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In 2021, White Trevally or Striped Jack cultured in the western part of Japan exhibited mild, but chronic mortalities from late September through early October. The cumulative mortality rate was approximately 0.02% per a net pen containing approximately 50,000 fish. Although the cumulative mortality rate was not high, most of the fish in net pens showed characteristic gross signs and an abnormal swimming behaviour. The body of diseased fish became pale and the yellow lines on the lateral sides of fish body became darken. In addition, silver lines along the dorsal fin became apparent. Loss of schooling behaviour was noted during the mortality event. In addition, affected fish became lethargic and failed to swim against current, or frequently stopped swimming and sank to the bottom of net pens after feeding. The goal of this study was to identify the cause of the mortality event. To achieve the goal, we used histopathology and metatranscriptome analysis. Histopathological examination revealed that xenoma of microsporidian were frequently observed in the nerve axon in the brain and spinal cord. Spores observed in the sections were stained with a fluorescent dye, Uvitex 2B, indicating those spores are microsporidian. The data from metatranscriptome analysis indicated that the microsporidian is Spraguea sp. The microsporidian was frequently detected from diseased fish with similar symptoms collected in the same region, suggesting that the microsporidian was highly associated with abnormal swimming behaviour of fish., (© 2024 John Wiley & Sons Ltd.)

Melanized focal changes (MFCs) in the fillet of farmed Atlantic salmon is a major quality concern. The changes are thought to initially appear as acute red focal changes (RFCs) that progress into chronic MFCs. Recent findings have indicated that hypoxia may be important in their development, possibly leading to necrosis affecting not only myocytes but also adipocytes. Thus, the aim of this study was to investigate possible hypoxic conditions in RFCs and the subsequent inflammatory responses and lesions in the adipose tissue in RFCs and MFCs. A collection of RFCs, MFCs and control muscle samples from several groups of farmed salmon was studied. Using immunohistochemistry, we found induction of the hypoxia-inducible factor 1 pathway in RFCs. Histological investigations of RFCs and MFCs revealed different stages of fat necrosis, including necrotic adipocytes, a myospherulosis-like reaction and the formation of pseudocystic spaces. Accumulations of foamy macrophages were detected in MFCs, indicating degradation and phagocytosis of lipids. Using in situ hybridization, we showed the presence of tyrosinase- and tyrosinase-related protein-1-expressing amelanotic cells in RFCs, which in turn became melanized in MFCs. In conclusion, we propose a sequence of events leading to the formation of MFCs, highlighting the pivotal role of adiposity, hypoxia and fat necrosis., (© 2024 The Author(s). Journal of Fish Diseases published by John Wiley & Sons Ltd.)

In the past decade, the increasing distribution of pollutants in the aquatic environment has been observed, causing integrative effects on fish. Likewise, due to anthropogenic activities, the southern gulf of Lake Tana is an impacted region, and the production of Nile tilapia fish is reduced. For this reason, the aim of this study was to conduct a histopathological-based study of 48 Nile tilapia fishes' health status at the southern gulf of Lake Tana and aquaculture using a cross-sectional study from February 2023 to May 2023. The study evaluated the histopathology of the gill, liver, gonads, and spleen organs using descriptive statistics accompanied by a 2 × 2 contingency table and t-test analysis. During the study, different histological alterations were detected, and the numbers of fish affected by a specific histological alteration were presented as percentage prevalence; hence, from the total fish examined, hyperplasia (54.15%), followed by pigment deposits (52%), hemorrhage (50%), and immune cell infiltration (50%), respectively, were the most frequently detected alterations. However, Nile tilapias from the southern gulf of Lake Tana were 1.4 (odds ratio) times more likely to show histopathological alterations than those from aquaculture, although statistically, was not significant (p > 0.05). In addition, the study found the mean value of the fish index (95.3) and regressive indices of the gill (13.6), liver (14.8), and gonad (12.3); moreover, the inflammatory indices of the spleen organ (11.3) and mean severity grade value of the gill (2.35) and gonad (1.7) organs, respectively, were obtained from the southern gulf of Lake Tana, and all those values were significantly higher (p < 0.05) from this site as compared to the aquaculture. In general, it has been found that tilapias from the southern gulf of Lake Tana showed higher pathological severity as compared with aquaculture. Among the four target organs evaluated, liver organs were observed to be the most damaged, while gonads were the least impacted organs. Therefore, it has been concluded that tilapia fish are living in abnormal conditions, so to ensure a sustainable fishery, water pollutant sources from Bahirdar city must receive proper attention, and future studies should consider age differences, seasonal variation, and the detection of specific pollutants., (© 2024. The Author(s).)

Gastric and intestinal mucosal hyperplasia and polyps are identified as a cause of morbidity and mortality in moray eels. This report describes the clinical presentations, diagnostic procedures, and therapeutic interventions in eight moray eels diagnosed with gastric polypoid hyperplasia. All described cases were humanely euthanized or found deceased, and multifocal adenomatous hyperplasia and polyps extending from the gastric mucosal epithelium were identified in all cases. The moray eels diagnosed with adenomatous hyperplasia and polyps often exhibited anorexia, regurgitation, and occasional changes in buoyancy, and supportive care was unsuccessful in alleviating or resolving these signs.

The recent discovery of disease caused by Nucleospora braziliensis in Nile tilapia (Oreochromis niloticus) is important as it has highlighted the high prevalence of infection and associated mortality in cultured fish. Thus, this study conducted an experimental infection of this microsporidium to evaluate pathological alterations and conduct proteomic analysis. For pathological observation, samples of brain, eyes, gall bladder, gut, heart, kidney, liver, muscle, skin, spleen, and stomach tissue, were collected, and liquid chromatography-mass spectrometry (LC-MS/MS) was performed for proteomic analysis. The most prevalent lesions were brownish color of the liver, gill filament fusion, gut ischemia, hemorrhage of the lips and fins, hepatomegaly, spleen atrophy, splenomegaly, and stomach congestion. The most common microscopic lesions were degeneration, hemorrhage, and inflammation in the brain, gills, gut, kidney, liver, muscle, spleen, and stomach. The digested peptides were identified by LC-MS/MS and the intersection of each group showed that in the spleen there were 121 exclusive proteins in the infected sample and 252 in the control, while in the kidney, 129 proteins were identified in the infected specimen compared to 83 in the control. In conclusion, this study demonstrates the proteome profile of O. niloticus kidney and spleen tissue in response to infection with N. braziliensis., (© 2024. The Author(s), under exclusive licence to Springer Nature Switzerland AG.)

Fish meal (FM) replacement is essential for the sustainable expansion of aquaculture. This study focussed on the feasibility of replacing FM with a single-cell protein (SCP) derived from methanotrophic bacteria (Methylococcus capsulatus, Bath) in barramundi fry (Lates calcarifer). Three isonitrogenous and isoenergetic diets were formulated with 0%, 6.4% and 12.9% inclusion of the SCP, replacing FM by 0%, 25% and 50%. Barramundi fry (initial body weight 2.5 ± 0.1 g) were fed experimental diets for 21 days to assess growth performance, gut microbiome composition and gut histopathology. Our findings revealed that both levels of SCP inclusion induced detrimental effects in barramundi fry, including impaired growth and reduced survival compared with the control group (66.7% and 71.7% survival in diets replacing FM with SCP by 25% and 50%, respectively; p < .05). Both dietary treatments presented mild necrotizing enteritis with subepithelial oedema and accumulation of PAS positive, diastase resistant droplets within hepatocytes (ceroid hepatopathy) and pancreatic atrophy. Microbiome analysis revealed a marked shift in the gut microbial community with the expansion of potential opportunistic bacteria in the genus Aeromonas. Reduced overall performance in the highest inclusion level (50% SCP) was primarily associated with reduced feed intake, likely related to palatability issues, albeit pathological changes observed in gut and liver may also play a role. Our study highlights the importance of meticulous optimization of SCP inclusion levels in aquafeed formulations, and the need for species and life-stage specific assessments to ensure the health and welfare of fish in sustainable aquaculture practices., (© 2024 The Author(s). Journal of Fish Diseases published by John Wiley & Sons Ltd.)

