Search

Type III secretion system (T3SS) is an important virulence system in Gram‐negative bacteria. In this investigation, different environmental conditions that regulate the expression of T3SS genes in Yersinia ruckeri were investigated aimed at obtaining a better understanding about its modulation after various environmental challenges. Four isolates of Y. ruckeri CSF007‐82, ATCC29473, A7959‐11 and YRNC10 were cultivated under the diverse in vitro challenges iron depletion, high salt, low pH and in the presence of fish serum or in the fish cell culture (Chinook Salmon Embryo – CHSE). The transcriptional modulation of the chromosomal genes ysaV, ysaC, ysaJ and prgH of ysa were investigated using quantitative real‐time PCR. The expression of prgH, ysaV, ysaC and ysaJ was differentially expressed in all four strains under evaluation. The highest gene expression levels were observed for Y. ruckeri YRNC10 AN after addition of 0.3 M NaCl in Luria Bertani broth. The results obtained from this study provide initial insights into T3SS responses in Y. ruckeri, which pave the way for further studies aimed at expanding our knowledge on the functional roles of the T3SS genes in Y. ruckeri. [ABSTRACT FROM AUTHOR]

ABSTRACT The objective of this study was to evaluate the effect of dietary supplementation with Citrus sinensis (L.) Osbeck essential oil on the growth, immune system, and resistance to Aeromonas hydrophila infection in Colossoma macropomum fingerlings. The experiment was conducted with five treatments (control diet, Tween80 diet, and diets supplemented with 200, 400, and 800 mg L−1 of C. sinensis essential oil) with four replicates. At the end of the experimental period, growth parameters were measured, and blood samples were collected for thrombogram, leukogram, and phagocytic activity analysis. A bacterial challenge with A. hydrophila was conducted for 96 h. C. macropomum fingerlings that were fed with 400 and 800 mg L−1 of C. sinensis essential oil had the highest growth parameters, with final weights of 533.18 ± 2.03 mg and 531.91 ± 2.67 mg, respectively, and an increase in the number of thrombocytes, lymphocytes, monocytes, and neutrophils, as well as higher phagocytosis rates compared to the control group. Regarding the challenge, fish in the 400 and 800 mg L−1 treatments also exhibited the lowest cumulative mortality rate (26.66% ± 3.33%). Therefore, supplementation with C. sinensis essential oil promotes growth, improves health, and enhances resistance to A. hydrophila in C. macropomum fingerlings. [ABSTRACT FROM AUTHOR]

ABSTRACT Haematological and plasma parameters are used in veterinary medicine as a diagnostic tool, allowing for the early detection of potential health and welfare issues. For the implementation of monitoring strategies, reference intervals (RIs) are needed to know when deviations in those parameters become clinically relevant. Here, haematological and plasma biochemical RIs of Norwegian Atlantic salmon (Salmo salar Linnaeus) are presented for 29 biomarkers. Two subgroups were defined from the reference population based on water type, referred to as pre‐adult (freshwater, 166.8 ± 118.4 g, N = 979) and adult (seawater, 2655 ± 2883 g, N = 4941), collected at 93 fish farms along the Norwegian coast. While the influence of various pre‐analytical variables on RIs is well documented, aquaculture practitioners cannot always control those variables in the field. Here, RIs were calculated from blood samples collected based on local pre‐analytical practices and then further processed under standardised conditions (analytical phase). RIs are therefore considered highly relevant for the application in the field. Finally, when taking blood samples in the field, it is advised to centrifuge whole blood as soon as possible, while plasma parameters investigated were stable for 168 h at 4°C or 72 h at room temperature. [ABSTRACT FROM AUTHOR]

ABSTRACT Chemokines play a crucial role in immune responses by facilitating the migration of cells expressing corresponding chemokine receptors along concentration gradients. Photobacterium damselae subsp. Damselae (PDD) and Nocardia seriolae (NS) are known to induce substantial mortality in silver pomfret populations, yet there exists a dearth of research regarding the immune response of CCLs in PDD‐ or NS‐infected silver pomfret. In our investigation, we identified 10 PaCCLs, which include one fish‐specific CCL (PaCCL44). Phylogenetic analysis revealed considerable diversity in CCL types and copy numbers among various teleost fishes. Notably, silver pomfret lacks specific CCL genes, with most PaCCLs exhibiting heightened expression levels in immune‐related organs such as the spleen and kidney, and some being expressed in mucosal immune‐related organs like the skin and gills. Transcriptome analysis conducted on silver pomfret infected with NS and PDD elucidated that the expression changes of PaCCLs primarily manifested in the spleen during the initial stages of NS infection, shifting to the kidney in later stages. Conversely, the expression changes of PaCCLs following PDD infection predominantly occurred in the kidney. In vitro studies using silver pomfret spleen cell lines demonstrated an early peak in PaCCLs expression during infection, followed by gradual decline with NS treatment and rapid diminishment with PDD treatment. These findings suggest that PaCCLs primarily support the innate immunity of silver pomfret, potentially exhibiting chemotactic effects in the early infection stages, such as the synergistic action of PaCCL4 and PaCCL25, and later serving as direct antibacterial agents. NS invasion is characterised by a chronic infection affecting multiple organs, whereas PDD primarily inflicts severe damage to the kidney. PaCCL19a and PaCCL19b are specific to PDD, and their expression levels may decrease in the later stages of infection due to PDD immune escape. These data offer initial insights into understanding the mechanism underlying the innate immune response of the CCL gene family in silver pomfret and provide theoretical underpinnings for fish culture practices. [ABSTRACT FROM AUTHOR]

Mucosal barriers are gatekeepers of health and exhibit homeostatic variation in relation to habitat and disease. Mucosal Mapping technology provides an in‐depth examination of the dynamic mucous cells (MCs) in fish mucosal barriers on tangential sections, about 90° from the view of traditional histology. The method was originally developed and standardized in academia prior to the establishment of QuantiDoc AS to apply mucosal mapping, now trademarked as Veribarr™ for the analysis of skin, gills and gastrointestinal tracts. Veribarr™ uses design‐based stereology for the selection and measurement of cell area (size) (μm2), the volumetric density of MCs in the epithelium (MCD, amount of the epithelia occupied by MCs, in %) and the calculated abundance of the MCs (barrier status or defence activity). MC production was mapped across the skin and gill epithelia in 12 species, discovering that gills consistently have two distinct groups of MCs, one on the lamellae where MCs are few and small and one on the filament where MCs are larger and more abundant. MCs were usually much larger in the skin than in the gills, with the latter requiring fewer and smaller cells for adequate respiration. The difference observed between MCs in gill lamella and gill filament is likely a result of functional demands. In addition, our findings also highlight a variation in the mucosal parameters between the species skin, which cannot be explained by the weight differences, and a potential link between MC distribution and species‐specific lifestyles in the gill lamella. This diversity necessitates the development of species and tissue site‐specific reference intervals for mucosal health evaluation. Mucosal bivariate reference intervals were developed for MC production, including size (trophy) and calculated defence activity (plasia) in the skin and gills of Atlantic salmon, to contrast new measurements against historical data patterns. The application of mucosal reference intervals demonstrates that stress from parasites and treatments can manifest as changes in mucosal architecture, as evidenced by MC hypertrophy and hyperplasia within the gill lamellae. These reference intervals also facilitate comparisons with wild Atlantic salmon, revealing a somewhat higher MC level in farmed salmon gill lamellae. These findings suggest that MC hyperplasia and hypertrophy in the gills are stress/environmental responses in aquaculture. They also advocate for developing specific mucosal bivariate homeostatic reference intervals in aquaculture to improve fish health and welfare across all farmed species. [ABSTRACT FROM AUTHOR]

