Salmonid Fish Farming Research Articles

Molecular mechanism of infectious hematopoietic necrosis G protein N-glycosylation escaping from rainbow trout immunity

Infectious Hematopoietic Necrosis (IHN) is a viral disease that causes great economic losses in salmonid fish farming industry. It is known that the N-glycosylation site on the G protein affects the immune evasion and pathogenicity of the Infectious Hematopoietic Necrosis Virus (IHNV). In this study, a mutation in the N-glycosylation site of the G protein (438 amino acids) of IHNV resulted in an increased the survival rate of rainbow trout from 17.1 % to 75.3 %, and a decreased serum antibody IgM level after virus infection. In addition, this mutation decreased the expression of CD83 and CD209 on dendritic cells and enhanced the binding of G protein to TCR. This suggests that the activation of CD4+ T-cells was enhanced. We hypothesize that IHNV evades the host's immune system by impairing antigen presentation and T-cell responses through the N-glycosylation of G protein. This finding could provide new insights for developing live attenuated vaccines.

Genetic and phenotypic diversity of Flavobacterium psychrophilum isolates from Czech salmonid fish farms

BackgroundThe salmonid pathogen Flavobacterium psychrophilum poses a significant economic threat to global aquaculture, yet our understanding of its genetic and phenotypic diversity remains incomplete across much of its geographic range. In this study, we characterise the genetic and phenotypic diversity of 70 isolates collected from rainbow trout (Oncorhynchus mykiss) and brown trout (Salmo trutta m. fario) from fish farms in the Czech Republic between 2012 and 2019 to compare their genomic content with all draft or complete genomes present in the NCBI database (n = 187).ResultsThe Czech isolates underwent comprehensive evaluation, including multiplex PCR-based serotyping, genetic analysis, antimicrobial resistance testing, and assessment of selected virulence factors. Multiplex PCR serotyping revealed 43 isolates as Type 1, 23 as Type 2, with sporadic cases of Types 3 and 4. Multi-locus sequence typing unveiled 12 sequence types (ST), including seven newly described ones. Notably, 24 isolates were identified as ST329, a novel sequence type, while 22 were classified as the globally-distributed ST2. Phylogenetic analysis demonstrated clonal distribution of ST329 in the Czech Republic, with these isolates lacking a phage sequence in their genomes. Antimicrobial susceptibility testing revealed a high proportion of isolates classified as non-wild type with reduced susceptibility to oxolinic acid, oxytetracycline, flumequine, and enrofloxacin, while most isolates were classified as wild type for florfenicol, sulfamethoxazole-trimethoprim, and erythromycin. However, 31 isolates classified as wild type for florfenicol exhibited minimum inhibitory concentrations at the susceptibility breakpoint.ConclusionThe prevalence of the Czech F. psychrophilum serotypes has evolved over time, likely influenced by the introduction of new isolates through international trade. Thus, it is crucial to monitor F. psychrophilum clones within and across countries using advanced methods such as MLST, serotyping, and genome sequencing. Given the open nature of the pan-genome, further sequencing of strains promises exciting discoveries in F. psychrophilum genomics.

Comparative safety and efficacy of autogenous vaccine administrated by different routes against furunculosis caused by Aeromonas salmonicida sub. salmonicida in large Rainbow trout (Oncorhynchus mykiss)

The development and growth of fish farming are hindered by viral and bacterial infectious diseases, which necessitate effective disease control measures. Furunculosis, primarily caused by Aeromonas salmonicida, stands out as a significant bacterial disease affecting salmonid fish farms, particularly rainbow trout. Vaccination has emerged as a crucial tool in combating this disease. The objective of this experiment was to assess and compare the efficacy and duration of different vaccine protocols against furunculosis in large trout under controlled rearing conditions, utilizing single and booster administrations via intraperitoneal, oral, and immersion routes. Among the various vaccination protocols tested, only those involving intraperitoneal injection, administered at least once, proved truly effective in preventing the expression of clinical signs of furunculosis and reducing mortality rates. A single intraperitoneal administration provided protection for up to 2352°-days, equivalent to approximately 5 months in water at 16 °C. However, intraperitoneal vaccination may lead to reduced growth in the fish due to resultant intraperitoneal adhesions. Additionally, protocols incorporating booster doses via intraperitoneal injection demonstrated efficacy regardless of the administration route of the primary vaccination. Nevertheless, the use of booster vaccinations via the intraperitoneal route did not confer any significant advantage over a single intraperitoneal injection in terms of efficacy.

