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Lactococcosis is a common bacterial fish disease caused by Lactococcus garvieae, L. petauri and L. formosensis. Although there are different PCR‐based techniques to identify the etiological agent, none of these can differentiate these two bacteria without sequencing PCR‐amplified fragments. In the present study, we developed a multiplex PCR assay for simultaneous detection and differentiation of L. garvieae and L. petauri. The specificity of the primers was validated against the bacterial DNA of the targeted and non‐targeted bacteria. The sizes of the PCR amplicons were obtained as 204 bp for the DUF1430 domain‐containing protein gene of L. garvieae, 465 bp for the Lichenan permease IIC component gene of L. petauri, and 302 bp for the teichoic acid biosynthesis protein F gene of both L. garvieae and L. petauri. The PCR amplicons were clearly separated by agarose gel electrophoresis. The multiplex PCR assay did not produce any amplification products with the DNA of the non‐targeted bacteria. The multiplex PCR detection limits for L. garvieae and L. petauri were 5 and 4 CFU in pure culture and 50 and 40 CFU/g in spiked tissue samples, respectively. It takes less than 2 h from plate‐cultured bacteria and 3 h from tissue samples to get results. In conclusion, the developed multiplex PCR assay is a rapid, specific, accurate, and cost‐effective method for the detection and differentiation of L. garvieae and L. petauri and is suitable to be used for routine laboratory diagnosis of L. garvieae and L. petauri. [ABSTRACT FROM AUTHOR]

ABSTRACT Hyperpigmented melanistic lesions (HPMLs) are a visual anomaly documented on the skin of smallmouth bass Micropterus dolomieu in the Susquehanna River Basin, Pennsylvania and in numerous other geographical locations. Currently, there is a lack of information on environmental and fish characteristics that may influence the prevalence of HPMLs associated with a recently described Adomavirus. The goal of this study was to understand potential drivers associated with HPMLs in socioeconomically and ecologically important riverine smallmouth bass populations. A total of 16,220 smallmouth bass were collected and examined for HPMLs between 2012 and 2022 in the Susquehanna River Basin. Overall, HPMLs were documented on 2.9% of fish collected. The interaction between temperature and fish size suggested differing relationships between shorter and longer fish with respect to temperature. Predicted probability of HPML prevalence ranged from 1.1% (95% CI = 0.3, 3.2) at 4°C to 0.01% (CI = 0.00, 0.04) at 26°C for an age‐0 (125 mm) fish. In contrast, predicted probability of HPML prevalence ranged from 10.5% (95% CI = 5.8, 18.9) at 4°C to 0.8% (CI = 0.4, 1.5) at 26°C for an adult (322 mm) fish. Overall, HPMLs were more common in longer fish during cooler temperature periods which also corresponds to key life history periods for smallmouth bass (e.g., pre‐spawn and overwintering) and could represent different exposure histories for juvenile and adult fish. [ABSTRACT FROM AUTHOR]

Alternatives to conventional chemical treatments for parasitic diseases in fish are needed. Microalgal‐sourced fatty acid ethyl esters (FAEEs) have shown an antiparasitic effect against Gyrodactylus turnbulli infection in guppies. Here, we tested a range of commercial FAEEs of various carbon chain lengths and unsaturation levels against two fish parasites. Guppies and barramundi infected with G. turnbulli and Trichodina sp., respectively, were used. The most effective FAEE, after excluding those toxic to fish, was ethyl laurate (12:0). For both parasites, the LD50 was 18.75 μM within 250 min of incubation. Ethyl eicosapentaenoate (20:5n3) was the next most effective FAEE against G. turnbulli, and dihomo‐γ‐linolenic acid ethyl ester (20:3n6) and ethyl α‐linolenate (18:3n3) were the next most effective against Trichodina sp. In addition, FAEEs prepared from the microalga Phaeodactylum tricornutum residue, after fucoxanthin extraction, were examined against Trichodina sp. infection in barramundi for the first time. LD85 and LD100 was achieved at 2.5 and 5 μL mL−1 of the FAEE preparation, respectively. In vivo, immersion of infected barramundi in 1.25 μL mL−1 of this preparation for 24 h reduced infection prevalence from 100% to 53% and was non‐toxic to fish. [ABSTRACT FROM AUTHOR]

This article discusses the discovery of a benign tumor in a turquoise killifish, which is commonly used in research on aging and toxicology. The tumor, known as a fibromyxoma, appeared as a small nodule on the fish's body and grew over time. Tests confirmed that the tumor was not caused by any infectious agents. This finding challenges previous beliefs about tumors in this species and provides valuable information for researchers studying aging. Additionally, another case of a tumor, specifically an adenocarcinoma, was found in a different species of fish used in aging research, highlighting the importance of considering neoplastic lesions in laboratory-reared fish. [Extracted from the article]