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. [ABSTRACT FROM AUTHOR]

The introduced salmonid ectoparasite Gyrodactylus salaris has been detected on Atlantic salmon in 53 Norwegian rivers and in 39 Norwegian fish farms. In affected rivers, the mortality of Atlantic salmon juveniles is very high, estimated to a mean of 86%. G. salaris has been considered one of the biggest threats to wild Norwegian Atlantic salmon stocks. With various measures, the authorities have reduced the potential for further spread of the parasite to new rivers and fish farms, and G. salaris has been eradicated from 43 rivers and all fish farms. Furthermore, the eradication process is almost completed in five affected rivers located at the Norwegian west coast, while preparations for the eradication in the remaining five rivers in the southeastern part of Norway have begun. The goal of Norwegian management is to eradicate the introduced pathogenic G. salaris strains from all occurrences in Norway. In fish farms, the parasite has been removed by mandatory slaughter of infected fish. In rivers, G. salaris has mostly been removed by killing all the fish hosts with rotenone. The indigenous genetic Atlantic salmon stocks are re‐established after eradication of the parasite. New methods are developed using chemicals that kill the parasite without killing fish in the rivers. Norwegian authorities have so far used more than NOK 1.5 billion on research, monitoring and combating G. salaris. However, the benefits are considered many times greater than the spending. Without control measures, G. salaris would likely have spread to new Atlantic salmon rivers where the same catastrophic outcome had to be expected. The Norwegian authorities seem to meet the goal in their long‐term work to halt the spread of G. salaris and to eradicate the parasite in affected rivers. [ABSTRACT FROM AUTHOR]

The emergence of antibiotic‐resistant bacteria (ARBs) and genes (ARGs) in aquaculture underscores the urgent need for alternative veterinary strategies to combat antimicrobial resistance (AMR). These measures are vital to reduce the likelihood of entering a post‐antibiotic era. Identifying environmentally friendly biotechnological solutions to prevent and treat bacterial diseases is crucial for the sustainability of aquaculture and for minimizing the use of antimicrobials, especially antibiotics. The development of probiotics with quorum‐quenching (QQ) capabilities presents a promising non‐antibiotic strategy for sustainable aquaculture. Recent research has demonstrated the effectiveness of QQ probiotics (QQPs) against a range of significant fish pathogens in aquaculture. QQ disrupts microbial communication (quorum sensing, QS) by inhibiting the production, replication, and detection of signalling molecules, thereby reducing bacterial virulence factors. With their targeted anti‐virulence approach, QQPs have substantial promise as a potential alternative to antibiotics. The application of QQPs in aquaculture, however, is still in its early stages and requires additional research. Key challenges include determining the optimal dosage and treatment regimens, understanding the long‐term effects, and integrating QQPs with other disease control methods in diverse aquaculture systems. This review scrutinizes the current literature on antibiotic usage, AMR prevalence in aquaculture, QQ mechanisms and the application of QQPs as a sustainable alternative to antibiotics. [ABSTRACT FROM AUTHOR]

Channel catfish (Ictalurus punctatus) are a food fish extensively reared in aquaculture facilities throughout the world and are also among the most abundant wild catfish species in North America, making them a popular target of anglers. Furthermore, channel catfish are important members of aquatic ecosystems; for example, they serve as a glochidial host for the endangered winged mapleleaf mussel (Quadrula fragosa), making them critical for conserving this species through hatchery‐based restoration efforts. During a routine health inspection, a novel aquareovirus was isolated from channel catfish used in mussel propagation efforts at a fish hatchery in Wisconsin. This virus was isolated on brown bullhead cells (ATCC CCL‐59) and identified through metagenomic sequencing as a novel member of the family Spinareoviridae, genus Aquareovirus. The virus genome consists of 11 segments, as is typical of the aquareoviruses, with phylogenetic relationships based on RNA‐dependent RNA polymerase and major outer capsid protein amino acid sequences showing it to be most closely related to golden shiner virus (aquareovirus C) and aquareovirus C/American grass carp reovirus (aquareovirus G) respectively. The potential of the new virus, which we name genictpun virus 1 (GNIPV‐1), to cause disease in channel catfish or other species remains unknown. [ABSTRACT FROM AUTHOR]

This article discusses the basibranchial structure and its impact on cardiac morphology in Atlantic salmon. The study finds that farmed salmon often experience structural abnormalities in their hearts, which may be caused by intensive breeding programs and stressful farming practices. These abnormalities can lead to cardiac collapse and increased mortality, particularly during treatments against sea lice. The article also highlights the importance of understanding optimal cardiac function in order to address these issues. The authors suggest that the basibranchial cartilaginous structure may play a role in altering the shape and function of the ventricle, negatively affecting the heart's performance. However, further research is needed to fully understand the causes and implications of these findings. [Extracted from the article]

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. [ABSTRACT FROM AUTHOR]

Flavobacterial infection associated with diseased fish is caused by multiple bacterial species within the family Flavobacteriaceae. In the present study, the Chilean isolate FP99, from the gills of a diseased, farmed rainbow trout (Oncorhynchus mykiss), was characterized using phenotypic and genomic analyses. Additionally assessed was pathogenic activity. Phylogenetic analysis based on 16S rRNA gene sequencing confirmed that isolate FP99 belonged to the genus Epilithonimonas, an average nucleotide identity value of 100% was detected with the Chilean isolate identified as Epilithonimonas sp. FP211‐J200. In silico genome analysis, mechanisms for toxins production, and superantigens, adhesion, or other genes associated with virulence were not detected. However, genes encoding proteins for antibiotic resistance were found, including the chrA gene and the nucleotide sequence that encodes for multiple antibiotic resistance MarC proteins. Furthermore, the blaESP‐1 gene (87.85% aminoacidic sequence identity), encoding an extended‐spectrum subclass B3 metallo‐β‐lactamase and conferring carbapenem‐hydrolysing activity, and the tet(X) gene, which encodes a monooxygenase that catalyses the degradation of tetracycline‐class antimicrobials were carried by this isolate. Phenotyping analyses also supported assignment as E. ginsengisoli. Challenge trials against healthy rainbow trout resulted in no observed pathogenic effect. Our findings identify for the first time the species E. ginsengisoli as associated with fish farming, suggesting that this isolate may be a component of the microbiota of the freshwater system. Notwithstanding, poor environmental conditions and any stressors associated with aquaculture situations or lesions caused by other pathogenic bacteria, such as F. psychrophilum, could favour the entry of E. ginsengisoli into rainbow trout. [ABSTRACT FROM AUTHOR]

This article provides a detailed analysis of gonadal neoplasia in Chinese gold barbs kept in home aquariums. The study examines the clinical, pathologic, and immunohistochemical characteristics of multiple gonadal neoplasms found in male and female gold barbs. The neoplasms identified include seminomas in males and fibrosarcoma in a female, as well as various other types of tumors in both sexes. The article also discusses the potential genetic and environmental factors that may contribute to the development of these neoplasms. This research emphasizes the need for further investigation into fish pathology and aquatic animal health in relation to neoplastic diseases. [Extracted from the article]