Forecasting ocean hypoxia in salmonid fish farms

IntroductionHypoxia is defined as a critically low-oxygen condition of water, which, if prolonged, can be harmful to fish and many other aquatic species. In the context of ocean salmon fish farming, early detection of hypoxia events is critical for farm managers to mitigate these events to reduce fish stress, however in complex natural systems accurate forecasting tools are limited. The goal of this research is to use a machine learning approach to forecast oxygen concentration and predict hypoxia events in marine net-pen salmon farms.MethodsThe developed model is based on gradient boosting and works in two stages. First, we apply auto-regression to build a forecasting model that predicts oxygen concentration levels within a cage. We take a global forecasting approach by building a model using the historical data provided by sensors at several marine fish farms located in eastern Canada. Then, the forecasts are transformed into binary probabilities that indicate the likelihood of a low-oxygen event. We leverage the cumulative distribution function to compute these probabilities.Results and discussionWe tested our model in a case study that included several cages across 14 fish farms. The experiments suggest that the model can detect future hypoxic events with a commercially acceptable false alarm rate. The resulting probabilistic predictions and oxygen concentration forecasts can help salmon farmers to prioritize resources, and reduce harm to crops.

Consequences of reduced effectiveness of salmon lice treatments for lice control

The effective control of ectoparasitic salmon lice, Lepeophtheirus salmonis, in fish farms is challenged by the salmon lice having developed resistance towards several antiparasitic drugs and by the effectiveness of non-medicinal treatments being limited by considerations of fish welfare. When new antiparasitics are introduced to the market, these should be used sparingly to slow resistance development. Using a population model for salmon lice parameterised for salmonid fish farms in Norway, we quantified how reduced treatment effectiveness influences treatment frequency and lice abundance. Furthermore, we investigated when in the production cycle a highly effective lice treatment leads to the largest reduction in the total number of treatments, mean lice abundance and lice larvae production. Results showed that reductions in treatment effectiveness to lower than 50% led to the steepest increases in treatment frequency and mean lice abundance, as well as to increased risk that lice abundance increased beyond control. The timing of the most effective treatment had only moderate effects on the total treatment need and the mean number of adult female lice through the production cycle, but large effect on the production of lice larvae in spring. These findings imply that farmers can optimise the timing of the most effective treatment to reduce the release of lice larvae in the period of year when wild salmonids are in coastal waters, without compromising total treatment need or mean lice levels.

Life history traits for Ophryotrocha craigsmithi (Wiklund, Glover & Dahlgren, 2009), a candidate species in integrated multitrophic aquaculture

Ophryotrocha craigsmithi (Wiklund, Glover & Dahlgren, 2009) has repeatedly been observed in high densities at salmonid fish farms in Norway and has been shown to feed on the organic waste released during fish production. This species has therefore been pointed out as a candidate for the benthic component of integrated multitrophic aquaculture (IMTA). Little is known about the life history traits of Ophryotrocha species found at greater depths in temperate waters. Insight into life traits is of key importance in evaluating the bio-mitigation and harvest potential of (benthic) species within IMTA systems. In this study, we aimed to describe key life history traits for O. craigsmithi, such as growth, reproductive pattern, and offset of reproduction. The main findings are as follows: (I) O. craigsmithi exhibits early sexual maturation and rapid growth, going from 5 to 16 mm in length in 8 weeks; (II) their estimated lifetime is approximately 15 weeks with a generation time of 4 weeks; (III) there are strong indications that they are gonochoric semicontinuous spawners; (IV) the majority of mature eggs ranged from 120 to 150 µm; and (V) the larvae developed into a free-swimming stage after approximately 1 week. To this end, we present a framework for assessing the suitability of deposit-feeding polychaetes as candidate species in IMTA, considering environmental adaptiveness, production potential, as well as carbon and nitrogen mitigation capacity. On the basis of this, O. craigsmithi shows a high suitability as an extractive farmed species in benthic IMTA.

Genomic features of Pseudomonas sp. RGM2144 correlates with increased survival of rainbow trout infected by Flavobacterium psychrophilum.

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.

Cyprinid fish farms in Vojvodina (North Serbia): Two decades of changes

Fish farming in Serbia takes place in cyprinid (carp) and salmonid (trout) fish farms that together account for over 95% of the total fish produced. Cyprinids are grown in warm water fish farms, and cold water farms are designed for trout. The great majority (97%) of cyprinid fish farms are located in Vojvodina, Serbia?s northern province. This paper examines how the traditional carp sector was developed during the recent 20- year period in Vojvodina, paying particular attention to the comparison of the number and condition of cyprinid fish farms in that interval according to the available sources. The comparison was made by comparing maps with data on fishponds from 1999 and 2021, and according to the data from the Statistical Office of Serbia. After the analysis of many data from other authors and our study, it is evident that there is a lot of room for improving the production of cyprinid fish in Vojvodina. This is especially because we noticed decreasing trends for two major parameters: the area of fishpond water surfaces in Serbia (from 7,190 ha to 6,299 ha), and the number of ponds in Vojvodina (from 72 to 60), from 1999 to 2021, respectively. On the contrary, the amount of fish produced has increased from 2,409 t in 2002 to 4,761 t in 2021. According to government planning documents, Serbia lacks a national strategy for development of aquaculture production in general, and especially of cyprinid fish species.