Dactylogyrus is one of the most common parasitic diseases in fish and causes huge losses to the aquaculture industry. With the advantages of safety, low toxicity and easy degradation, plant‐derived drugs are ideal for the creation of green aquatic ingredients. The use of plant‐derived drugs in aquaculture is limited by their low content and high processing costs, which is a challenge that can be solved by the chemical synthesis of plant‐derived drugs. Eleven new coumarin derivatives were synthesized and assessed for their anthelmintic activity in this study. Among them, the derivative 7‐((1‐tosyl‐1H‐1,2,3‐triazol‐4‐yl)methoxy)‐2H‐chromen‐2‐one (N11) has good anthelmintic activity and its mean anthelmintic efficacy against D. intermedius at a concentration of 10 μM reached 99.84%, which is even better than the anthelmintic activity of the positive control mebendazole. Further studies showed that N11 had concentration values of 3.31 and 1.94 μM for 50% maximal effect (EC50) against D. intermedius at 24 and 48 h, respectively. Furthermore, scanning electron microscopy revealed that N11 caused damage to D. intermedius. What is more noteworthy is that a substantial reduction in the ATP content of the parasite was observed following in vitro and in vivo administration of N11. Moreover, it was also found that N11 was able to inhibit the horizontal transmission of D. intermedius. Furthermore, real‐time quantitative PCR analysis was utilized to determine the expression profile of genes associated with anti‐inflammatory cytokines (IL‐10, TGF‐β and IL‐4) in goldfish. In all examined organs, it was observed that the expression of anti‐inflammatory cytokines increased subsequent to treatment with N11, according to the results. Thus, these results all suggest that N11 possesses good anthelmintic activity and is a potentially effective agent for the control of D. intermedius. [ABSTRACT FROM AUTHOR]

The skin represents an indicator of an animal's health status. Causes of cutaneous diseases in fish most often trace back to biological agents. However, fish skin diseases can also arise from a complex interaction of infectious and non‐infectious causes, making it more difficult to identify a specific aetiology. In the period between April and September of the years 2019–2022, four koi carp (Cyprinus carpio koi) from two European countries presented with multifocal, irregularly round, few mm to 1 cm, variably raised cutaneous reddened areas. The fish displayed good general condition. Cutaneous samples, investigated by microbiological and molecular methods and microscopy, did not indicate a primary pathogenic agent. Gross and histological findings of the cutaneous biopsies were consistent with a multifocal/reactive process centred on dermal vessels. The histological features were reminiscent of angiomatosis, a benign proliferative condition affecting the dermal vessels of mammals, including human patients. The clinical–pathological presentation and the dermatologic condition that affected the koi carp are discussed and compared with the veterinary and human literature. [ABSTRACT FROM AUTHOR]

Gambusia affinis is regarded as an important animal model. Edwardsiella tarda is one of the most serious pathogens affecting aquaculture. The study explores the effects of TLR2/4 partial signalling pathway in G. affinis of E. tarda infection. The study collected the brain, liver, and intestine after E. tarda LD50 and 0.85% NaCl solution challenge at different times (0 h, 3 h, 9 h, 18 h, 24 h, and 48 h). In these three tissues, the mRNA levels of PI3K, AKT3, IRAK4, TAK1, IKKβ, and IL‐1β were substantially enhanced (p <.05) then returned to normal levels. Additionally, Rac1 and MyD88 in liver had different trend with other genes in brain and intestine, which displayed significantly indifference. The overexpression of IKKβ, and IL‐1β indicated that E. tarda also caused immune reaction in intestine and liver, which would be consistent with delayed edwardsiellosis, which causes intestinal lesions and liver and kidney necrosis. Additionally, MyD88 plays a smaller role than IRAK4 and TAK1 in this signalling pathways. This study could enrich the understanding of the immune mechanism of the TLR2/4 signalling pathway in fish and might help to prescribe preventive measures against E. tarda to prevent infectious diseases in fish. [ABSTRACT FROM AUTHOR]

Granulomatous diseases caused by Nocardia seriously endanger the health of cultured fish. These bacteria are widely distributed, but prevention and treatment methods are very limited. Chronic granulomatous inflammation is an important pathological feature of Nocardia infection. However, the molecular mechanisms of granuloma formation and chronic inflammation are still unclear. Constructing a granuloma infection model of Nocardia is the key to exploring the pathogenesis of the disease. In this study, we established a granuloma model in the liver of largemouth bass (Micropterus salmoides) and assessed the infection process of Nocardia seriolae at different concentrations by analysing relevant pathological features. By measuring the expression of pro‐inflammatory cytokines, transcription factors and a pyroptosis‐related protein, we revealed the close relationship between pyroptosis and chronic inflammation of granulomas. We further analysed the immunofluorescence results and the expression of pyroptosis‐related protein of macrophage infected by N. seriolae and found that N. seriolae infection induced macrophage pyroptosis in vitro. These results were proved by flow cytometry analysis of infection experiment in vivo. Our results indicated that the pyroptosis effect may be the key to inducing chronic inflammation in the fish liver and further mediating granuloma formation. In this study, we explored the molecular mechanism underlying chronic inflammation of granulomas and developed research ideas for understanding the occurrence and development of granulomatous diseases in fish. [ABSTRACT FROM AUTHOR]