Golden pompano (Trachinotus blochii) is a carnivorous teleost cultured in the Asia‐Pacific region. Fish culture in high densities and numbers results in disease outbreaks, causing huge economic losses. Here, we collected cultured golden pompanos from 2021 to 2022 and identified the pathogens isolated from the diseased fish. Out of a total of 64 clinical cases observed in both sea cages and fish ponds, it was found that Nocardia seriolae was the predominant pathogen (26%), followed by Lactococcus garvieae (13%). Trichodina spp. was the most prevalent parasite in sea cages and earthen ponds (21%), while Neobenedenia spp. was the primary parasitic pathogen (16%) in sea cages. Given these findings, further investigations were conducted, including antibiotic susceptibility and pathogenicity tests specific to N. seriolae in golden pompanos. Antibiotic susceptibility tests of N. seriolae revealed that all strains were susceptible to doxycycline, oxytetracycline, florfenicol and erythromycin but resistant to amoxicillin and ampicillin. Additionally, a pathogenicity assessment was carried out by administering an intraperitoneal injection of 0.1 mL containing 107 CFU of N. seriolae per fish. The mortality rates observed varied between 40% and 90%, with the P2 strain exhibiting the highest level of virulence, resulting in a cumulative mortality of 90%. Therefore, disease outbreaks in fish can be minimized by developing effective treatments and prevention methods. [ABSTRACT FROM AUTHOR]

Viral diseases are a serious problem in Atlantic salmon (Salmo salar L.) farming in Norway, often leading to reduced fish welfare and increased mortality. Disease outbreaks in salmon farms may lead to spread of viruses to the surrounding environment. There is a public concern that viral diseases may negatively affect the wild salmon populations. Pancreas disease (PD) caused by salmonid alphavirus (SAV) and heart and skeletal muscle inflammation (HSMI) caused by piscine orthoreovirus‐1 (PRV‐1) are common viral diseases in salmon farms in western Norway. In the current study, we investigated the occurrence of SAV and PRV‐1 infections in 651 migrating salmon post‐smolt collected from three fjord systems (Sognefjorden, Osterfjorden and Hardangerfjorden) located in western Norway in 2013 and 2014 by real‐time RT‐PCR. Of the collected post‐smolts, 303 were of wild origin and 348 were hatchery‐released. SAV was not detected in any of the tested post‐smolt, but PRV‐1 was detected in 4.6% of them. The Ct values of PRV‐1 positive fish were usually high (mean 32.0; range: 20.1–36.8). PRV‐1 prevalence in post‐smolts from the three fjords was 6.1% in Sognefjorden followed by 4.8% in Osterfjorden and 2.3% in Hardangerfjorden. The prevalence PRV‐1 was significantly higher in wild (6.9%) compared to hatchery‐released post‐smolt (2.6%). The occurrence of PRV‐1 infection in the fish was lowest in the Hardangerfjorden which has the highest fish farming intensity. Our results suggest that SAV infection are uncommon in migrating smolt while PRV‐1 infection can be detected at low level. These findings suggest that migrating smolts were at low risk from SAV or PRV‐1 released from salmon farms located in their migration routes in 2013 and 2014. [ABSTRACT FROM AUTHOR]

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. [ABSTRACT FROM AUTHOR]

In vivo and in vitro treatments against Sparicotyle chrysophrii (Monogenea: Microcotylidae) parasitizing the gills of gilthead sea bream (Sparus aurata L.). Strong effect of long-term Sparicotyle chrysophrii infection on the cellular and innate immune responses of gilthead sea bream, Sparus aurata. Keywords: antiparasitics; efficacy; gilthead seabream; praziquantel; Sparicotyle chrysophrii; Sparus aurata EN antiparasitics efficacy gilthead seabream praziquantel Sparicotyle chrysophrii Sparus aurata 1439 1443 5 11/10/23 20231201 NES 231201 INTRODUCTION Praziquantel (PZQ) is a synthetic broad-spectrum antiparasitic, widely used in veterinary and human medicine (Andrews et al., [1]). [Extracted from the article]

Streptococcus iniae is a bacterial pathogen that causes streptococcosis, leading to significant losses in fish aquaculture globally. This study reported a newly developed probe‐based quantitative polymerase chain reaction (qPCR) method for the detection of S. iniae. The primers and probes were designed to target the lactate oxidase gene. The optimized method demonstrated a detection limit of 20 copies per reaction and was specific to S. iniae, as evidenced by no cross‐reactivity when assayed against genetic materials extracted from 23 known aquatic animal pathogens, and fish samples infected with Streptococcus agalactiae or Streptococcus dysgalactiae. To validate the newly developed qPCR protocol with field samples, fish specimens were systematically investigated following the Food and Agriculture Organization of the United Nations & Network of Aquaculture Centres in Asia‐Pacific three diagnostic levels approach, which integrated basic and advanced techniques for disease diagnosis, including observation of gross signs (level I), bacterial isolation (level II), qPCR and 16S rDNA sequencing (level III). The result showed that 7/7 affected farms (three Asian seabass farms and four tilapia farms) experiencing clinical signs of streptococcosis were diagnosed positive for S. iniae. qPCR assays using DNA extracted directly from fish tissue detected S. iniae in 11 out of 36 fish samples (30.6%), while 24 out of 36 samples (66.7%) tested positive after an enrichment step, including apparently healthy fish from affected farms. Bacterial isolation of S. iniae was only successful in a proportion of clinically diseased fish but not in healthy‐looking fish from the same farm. Overall, the newly developed qPCR protocol combined with enrichment would be a useful tool for the diagnosis and surveillance of S. iniae infections in fish populations, thereby aiding in the disease control and prevention. [ABSTRACT FROM AUTHOR]

Cryptocaryoniasis (marine white spot disease), caused by Cryptocaryon irritans, is a major threat to marine fish cultures in tropical and subtropical waters, and a serious nuisance to hobbyists with saltwater fish tanks. With only classical treatment schedules such as copper salts or hyposaline baths being available, control of the disease remains a challenge. In this study, we investigated the effect of Biokos, a viscosin‐like lipopeptide surfactant extracted from a bacterium of the Pseudomonas genus, on the external life stages of C. irritans, including theronts, protomonts and tomonts. The present study demonstrated that the compound has an antiparasitic effect on all tested external stages of the parasite. In particular, when Biokos was used at 48 mg/L, it was able to kill almost all theronts and protomonts within 1 h in in vitro experiments, and using the same concentration in an in vivo challenge experiment, the parasitic load was reduced by more than 95% compared to the control group with no Biokos. Additionally, cultured fish cells were able to proliferate, and fish showed no adverse signs at Biokos concentrations that were effective in killing the parasite. Thus, Biokos may be a promising way for preventing or reducing the burden of this parasitic disease in the future. [ABSTRACT FROM AUTHOR]