Edwardsiella anguillarum, a highly virulent species within the Edwardsiella genus, causes significant mortality in milkfish farms in Taiwan. This study aimed to investigate the comparison of milkfish susceptibility, a newly identified host species in Taiwanese aquaculture, with other species Nile tilapia (Oreochromis niloticus) and Asian seabass (Lates calcarifer), to E. anguillarum, elucidating its pathogenicity across both seawater and freshwater aquaculture environments. The results showed milkfish exhibited the highest mortality rate of 85% within 48 h of infection, whereas Nile tilapia exhibited a mortality rate of 70% between the second- and tenth-day post challenge, and seabass exhibited a mortality rate of 25% between the second- and sixth-day post challenge. Gross lesions observed in milkfish included splenomegaly and haemorrhage, whereas Nile tilapia exhibited signs of ascites, exophthalmia and brain haemorrhage. Seabass displayed spleen granulomas and haemorrhage at the injection site. Histopathological analysis revealed common features across all three species, including multifocal necrosis, bacterial presence in the necrotic areas, serositis and oedema. Asian seabass also exhibited chronic lesions in the form of splenic granulomas. This study highlights the high susceptibility of milkfish and Nile tilapia to E. anguillarum, emphasizing the urgent need for further investigation into targeted vaccine development for these fish species. These results not only deepen our understanding of the differing levels of pathogenicity among the three species but also offer valuable insights for improving disease prevention and management strategies in aquaculture, including those applied within polyculture systems and for the maintenance of aquaculture water environments., (© 2024 John Wiley & Sons Ltd.)

Amyloodiniosis, caused by the ectoparasite Amyloodinium ocellatum, affects the healthy development of mariculture. This study used a local infection method to identify the pathogenic target organ responsible for the death of infected fish. Comparing the relationship between the abundance of trophonts in gills and skin with the mortality of infected fish using local infection showed that severe gill infections cause the mortality of infected fish. At the 40 % survival rate of infected fish, the parasite abundance in the gill was 14,167 ± 4371. The gill filaments of the infected fish were structurally disordered, with pronounced lesions associated with the presence of trophonts, such as epithelial cell degeneration and massive lymphocytic infiltration. However, the skin showed no obvious pathological changes. The TUNEL assay showed a significant presence of apoptotic cells concentrated in the area of A. ocellatum infection. The trophonts on the gills developed faster than those parasitising the skin and fins. Microbiome analysis revealed that at the phylum level, Proteobacteria, Bacteroidota, and Firmicutes are abundant in the skin, while Verrucomicrobiota, Bacteroidota, and Proteobacteria are abundant in the gills of A. latus. Furthermore, A. ocellatum infection significantly reduced (p < 0.05) the richness and diversity of the gill microbial community of A. latus. Infection by A. ocellatum increased the relative abundance of several putative pathogenic bacteria (Flavobacterium and Nocardia) in the gill and skin of A. latus, possibly increasing the likelihood of disease in the host. In conclusion, these results evidenced that severe gill infections by A. ocellatum cause mortality in infected fish, which clarifies the direction for exploring the pathogenesis of amyloodiniosis., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024. Published by Elsevier Ltd.)

There is a claimed increase in the global prevalence and incidence of emerging diseases observed in many organisms. Myxozoa represents an essential group of metazoan parasites that hold both economic and ecological significance. In the current study, 1% of the fish population at two commercial goldfish (Carassius auratus) farms in Tehran and Ghom province, Iran, developed cavitating muscular lesions resembling humps in February 2021 and January 2022. Fish displaying pathological abnormalities were transported to the Ornamental Fish Clinic and subjected to clinical examination. Light microscopy was subsequently used to examine wet smears of skin and gills, as well as whitish exudate. In addition, tissue homogenates were collected for more precise identification and molecular confirmation. The study discovered that individuals from the goldfish farms were infected with the pathogenic myxozoan Myxobolus lentisuturalis, which caused significant damage to the epaxial muscles. The spores collected from the humps had a lack of uniformity and were primarily ellipsoidal in shape. Histopathological analysis also revealed parasites in various stages of development, such as plasmodia and spores, as well as inflammatory cell infiltration (macrophage, giant cell and lymphoplasmacytic infiltration) between skeletal muscle fibers. Phylogenetic analysis of M. lentisuturalis was performed by using MEGA 11 and the maximum likelihood method. M. lentisuturalis is a myxozoan parasite that has been sparsely recorded and lacks widespread recognition. The current study is the first clinical, histopathological, and molecular characterization of M. lentisuturalis isolated from the skeletal musculature of goldfish (C. auratus) in Iran., (© 2024. The Author(s).)

Choline is recognized as an essential nutrient for Atlantic salmon at all developmental stages. However, its dietary requirement is not well defined. Choline plays a critical role in lipid transport, and the clearest deficiency sign is intestinal steatosis. The present work, aiming to find whether lipid source and fish size may affect steatosis symptoms, was one of a series of studies conducted to identify which production-related conditions may influence choline requirement. Six choline-deficient diets were formulated varying in ratios of rapeseed oil to fish oil and fed to Atlantic salmon of 1.5 and 4.5 kg. After eight weeks, somatic characteristics were observed, and the severity of intestinal steatosis was assessed by histological, biochemical, and molecular analyses. Fatty acid composition in pyloric intestine, mesenteric tissue, and liver samples was also quantified. The increasing rapeseed oil level increased lipid digestibility markedly, enhancing lipid supply to the fish. Moreover, small fish consumed more feed, and consequently had a higher lipid intake. In conclusion, the results showed that choline requirement depends on dietary lipid load, which depends on the fatty acid profile as well as the fish size., (© 2024. This is a U.S. Government work and not under copyright protection in the US; foreign copyright protection may apply.)

Background: Understanding the relationship between resident microbiota and disease in cultured fish represents an important and emerging area of study. Marine gill disorders in particular are considered an important challenge to Atlantic salmon (Salmo salar) aquaculture, however relatively little is known regarding the role resident gill microbiota might play in providing protection from or potentiating different gill diseases. Here, 16S rRNA sequencing was used to examine the gill microbiome alongside fish health screening in farmed Atlantic salmon. Results were used to explore the relationship between microbial communities and gill disease., Results: Microbial community restructuring was observed throughout the sampling period and linked to varied drivers of change, including environmental conditions and severity of gill pathology. Taxa with significantly greater relative abundance on healthier gills included isolates within genus Shewanella, and taxa within family Procabacteriaceae. In contrast, altered abundance of Candidatus Branchiomonas and Rubritalea spp. were associated with damaged gills. Interestingly, more general changes in community richness and diversity were not associated with altered gill health, and thus not apparently deleterious to fish. Gross and histological gill scoring demonstrated seasonal shifts in gill pathology, with increased severity of gill damage in autumn. Specific infectious causes that contributed to observed pathology within the population included the gill disorder amoebic gill disease (AGD), however due to the uncontrolled nature of this study and likely mixed contribution of various causes of gill disease to observed pathology results do not strongly support an association between the microbial community and specific infectious or non-infectious drivers of gill pathology., Conclusions: Results suggest that the microbial community of farmed Atlantic salmon gills undergo continual restructuring in the marine environment, with mixed influences upon this change including environmental, host, and pathogenic factors. A significant association of specific taxa with different gill health states suggests these taxa might make meaningful indicators of gill health. Further research with more frequent sampling and deliberate manipulation of gills would provide important advancement of knowledge in this area. Overall, although much is still to be learnt regarding what constitutes a healthy or maladapted gill microbial community, the results of this study provide clear advancement of the field, providing new insight into the microbial community structure of gills during an annual production cycle of marine-stage farmed Atlantic salmon., (© 2024. The Author(s).)

During recent years, there has been a renewed interest in establishing farming of Atlantic cod (Gadus morhua) in Norway. However, a fatal abdominal disorder compromises animal welfare and causes economic losses. A similar problem was present during a previous attempt to establish Atlantic cod farming more than a decade ago. In this paper, we provide the first in-depth description of this intestinal disorder, which is correctly denoted 'strangulating obstruction'. In affected fish, part of the intestine is permanently entrapped (incarcerated) under fibrous strands in the mesentery. The entrapment interferes with blood flow and physically blocks the intestine, causing a strangulating obstruction with severe venous congestion and ischemia of the intestinal wall. Furthermore, comparison of macroscopical and histological anatomy of farmed and wild Atlantic cod is presented and risk factors associated with the anatomical differences are discussed., (© 2024 The Authors. Journal of Fish Diseases published by John Wiley & Sons Ltd.)