Viral diseases of fish cause significant economic losses in the aquaculture industry. Viral haemorrhagic septicemia virus (VHSV) is one of the most important viral diseases that affects more than 80 fish species. Detection of the disease, especially in the field, is critical to managing disease prevention and control programmes. Recombinase polymerase amplification (RPA) is an isothermal method with a very short amplification period and a single incubation temperature ranging from 37 to 42°C, which is a good alternative to the polymerase chain reaction (PCR). This study aimed to develop an RPA assay as sensitive as a real‐time RT‐PCR to detect VHSV. For this purpose, primers and probes are designed for the same targeted region of gG of VHSV. The ssRNA standards were prepared to find the detection limits of the assay. Detection limits were found ten‐fold differences between real‐time RT‐PCR and real‐time RT‐RPA. While the detection limit of the RT‐PCR was found as 95.5 viral RNA molecules/reaction in 95% probit value, the detection limit of RT‐RPA was found as 943.75 viral RNA molecules/reaction in 95% probit value using ssRNA standards. These results show that RPA is a suitable test for VHSV Ie detection. [ABSTRACT FROM AUTHOR]

Previously, rainbow trout fed deoxynivalenol (DON) or partially fed (pair‐fed) for 4 weeks before and during experimental infection with Flavobacterium psychrophilum had significantly decreased mortality rates. Similar results were obtained in the present study after 12 days, but not after 6 days, feeding 5 ppm DON or pair‐fed before infection. Furthermore, feeding 250 ppm chloroquine (CQ) also reduced mortality (p =.052) compared with controls and may have promise for treatment of some fish disease. Parallel groups of fish were maintained on the respective treatments for 15 days, with an additional group that was fasted, but were not infected to monitor autophagy. Fish that were fasted or fed DON had significantly increased LC3II in the liver and fasted fish had significantly decreased LC3II in muscle compared with controls using western blot. There was no difference in LC3II signal in the spleen of any treatment group. Fish that were fasted or pair‐fed had significant up‐regulation of the Atg genes atg4, atg7, lc3, gabarap and atg12 in muscle using quantitative PCR. Less alteration of Atg expression was seen in liver. Fish treated with CQ had significantly increased expression of atg4, becn1, lc3 and atg12 in the liver. Fish fed DON for 15 days had few alterations of Atg genes in either the liver or muscle. It is still not clear if autophagy is responsible for the resistance of rainbow trout fed DON, CQ or pair‐fed before F. psychrophilum infection. [ABSTRACT FROM AUTHOR]

The half‐smooth tongue sole, Cynoglossus semilaevis, is an important cultured flatfish species. Vibrio harveyi is a common pathogen to this fish, which may result in great economic loss to C. semilaevis culture industry. piRNAs, a non‐coding RNAs with 26–32 nt, have been regarded as promising biomarkers for cancer diagnosis and fish diseases. Here, we extracted the RNA from mucus of C. semilaevis and constructed the differential expression profiles of piRNAs between the sick fish (MS) and healthy fish (MC). We identified 45,696 differentially expressed piRNAs including 22,735 up‐regulated piRNAs and 22,961 down‐regulated piRNAs in MS group compared with MC group. The GO enrichment and KEGG pathway enrichment analyses of the differential piRNAs were carried out. The result showed immunity‐related target genes mainly involved in immune system process, response to stimulus, cell killing, immune system, infectious diseases and cell growth and death. The 10 most differentially expressed piRNAs were chosen to perform the qRT‐PCR, while only seven piRNAs were consistent with the sequence result. Compared with MC group, the expression levels of piR‐mmu‐72173>piR‐rno‐62831>piR‐xtr‐704880, piR‐dme‐15546979, piR‐mmu‐49941660, piR‐mmu‐29283297 and piR‐mmu‐1758399 were significantly lower, and piR‐gga‐10574 and piR‐gga‐134812 were significantly higher in MS group. These piRNAs may be potential biomarkers during the V. harveyi infection of C. semilaevis. This study could provide a new method to identify the infection status of C. semilaevis and understand better about the innate and adaptive immune system in C. semilaevis during bacterial infection. [ABSTRACT FROM AUTHOR]

To make crucial prevention, reduce fish losses and minimize the economic damage of diseases on the fish farm owners, a rapid detection of fish pathogens is mandatory. In this study, a loop‐mediated isothermal amplification assay combined with hydroxynaphthol blue dye (LAMP‐HNB) was developed and used for the rapid detection of Aeromonas salmonicida that caused significant economic losses in fish farming. Firstly, a pair of outer and inner primers specific for conserved fragment of vapA gene in A. salmonicida were designed and synthesized. Secondly, by optimizing the reaction conditions including reaction temperature, time, Mg2+ concentration, dNTP concentration and primer ratio, a LAMP‐HNB assay was successfully established for the detection of A. salmoncida. Thirdly, the assay showed good specificity with no false‐positive and false‐negative results, and good sensitivity with the detection limit of 3.077 × 10–6 ng/μl, which was 102 times more sensitive than the conventional PCR. Finally, the LAMP‐HNB assay was validated by the fish samples inoculated with different concentrations of A. salmoncida. This is the first development of rapid visual detection of A. salmonicida based on LAMP‐HNB assay, which has great application prospect and market for diagnostic testing, health certification and active surveillance programmers. [ABSTRACT FROM AUTHOR]