Flavobacterium psychrophilum affects many cultured fish species and is considered one of the most important bacterial pathogens causing substantial economic losses in salmonid aquaculture worldwide. Here, F. psychrophilum was identified by matrix‐assisted laser desorption ionization time‐of‐flight mass spectrometry (MALDI‐TOF MS) and nested PCR as the aetiological agent causing mortality in diseased juvenile Siberian sturgeons (Acipenser baerii) reared on a freshwater fish farm. Diseased sturgeons were lethargic and displayed dark skin pigmentation, increased mucus production and the presence of skin ulcerations and haemorrhages specially on the ventral side and the base of fins. The histological examination of fish revealed proliferative branchitis, ulcerative and necrotizing dermatitis and myositis, lymphoid tissue atrophy, liver and kidney degeneration and thrombosis. To the best of our knowledge, this is the first report describing the infection of Siberian sturgeons by F. psychrophilum. The detection of F. psychrophilum in diseased Siberian sturgeons and the description of the pathological findings observed during the outbreak may contribute to a better understanding of the bacterium pathogenicity and the range of fish species susceptible to infection. [ABSTRACT FROM AUTHOR]

Acanthocephalosis is caused by the endoparasite Neoechynorhyncus buttnerae and affects fish farmed in the Amazon. This study assessed the efficacy of therapeutic levamisole hydrochloride (LVC) baths against N. buttnerae and its effects on juvenile tambaqui blood parameters. In vitro and in vivo tests were carried out, the latter employing two experimental therapeutic LVC bath protocols. Concerning in vitro efficacy, the T75 (75 mg.L−1 LVC) and T100 (mg.L−1 LVC) treatments were 100% effective in 15 min, while the T50 (50 mg.L−1 LVC) and T25 (25 mg.L−1 LVC) treatments required parasite exposure for 45 and 60 min, respectively. During exposure, the parasites displayed reduced motility, proboscis retraction, coiling into a spiral shape, body rigidity and swelling. The LVC LC50–72h for juvenile tambaqui was 115 mg.L−1. Regarding in vivo efficacy for Protocol I (8‐h bath), the T125 resulted in 82% effectiveness, while in Protocol II (two 8‐h baths with a 24‐h interval), the T115 treatment (115 mg.L−1 LVC) achieved 95.6% effectiveness without clinical intoxication signs, despite behavioural changes. No significant changes were observed in fish blood parameters. LVC was, therefore, highly effective both in vitro and in vivo in controlling the acanthocephalan N. buttnerae without compromising tambaqui juvenile homeostasis. [ABSTRACT FROM AUTHOR]

Oomycete infections in farmed fish are one of the most significant disease issues in salmonid aquaculture worldwide. In the present study, Saprolegnia spp. in different farmed fish species in Finland were identified, and the molecular epidemiology of especially Saprolegnia parasitica was examined. We analysed tissue samples from suspected oomycete‐infected salmonids of different life stages from a number of fish farms, as well as three wild salmonids. From collected oomycete isolates, the ITS1, 5.8S and ITS2 genomic regions were amplified, analysed phylogenetically and compared with corresponding sequences deposited in GenBank. Of the sequenced isolates, 91% were identified as S. parasitica. Isolates of yolk sac fry were identified as different Saprolegnia spp. Among the isolates from rainbow trout eggs Saprolegnia diclina dominated. In order to determine potential dominating clones among the S. parasitica, isolates were analysed using Multi Locus Sequence Typing (MLST). The results showed that one main clone contained the majority of the isolates. The MLST analysis showed four main sequence types (ST1–ST4) and 13 unique STs. This suggests that the Saprolegnia infections in farmed fish in Finland are not caused by different strains originating in the farm environment. Instead, one main clone of S. parasitica is present in Finnish fish farms. [ABSTRACT FROM AUTHOR]

Chronic subclinical infection with the aetiological agent of bacterial kidney disease (BKD), Renibacterium salmoninarum, presents challenges for the clinical management of disease in farmed salmonids and for prevalence estimation. Harvested salmon sampled at processing plants provide the opportunity to describe subclinical outcomes of BKD using gross necropsy observations and diagnostic test results in farmed Atlantic salmon (Salmo salar L.) populations that are apparently healthy (i.e. alive at harvest) but naturally exposed to R. salmoninarum infection. Sampling of farmed salmon (Population A, n = 124 and Population B, n = 160) was performed immediately post‐slaughter as fish were being processed at a plant in New Brunswick, Canada. Populations were selected based on planned harvests from sites with histories of recent exposure events related to clinical BKD as evidenced by the site veterinarian's diagnosis of mortality attributable to BKD: One site (Pop A) had recently increasing mortalities attributed to BKD, and the other site (Pop B) had ongoing low‐level mortalities with BKD pathology. As expected with the different exposure histories, Pop A had a higher percentage (57.2%) of R. salmoninarum culture‐positive kidney samples compared with similar fish samples in Pop B (17.5%). Diagnosis of R. salmoninarum by gross granulomatous lesions in internal visceral organs, bacterial culture and identification by matrix‐assisted laser desorption/ionization time‐of‐flight mass spectrometry (MALDI‐TOF MS) using different swab transport methods, and molecular detection methods (quantitative PCR, qPCR) were compared. Agreement of culture‐positive percentages at the sample level was moderate (kappa: 0.61–0.75) among specimens collected using different kidney sampling methods in Pop A and Pop B. The highest proportion of R. salmoninarum‐positive cultures occurred when kidney tissues were transported to the laboratory and inoculated directly onto agar using a swab (94% of cultures from Pop A and 82% from Pop B when fish were positive by any culture method). Fish with cumulative lesion scores (severity of granulomatous lesions in 3 different visceral organs) of >4 were all culture positive, and when compared with non‐lesioned fish, had substantially higher odds of being culture positive: Pop A: odds ratio (OR) = 73, 95% confidence interval (CI) (7.91, 680.8); Pop B: OR = 66, 95% CI (6.12, 720.7). Our study found that onsite postmortem examinations with severity scores of gross granulomatous lesions were predictive of positive culture results for R. salmoninarum, and they were a useful proxy for assessing prevalence in apparently healthy populations with subclinical infection. [ABSTRACT FROM AUTHOR]

Mycobacterium infection in fish is a well‐known disease problem globally, mainly in the farming of ornamental fish or fish for food. Less is known about the prevalence, distribution and the effects such infections have on wild fish species. Presumptive mycobacteriosis has previously been observed in Atlantic mackerel (Scomber scombrus). Since 2018, there has been an increase in reports of granulomatous kidney disease in Atlantic mackerel with the suspicion of this being mycobacteriosis. A total of six individuals were sent to the Institute of Marine Research for further examination. They were caught in the Nordic Sea by either commercial fishing vessels or during the International Ecosystem Summer Survey in the Nordic Seas (IESSNS research cruise) between 2018 and 2020. Samples for both histological and molecular analysis were collected. Here, we detect a likely novel Mycobacterium species in tissue samples from Atlantic mackerel with this condition, on the basis of rDNA and protein gene sequences. The same unnamed bacterium seems to have been found in some Pacific marine fishes. The macroscopic and histological manifestation of the disease is described. Over the past years, there has been an increase in reports of mycobacteriosis worldwide and climate change has been suggested as one of the driving forces as these bacteria prefer warm water. [ABSTRACT FROM AUTHOR]