Mycotoxins, produced by fungi, can contaminate fish food and harm their health. Probiotics enhance immune balance and primarily function in the animal intestine. This study aimed to assess aflatoxin's impact on Piaractus mesopotamicus and explore probiotic-based additive (PBA) benefits in mitigating these effects, focusing on antioxidant activity, biochemical indices, and hepatic histopathology. Two experiments were conducted using P. mesopotamicus fry. The first experimental assay tested various levels of aflatoxin B1 (0.0, 25.0, 50.0, 100.0, 200.0, and 400.0 µg kg -1 ) over a 10-day period. The second experimental assay examined the efficacy of the probiotic (supplemented at 0.20%) in diets with different levels of aflatoxin B1 (0.0, 25.0, and 400.0 µg kg -1 ) for 15 days. At the end of each assay, the fish underwent a 24-hour fasting period, and the survival rate was recorded. Six liver specimens from each treatment group were randomly selected for metabolic indicator assays, including superoxide dismutase, catalase, alanine aminotransferase, aspartate aminotransferase, and albumin. Additionally, histopathological analysis was performed on six specimens. The initial study discovered that inclusion rates above 25.0 µg kg -1 resulted in decreased activity of AST (aspartate aminotransferase), ALT (alanine aminotransferase), ALB (albumin), CAT (catalase), and SOD (superoxide dismutase), accompanied by liver histopathological lesions. In the second study, the inclusion of PBA in diets contaminated with AFB1 improved the activity of AST and ALT up to 25.0 µg kg -1 of AFB1, with no histopathological lesions observed. The study demonstrated the hepatoprotective effects of PBA in diets contaminated with AFB1. The enzyme activity and hepatic histopathology were maintained, indicating a reduction in damage caused by high concentrations of AFB1 (400.0 µg kg -1 of AFB1). The adverse effects of AFB1 on biochemical and histopathological parameters were observed from 25.0 µg kg -1 onwards. Notably, PBA supplementation enhanced enzymatic activity at a concentration of 25 µg kg -1 of AFB1 and mitigated the effects at 400.0 µg kg -1 of AFB1. The use of PBAs in pacu diets is highly recommended as they effectively neutralize the toxic effects of AFB1 when added to diets containing 25.0 µg kg -1 AFB1. Dietary inclusion of aflatoxin B1 at a concentration of 25.0 µg kg -1 adversely affects the liver of Piaractus mesopotamicus (Pacu). However, the addition of a probiotic-based additive (PBA) to the diets containing this concentration of aflatoxin neutralized its toxic effects. Therefore, the study recommends the use of PBAs in Pacu diets to mitigate the adverse effects of aflatoxin contamination., (© 2024. The Author(s), under exclusive licence to Springer Nature B.V.)

Diagnostic imaging techniques provide a new aspect of the ante-mortem and post-mortem diagnostics in fish medicine. Ultrasonography, computed tomography (CT) and magnetic resonance imaging (MRI) can provide more information about the internal organs and pathognomic lesions. The authors used diagnostic imaging techniques to evaluate and describe the neoplastic malformation in a 3-year-old female rainbow trout (Oncorhynchus mykiss). The fish was examined with Siemens Somatom Definition AS + CT scanner and Siemens Biograph mMR scanner. The animal was lethargic and showed anorectic signs and muscular dystrophy. During the post-mortem investigation, histopathology and immunohistochemistry were also performed allowing us to identify the neoplasms. The results showed a large soft tissue mass in the first mid-intestine segment, which proved to be an adenocarcinoma. This subsequently led to digestion problems and absorption disorders. Immunohistochemically, neoplastic cells of carcinoma revealed E-cadherin and pancytokeratin positivity. This is the first study to report the use of MRI and CT for studying gastrointestinal adenocarcinoma in rainbow trout., (© 2024 The Authors. Journal of Fish Diseases published by John Wiley & Sons Ltd.)

Koi(Cyprinus carpio) is an ornamental variety of common carp frequently kept as pets. Given their long lifespan, neoplasia, albeit uncommon, may occur in these animals, and only a few studies have faced their cytological diagnosis. In the present case, a koi carp was referred to the clinicians due to coelomic swelling. The carp underwent surgery, which revealed an enlargement of both testes. Testicular samples were cytologically and histologically examined. The lesion was diagnosed as a seminoma since it was composed of round, large, atypical, and often multinucleated cells with round central nuclei and moderate cytoplasm. These tumors had the same appearance as seminomas in mammals and should be considered among differential diagnoses when coelomic swelling occurs in koi carp. Seminomas in koi carp are diagnosed histologically, but cytology, a rapid and cheap exam executable in all veterinary clinical facilities, could be a relevant preliminary diagnostic tool that may influence the entire diagnostic process., (© 2024. The Author(s).)

This study investigated the prevalence, morphology, molecular identification, and histopathological effects of larval tapeworms (plerocercoids) infecting the skeletal muscles of the Indian halibut (Psettodes erumei) collected from the coastal waters of the Arabian Gulf. Numerous oval or round blastocysts, measuring 13-26 mm, were found embedded within the muscular tissues of the Indian halibut, rendering the fish unsuitable for human consumption. Morphological and molecular analyses identified the plerocercoids as Dasyrhynchus giganteus (family Dasyrhynchidae), with an overall prevalence of 15.4%. The seasonal prevalence was the highest in summer (14.6%), followed by spring (10.6%), winter (4.4%), and autumn (3.5%). Infection rates increased with fish size. Histopathological examination revealed fibrous connective tissue capsules surrounding the larvae, causing muscular atrophy and degenerative changes, with few inflammatory eosinophilic cells. Molecular and phylogenetic analysis of the 28S rDNA gene sequences confirmed the specimens as D. giganteus, clustered closely with other sequences of D. giganteus with 100% bootstrap values. This study provided valuable insights into the parasitic infection dynamics, seasonal variation, molecular identification, and histopathological effects, highlighting the importance of monitoring fish for food safety and public health implications., (© 2024. The Author(s).)

Eustrongylides excisus is a fish-borne zoonotic parasite known to infect various fish species, including Northern pike (Esox Lucius). This nematode, belonging to the family Dioctophymatidae, has a complex life cycle involving multiple hosts. This study aimed to investigate the occurrence of Eustrongylides nematodes in Northern pike (E. Lucius) collected from Mijran Dam (Ramsar, Iran). Between June and October 2023, an investigation was conducted on Northern pike from Mijran Dam in Ramsar, Iran, following reports of reddish parasites in their muscle tissues. Sixty fish were examined at the University of Tehran, revealing live parasites in the muscles, which were then analyzed microscopically and preserved for a multidisciplinary study. The skeletal muscle tissues of 85% (51/60) of fish specimens were infected by grossly visible larvae which were microscopically identified as Eustrongylides spp. In histopathological examination, the lesion was composed of encapsulated parasitic granulomatous myositis. Microscopically, the cystic parasitic granulomas compressed the adjacent muscle fibers, leading to their atrophy and Zenker's necrosis. Moreover, epithelioid macrophages, giant cells and mononuclear inflammatory cells were present around the larvae and between the muscle fibers. Finally, a molecular analysis by examining the ITS gene region, revealed that they belong to the species E. excisus. Eustrongylidiasis in northern Iran necessitates further research into the biology, epidemiology, and control of Eustrongylides nematodes, focusing on various hosts. This study is the first to comprehensively characterize E. excisus in Northern pike in Ramsar, Iran, raising concerns about possible zoonotic transmission., (© 2024. The Author(s).)

Nocardiosis, caused by Nocardia seriolae, has been a prominent disease in Southeast Asian aquaculture in the last three decades. This granulomatous disease reported in various fish species is responsible for significant economic losses. This study investigated the pathogenicity of N. seriolae in three cultured species in Taiwan: Nile tilapia (omnivore), milkfish (herbivore) and Asian seabass (carnivore). Administration of an infective dose of 1 × 10 6  CFU/ fish in tilapia, seabass and milkfish demonstrated mortalities of 100%, 90% and 75%, respectively. Additionally, clinical signs namely, granuloma and lesions displayed varying intensities between the groups and pathological scores. Polymerase chain reaction (PCR) amplification specific for N. seriolae was confirmed to be positive (432 bp) using NS1/NG1 primers. Post-mortem lesions revealed the absence of granulomas in tilapia and milkfish and their presence in the seabass. Interestingly, the gut in tilapia showed an influx of eosinophils suggesting its role during the acute stages of infection. However, post-challenge, surviving milkfish exhibited granulomatous formations, while surviving seabass progressed toward healing and tissue repair within sampled tissues. Overall, in conclusion, these results demonstrate the versatility in the immunological ability of individual Perciformes to contain this pathogen as a crucial factor that influences its degree of susceptibility., (© 2024 John Wiley & Sons Ltd.)