A better understanding of unique anatomical and functional features of the visual systems of teleost fish could provide key knowledge on how these systems influence the health and survival of these animals in both wild and culture environments. We took a systematic approach to assess some of the visual systems of spotted wolffish (Anarhichas minor), a species of increasing importance in North Atlantic aquaculture initiatives. The lumpfish (Cyclopterus lumpus) was included in these studies in a comparative manner to provide reference. Histology, light and electron microscopy were used to study the spatial distribution and occurrence of cone photoreceptor cells and the nature of the retinal tissues, while immunohistochemistry was used to explore the expression patterns of two photoreceptor markers, XAP-1 and XAP-2, in both species. A marine bacterial infection paradigm in lumpfish was used to assess how host-pathogen responses might impact the expression of these photoreceptor markers in these animals. We define a basic photoreceptor mosaic and present an ultrastructural to macroscopic geographical configuration of the retinal pigment tissues in both animals. Photoreceptor markers XAP-1 and XAP-2 have novel distribution patterns in spotted wolffish and lumpfish retinas, and exogenous pathogenic influences can affect the normal expression pattern of XAP-1 in lumpfish. Live tank-side ophthalmoscopy and spectral domain optical coherence tomography (SD-OCT) revealed that normal cultured spotted wolffish display novel variations in the shape of the retinal tissue. These two complementary imaging findings suggest that spotted wolffish harbour unique ocular features not yet described in marine teleosts and that visual function might involve specific retinal tissue shape dynamics in these animals. Finally, extensive endogenous biofluorescence is present in the retinal tissues of both animals, which raises questions about how these animals might use retinal tissue in novel ways for visual perception and/or communication. This work advances fundamental knowledge on the visual systems of two economically important but now threatened North Atlantic teleosts and provides a basic foundation for further research on the visual systems of these animals in health versus disease settings. This work could also be useful for understanding and optimizing the health and welfare of lumpfish and spotted wolffish in aquaculture towards a one health or integrative perspective., (© 2024 John Wiley & Sons Ltd.)

Ferritin, transferrin, and transferrin receptors I and II play a vital role in iron metabolism, health, and indication of iron deficiency anaemia in fish. To evaluate the use of high-iron diets to prevent or reverse channel catfish (Ictalurus punctatus) anaemia of unknown causes, we investigated the expression of these iron-regulatory genes and proteins in channel catfish fed plant-based diets. Catfish fingerlings were fed five diets supplemented with 0 (basal), 125, and 250 mg/kg of either inorganic iron or organic iron for 2 weeks. Ferritin, transferrin, and transferrin receptor I and II mRNA and protein expression levels in fish tissues (liver, intestine, trunk kidney, and head kidney) and plasma were determined. Transferrin (iron transporter) and TfR (I and II) genes were generally highly expressed in fish fed the basal diet compared to those fed the iron-supplemented diets. In contrast, ferritin (iron storage) genes were more expressed in the trunk kidney of fish fed the iron-supplemented diets than in those fed the basal diet. Our results demonstrate that supplementing channel catfish plant-based diets with iron from either organic or inorganic iron sources affected the expression of the iron-regulatory genes and increased body iron status in the fish., (© 2024 John Wiley & Sons Ltd.)

Piscirickettsiosis is a fish disease caused by the facultative intracellular bacterium, Piscirickettsia salmonis. Even though entry routes of P. salmonis in fish are not fully clear yet, the skin seems to be the main portal in some salmonid species. Despite the importance of fish mucous skin barrier in fighting waterborne pathogens, the interaction between salmonid skin mucus and the bacterium is unknown. This study seeks to determine the in vitro changes in the growth of two Chilean P. salmonis strains (LF‐89‐like and EM‐90‐like genotypes) and the type strain LF‐89T under exposures to skin mucus from Salmo salar and Oncorhynchus mykiss, as well as changes in the cytotoxic effect of P. salmonis on the SHK‐1 cells following exposures. The results suggest that the growth of three P. salmonis strains was not significantly negatively affected under exposures to skin mucus (adjusted at 100 μg total protein ml−1) of O. mykiss (69 ± 18 U lysozyme ml−1) and S. salar (48 ± 33 U lysozyme ml−1) over time. However, the cytotoxic effect of P. salmonis, pre‐exposed to salmonid skin mucus, on the SHK‐1 cell line was reliably identified only towards the end of the incubation period, suggesting that the mucus had a delaying effect on the cytotoxic response of the cell line to the bacterium. These results represent a baseline knowledge to open new avenues of research intended to understand how P. salmonis faces the fish mucous skin barrier. [ABSTRACT FROM AUTHOR]

Sixteen whitespotted bamboo sharks (Chiloscyllium plagiosum) with histologically similar bacterial abscesses were submitted to Northwest ZooPath from nine zoo and aquarium institutions over a 17-year period. These abscesses were characterized by inflammatory cell infiltrates and necrosis with intralesional small, Gram-positive, acid-fast negative, cocci bacteria. The clinical presentation, histologic findings, and culture results indicate that Enterococcus faecalis is a relatively common cause of these lesions in whitespotted bamboo sharks. This organism also provides a treatment challenge due to its inherent antibiotic-resistant properties and ability to form biofilms, confounding the host's immune response. Enterococcus faecalis represents an important cause for abscess formation and cellulitis in captive whitespotted bamboo sharks., (© 2024 John Wiley & Sons Ltd.)