Ichthyophthiriasis, caused by the parasitic ciliate Ichthyophthirius multifiliis (Ich), is considered one of the most harmful diseases affecting freshwater fish globally. It can cause mass mortalities of fish in intensive farming systems. In such systems, it is thus necessary to detect and quantify the number of Ich in the water so that control measures can be implemented before Ichthyophthiriasis breaks out. In recent years, molecular diagnostic methods have become increasingly important in aquaculture. Real‐time quantitative polymerase chain reaction (qPCR) and droplet digital polymerase chain reaction (ddPCR) have become robust assays for detecting pathogens. In this study, a set of specific primers and a TaqMan‐minor groove binder probe targeting the small‐subunit rDNA (SSU rDNA) of Ich were developed. They were used in qPCR and ddPCR assays to compare the performance of these two different methods in quantitatively detecting Ich. After optimizing the reaction conditions, both qPCR and ddPCR assays were found to have high linearity and quantitative correlations for standard plasmid DNA. When used for the detection of Ich eDNA in water samples, the qPCR assay had a wider detection range, making it a suitable method to screen for the prevalence of Ichthyophthiriasis. However, the ddPCR approach had higher sensitivity, which would help provide advance notice of the disease in complex water environmental samples. [ABSTRACT FROM AUTHOR]

Cryptocaryon irritans causes one of the most serious diseases in various wild and cultured marine fish, leading to mass mortality and economic loss. In this study, hydroxyl radical (•OH) solution produced by strong ionization discharge combined with water jet cavitation effect was injected into orange‐spotted grouper (Epinephelus coioides) aquaculture tanks for C. irritans control. The results showed that all C. irritans theronts were inactivated by •OH solution at concentrations of 0.5 mg/L within 2 min. •OH could induce alteration of shape, the absence of motility and macronucleus dispersion in theronts. A possible explanation was that the macronucleus of C. irritans might be damaged by •OH; as a result, its metabolism and life activities were disturbed. The •OH treatment increased the survival rate of E. coioides challenged with C. irritans from 64.7 ± 8.0% (mean ± SD) to 100% and reduced their infection intensity significantly. Stress response biomarkers such as malonaldehyde, glutathione, glutathione peroxidase, superoxide dismutase (SOD) and catalase levels in the gills of E. coioides at different time points were analysed. The SOD activity in the •OH group first decreased and then recovered to the initial level at the end of the experiment. The other stress response biomarkers had no significant difference from that in the uninfected control group after •OH treatment. Additionally, the gill of E. coioides in the •OH group exhibited slight and reversible transformation compared with the uninfected control group. Compared with •OH treatment, chlorine dioxide and formalin treatment reduced the survival rate, induced oxidative damage and changed the histological gill structure in E. coioides. In conclusion, •OH could be applied effectively to control C. irritans infection without affecting the normal physiological condition of E. coioides. [ABSTRACT FROM AUTHOR]

This study evaluated the probiotic potential of the biofilm formed by the strain Pseudomonas sp. RGM2144 on rainbow trout survival. When challenged with the fish pathogen Flavobacterium psychrophilum, Pseudomonas sp. RGM2144 increased rainbow trout survival to 92.7 ± 1.2% (control: 35.3 ± 9.5%, p <.0001). The draft genome of Pseudomonas sp. RGM2144 is 6.8 Mbp long, with a completeness 100% and a contamination of 0.4%. The genome contains 6122 protein‐coding genes of which 3564 (~60%) have known functions. The genome and phylogeny indicate that Pseudomonas sp. RGM2144 is a new species in the Pseudomonas genus, with few virulence factors, plasmids, and genes associated with antimicrobial resistance, suggesting a non‐pathogenic bacterium with protective potential. In addition, the genome encodes for 11 secondary metabolite biosynthetic gene clusters that could be involved in the inhibition of F. psychrophilum. We suggest that Pseudomonas sp. RGM2144 may be applied as a probiotic in salmonid fish farming. [ABSTRACT FROM AUTHOR]

Keywords: digital droplet PCR; Midichloria-like organism; rainbow trout; recirculating aquaculture system; red mark syndrome EN digital droplet PCR Midichloria-like organism rainbow trout recirculating aquaculture system red mark syndrome 1889 1892 4 11/11/22 20221201 NES 221201 Red mark syndrome is a non-lethal skin disease, reported to affect farmed rainbow trout over 50 g worldwide (Metselaar et al., 2022). The manuscript by Oh et al. (2019) describes several cases of the disease in rainbow trout in South Korea, in which macroscopic lesions resembling RMS can be seen in juvenile rainbow trout weighed less than 30 g. [Extracted from the article]

Variants of perch rhabdovirus (PRV) circulate across European percid farms via the fish trade. To trace their circulation, they are usually isolated by cell culture and subsequently identified genetically by sequencing partial or complete genes. Here, a newly developed nested PCR‐based method was used to amplify and sequence the complete N and P genes directly from clinical samples obtained during an outbreak on a farm as well as from four batches of fish sampled from two other farms in another country. In an attempt to trace the origin of the five detected viruses, their N and P sequences were concatenated and compared with related viruses. One virus found in pike‐perch was highly related to a virus isolated in 2016 in Belgium. Two other viruses detected on a single farm were distinct from one another, with one being almost identical to another virus isolated in 2016 in Belgium and the other being more closely related to a subgroup with different origins, France and Belgium. Two other viruses found in perch from a third farm were identical and were more related to a subgroup of viruses isolated in France. Identifying variants by a direct PCR approach will help to prevent further dissemination in farms. [ABSTRACT FROM AUTHOR]

The impact that escaped farmed fish may have on wild populations is of major concern for Atlantic salmon (Salmo salar) farming. Triploid fish, being infertile, were originally introduced to mitigate the genetic impact of escaped fish. In the recent years, an increase in the number of infectious salmon anaemia (ISA) outbreaks in Norway has been observed, mainly in the northern parts, which is also where farming of triploid fish has been licensed. The present study investigated the susceptibility of triploid Atlantic salmon to ISA both by field observations and experimental infections. Based on field observations, we found an increased susceptibility, with 9.4 increased odds to primary ISA outbreaks in triploid fish versus diploid fish at production‐site level, and a tendency of increased odds (3.4) of ISA in triploid fish at individual cage level at sited with primary outbreaks. At some sites, ISA outbreaks were only diagnosed in cages with triploid fish and not in cages with diploid fish. Primary ISA outbreaks are the source for further spread of the disease, and it is noteworthy that in an experimental trial we found significantly more viral RNA in non‐ISA‐vaccinated triploid than in non‐ISA‐vaccinated diploid fish at the peak of the infection. Interestingly, the notable differences of susceptibility to ISA for non‐ISA vaccinated diploid and triploid fish observed in field were not repeated experimentally. The possible increased risk of ISA should be considered when evaluating the costs and benefits of triploid salmon in farming. It is recommended to keep triploid and diploid fish in biosecure separated sites, or that triploid fish are not farmed at all. [ABSTRACT FROM AUTHOR]

Epitheliocystis, an intracellular bacterial infection in the gills and skin epithelium, has been frequently reported in Atlantic salmon (Salmo salar) during freshwater production in a number of countries. This study describes the prevalence and intensity of a natural epitheliocystis infection present in the gills of two strains of Atlantic salmon reared in either a flow‐through (FT) or a recirculation aquaculture system (RAS) in Ireland. Repeated sampling of gills prior to and throughout seawater transfer, histology and quantitative real‐time PCR were used to determine infection prevalence and intensity. Despite no clinical gill disease, and minor histopathological changes, epitheliocystis lesions were identified in histology at all time points. Specific PCR confirmed the presence of Candidatus Clavichlamydia salmonicola in both strains and its number of copies was correlated with intensity of epitheliocystis lesions. A significant interaction between hatchery system and fish strain on the prevalence and intensity of gill epitheliocystis was found both using histological and molecular methods. Specifically, fish from FT had higher prevalence and intensity than RAS reared fish and within FT, the Irish cohort were more affected than Icelandic. [ABSTRACT FROM AUTHOR]