An adult female Brazilian cownose ray (Rhinoptera brasiliensis), housed in Ubatuba Aquarium died after loss of appetite period. During necropsy, an enterolith was discovered partially obstructing the intestinal lumen. Examination of the enterolith revealed a bonefish spine nidus. Enterolithiasis has been linked to multiple factors including diet, genetics, alkaline intestinal environments and, as in this particular case, the ingestion and retention of foreign bodies. The composition of this enterolith from a cownose ray was primarily monohydrocalcite. This short communication, apparently the first to report enterolithiasis in stingrays, emphasizes the need for post-mortem examinations of carcasses on macroscopic and microscopic levels., (© 2024 John Wiley & Sons Ltd.)

Monogenean trematodes, particularly those belonging to the Diplectanidae family, are significant metazoan parasites with substantial implications for aquaculture expansion. This study, investigatied the occurrence, prevalence, and pathological impact of Diplectanum spp. in European seabass (Dicentrarchus labrax) across three distinct Egyptian fish farms. During 2021-2022, we sampled 1800 European seabass (Dicentrarchus labrax) from three Egyptian fish farms (600 fish per farm). Farms 1 and 2 used semi-intensive earthen pond systems, while Farm 3 utilized an intensive floating cage system. Employing Clinical, post-mortem, parasitological, and molecular examination technique. Pathological lesions were identified, including skin and gill discoloration, emaciation, and internal organ abnormalities. Seasonal prevalence exhibited significant variations between farms, with highest rates observed in spring and Farm 3 reached an overall peak prevalence of 84.67 %. Parasitological examination distinguished two Diplectanum species morphologically, while molecular techniques exhibited limited specificity. Histopathology unveiled damage to gill, liver, spleen, kidney, and intestine, attributed to Diplectanum haptors including inflammation and internal bleeding, potentially leading to secondary infections. Molecular identification via PCR targeting ITS and 28SrDNA genes, revealing similar band sizes for the two Diplectanum species, indicating limited intraspecific genetic diversity. The study emphasizes investigating parasitic infections' prevalence and impact in aquaculture, necessitating robust molecular techniques for species differentiation. This study underscores the importance of investigating the prevalence and impact of parasitic infections in aquaculture. It highlights the need for robust molecular techniques to differentiate species. By focusing on Diplectanum spp. infections in D. labrax, the study offers valuable insights into managing parasites in aquaculture effectively., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper, (Copyright © 2024 Elsevier B.V. All rights reserved.)

Over the course of an approximately 11-month period, an outdoor, freshwater, mixed species, recirculating, display system at a public aquarium experienced intermittent mortalities of channel catfish ( Ictalurus punctatus ) and blue catfish ( I. furcatus) . Catfish acutely presented for abnormal buoyancy, coelomic distention, and protein-rich coelomic effusion. Gross lesions typically involved massive coelomic distension with protein-rich effusion, generalized edema, and gastric hemorrhage and edema. Microscopically, primary lesions included renal tubular necrosis, gastric edema with mucosal hemorrhages, and generalized edema. Aerobic culture and virus isolation could not recover a consistent infectious agent. Intracoelomic injection of coelomic effusion and aspirated retrobulbar fluid from a catfish into naïve zebrafish (bioassay) produced peracute mortality in 3 of 4 fish and nervous signs in the fourth compared with 2 saline-injected control zebrafish that had - no mortality or clinical signs. Kidney tissue and coelomic effusion were submitted for gas chromatography tandem mass spectrometry by multiple reaction monitoring against laboratory standards, which detected the presence of multiple pyrethroid toxins, including bioallethrin, bifenthrin, trans-permethrin, phenothrin, and deltamethrin. Detection of multiple pyrethroids presumably reflects multiple exposures with several products. As such, the contributions of each pyrethroid toward clinical presentation, lesion development, and disease pathogenesis cannot be determined, but they are suspected to have collectively resulted in disrupted osmoregulation and fluid overload due to renal injury. Pesticide-induced toxicoses involving aquarium fish are rarely reported with this being the first description of pyrethroid-induced lesions and mortality in public aquarium-held fish., Competing Interests: Declaration of Conflicting InterestsThe authors declared no potential conflicts of interest with respect to the research, authorship, and/or publication of this article.

Ulcerative dermatitis (UD) is common in ornamental fish collections and is typically associated with a wide range of bacterial aetiologies. Clinical reports describing Shewanella xiamenensis-associated UD are limited, however, despite growing attention to pathogenic Shewanella species in fish. Two out of 95 koi carp with UD were presented for clinical assessment by a commercial collection (n = 3000 fish) and subsequently killed on welfare grounds for necropsy. Both specimens exhibited extensive cutaneous ulcers and coelomic fat necrosis with petechial haemorrhages on post-mortem examination. Shewanella xiamenensis was cultured from ulcerated skin tissues taken from both fish, with consistent intralesional gram-negative rod-like bacteria seen on skin scrape cytology. Histology also confirmed intralesional gram-negative rod-like bacteria within multiple ulcerative and erosive dermatitis lesions, plus myofibre necrosis and necrotising coelomic steatitis, in both specimens. Features associated with impaired generalised osmoregulation secondary to UD were detected within the striated muscle underlying the ulcers, the gills, and the caudal aspects of the kidneys. Additional histological features suggestive of sepsis were also seen in one of the fish. In the interim period, morbidity had increased from 3.2% to around 30% of the entire stock. Following culture results, increased pond water changes were implemented (q.2-3d) and the remaining stock was treated with florfenicol, resulting in complete resolution of UD in the collection (as per client). This article highlights the first description of S. xiamenensis-associated UD in koi carp/diseased ornamental fish in the United Kingdom., (© 2024 John Wiley & Sons Ltd.)

Hypoxia, a major issue in aquatic ecosystems, in special reference to climate change, and exacerbated by anthropogenic activities. It is causing slow growth, disease outbreaks, and mortality in finfish and shellfish. Therefore, adaptation to lowering oxygen levels through supplementation of herbs or their extracts in diets is imperative. In this study, hypoxia was simulated in controlled conditions with quercetin-enriched diets. Quercetin is a plant pigment (flavonol) possessing anti-oxidant property and is present in vegetables, leaves, seeds, pulses, and fruits. The experiment was conducted on rohu Labeo rohita, which is most widely cultured in India. There were four treatments including T1 (Normoxia: > 5 ppm dissolved oxygen; DO 2 ), T2 (hypoxia: 3-4 ppm DO 2 ), T3 (hypoxia + 50 mg quercetin/kg diet), and T4 (hypoxia + 100 mg quercetin/kg diet). The study was conducted for 30 days, and water quality was measured regularly. The results revealed that the hematological parameters were negatively affected. The tissue micro-architecture illustrated the impairment through degeneration of neurons in the brain, increased pigmentation as melanosis in the kidney, increased thickness of primary lamellae in the gills, and dilatations of sinusoids in the liver in hypoxia groups, while quercetin-enriched diets improved the hematological and histomorphological parameters. The results confirm the utility of hematological and histopathological tools as biomarkers and reflect the possible threats of hypoxia on fish. In conclusion, quercetin in diets appeared to show resistance towards chronic hypoxia by restoring the structure and functions of the vital organs towards normalcy and could be recommended as a potential ameliorative agent., (© 2024. The Author(s), under exclusive licence to Springer Nature B.V.)

The guppy, Poecilia reticulata, is one of the most common cultured ornamental fish species, and a popular pet fish highly desired by hobbyists worldwide due to its availability of many brilliantly coloured fish of many varieties. The susceptibility of guppies to diseases presents a remarkable concern for both breeders and hobbyists. In this study, we report the emergence of disease in fancy guppies caused by a previously uncharacterized virus in the USA. This virus was isolated from moribund guppies in two separate outbreaks in California and Alabama, from December 2021 to June 2023. The infected guppies presented with acute morbidity and mortality shortly after shipping, displaying nonspecific clinical signs and gross changes including lethargy, anorexia, swimming at the water surface, gill pallor, mild to moderate coelomic distension and occasional skin lesions including protruding scales, skin ulcers and hyperaemia. Histological changes in affected fish were mild and nonspecific; however, liver and testes from moribund fish were positive for Tilapia lake virus (TiLV), the single described member in the family Amnoonviridae, using immunohistochemistry and in situ hybridization, although the latter was weak. A virus was successfully recovered following tissue inoculation on epithelioma papulosum cyprini and snakehead fish cell lines. Whole genome sequencing and phylogenetic analyses revealed nucleotide and amino acid homologies from 78.3%-91.2%, and 78.2%-97.7%, respectively, when comparing the guppy virus genomes to TiLV isolates. Based on the criteria outlined herein, we propose the classification of this new virus, fancy tailed guppy virus (FTGV), as a member of the family Amnoonviridae, with the name Tilapinevirus poikilos (from the Greek 'poikilos', meaning of many colours; various sorts, akin to 'poecilia')., (© 2024 The Authors. Journal of Fish Diseases published by John Wiley & Sons Ltd.)