Teratoma is a rare tumour in fish consisting of tissues from more than one germ layer, that may be located in either the gonads or extragonadal sites. Teratomas in many fish species remain poorly understood. In this work, we performed the first histological examinations of extragonadal teratomas in Poecilia wingei and also examined the influence of a large teratoma on male sexual activity. The studied teratomas shared general organizational features, but they also had variations in both external and internal features. In teratomas, the most common and highly differentiated tissues were striated muscle and nervous tissue. Despite the tumour, the male P. wingei exhibited normal mating behaviour and retained the ability for successful copulation. The structural features of extragonadal teratomas in guppy fish indicate a possible connection between these tumours and a failure of conserved processes operating in the embryonic germline., (© 2024 John Wiley & Sons Ltd.)

A permanent cell line, SPB (Snubnose pompano brain) was established from Trachinotus blochii by the explant culture method. It has been sub-cultured more than 75 passages and showed optimal growth at 28°C using L-15 medium supplemented with 15% to 20% FBS. The SPB cells were cryopreserved at different passage levels for various applications. SPB cells were composed of fibroblastic and epithelial-like cells. The SPB cells were tested for mycoplasma contamination which was found to be negative. The origin of the SPB cell line from T. blochii was confirmed by amplification of the mitochondrial cytochrome oxidase I (COI) gene. The transfection efficiency of SPB cell line is 15% assessed by expression of green fluorescent protein using pEGFP-N1 plasmid. In addition, two CMV promotor plasmids pFNCPE42-DNA and pcDNAVP28 were transfected to SPB cells and it shows high expression levels of FNCP of fish nodavirus and VP28 protein of white spot syndrome virus by immunostaining. The SPB cells showed susceptibility to SJNNV and the infection was confirmed by RT-PCR, Western blot, ELISA, TCID 50 and RT-qPCR. Experimental infection was carried out in T. blochii using SJNNV propagated in SPB cell line and found 100% mortality with clinical signs. The infection was confirmed by RT-PCR. The SPB cell line can be used for propagation of fish viral pathogens and production of the recombinant proteins., (© 2024 John Wiley & Sons Ltd.)

Lobsters and crayfish in Australasia can develop a condition known as Tail Fan Necrosis (TFN syndrome). Many attempts have been made to find a primary pathogen or link the syndrome to commercial activities, but a solution remains elusive. TFN syndrome is a 'wicked problem', a problem difficult or impossible to solve because of incomplete and contradictory information forming a matrix of potential outcomes with no simple solution. Reviewing the literature shows TFN syndrome is sometimes reported to develop in association with sterile blisters on the telson and uropods which may rupture permitting invasion by environmental fungal and/or bacterial flora. Whether blisters form prior to, or because of, infection is unknown. TFN syndrome sometimes develops in captivity, sometimes requires a previous insult to the telson and uropods, and prevalence is patchy in the wild. The literature shows the cause of blisters associated with TFN syndrome remains an enigma, for which we suggest several possible initiating factors. We strongly urge that researchers not 'jump to conclusions' as to the aetiology of TFN syndrome. It cannot be explained without carefully exploring alternative aetiologies whilst being cognisant of the age-old lesson that 'correlation does not equal causation'., (© 2024 The Authors. Journal of Fish Diseases published by John Wiley & Sons Ltd.)

Nanotechnology is an expanding and new technology that prompts production with nanoparticle-based (1-100 nm) organic and inorganic materials. Such a tool has an imperative function in different sectors like bioengineering, pharmaceuticals, electronics, energy, nuclear energy, and fuel, and its applications are helpful for human, animal, plant, and environmental health. In exacting, the nanoparticles are synthesized by top-down and bottom-up approaches through different techniques such as chemical, physical, and biological progress. The characterization is vital and the confirmation of nanoparticle traits is done by various instrumentation analyses like UV-Vis spectrophotometry (UV-Vis), Fourier transform infrared spectroscopy, scanning electron microscope, transmission electron microscopy, X-ray diffraction, atomic force microscopy, annular dark-field imaging, and intracranial pressure. In addition, probiotics are friendly microbes which while administered in sufficient quantity confer health advantages to the host. Characterization investigation is much more significant to the identification of good probiotics. Similarly, haemolytic activity, acid and bile salt tolerance, autoaggregation, antimicrobial compound production, inhibition of pathogens, enhance the immune system, and more health-beneficial effects on the host. The synergistic effects of nanoparticles and probiotics combined delivery applications are still limited to food, feed, and biomedical applications. However, the mechanisms by which they interact with the immune system and gut microbiota in humans and animals are largely unclear. This review discusses current research advancements to fulfil research gaps and promote the successful improvement of human and animal health., (© 2024 John Wiley & Sons Ltd.)