Nervous necrosis virus (NNV) is one of the most destructive pathogens in marine fish aquaculture and is capable of infecting more than 50 fish species worldwide, which resulted in great economic losses. Effective drugs for managing NNV infection are urgently required. Medicinal plants have been known for thousands of years and benefit of medicinal plants against pathogens in aquaculture have emerged. Nowadays, the most commonly used method for detecting virus infection and assessing antiviral drugs efficacy is reverse transcription‐quantitative real‐time PCR. However, the application is limited on account of high reagent costs, complex time‐consuming operations and long detection time. Aptamers have been widely applied in application of pathogens or diseases diagnosis and treatments because of high specificity, strong affinity, good stability, easy synthesized and low costs. This study aimed to establish an aptamer (GBN34)‐based high‐throughput screening (GBN34‐AHTS) model for efficient selection and evaluation of natural ingredients against NNV infection. GBN34‐AHTS is an expeditious rapid method for selecting natural ingredients against NNV, which is characterized with high‐speed, dram, sensitive and accurate. AHTS strategy could reduce work intensity and experimental costs and shorten the whole screening cycle of effective ingredients. AHTS should be suitable for rapid selection of effective ingredients against other viruses, which is important for improving the prevention and controlling of aquatic diseases. [ABSTRACT FROM AUTHOR]

Haematological parameters are frequently used as physiological indicators in aquaculture studies. These parameters also have extended applications in clinical evaluation, diagnosis and prognosis in fish health status. However, no normal reference range of values has been demonstrated in depth for any of these haematological parameters for the European sea bass (Dicentrarchus labrax) or gilthead seabream (Sparus aurata). The main objective of this article is to present for the first time through extended literature review, the haematological parameters normal range values for the two most important aquaculture fish species farmed in Mediterranean Sea, D. labrax and S. aurata, and to demonstrate their similarities and their differences. In this article, we also discuss the environmental and external factors affecting their normal blood parameters values and we propose fundamental guidelines on the reporting units. [ABSTRACT FROM AUTHOR]

Tilapia Lake Virus (TiLV) is reported as a threat to tilapia aquaculture in 16 countries on four continents with outbreaks causing up to 90% mortality. This research is one of the first studies on TiLVs from Vietnam. We propagated successfully one TiLV isolate HB196‐VN‐2020 from a diseased tilapia sample using an E‐11 cell line and evaluated its virulence in two different weights of red hybrid tilapia and three serial 10‐fold diluted viral titers. Smaller fish (4.5 ± 1.98 g) were proved to be more susceptible to TiLV infection at the viral titre of 9.1 × 105 TCID50 fish−1 than larger fish (20.8 ± 7.5 g) with the mortalities of 92.5% and 12.5%, respectively. Reassortant detection analysis revealed seven potential reassortment events among 23 TiLV genomes, indicating the mixed infection of multiple TiLV isolates at the farms and the fish movement among different regions. Seven maximum likelihood phylogenetic trees based on the individual segments or the concatenated coding regions of some segments showed the genetically distant relationship of the Southern Vietnamese isolate RIA2‐VN‐2019 with the 21 reference isolates, and suggest the different origins of two Vietnamese TiLV isolates (RIA2‐VN‐2019 and HB196‐VN‐2020). However, additional sequences from various sampling locations and times are required to better understand the impacts of genetic diversity and reassortments on the evolution, migration and natural selection of TiLVs in Vietnam and other countries. [ABSTRACT FROM AUTHOR]

Dilution series were made from I K. thrysites i genomic DNA diluted in both dH SB 2 sb O and a pooled sample of DNA eluate from I K. thyrsites- i negative filtration substrates (File S1). Keywords: actinospore; aquaculture; fish parasite; myxospore; PCR optimization; QuantStudio 3D EN actinospore aquaculture fish parasite myxospore PCR optimization QuantStudio 3D 1403 1407 5 08/12/22 20220901 NES 220901 INTRODUCTION I Kudoa thyrsites i (Gilchrist, 1924) (Myxozoa) forms intracellular cysts within muscle tissues of at least 37 species of fish (Moran, Kent, & Whitaker, 1999; Whipps & Kent, 2006), resulting in decreased flesh quality in commercial harvests. Jones et al. (2016) used a real-time quantitative PCR (qPCR) (Funk et al., 2007) to measure I K. thyrsites i eDNA from PBS intake sea water where results suggested higher abundance in summer relative to winter. Aquaculture, fish parasite, actinospore, myxospore, PCR optimization, QuantStudio 3D. [Extracted from the article]

This method generates a negative image of fish skeleton, and it is also applicable to alive fish, provided that suitable X-ray equipment (X-ray generator and detector) is available in the farming facility. Larval performance and skeletal deformities in farmed gilthead sea bream (Sparus aurata) fed with graded levels of Vitamin A enriched rotifers (Brachionus plicatilis). Keywords: juvenile fish; microradiography; sekeletal anomalies EN juvenile fish microradiography sekeletal anomalies 1237 1241 5 07/15/22 20220801 NES 220801 INTRODUCTION As for any industrial sector, the aquaculture industry needs to minimize production costs to increase business profit and competitiveness. Skeletal descriptors and quality assessment in larvae and post-larvae of wild-caught and hatchery-reared gilthead sea bream (Sparus aurata L. 1758). [Extracted from the article]

Piscine lactococcosis is an emergent bacterial disease that is associated with high economic losses in many farmed and wild aquatic species worldwide. Early and accurate detection of the causative agent of piscine lactococcosis is essential for management of the disease in fish farms. In this study, a TaqMan quantitative polymerase chain reaction (qPCR) targeting the 16S–23S rRNA internal transcribed spacer region was developed and validated. Validation of the qPCR was performed with DNA of previously typed L. petauri and L. garvieae recovered from different aquatic hosts from distinct geographical locations, closely related bacterial species and common pathogens in trout aquaculture. Further diagnostic sensitivity and specificity was investigated by screening of fish, water and faecal samples. The developed qPCR assay showed high specificity, sensitivity and accuracy in detection of L. petauri and L. garvieae with lack of signals from non‐target pathogens, and in screening of rainbow trout (Oncorhynchus mykiss) posterior kidney and environmental samples. The detection limit of the qPCR was four amplicon copies. Moreover, the sensitivity of the qPCR assay was not affected by presence of non‐target DNA from either fish or environmental samples. The robustness, specificity and sensitivity of the developed qPCR will facilitate fast and accurate diagnosis of piscine lactococcosis to establish appropriate control measures in fish farms and aquaria. [ABSTRACT FROM AUTHOR]

Viral nervous necrosis (VNN) is now endemic in the Mediterranean basin and the RGNNV genotype betanodavirus has caused frequent epidemics in European sea bass for a long time. Unexpected and increasing VNN epidemics have been reported in gilthead sea bream (GSB) farms in the last few years, from which the RGNNV/SJNNV genotype has been mostly isolated. The aim of this study was to perform a molecular characterization of the betanodavirus isolated from GSB (weighing 90–100 g) in a marine fish farm in the Aegean Sea and also, as an early warning exercise, to investigate the presence/absence of the virus in associated nearby farms (n:20) and in hatcheries (n:3). No virus was detected in any of the nearby farms or two hatcheries. However, in one hatchery, betanodavirus was detected in a 160‐day‐old GSB. The identified betanodavirus was genotyped as reassortant RGNNV/SJNNV and was phylogenetically related to the virus detected in the farm located in the Aegean sea. There have been multiple detections of the RGNNV genotype in Turkish coastal waters; however, the RGNNV/SJNNV genotype has been detected for the first time and it should be an early warning to focus attention on betanodaviruses in Turkish aquaculture. [ABSTRACT FROM AUTHOR]