Salmonid fish include some of the most valued cultured fish species worldwide. Unlike most other fish, the hearts of salmonids, including Atlantic salmon and rainbow trout, have a well-developed coronary circulation. Consequently, their hearts' reliance on oxygenation through coronary arteries leaves them prone to coronary lesions, believed to precipitate myocardial ischemia. Here, we mimicked such coronary lesions by subjecting groups of juvenile rainbow trout to coronary ligation, assessing histomorphological myocardial changes associated with ischemia and scarring in the context of cardiac arrhythmias using electrocardiography (ECG). Notable ECG changes resembling myocardial ischemia-like ECG in humans, such as atrioventricular blocks and abnormal ventricular depolarization (prolonged and fragmented QRS complex), as well as repolarization (long QT interval) patterns, were observed during the acute phase of myocardial ischemia. A remarkable 100% survival rate was observed among juvenile trout subjected to coronary ligation after 24 wk. Recovery from coronary ligation occurred through adaptive ventricular remodeling, coupled with a fast cardiac revascularization response. These findings carry significant implications for understanding the mechanisms governing cardiac health in salmonid fish, a family particularly susceptible to cardiac diseases. Furthermore, our results provide valuable insights into comparative studies on the evolution, pathophysiology, and ontogeny of vertebrate cardiac repair and restoration. NEW & NOTEWORTHY Juvenile rainbow trout exhibit a remarkable capacity to recover from cardiac injury caused by myocardial ischemia. Recovery from cardiac damage occurs through adaptive ventricular remodeling, coupled with a rapid cardiac revascularization response. These findings carry significant implications for understanding the mechanisms governing cardiac health within salmonid fishes, which are particularly susceptible to cardiac diseases.

The relationship of histopathological changes and the infection of Piscine orthoreovirus 2 (PRV-2) was investigated in coho salmon that were suffering from the erythrocytic inclusion body syndrome (EIBS). Immunohistochemical observations revealed abundant σ1 protein of PRV-2 in the spongy layer of the ventricle of the heart, where severe myocarditis was observed. In the spleen, the virus protein was detected in many erythrocytes, some of which were spherical-shaped and apparently dead. The number of erythrocytes was decreased in the spleen compared to the apparently healthy fish. The virus protein was also detected in some erythrocytes in blood vessels. The viral protein was often detected in many macrophages ingesting erythrocytes or dead cell debris in the spleen or in the kidney sinusoids. Large amounts of the viral genomic segment L2 were also detected in these organs by RT-qPCR. Many necrotic foci were found in the liver, although the virus protein was not detected in the hepatocytes. These results suggest that the primary targets of PRV-2 are myocardial cells and erythrocytes and that clinical symptoms such as anaemia or jaundice and histopathological changes such as myocarditis in EIBS-affected coho salmon are caused by PRV-2 infection., (© 2024 John Wiley & Sons Ltd.)

A series of 12 contiguous caudal vertebrae of an ichthyodectiform fish from the Smoky Hill Chalk Member of the Niobrara Formation is described. The vertebral centra exhibit extensive overgrowth of pathological bone and there is additional pathological bone within the centra and intervertebral spaces, which together resulted in the coossification of most centra. The extent of the pathology is greatest on preural vertebrae 1-3 and decreases anteriorly, which suggests that the pathology began posteriorly and progressed anteriorly. In addition to the pathological overgrowth on bones, the specimen preserves features interpreted as calcified and/or ossified soft tissues associated with the neural and haemal canals. The pathologies are unlike previously described examples of bony pathologies in fish, and it is suggested that they resulted from combined bacterial and fungal infections. As the pathologies developed, they would have adversely impacted the fish's swimming and feeding abilities, and presumably eventually led to the fish's death., Competing Interests: The author declares that they have no competing interests., (©2024 Bennett.)

Captive fish populations, such as those encompassing aquarium and pet fish, offer significant economic value and are integral to conservation, research, and education. However, these ornamental fish exhibit a reduced ability to protect their ocular surfaces, and our understanding of the ocular diseases that affect them remains limited. Although corneal neoplasms in carp are uncommon, identifying their distinct characteristics is crucial in selecting appropriate therapeutic interventions that aim to preserve vision, prevent the ocular loss, and ultimately ensure the survival of the affected fish. This study provides clinical and histopathological details of various proliferative corneal masses in Cyprininae species, including five koi (Cyprinus carpio) and four goldfish (Carassius auratus). It discusses a spectrum of neoplasms, including soft tissue sarcoma, spindle cell sarcoma, chromatophoroma, and papilloma, in addition to conditions like exuberant granulation tissue and proliferative carp pox. These findings bear significant implications for clinical decision-making and treatment, offering valuable insights into the incidence and characteristics of corneal tumors in captive fish, which could inform further studies in this area., (© 2023 The Authors. Veterinary Ophthalmology published by Wiley Periodicals LLC on behalf of American College of Veterinary Ophthalmologists.)

Fish maintained in managed care may have longer lifespans as a result of advances in veterinary medicine and husbandry and reduced risk of predation. Neoplasia is of increasing interest in managed aquarium populations. However, few studies have systematically evaluated neoplasia in managed fish populations. Our objective in this retrospective study was to review and describe neoplasia diagnosed in fish at a large public display aquarium between 2005 and 2021. Any fish diagnosed with neoplasia on either antemortem or postmortem evaluation during the study period was included, and all medical records, biopsy, and autopsy reports were reviewed. Sixty-two fish met the inclusion criteria; 37 species were included in the study population, most of which were tropical freshwater fish ( n  = 34 fish). Thirty-two types of neoplasia were identified. Ten fish had benign neoplasms, and 53 fish had malignant neoplasms. The most common neoplasms were of epithelial and neuroectodermal origin. The most common site of tumor origin was the skin. Our data suggest that mesenchymal neoplasms may be more common in cold saltwater fish than in tropical freshwater and saltwater fish. Malignant neoplasms were most commonly diagnosed in the study population and should be a top differential when neoplasms are identified in fish managed under human care. Our study contributes to the overall knowledge of the health of aquarium fish and may aid clinicians in characterizing neoplasia that may be present in fish under human care., Competing Interests: Declaration of conflicting interestsHeindrich Snyman is a guest editor of the JVDI special section on “Aquatic animal health and disease” but was not involved in the editorial evaluation or decision to accept this article for publication

Spinal deformities in finfish have the potential to impact aquaculture industries and wild populations by increasing morbidity, mortality, and reducing growth rates. Myxobolus acanthogobii has been implicated in causing scoliosis and lordosis in various aquatic species in Japan. We investigated 4 cases of spinal deformity in 2 flathead ( Platycephalus ) species that were submitted to the Elizabeth Macarthur Agricultural Institute (EMAI) in New South Wales (NSW), Australia, between 2015 and 2021. Flathead are commercially significant species that are popular among Australian consumers, and are also sought-after species targeted by recreational fishers. Gross deformities are concerning to the community and may impact the quality and quantity of specimens available for consumption. Three blue-spotted flathead ( P. caeruleopunctatus ) and one marbled flathead ( P. marmoratus ) were submitted, all with marked scoliosis and kyphosis; 1-2-mm cysts were present on the dorsum of the brain, most often over the optic lobe or cerebellum. Cytology and differential interference microscopy of cyst material revealed numerous oval spores, x̄ 14 ± SD 0.75 µm × x̄ 11.5 ± SD 0.70 µm, with 2 pyriform polar capsules, the morphology of which is consistent with a Myxobolus sp. PCR assay and 18S rDNA sequencing of the cyst material identified a Myxobolus sp. with 96% identity to M. acanthogobii. The identification of this Myxobolus sp. confirms the presence of parasites with the potential to cause spinal deformity in significant aquatic species in NSW waterways., Competing Interests: Declaration of conflicting interestsThe authors declared no potential conflicts of interest with respect to the research, authorship, and/or publication of this article.