Aeromonas jandaei is a gram-negative bacterium commonly found in aquatic environments and can induce illnesses in amphibians, reptiles and aquatic animals. In this study, a strain of bacteria was isolated from the diseased Chinese soft-shell turtle (Pelodiscus sinensis), then named strain JDP-FX. This isolate was identified as A. jandaei after analysis of morphological, physiological and biochemical characteristics, as well as 16S rRNA and gyrB gene sequences. Virulence genetic testing further detected temperature-sensitive protease (eprCAI), type III secretion system (TTSS) (ascv), nuclease (nuc), cytotonic enterotoxin (alt) and serine proteinase (ser) in JDP-FX. Compared with healthy Chinese soft-shell turtle, the serum levels of total protein (TP), albumin (ALB) and globulin (GLB) were significantly decreased in the diseased Chinese soft-shell turtle, while, the levels of alanine aminotransferase (ALT), aspartate aminotransferase (AST) and alkaline phosphatase (ALP) were significantly increased. Histopathological observations showed that multiple tissues, including intestinal mucosa, liver and kidney, were severely damaged in the diseased Chinese soft-shell turtle. Moreover, the diseased Chinese soft-shell turtle had significant cell degeneration, necrosis, sloughing and interstitial inflammatory cell infiltration. The pathogenicity of JDP-FX was tested via artificial infection. The median lethal dosage (LD 50 ) of the strain was 1.05 × 10 5 colony forming units (CFU/g) per weight of Chinese soft-shell turtle. Drug susceptibility analysis revealed that JDP-FX was susceptible to ceftazidime, minocycline, cefoperazone, ceftriaxone and piperacillin. In addition, JDP-FX was resistant to doxycycline, florfenicol, sulfonamides, gentamicin, ampicillin and neomycin. Therefore, this study may provide guidance for further research into the diagnosis, prevention and treatment of JDP-FX infection., (© 2024 John Wiley & Sons Ltd.)

The live attenuated vaccine P7-P8 strain against herpesviral haematopoietic necrosis, which is caused by cyprinid herpesvirus 2 (CyHV-2), exhibits high protective efficacy in goldfish at 25°C, the predominant temperature for this disease; however, the effect of water temperature during the vaccination period on efficacy has not been determined. In this study, an in vitro experiment revealed that the vaccine strain grew between 15 and 30°C in the goldfish cell line RyuF-2. Subsequent in vivo efficacy tests were conducted with vaccination temperatures ranging from 15 to 30°C. During the vaccination period, organs were sampled to determine the vaccine growth dynamics. Blood plasma was collected to assess anti-CyHV-2 antibody titres. The protective efficacy of the vaccine at 15, 20, 25, and 30°C after subsequent virulent CyHV-2 challenge resulted in a relative percentage survival of 73.3%, 77.8%, 100%, and 77.8%, respectively, which indicated that the vaccine is effective over this temperature range. The vaccine virus load in the spleen was lowest at 15°C (10 3.7 DNA copies/mg) and highest at 25°C (10 6.5 DNA copies/mg). This indicates that the vaccine virus load over 10 4 DNA copies/mg may elicit sufficient acquired immunity. No significant differences in antibody titre were observed between groups, which suggests that cell-mediated immunity can be fundamentally involved in protection., (© 2023 John Wiley & Sons Ltd.)

In three water-supply reservoirs in the catchment area of the Odra River (Czech Republic), a special fish stock was monitored for control of health to estimate the mutagenic effect of chemicals. The results contribute to obtaining initial information about the morphology of erythrocyte abnormalities classified in 21 categories in 16 fish species in reservoirs with abundant salmonids (the Morávka Reservoir) or with the prevalence of cyprinids (the Kružberk and Šance Reservoirs), not directly exposed to the adverse environmental effects such as industrial, urban, agricultural and intensive farming activities. The different intensities and prevalence of nuclear abnormalities (NA) and cytoplasmic abnormalities (CA) in fish from the same reservoir habitat show that to be able to obtain an objective view of the genotoxic risk of chemicals, it is necessary to respect the different requirements of the fish for the exploitation of the food available in the biotope and to subject all representatives of piscivorous, omnivorous and benthophagous fishes in the reservoir to cytogenetic analysis. The occurrence of certain categories of erythrocyte abnormalities in diseased fish draws attention to the need to know the state of health of the fish and to employ this knowledge to exclude parasitological, viral and other infectious agents. These results are the first report of the frequencies of erythrocyte abnormalities in native fish. They should serve to check which of the categories examined could be of use in assessing the genotoxic risk in other stagnant and running aquatic ecosystems affected by anthropogenic activities., (© 2023 John Wiley & Sons Ltd.)