Epitheliocystis was present in the gills of all three fish species including armoured catfish, anabas and Nile tilapia from all four water bodies in central Vietnam. While armoured catfish has no economic value, anabas is a species target for artesian fishing and tilapia is one of two main fish species farmed in Vietnam. Keywords: anabas; armoured catfish; epitheliocystis; southeast Asia; tilapia; wild fish EN anabas armoured catfish epitheliocystis southeast Asia tilapia wild fish 755 760 6 04/08/22 20220501 NES 220501 Epitheliocystis is a condition affecting fish gill or skin epithelium and caused by intracellular Gram-negative bacteria including either Chlamydia or Proteobacteria (Blandford et al., 2018; Nowak & LaPatra, 2006; Polkinghorne et al., 2010; Stride et al., 2014). [Extracted from the article]

Tilapia tilapinevirus, or Tilapia Lake Virus (TiLV), is a RNA virus associated with mass morbidity and mortality in tilapia, leading to severe economic losses for global tilapia aquaculture. In this study, we investigated the persistence of TiLV in water by spiking sterile distilled water (SDW), freshwater collected from rearing fish tanks (FW) and natural pond water (PW) at 27°C as a representative of environmental water conditions with 0.6 ml of stock virus (3.18 × 107 viral copies/ml of water). The water samples were filtered through an electronegative charge membrane and quantified using reverse transcriptase quantitative PCR at 0, 3, 5, 7, 10 and 14 days post‐inoculation. The results revealed that TiLV RNA in SDW was reduced by 1.34 log10 in 14 days. A similar approximately 4 log10 removal of the virus in FW and PW was observed at 3 and 7 days, respectively. Moreover, the infectivity of TiLV was further studied; the virus lost its infectivity in E‐11 cells after 1 day in SDW, FW and PW water samples, even though the virus was spiked 10 more times than in the viral persistence study. Taken together, the results could be applied to improving biosecurity practices in tilapia farms by disinfecting or resting reservoir water for at least three to five days prior to stocking tilapia, to limit the spread of TiLV. [ABSTRACT FROM AUTHOR]

The protozoan Cryptocaryon irritans is one of the most important ectoparasites of marine fish, causing 'white spot disease' and mass mortality in aquaculture. To accurately predict disease outbreaks and develop prevention strategies, improved detection methods are required that are sensitive, convenient and rapid. In this study, a pair of specific primers based on the C. irritans 18S rRNA gene was developed and used in a real‐time PCR (qPCR) assay. This assay was able to detect five theronts in 1 L of natural seawater. Furthermore, a linear model was established to analyse the log of Ct value and parasite abundance in seawater (y = –2.9623x + 24.2930), and the coefficient of determination (R2) value was 0.979. A lysis buffer was optimized for theront DNA extraction and used for storage sample. This method was superior to the commercial water DNA kit, and there was no significant degradation of DNA at room temperature for 24–96 hr. A dilution method was developed to manage qPCR inhibitors and used to investigate natural seawater samples in a net cage farm with diseased fish, and the findings were consistent with the actual situation. This study provides a valuable tool for assisting in the early monitoring and control of cryptocaryoniasis in aquaculture. [ABSTRACT FROM AUTHOR]

Acanthocephalosis in tambaqui Colossoma macropomum culture is a limiting factor for the production of the species, being the most recurrent, which makes it necessary to develop strategies to control parasitosis even in the early stages of parasite development. This study aimed to elucidate the morphological characteristics of eggs of the acanthocephalan Neoechinorhynchus buttnerae and the ovicidal effect of the essential oils of Lippia alba, Lippia sidoides, and Lippia gracilis. The parasites were collected from hosts naturally infected with N. buttnerae, and the eggs obtained from the female 'parasites' abdominal cavity were added to 12‐well plates containing the essential oils at different dilutions. After the eggs (N = 300) were separated and measured (length and width) with the aid of an optical microscope coupled to a digital program, the viability of the treated eggs was monitored 24 h after treatments, L. gracilis essential oil was 100% effective at the highest concentration, whereas L. sidoides essential oil showed over 50% efficacy at 2 mg/ml, reaching 100% efficacy at all higher concentrations. Lippia. alba essential oil did not show satisfactory efficacy in preventing N. buttnerae egg hatching. [ABSTRACT FROM AUTHOR]

Clarias gariepinus (Burchell, 1822) is one of the two most actively cultured freshwater fish in Africa and therefore, economically important. Specimens of this species were purchased from a fish farm near Hartbeespoort Dam (North West, South Africa) and introduced into the tanks of the research aquarium in the Department of Zoology at the University of Johannesburg. However, the skin of these fish was infected with Macrogyrodactylus congolensis (Prudhoe, 1957), which proliferated profusely in the favourable conditions of the aquarium, posing a potential threat to its host. The current study was aimed at examining the pathology caused by M. congolensis on the skin of C. gariepinus. Species identification of the parasite was confirmed using light microscopy (LM), scanning electron microscopy (SEM) and DNA barcoding of the internal transcribed spacer region. Examination of the pathology was studied using LM of haematoxylin and eosin–stained sections (epoxy embedded) and SEM of parasites attached to the hosts' skin. Infected skin exhibited excessive mucus production, corroborated by an increased number of mucus cells alongside proliferated and abnormally enlarged club cells, resulting in varying thickness of the epidermal layer. At the site of attachment, the basement membrane detached from the dermis. Hamulus points and marginal hooks of the parasite pierce through the hosts' skin resulting in tearing. Epidermal cells and melanin granules were observed in the intestinal lumen of the parasite. Melanin granules were absorbed by the parasite's intestinal epithelium confirming that the parasite feeds on host skin tissue. [ABSTRACT FROM AUTHOR]

In November 2020 a mortality episode (30%) in juvenile Siberian and Russian sturgeons (Acipenser baerii, Brandt, and A. gueldenstaedtii, Brandt & Ratzeburg) and GUBA hybrid sturgeons (A. gueldenstaedtii × A. baerii) occurred in a hatchery in Northern Italy, associated with severe coelomic distension and abnormal reverse surface swimming. The fish were reared in concrete tanks supplied by well water, fed at 0.4% of body weight (b.w.) per day. Thirty sturgeon specimens were collected for necropsy, histological, bacteriological and virological examination. Macroscopic findings included diffuse and severe bloating of gastrointestinal tracts due to foamy contents with thinning and stretching of the gastrointestinal walls. Histological analysis revealed variable degrees of sloughing and necrosis of the intestinal epithelium, and the presence of bacterial aggregates. Anaerobic Gram‐positive bacteria were investigated, and Clostridium perfringens was isolated from the gut. Specific PCRs identified the toxinotype A and the β2 toxin gene. The daily feed administration was increased to 1.5% b.w. and after 5 days, the mortality ceased. A new animal cohort from the same groups was examined after 12 weeks, showing neither gut alterations nor isolation of C. perfringens. The imbalance of intestinal microbiota, presumably caused by underfeeding, favoured C. perfringens overgrowth and severe gas formation. The diet increase possibly restored the normal microbiota. [ABSTRACT FROM AUTHOR]