Viral nervous necrosis (viral encephalopathy and retinopathy) is caused by piscine nodavirus ( Nodaviridae , Betanodavirus ). Since 1986, this highly infectious virus has caused mass mortalities of up to 100% in farmed saltwater and freshwater fish around the world (with the exception of South America and Antarctica), affecting >60 species across 10 orders. The Atlantic blue marlin ( Makaira nigricans Lacépède, 1802) is a top-level predator found throughout the tropical waters of the Atlantic and Indo-Pacific oceans. Despite their popularity as a sportfish, relatively little is known about the Atlantic blue marlin and other billfish. We describe here chronic betanodavirus infection in a juvenile Atlantic blue marlin, which is, to our knowledge, the first report of disease in M. nigricans ., Competing Interests: Declaration of conflicting interestsThe authors declared no potential conflicts of interest with respect to the research, authorship, and/or publication of this article.

We performed a diagnostic disease investigation on a cohort of coho salmon ( Oncorhynchus kisutch ) fingerlings in Alaska exhibiting anorexia, gaping mouths, anemia, and increased mortality. Histologic examination revealed mild-to-severe myocardial degeneration and lymphohistiocytic and neutrophilic myocarditis, moderate splenic histiocytosis, and mild renal histiocytosis. Piscine orthoreoviruses 1 and 3 were not detected by molecular methods, and no other viruses could be cultured on 3 common diagnostic fish cell lines. De novo assembly produced a viral genome of 10 linear segments with >80% homology to piscine orthoreovirus 2 (PRV2) encoding all 11 PRV2 proteins. An in situ hybridization probe using RNAscope was developed against 697 viral nucleotides identified by sequencing, which revealed viral genome in heart, spleen, gill, kidney, liver, blood, and the lamina propria of the intestines. Our findings are supportive of a novel piscine orthoreovirus most closely related to PRV2 associated with morbidity and mortality of coho salmon in the northeastern Pacific., Competing Interests: Declaration of conflicting interestsThe authors declared no potential conflicts of interest with respect to research, authorship, and/or publication of this article.

Non-neoplastic thyroid hyperplasia is common in terrestrial animals, secondary to nutritional imbalances or other goitrogenic compounds. Thyroid hyperplasia is relatively common in teleost fish; however, malignant thyroid neoplasia is rarely reported. We diagnosed cases of thyroid neoplasia in a population of jade perch ( Scortum barcoo ). The 3,000 affected fish had grossly apparent, bilateral pharyngeal swellings. Histologic examination confirmed proliferative thyroid lesions ranging from hyperplasia to well-differentiated follicular cell carcinoma. In addition, the younger population of animals on the farm also had bacterial septicemia and mild Dactylogyrus sp. gill infections. Feed analysis revealed a severe deficiency of iodine and vitamin C in the homemade fish diet used on the farm. The concentrations of other minerals, such as zinc, were also on the lower end of the recommended requirements for freshwater fish. The farm was using surface water in its recirculating aquaculture system. We recommended a switch to a commercial aquaculture diet, as well as to use well water rather than surface water to avoid any contaminants, and to treat the younger fish with an antibiotic for bacterial septicemia. Our case provides evidence of progression from nutritional-associated thyroid hyperplasia to neoplasia in farmed teleost fish., Competing Interests: Declaration of conflicting interestsThe authors declared no potential conflicts of interest with respect to the research, authorship, and/or publication of this article.

Ichthyophthirius multifiliis, the causative agent of white spot disease, is a ciliated protozoan parasite that infects freshwater fish and induces high mortality. Outbreaks occur both in natural and production sites. The aim of the present study was to describe the lesions caused by chronic infection by I. multifiliis in goldfish (Carassius auratus) from an ornamental fish farm, highlighting important ultrastructural aspects of this protozoan. Damaged skin and gills, collected from fish with white or ulcerative skin lesions, were routinely processed for histological analysis and transmission electron microscopy. The parasitic forms present in the skin were associated with an inflammatory infiltrate consisting of macrophages, lymphocytes and other polymorphonuclear cells. The lesions associated with the presence of the parasite were organized in the form of granulomas, with macrophages in the layers closest to the parasites. A trophont-thickened membrane and induction of granulomatous inflammation were identified in this study as mechanisms for evasion of the immune response. We concluded that the presence of I. multifiliis trophonts resulted in the formation of granulomatous inflammation, whether associated or not with pathogen lysis, suggesting that the parasite can use an inflammatory response to evade the immune response., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

We investigated disease trends of concern for fish or public health in a 5-y (2017-2021) prospective survey of fish in Cook County, IL, inland lakes. Lesions were assessed in relation to fish species, lake type and location, season and collection year, and lake water chemistry parameters. Fish included bullheads ( n  = 98), common carp ( n  = 99), bluegill ( n  = 114), and largemouth bass ( n  = 118). Annually, fish collection and point-source water sampling were conducted in spring, summer, and fall from both seepage and impoundment lakes. Examinations included autopsy, wet-mount cytologic assessment for ectoparasites, and histopathology. No lesions of public health concern were detected. The most common abnormalities were branchitis (261 of 429; 60.8%) and endoparasitism (312 of 429; 72.7%). Branchitis was mild in most cases (189 of 261; 72.4%) and concurrent with branchial parasitism in 175 of 261 (67%) cases. Monogeneans were the most common gill parasites but did not influence branchitis severity (Kruskal-Wallis, p  = 0.484). Using generalized ordered logistic regression, predictors of branchitis severity included fish species ( p  < 0.001), the interaction of lake or location and alkalinity ( p  < 0.001), and water temperature or season ( p  < 0.001). Endoparasites included tissue larval trematodes (metacercaria), nematodes, and cestodes (plerocercoids), enteric acanthocephalans, gastric trematodes, renal myxosporidia, biliary and gall bladder myxosporidia, enteric cestodes, and tissue microsporidia. Using generalized ordered logistic regression, variables influencing endoparasitism severity included species ( p  < 0.001), year ( p  < 0.001), chloride level ( p  = 0.009), and the interaction of year and chloride level ( p  < 0.001). Our results suggested overall good health of fish in sampled Cook County inland lakes and provide a foundation for continued monitoring of ecosystem and public health in the urban environment., Competing Interests: Declaration of conflicting interestsThe authors declared no potential conflicts of interest with respect to the research, authorship, and/or publication of this article.

Grass carp reovirus (GCRV), particularly the highly prevalent type II GCRV (GCRV-II), causes huge losses in the aquaculture industry. However, little is known about the mechanisms by which GCRV-II invades grass carp and further disseminates among tissues. In the present study, monocytes/macrophages (Mo/Mφs) were isolated from the peripheral blood of grass carp and infected with GCRV-II. The results of indirect immunofluorescent microscopy, transmission electron microscopy, real-time quantitative RT-PCR (qRT-PCR), western blot (WB), and flow cytometry analysis collectively demonstrated that GCRV-II invaded Mo/Mφs and replicated in them. Additionally, we observed that GCRV-II induced different types (M1 and M2) of polarization of Mo/Mφs in multiple tissues, especially in the brain, head kidney, and intestine. To assess the impact of different types of polarization on GCRV-II replication, we recombinantly expressed and purified the intact cytokines CiIFN-γ2, CiIL-4/13A, and CiIL-4/13B and successfully induced M1 and M2 type polarization of macrophages using these cytokines through in vitro experiments. qRT-PCR, WB, and flow cytometry analyses showed that M2 macrophages had higher susceptibility to GCRV-II infection than other types of Mo/Mφs. In addition, we found GCRV-II induced apoptosis of Mo/Mφs to facilitate virus replication and dissemination and also detected the presence of GCRV-II virus in plasma. Collectively, our findings indicated that GCRV-II could invade immune cells Mo/Mφs and induce apoptosis and polarization of Mo/Mφs for efficient infection and dissemination, emphasizing the crucial role of Mo/Mφs as a vector for GCRV-II infection.IMPORTANCEType II grass carp reovirus (GCRV) is a prevalent viral strain and causes huge losses in aquaculture. However, the related dissemination pathway and mechanism remain largely unclear. Here, our study focused on phagocytic immune cells, monocytes/macrophages (Mo/Mφs) in blood and tissues, and explored whether GCRV-II can invade Mo/Mφs and replicate and disseminate via Mo/Mφs with their differentiated type M1 and M2 macrophages. Our findings demonstrated that GCRV-II infected Mo/Mφs and replicated in them. Furthermore, GCRV-II infection induces an increased number of M1 and M2 macrophages in grass carp tissues and a higher viral load in M2 macrophages. Furthermore, GCRV-II induced Mo/Mφs apoptosis to release viruses, eventually infecting more cells. Our study identified Mo/Mφs as crucial components in the pathway of GCRV-II dissemination and provides a solid foundation for the development of treatment strategies for GCRV-II infection., Competing Interests: The authors declare no conflict of interest.