The ISAV has a genome composed of eight segments of (–)ssRNA, segment 6 codes for the hemagglutinin–esterase protein, and has the most variable region of the genome, the highly polymorphic region (HPR), which is unique among orthomyxoviruses. The HPR has been associated with virulence, infectivity and pathogenicity. The full length of the HPR is called HPR0 and the strain with this HPR is avirulent, in contrast to strains with deleted HPR that are virulent to varying degrees. The molecular mechanism that gives rise to the different HPRs remains unclear. Here, we studied in vitro the evolution of reassortant recombinant ISAV (rISAV) in Atlantic salmon head kidney (ASK) cells. To this end, we rescued and cultivated a set of rISAV with different segment 6‐HPR genotypes using a reverse genetics system and then sequencing HPR regions of the viruses. Our results show rapid multiple recombination events in ISAV, with sequence insertions and deletions in the HPR, indicating a dynamic process. Inserted sequences can be found in four segments of the ISAV genome (segments 1, 5, 6, and 8). The results suggest intra‐segmental heterologous recombination, probably by class I and class II template switching, similar to the proposed segment 5 recombination mechanism. [ABSTRACT FROM AUTHOR]

Viral encephalopathy and retinopathy (VER) is a serious neuropathological fish disease affecting in the Mediterranean aquaculture mainly European sea bass, Dicentrarchus labrax. It is well known that betanodaviruses are neurotropic viruses that replicate in nerve tissues, preferentially brain and retina. However, routes of entry and progression of the virus in the central nervous system (CNS) remain unclear. The role of four tissues—eye, oesophagus, gills and skin—as possible gateways of a betanodavirus, the redspotted grouper nervous necrosis virus (RGNNV), was investigated after experimental challenges performed on European seabass juveniles. The dispersal pattern of Betanodavirus at primarily stages of the disease was also assessed, using a real‐time qPCR assay. The development of typical clinical signs of VER, the presence of characteristic histopathological lesions in the brain and retina and the detection of viral RNA in the tissues of all experimental groups ascertained that successful invasion of RGNNV under all experimental routes was achieved. Transneuronal spread along pathways known to be connected to the initial site of entry seems to be the predominant scenario of viral progression in the CNS. Furthermore, viraemia appeared only after the installation of the infection in the brain. [ABSTRACT FROM AUTHOR]

The marine leech, Zeylanicobdella arugamensis, is a major threat to aquaculture in grouper‐producing countries including Indonesia. This study aimed at investigating prevalence, intensity and histopathology of the ectoparasite in humpback and hybrid groupers cultured in different rearing systems. A total of 260 groupers (60 humpback groupers and 200 hybrid groupers) were used for samples. The marine leech was observed on skin, fins, gills and mouth, followed by histopathological assay on the skin tissue. The results showed that prevalence of the leech in both groupers was higher when they were cultured in the floating net cages compared with the hatchery, p <.05. Furthermore, humpback grouper had a higher prevalence than hybrid grouper when they were cultured in a similar system, p <.05. Meanwhile, there was no significant difference in intensity between the two groupers, p >.05. Within the hybrid groupers, the highest prevalence was obtained from hybrid groupers reared in the earthen ponds. Histopathological studies showed that the infected groupers exhibited inflammation, congestion and erosion of the epidermis layer. Hybrid grouper had more severe histopathological lesions in the skin tissues. These results suggested that species and type of aquaculture system had significantly determined the prevalence, intensity and severity of lesion in Z. arugamensis infestation. [ABSTRACT FROM AUTHOR]

The use of zebrafish (Danio rerio) in biomedical research has expanded at a tremendous rate over the last two decades. Along with increases in laboratories using this model, we are discovering new and important diseases. We review here the important pathogens and diseases based on some 20 years of research and findings from our diagnostic service at the NIH‐funded Zebrafish International Resource Center. Descriptions of the present status of biosecurity programmes and diagnostic and treatment approaches are included. The most common and important diseases and pathogens are two parasites, Pseudoloma neurophilia and Pseudocapillaria tomentosa, and mycobacteriosis caused by Mycobacterium chelonae, M. marinum and M. haemophilum. Less common but deadly diseases are caused by Edwardsiella ictaluri and infectious spleen and kidney necrosis virus (ISKNV). Hepatic megalocytosis and egg‐associated inflammation and fibroplasia are common, apparently non‐infectious, in zebrafish laboratories. Water quality diseases include supersaturation and nephrocalcinosis. Common neoplasms are spindle cell sarcomas, ultimobranchial tumours, spermatocytic seminomas and a small‐cell carcinoma that is caused by a transmissible agent. Despite the clear biosecurity risk, researchers continue to use fish from pet stores, and here, we document two novel coccidia associated with significant lesions in zebrafish from one of these stores. [ABSTRACT FROM AUTHOR]

