis for phenotypic heterogeneity.

Streptococcus iniae is a Gram-positive, opportunistically zoonotic bacterium infective to a wide variety of farmed and wild fish species worldwide. Outbreaks in wild fish can have detrimental environmental and cultural impacts, and mortality events in aquaculture can result in significant economic losses. As an emerging or re-emerging pathogen of global significance, understanding the coalescing factors contributing to piscine streptococcosis is crucial for developing strategies to control infections. Intraspecific antigenic and genetic variability of S. iniae has made development of autogenous vaccines a challenge, particularly where the diversity of locally endemic S. iniae strains is unknown. This study genetically and phenotypically characterized 11 S. iniae isolates from diseased wild and farmed fish from North America, Central America, and the Caribbean. A multilocus sequence analysis (MLSA) scheme was developed to phylogenetically compare these isolates to 84 other strains of Streptococcus spp. relevant to aquaculture. MLSA generated phylogenies comparable to established genotyping methods, and isolates formed distinct clades related to phenotype and host species. The endothelial Oreochromis mossambicus bulbus arteriosus cell line and whole blood from rainbow trout Oncorhynchus mykiss, Nile tilapia Oreochromis niloticus, and white sturgeon Acipenser transmontanus were used to investigate the persistence and virulence of the 11 isolates using in vitro assays. In vivo challenges using an O. niloticus model were used to evaluate virulence by the intragastric route of infection. Isolates showed significant differences (p < 0.05) in virulence and persistence, with some correlation to genogroup, establishing a basis for further work uncovering genetic factors leading to increased pathogenicity.

Mycobacteriosis in Aquatic Invertebrates: A Review of Its Emergence.

Mycobacteriosis is a chronic bacterial disease reported in aquatic and terrestrial animals, including humans. The disease affects a wide range of cultured and wild organisms worldwide. Mycobacteriosis is well-known in aquatic vertebrates (e.g., finfish, marine mammals), while in the last few years, reports of its presence in aquatic invertebrates have been on the rise, for both freshwater and marine species. The number of cases is likely to increase as a result of increased awareness, surveillance and availability of diagnostic methods. Domestication of wild aquatic species and the intensification of modern aquaculture are also leading to an increase in the number of reported cases. Moreover, climate changes are affecting fresh and marine aquatic ecosystems. The increasing reports of mycobacteriosis in aquatic invertebrates may also be influenced by global climate warming, which could contribute to the microbes' development and survival rates, pathogen transmission and host susceptibility. Several species of the genus Mycobacterium have been diagnosed in aquatic invertebrates; a few of them are significant due to their wide host spectrum, economic impact in aquaculture, and zoonotic potential. The impact of mycobacteriosis in aquatic invertebrates is probably underestimated, and there is currently no effective treatment other than facility disinfection. In this review, we provide an overview of the diversity of mycobacterial infections reported in molluscs, crustaceans, cnidarians, echinoderms and sponges. We highlight important issues relating to its pathological manifestation, diagnosis and zoonotic considerations.

Correction: Regev, Y., et al. Molecular Identification and Characterization of Vibrio Species and Mycobacterium Species in Wild and Cultured Marine Fish from the Eastern Mediterranean Sea. Microorganisms 2020, 8, 863.

We would like to change the authors' affiliation of paper [...].

Molecular Identification and Characterization of Vibrio Species and Mycobacterium Species in Wild and Cultured Marine Fish from the Eastern Mediterranean Sea.

In contrast to numerous documented pathogens and infectious diseases of aquaculture, there is a lack of baseline data and information regarding pathogenic agents' prevalence in wild marine fish populations. This study focused on two common fish pathogenic microorganisms, namely Mycobacterium species and Vibrio species, both of which are known to be major causes of fish loss, occasionally to the extent of being a limiting factor in fish production. Both microorganisms are known as zoonotic agents. In total, 210 wild marine indigenous and Lessepsian fish from four different species from the eastern Mediterranean Sea were sampled and tested for Vibrio species and Mycobacterium species during a two-year period (2016-2017). Using PCR with 16S rRNA primers, we detected different strain variations of Mycobacterium species and Vibrio species and, based on the sequencing results, the overall prevalence for Vibrio species in wild fish in 2016 was significantly higher compared to 2017. No significant difference was detected for Mycobacterium species prevalence in wild fish between 2016 and 2017. In addition, 72 gilthead seabream (Sparus aurata) from an Israeli offshore marine farm were also examined during the two-year period (2017-2018). The results suggest that Mycobacterium species prevalence was significantly higher in 2018, while in 2017 there was no positive results for Mycobacterium species. In addition, there was no significant difference between both years in regard to the prevalence of Vibrio species for maricultured fish. These results highlight the necessity of continuous molecular monitoring in order to evaluate the prevalence of pathogenic microorganisms in both wild and cultured fish populations.

Indigenous versus Lessepsian Hosts: Nervous Necrosis Virus (NNV) in Eastern Mediterranean Sea Fish.