Sturgeon farming is rapidly expanding in Europe, where Italy ranks first in farmed caviar production. A major threat to sturgeon health in captivity is infection with Acipenser European Iridovirus (AcIV-E), a viral disease definitively identified in 2016. Here we present data on the occurrence of AcIV-E in 482 sturgeons (age ≤ 12 months, species of the genus Acipenser and the species Huso huso ) collected from sturgeon farms in northern Italy between January 2021 and December 2023. The health status of each specimen was determined by necroscopy and virological assay. Virological analysis was performed on gill samples and real-time PCR specific to the MCP gene of the iridovirus viral capsid. Molecular analysis revealed positivity to the virus in 204 samples (42.68% of the total), while anatomopathological examination of nearly all fish with positive real-time PCR disclosed swollen abdomen, hepatic steatosis, splenomegaly, and increased gill volume. Two challenges to timely diagnosis are the absence of pathognomonic symptoms and the inability to isolate the virus on cell monolayers. Continuous and widespread health monitoring is therefore crucial for disease management and to effectively control spread of the virus.

The mechanism by which Y. ruckeri infection induces enteritis in Chinese sturgeon remains unclear, and the efficacy of drug prevention and control measures is not only poor but also plagued with numerous issues. We conducted transcriptomic and 16 S rRNA sequencing analyses to examine the differences in the intestinal tract of hybrid sturgeon before and after Y. ruckeri infection and florfenicol intervention. Our findings revealed that Y. ruckeri induced the expression of multiple inflammatory factors, including il1β, il6, and various chemokines, as well as casp3, casp8, and multiple tumor necrosis factor family members, resulting in pathological injury to the body. Additionally, at the phylum level, the relative abundance of Firmicutes and Bacteroidota increased, while the abundance of Plesiomonas and Cetobacterium decreased at the genus level, altering the composition of the intestinal flora. Following florfenicol intervention, the expression of multiple apoptosis and inflammation-related genes was down-regulated, promoting tissue repair. However, the flora became further dysregulated, increasing the risk of infection. In conclusion, our analysis of the transcriptome and intestinal microbial composition demonstrated that Y. ruckeri induces intestinal pathological damage by triggering apoptosis and altering the composition of the intestinal microbiota. Florfenicol intervention can repair pathological damage, but it also exacerbates flora imbalance, leading to a higher risk of infection. These findings help elucidate the molecular mechanism of Y. ruckeri-induced enteritis in sturgeon and evaluate the therapeutic effect of drugs on intestinal inflammation in sturgeon., Competing Interests: Declaration of Competing Interest The authors declare that they have no conflict of interest., (Copyright © 2024 The Authors. Published by Elsevier Inc. All rights reserved.)

Objective: During routine histological examination of tissues from mortality events of anadromous Brook Trout Salvelinus fontinalis from Prince Edward Island (PEI), Canada, myxospores consistent with Myxobolus were observed infecting the central nervous system. The objective of this study was to identify the species of Myxobolus infecting the nervous system of anadromous Brook Trout from PEI, Canada., Methods: Myxospore morphology, small subunit (SSU) ribosomal DNA (rDNA) sequence data, and histology were used to identify myxospores isolated from infected Brook Trout., Result: Myxospore measurements from the PEI samples matched those reported in the description of Myxobolus neurofontinalis from North Carolina. A 1057-bp fragment of the SSU rDNA from myxospores collected from Brook Trout in PEI was identical to an isolate of M. neurofontinalis (MN191598) collected previously from the type locality, New River basin, North Carolina. Histological sections confirmed infections were intercellular in the central nervous system. Minimal host response was observed, with only sparse mononuclear inflammatory infiltrates present at the periphery of and within dispersed myxospores, suggesting that infections are not pathogenic to Brook Trout., Conclusion: Myxospores were identified as M. neurofontinalis, which was previously described from the central nervous system of Brook Trout from the New River basin, North Carolina, USA. This constitutes the first time M. neurofontinalis has been documented outside of the New River basin in North Carolina., (© 2024 American Fisheries Society.)

Chinook salmon (Oncorhynchus tshawytscha) farmed in New Zealand are known to develop abnormal spinal curvature late in seawater production. Its cause is presently unknown, but there is evidence to suggest a neuromuscular pathology. Using magnetic resonance imaging (MRI), we evaluated the relationship between soft tissue pathology and spinal curvature in farmed Chinook salmon. Regions of interest (ROIs) presenting as pathologic MRI signal hyper-intensity were identified from scans of 24 harvest-sized individuals: 13 with radiographically-detectable spinal curvature and 11 without. ROIs were excised from individuals using anatomical landmarks as reference points and histologically analysed. Pathologic MRI signal was observed more frequently in individuals with radiographic curvature (92%, n = 12) than those without (18%, n = 2), was localized to the peri-vertebral connective tissues and musculature, and presented as three forms: inflammation, fibrosis, or both. These pathologies are consistent with a chronic inflammatory process, such as that observed during recovery from a soft tissue injury, and suggest spinal curvature in farmed Chinook salmon may be associated with damage to and/or compromised integrity of the peri-vertebral soft tissues. Future research to ascertain the contributing factors is required., (© 2023 The Authors. Journal of Fish Diseases published by John Wiley & Sons Ltd.)

The present work is the first comprehensive study of fungus-like stramenopilous organisms (Oomycota) diversity in Lithuanian fish farms aimed at proper identification of saprolegniasis pathogens, which is important for water quality control, monitoring infection levels and choosing more effective treatments for this disease in aquaculture. Pathogenic to fish, Saprolegnia and other potentially pathogenic water moulds were isolated from adult fish, their eggs, fry and from water samples. All detected isolates were examined morphologically and confirmed by sequence-based molecular methods. A total of eight species belonging to the genera Saprolegnia, Achlya, Newbya and Pythium were identified. Four species (S. parasitica, S. ferax, S. australis and S. diclina) were found to be the main causative agents of saprolegniasis in Lithuania. S. parasitica and S. ferax dominated both in hatcheries and open fishponds, accounting for 66.2% of all isolates. S. parasitica was isolated from all farmed salmonid fish species as well as from the skin of Cyprinus carpio, Carassius carassius and Perca fluviatilis. S. australis was isolated from water and once from the skin of Oncorhynchus mykiss, and S. diclina was detected only once on the skin of Salmo salar fish. In addition, Achlya ambisexualis, Saprolegnia anisospora and Newbia oligocantha isolated during this study are noted as a possible source of saprolegniasis. The results of this study are relevant for assessing the risk of potential outbreaks of saprolegniasis or other saprolegnia-like infection in Lithuanian freshwater aquaculture., (© 2023 John Wiley & Sons Ltd.)

Lumpfish (Cyclopterus lumpus L) are highly prone to cataract development in the wild and in culture. There is evidence that cataract in farmed fish is related to nutrition. However, both the nutrients and the mechanisms involved in cataract development in lumpfish are not clear. Here we investigated the mechanisms involved and the role of dietary vitamin A in cataract development in a cultured lumpfish population. Cultured lumpfish were fed three diets differing only in vitamin A supplementation level (5000, 15,000 and 120,000 IU/kg) over an 18-month period, and fish weight, cataract frequencies and severities were determined. Western blotting and immunohistochemistry were performed on lens tissue to measure the levels of oxidative stress, and apoptosis. The lowest levels of vitamin A significantly reduced cataract frequencies in adult lumpfish and resulted in less severe cataract and increased weight in males. Oxidative stress levels in the lens were positively correlated with vitamin A intake. Apoptosis was observed at high levels in lenses with severe cataract. Oxidative stress and apoptosis levels were the highest in regions of the lens with severe, advanced cataract pathology when compared to regions with no visible pathology. These results suggest that higher vitamin A intake contributes to cataract development through an oxidative stress pathway, and that both oxidative stress and apoptosis are involved in advanced stages of cataract in lumpfish., (© 2023 John Wiley & Sons Ltd.)