Keywords: Atlantic salmon; cardiomyopathy syndrome; piscine myocarditis virus; vertical transmission EN Atlantic salmon cardiomyopathy syndrome piscine myocarditis virus vertical transmission 715 718 4 05/27/20 20200601 NES 200601 INTRODUCTION Piscine myocarditis virus (PMCV) is the causal agent of cardiomyopathy syndrome (CMS), a disease affecting the heart of Atlantic salmon (Haugland et al., [7]). RNA extraction and real-time PCR PatoGen AS (Norway) performed RNA extraction and real-time PCR analysis for PMCV RNA in all samples. Wiik-Nielsen et al. screened eggs before any disinfection with resulting PMCV RNA detection prevalence of 10%. This showed a clear reducing effect of disinfection on the prevalence of eggs with detectable PMCV RNA, including 8 of 14 egg batches where the disinfection procedure completely removed the virus. [Extracted from the article]

Co‐infections commonly arise when two or multiple different pathogens infect the same host, either as simultaneous or as secondary concurrent infection. This potentiates their pathogenic effects and leads to serious negative consequences on the exposed host. Numerous studies on the occurrence of the bacterial, parasitic, fungal and viral co‐infections were conducted in various tilapia species. Co‐infections have been associated with serious negative impacts on susceptible fish because they increase the fish susceptibility to diseases and the likelihood of outbreaks in the affected fish. Co‐infections can alter the disease course and increase the severity of disease through synergistic and, more rarely, antagonistic interactions. In this review, reports on the synergistic co‐infections and their impacts on the affected tilapia species are highlighted. Additionally, their pathogenic mechanisms are briefly discussed. Tilapia producers should be aware of the possible occurrence of co‐infections and their effects on the affected tilapia species and in particular of the clinical signs and course of the disease. To date, there is still limited information regarding the pathogenicity mechanisms and pathogen interactions during these co‐infections. This is generally due to low awareness regarding co‐infections, and in many cases, a dominant pathogen is perceived to be of vital importance and hence becomes the target of treatment while the treatment of the co‐infectious agents is neglected. This review article aimed at raising awareness regarding co‐infections and helping researchers and fish health specialists pay greater attention to these natural cases, leading to increased research and more consistent diagnosis of co‐infectious outbreaks in order to improve control strategies to protect tilapia when infected with multiple pathogens. [ABSTRACT FROM AUTHOR]

Vibrio alginolyticus is one of the most serious causative agents of diseases in cultured marine fish and shellfish. However, the characteristics of virulence factors in pathogenic V. alginolyticus are poorly known. To gain insight into fish diseases caused by V. alginolyticus, we carried out two‐dimensional gel electrophoresis (2‐DE) combined with MALDI‐TOF mass spectrometry to identify uniquely expressed proteins in the disease‐causing V. alginolyticus. V. alginolyticus strains were isolated from marine environments and diseased fish obtained from southern Thailand. We identified seven unique proteins in the disease‐causing V. alginolyticus strain. Among those, the outer membrane protein A (OmpA) had the strongest expression. Therefore, the function of this protein was further analysed. To investigate the role of OmpA protein, an in‐frame deletion mutant of ompA was constructed using the homologous recombination method. Although the ompA mutant V. alginolyticus strain (ΔompA) grew normally, the mutant exhibited a significant defect in the swarming ability and the biofilm formation. Furthermore, Galleria mellonella larvae injected with the mutant bacteria had a significantly greater survival percentage than those injected with the wild‐type strain, demonstrating that OmpA protein is required for the pathogenicity of V. alginolyticus. Together, this study suggests a potential target for vaccine development against pathogenic V. alginolyticus strain. [ABSTRACT FROM AUTHOR]

Betanodaviruses are small ssRNA viruses that cause viral encephalopathy and retinopathy, a severe neuropathological infectious disease in marine fish species worldwide. In the present study, the occurrence of betanodaviruses was investigated in wild and cultured populations of fishes and invertebrates of the Greek territorial waters. Betanodaviruses were detected in 35 species belonging to 21 families and 12 orders. To our knowledge, 23 of those are reported for the first time in Greek waters, while 11 of them are reported for the first time globally. The positive samples were subjected to sequencing and phylogenetic analysis of partial segments of RNA1 and RNA2 genes. Almost all the viruses circulating in Greece fell within RGNNV genotype, while reassortant viruses were detected in three samples, namely two inter‐RGNNV and one RGNNV/SJNNV. A novel unclassified Betanodavirus sequence was also identified. Most of the Greek sequence types have a restricted geographic distribution except for two RNA1 and one RNA2 sequence types that are widespread throughout the Mediterranean basin. The results of this study indicate the range of reservoirs/hosts of betanodaviruses and also their wide spread in the Greek territorial waters and reinforce the hypothesis that wild fish species transmit the virus to cultured ones and vice versa. [ABSTRACT FROM AUTHOR]

While the book provides a comprehensive overview of bacterial fish diseases, it would have been more beneficial if it had included more information on the ecological, economic and social factors involved in preventing and treating bacterial diseases in aquaculture. They also highlight the impact of pollution on fish and the need to understand the epidemiology and microbiology of fish illnesses. Aquaculture plays a crucial role in meeting the growing demand for food worldwide, with fish providing approximately 17% of the global population's animal protein intake (FAO, [2]). [Extracted from the article]