Viruses are among the most abundant and diverse biological components in the marine environment. In finfish, viruses are key drivers of host diversity and population dynamics, and therefore, their effect on the marine environment is far-reaching. Viral encephalopathy and retinopathy (VER) is a disease caused by the marine nervous necrosis virus (NNV), which is recognized as one of the main infectious threats for marine aquaculture worldwide. For over 140 years, the Suez Canal has acted as a conduit for the invasion of Red Sea marine species into the Mediterranean Sea. In 2016-2017, we evaluated the prevalence of NNV in two indigenous Mediterranean species, the round sardinella (Sardinella aurita) and the white steenbras (Lithognathus mormyrus) versus two Lessepsian species, the Randall's threadfin bream (Nemipterus randalli) and the Lessepsian lizardfish (Saurida lessepsianus). A molecular method was used to detect NNV in all four fish species tested. In N. randalli, a relatively newly established invasive species in the Mediterranean Sea, the prevalence was significantly higher than in both indigenous species. In S. lessepsianus, prevalence varied considerably between years. While the factors that influence the effective establishment of invasive species are poorly understood, we suggest that the susceptibility of a given invasive fish species to locally acquired viral pathogens such as NVV may be important, in terms of both its successful establishment in its newly adopted environment and its role as a reservoir 'host' in the new area.

Specific pathogens and microbial abundance within liver and kidney tissues of wild marine fish from the Eastern Mediterranean Sea.

This study is an initial description and discussion of the kidney and liver microbial communities of five common fish species sampled from four sites along the Eastern Mediterranean Sea shoreline. The goals of the present study were to establish a baseline dataset of microbial communities associated with the tissues of wild marine fish, in order to examine species-specific microbial characteristics and to screen for candidate pathogens. This issue is especially relevant due to the development of mariculture farms and the possible transmission of pathogens from wild to farmed fish and vice versa. Although fish were apparently healthy, 16S rRNA NGS screening identified three potential fish bacterial pathogens: Photobacterium damselae, Vibrio harveyi and Streptococcus iniae. Based on the distribution patterns and relative abundance, 16 samples were classified as potential pathogenic bacteria-infected samples (PPBIS). Hence, PPBIS prevalence was significantly higher in kidneys than in liver samples and variation was found between the fish species. Significant differences were observed between fish species, organs and sites, indicating the importance of the environmental conditions on the fish microbiome. We applied a consistent sampling and analytical method for monitoring in long-term surveys which may be incorporated within other marine fish pathogens surveys around the world.

Fatal Infection in a Wild Sandbar Shark (Carcharhinus plumbeus), Caused by Streptococcus agalactiae, Type Ia-ST7.

Streptococcus agalactiae is one of the most important fish pathogenic bacteria as it is responsible for epizootic mortalities in both wild and farmed species. S. agalactiae is also known as a zoonotic agent. In July 2018, a stranded wild sandbar shark (Carcharhinus plumbeus), one of the most common shark species in the Mediterranean Sea, was found moribund on the seashore next to Netanya, Israel, and died a few hours later. A post-mortem examination, histopathology, classical bacteriology and advanced molecular techniques revealed a bacterial infection caused by S. agalactiae, type Ia-ST7. Available sequences publicly accessible databases and phylogenetic analysis suggest that the S. agalactiae isolated in this case is closely related to fish and human isolates. To the best of our knowledge, this is the first description of a fatal streptococcosis in sandbar sharks.

Prevalence of nervous necrosis virus (NNV) and Streptococcus species in wild marine fish and crustaceans from the Levantine Basin, Mediterranean Sea.

Infectious diseases in marine animals have ecological, socio-economic and environmental impacts. Nervous necrosis virus (NNV) and Streptococcus iniae have become major threats to marine aquaculture and have been detected in morbid marine organisms worldwide. However, despite their importance, there is a lack of knowledge regarding the prevalence of these pathogens in wild fish species. Here we sampled indigenous and Lessepsian species from different trophic levels and different biological niches in the eastern Mediterranean. A total of 174 fish and 32 crustaceans were tested for S. iniae and a total of 195 fish and 33 crustaceans were tested for NNV. We found an overall prevalence of 9.71% Streptococcus spp. and 21.49% NNV in selected marine fish and crustaceans by PCR and qPCR. In fish, the zoonotic agent S. iniae was detected at a higher prevalence in kidney compared to liver tissue. Co-infection by both pathogens was detected only in 5 specimens. We also examined gilthead sea bream Sparus aurata from an Israeli offshore marine farm during the grow-out period, in order to assess the possibility of horizontal pathogen transmission from wild to maricultured fish. Three out of 15 (20%) fish were found to be NNV positive after 120 d in the sea, suggesting spontaneous transmission from wild to farmed fish. Our findings suggest that more surveys should be conducted, especially in areas were mariculture farms are planned to be established.