Spatial transcriptomics is a technique that provides insight into gene expression profiles in tissue sections while retaining structural information. We have employed this method to study the pathological conditions related to red and melanized focal changes in farmed Atlantic salmon (Salmo salar). Our findings support a model where similar molecular mechanisms are involved in both red and melanized filet discolorations and genes associated with several relevant pathways show distinct expression patterns in both sample types. Interestingly, there appears to be significant cellular heterogeneity in the foci investigated when looking at gene expression patterns. Some of the genes that show differential spatial expression are involved in cellular processes such as hypoxia and immune responses, providing new insight into the nature of muscle melanization in Atlantic salmon., (© 2024. The Author(s).)

A new Myxobolus species, Myxobolus nekrasovi n. sp., was found in the gill arch of the gibel carp Carassius auratus gibelio during investigation of fish myxosporean fauna of ponds of Lake Baikal basin. The parasites were studied on the basis of spore morphology, as well as with histological and molecular methods. Mature spores of M. nekrasovi n. sp. are ellipsoidal in frontal view and lemon-shaped in lateral view, measuring 13.84 ± 0.4 (12.2-15) μm in length, 9.73 ± 0.2 (8.5-10.7) μm in width, 6,75 ± 0.1 (6.0-7.6) μm in thickness. Polar capsules are unequal and pyriform, measuring: length 6.31 ± 0.1 (5.4-7.4), width 3.49 ± 0.04 (3.12-4) μm and length 2.88 ± 0.1 (2.1-3.5), width 1.4 ± 0.03 (1-1.6) μm. Phylogenetic analysis with the SSU rDNA gene shows Myxobolus nekrasovae n. sp. as a sister species of the subclade formed by Thellohanellus sinensis, Myxobolus acutus, M. zhaltsanovae that infect gibel carp Carassius auratus gibelio., Competing Interests: Declaration of competing interest Ethics approval Not applicable. Consent to participate Not applicable. Consent for publication All authors agreed to the publication of the manuscript. Competing interests The authors declare no competing interests., (Copyright © 2023 Elsevier Ltd. All rights reserved.)

Selective breeding plays a vital role in the production of farmed Atlantic salmon and has shown success in many aspects. Still, challenges related to fish health and welfare continue to result in significant economic losses. One such challenge is red and melanized focal changes (RFC/MFC), which result from acute and chronic inflammation, respectively, in the skeletal muscle. Importantly, RFC/MFC has not been observed in wild Atlantic salmon, suggesting that both external and genetic factors may contribute to the development of inflammation. To investigate the underlying cause of RFC/MFC, we conducted a study involving 1854 Atlantic salmon of farmed, wild and hybrid origin. All fish were reared under identical conditions to minimize the influence of external factors. Throughout the production cycle, the fish was monitored for growth parameters and examined for RFC/MFC using macroscopic and histological analysis. We found no association between the experimental groups and the presence of RFC/MFC. Histological investigations revealed melano-macrophages in the soft tissue in freshwater smolt, although no macroscopic discoloration was observed. MFC showed granulomas in various stages, suggesting a complex progression of the condition. In summary, we conclude that RFC/MFC is primarily caused by external factors found in the rearing facilities of farmed Atlantic salmon., (© 2023 The Authors. Journal of Fish Diseases published by John Wiley & Sons Ltd.)

Several parasites infecting the Asian carp species have been broadly spread along with the global fish trade. However, the diversity of specific parasite groups and their pattern of parasitism remain insufficiently elucidated even in their native regions. Here, we conducted a holistic identification and histological analysis of three Myxobolus species. The oblate myxospores of the first isolates were found infecting the spleen of silver carp Hypophthalmichthys molitrix, measuring 6.4 ± 0.4 (5.4-7.1) μm in length, 8.2 ± 0.4 (7.5-9.0) in width, and 5.9 ± 0.3 (5.2-6.2) in thickness. They were morphologically distinct from other congeners and regarded as a novel species, Myxobolus xiaoganensis n. sp. For the second isolates, we associated the formation of round plasmodia on the gill raker of silver carp with Myxobolus allotypica Chen, 1998. The third isolates, encapsulated in the intestinal serosa membrane of the grass carp Ctenopharyngodon idella, was proved to be conspecific to Myxobolus huasaensis Chen, 1998. While M. xiaoganensis n. sp. and M. huasaensis exhibited distinct origins, with genetic differences exceeding 4% from other congeners, M. allotypica displayed complete genetic congruence with an item available in Genbank. Histologically, myxospores of M. xiaoganensis n. sp. were scattered in the spleen, and the branchial and intestinal infections of M. allotypica and M. huasaensis were determined, respectively. Phylogenetic analysis demonstrated a non-monophyletic origin of both the Thelohanellus and Myxobolus genera, with a remarkable association of host affinity with myxosporean clustering., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier Ltd. All rights reserved.)

Nodular Gill Disease (NGD) is an emerging pathogenic condition that causes gill damage and mainly affects farmed freshwater fish, rainbow trout (Oncorhynchus mykiss) in particular, and this inevitably generates noticeable economic losses for the industry. The present study aimed to assess the prevalence of NGD in the Autonomous Province of Trento, a highly productive area located in Northeastern Italy, traditionally suited to rainbow trout production, and to identify possible risk factors for the introduction of this disease in trout farms. The necessary data were obtained through a questionnaire and the collection of fish samples. According to the data analysis, 42% of the investigated farms tested positive for NGD. The two possible risk factors identified for its introduction in farms are the presence of other diseases in the same farm (OR = 17.5; 95% CI = 2.7; 111.5) and having farms located 5 km upstream (OR = 24.8; 95% CI = 2.9; 211.1). These results highlight (i) a possible impairment of the immune system caused by other diseases as a predisposing factor to the manifestation of the pathology and (ii) the role of water in spreading pathogens., (© 2023 The Authors. Journal of Fish Diseases published by John Wiley & Sons Ltd.)

Nodular gill disease (NGD) is an infectious condition characterized by proliferative gill lesions leading to respiratory problems, oxygen deficiency and mortality in fish. Globally, NGD primarily impacts freshwater salmonids in intensive aquaculture systems. In recent years, numerous outbreaks of severe gill disease have affected more than half of the larger rainbow trout (Oncorhynchus mykiss) farms in Switzerland, mainly during spring and early summer. Mortality has reached up to 50% in cases where no treatment was administered. Freshwater amoeba are the presumed aetiologic agent of NGD. The gross gill score (GS) categorising severity of gill pathology is a valuable first-line diagnostic tool aiding fish farmers in identifying and quantifying amoebic gill disease (AGD) in farmed marine salmonids. In this study, the GS was adapted to the NGD outbreak in farmed trout in Switzerland. In addition to scoring disease severity, gill swabs from NGD-affected rainbow trout were sampled and amoeba were cultured from these swabs. Morphologic and molecular methods identified six amoeba strains: Cochliopodium sp., Naegleria sp., Vannella sp., Ripella sp., Saccamoeba sp. and Mycamoeba sp. However, the importance of the different amoeba species for the onset and progression of NGD still has to be evaluated. This paper presents the first description of NGD with associated amoeba infection in farmed rainbow trout in Switzerland., (© 2023 The Authors. Journal of Fish Diseases published by John Wiley & Sons Ltd.)

Lates calcarifer herpesvirus (LCHV) causes an emerging serious disease in aquaculture. Sudden drops in feed rates and mortality spikes exceeding 40%-50% often accompany LCHV infections in juvenile L. calcarifer, soon after transfer into sea cages. Affected fish have patchy white skin and fins, corneal opacity and frequently hang in surface water column like 'ghost' or 'zombie' fish. Fish have pale gills, fluid-filled intestines with yellowish casts, lipid depleted liver, enlarged spleen and kidney and reddened brain. Epithelial hyperplasia, apoptosis, marginated nuclear chromatin, amphophilic intranuclear inclusion bodies and the occasional multinucleated cells are observed in gills, skin, intestines, liver and kidney. These are often accompanied by lymphocytic-monocytic infiltration and extensive necrosis in gills, skin, kidney and intestines. Martius scarlet blue stains indicate presence of fibrin in vasculature in brain, gills, intestines, kidney and liver, or disseminated intravascular coagulation (DIC). DIC has been reported in human herpesviral infections. Multifocal lifting of intestinal epithelium with proteinaceous exudate and necrosis of several adjacent villi often progress to involve entire gut sections. Atrophied livers with accentuated lobules may progress to marked loss of hepatic acini. Multifocal dilated attenuated renal tubules are often accompanied by casts and marked protein loosing renopathy. This study on LCHV demonstrates that it can cause significant pathology and mortality., (© 2023 The Authors. Journal of Fish Diseases published by John Wiley & Sons Ltd.)