Keywords: Flavobacterium psychrophilum; immersion challenge; immersion vaccination; proteomics; rainbow trout; skin mucus EN Flavobacterium psychrophilum immersion challenge immersion vaccination proteomics rainbow trout skin mucus 491 495 5 02/10/22 20220301 NES 220301 I Flavobacterium psychrophilum i is one of the most important pathogens affecting rainbow trout ( I Oncorhynchus mykiss i ) worldwide at the fry stage of their life cycle. Briefly, skin mucus was sampled by placing three fish (from each duplicate tank giving 2 pools of mucus/group) into a plastic bag containing 5 ml of Tris-buffered saline (TBS: 10 mM Tris base and 0.5 M NaCl, pH 7.5) and gently massaging for 2 min. In the current study, we examined the proteins of skin mucus of rainbow trout following immersion vaccination with I F. psychrophilum i . [Extracted from the article]

Scale drop disease virus (SDDV) is a major pathogen of Asian sea bass that has emerged in many countries across the Asia Pacific since 1992 and carries the potential to cause drastic economic losses to the aquaculture sector. The lack of an approved vaccine for SDDV necessitates timely prevention as the first line of defence against the disease, but current diagnostic platforms still face challenges that render them incompatible with field applications, particularly in resource‐limited settings. Here, we developed a novel detection platform for SDDV based on a CRISPR‐Cas12a‐based nucleic acid detection technology combined with recombinase polymerase amplification (RPA‐Cas12a). Using the viral adenosine triphosphatase (SDDV‐ATPase) gene as a target, we achieved the detection limit of 40 copies per reaction and high specificity for SDDV. The coupling with fluorescence and lateral flow readouts enables naked‐eye visualization and straightforward data interpretation requiring minimal scientific background. Compared with semi‐nested PCR in field sample evaluation, our RPA‐Cas12a assay is more sensitive and capable of detecting SDDV in asymptomatic fish. Importantly, the entire workflow can be carried out at a constant temperature of 37°C within an hour from start to finish, thus removing the need for an expensive thermal cycling apparatus and long turnaround times associated with PCR‐based methods. Therefore, owing to its high accuracy, rapidity and user‐friendliness, the developed RPA‐Cas12a platform shows the potential for diagnosis of SDDV at point of need and could be a valuable tool to help protect fish farming communities from large‐scale epidemics. [ABSTRACT FROM AUTHOR]

The ISA virus has two main phenotypic variants based on the sequence of the highly polymorphic region (HPR) of gene segment 6: non-pathogenic ISAV-HPR0 and pathogenic ISAV-HPR . Primary cases of ISA are thought to occur from ISAV-HPR0 transitioning to ISAV-HPR , causing ISA outbreaks (Christiansen et al., 2017; Nylund et al., 2019). Keywords: Atlantic salmon; infectious salmon anaemia virus; surveillance EN Atlantic salmon infectious salmon anaemia virus surveillance 225 229 5 12/13/21 20220101 NES 220101 INTRODUCTION Infectious salmon anaemia (ISA) is a serious disease that primarily affects farmed Atlantic salmon ( I Salmo salar i ), although some other salmonids are also susceptible (OIE, 2019a). Atlantic salmon, infectious salmon anaemia virus, surveillance. [Extracted from the article]

Amyloodinium ocellatum (AO) infection in silver pomfret (Pampus argenteus) causes extensive mortality. Insufficient information exists on the molecular immune response of silver pomfret to AO infestation, so herein we simulated the process of silver pomfret being infected by AO. Translucent trophosomes were observed on the gills of AO‐infected fish. Transcriptome profiling was performed to investigate the effects of AO infection on the gill, kidney complex and spleen. Overall, 404,412,298 clean reads were obtained, assembling into 96,341 unigenes, which were annotated against public databases. In total, 2730 differentially expressed genes were detected, and few energy‐ and immune‐related genes were further assessed using RT‐qPCR. Moreover, activities of three immune‐related (SOD, AKP and ACP) and three energy‐related (PKM, LDH and GCK) enzymes were determined. AO infection activated the immune system and increased interleukin‐1 beta and immunoglobulin M heavy chain levels. Besides, the PPAR signalling pathway was highly enriched, which played a role in improving immunity and maintaining homeostasis. AO infection also caused dyspnoea, leading to extensive lactic acid accumulation, potentially contributing towards a strong immune response in the host. Our data improved our understanding regarding the immune response mechanisms through which fish coped with parasitic infections and may help prevent high fish mortality in aquaculture. [ABSTRACT FROM AUTHOR]

Intensive fish farming at high densities results in a wide range of adverse consequences on fish welfare, including pathogen spreading, stress and increased mortality rates. In this work, we have assessed whether the survival of Senegalese sole infected with the nervous necrosis virus (NNV), a pathogen responsible for severe disease outbreaks, is affected by rearing density. Based on the different fish ratios per surface area (g cm−2) and water volume (g L−1), our research showed an earlier mortality onset in the tanks containing NNV‐infected fish reared at medium density (MD: 0.071 g cm−2/5 g L−1) and high density (HD: 0.142 g cm−2/10 g L−1), as well as higher cumulative mortality values. However, transcription analysis of hsp70, gr1 and pepck genes, well‐known stress biomarkers, seems to indicate that none of the challenged fish were under high stress conditions. NNV load was slightly higher both in dead and in sampled fish from MD and HD groups, and especially in the rearing water from these groups, where peaks in mortality seemed to correlate with increasing NNV load in the water. In conclusion, our results suggest that rearing NNV‐infected Senegalese sole at high densities resulted in an earlier mortality onset and higher cumulative values and viral load. [ABSTRACT FROM AUTHOR]

Cardiomyopathy syndrome (CMS) is the most common viral cardiac disease in Norwegian Atlantic salmon farming and typically affects large, market size fish. Only six months after seawater transfer, Atlantic salmon were diagnosed with CMS at a fish farm in the south‐western part of Norway. Due to the unexpected young age and the remarkable large amounts of virus‐specific RNA (Ct <10), the fish group was monitored with five additional samplings until slaughtered almost 10 months later. At three weeks after the first CMS diagnosis (weeks post‐diagnosis, wpd) and at slaughter (39 wpd), more comprehensive samplings were performed of the study cage, with specific focus on three different cardiac compartments. The clinical, autopsy and histopathological findings at first diagnosis and at all succeeding samplings were similar to previous descriptions of typical CMS. A slightly elevated mortality was observed in the cage with diseased fish at the time of the first CMS diagnosis and continued throughout the study. The prevalence and load of PMCV‐specific RNA in the fish remained high until slaughtering, with similar amounts in all sampled cardiac compartments. No fish from the other five cages at the site were diagnosed with CMS, until fish sampled from the last cage at the site were diagnosed 10 weeks after slaughtering of the study cage (49 wpd). Sequence analysis of the PMCV on the site showed that the outbreak virus was similar to PMCV variants previously sequenced from Norwegian field outbreaks. In conclusion, CMS in young Atlantic salmon had clinical signs and histopathological cardiac lesions typical for the disease, and diseased fish could be found in the study cage until slaughtering. [ABSTRACT FROM AUTHOR]