A statistical mechanistic approach including temperature and salinity effects to improve salmon lice modelling of infestation pressure

Salmon lice Lepeophtheirus salmonis pose a major threat to the sustainable development of salmonid farming. To investigate effects of farm-origin salmon lice on wild salmonids, salmon lice dynamics are typically simulated using models that depend on experimentally determined rates of development, reproduction, mortality and infestation. Several recent studies provide new estimates of how these demographic rates depend on temperature and salinity. Here, we review and synthesize these studies and test if updating a salmon lice infestation model based on the new insights improves predictions of salmon lice infestations on salmon post-smolts in experimental cages in the sea. This model predicts spatiotemporal variation in infestation pressure based on weekly monitoring data of salmon lice and sea temperature in all salmonid fish farms in Norway, here supplemented by temperature and salinity data from a regional ocean model. Using data from 2012-2017 to select model formulation, we found the largest improvement in explanatory power by incorporating a salinity-dependent infestation rate. Updating functions for temperature-dependent egg production and infestation rates led to smaller improvements. Moreover, results suggest additional effects of temperature and a possible temperature-salinity interaction effect, not captured by the modelled processes. Out-of-sample predictions for experimental cage data from 2018-2020 confirmed that the uncertainty was realistically quantified, but also showed that associations of salmon lice infestations with salinity and temperature had changed. These results provide a field evaluation of experimental data and point to a knowledge gap regarding the combined effects of temperature and salinity on salmon lice infestations.

Phenylalanine Hydroxylase RNAi Knockdown Negatively Affects Larval Development, Molting and Swimming Performance of Salmon Lice

Phenylalanine hydroxylase (PAH) is a crucial enzyme involved in tyrosine biosynthesis, having roles in neurological and physiological processes. The purpose of PAH has received little attention in crustaceans despite extensive investigations in other arthropods. Here, we characterize the PAH gene for the first time in the parasite Lepeophtheirus salmonis, a copepod that is responsible for huge economic losses in salmonid fish farming. Phylogenetic and sequence analyses confirmed that LsPAH is closely related to the metazoan PAH with conserved ACT regulatory and catalytic domains. Temporal expression patterns revealed that LsPAH is expressed throughout all developmental stages peaking during the copepodite stages, suggesting an essential role in developmental physiology. We used RNAi to knockdown LsPAH expression in the nauplius I stage to study developmental function during the larval stages. PAH knockdown impaired larval development, molting and swimming ability with severe morphological defects. This study provides insight into the role of PAH in copepods and demonstrates the importance of this metabolic gene in salmon louse growth and development.

Serological diversity in Flavobacterium psychrophilum: A critical update using isolates retrieved from Chilean salmon farms.

Chile is currently the second largest producer of farmed salmon worldwide, but Flavobacterium psychrophilum, as one of the most detrimental pathogens, is responsible for major losses during the freshwater culturing step in salmonid fish farms. An antigenic study conducted 10years ago reported four serological groups using 20 F.psychrophilum Chilean strains. To reduce disease outbreaks and to develop vaccine candidates, antigenic knowledge needs to be regularly updated using a significant number of additional recent F.psychrophilum isolates. The present study aimed at investigating the serological diversity of 118 F.psychrophilum isolates collected between 2006 and 2018 from farmed Atlantic salmon (Salmo salar), rainbow trout (Oncorhynchus mykiss) and coho salmon (Oncorhynchus kisutch). The current study supports an expansion of the known antigenic groups in Chile from 4 to 14. However, the use of the slide-agglutination technique for serotyping is costly, is labour-intensive and requires significant technical expertise. Addressing these points, the mPCR-based procedure was a very useful tool for serotyping the collected Chilean F.psychrophilum isolates. This technique revealed the presence of diverse mPCR serotypes (i.e. types 0, 1, 2 and 4). Therefore, mPCR should be employed to select the bacterial strain(s) for vaccine development and to conduct follow-up, selective breeding or epidemiological surveillance in Chilean fish farms. Given the presented findings, changes to Chilean fish-farming practices are vital for ensuring the continued productivity and well-being of farmed salmonids.

Chilean aquaculture and the new challenges: Pathogens, immune response, vaccination and fish diversification

In Chile, the salmon and trout farmed fishing industries have rapidly grown during the last years, becoming one of the most important economic sources for the country. However, infectious diseases caused by bacteria, virus, mycoses and parasites, result in losses of up to 700 million dollars per year for the Chilean aquaculture production with the consequent increase of antibiotic and antiparasitic usage. After 30 years of its first appearance, the main salmon health problem is still the salmonid rickettsial septicaemia (SRS), which together with other disease outbreaks, reveal that vaccines do not provide acceptable levels of long-lasting immune protection in the field. On the other hand, due to the large dependence of the industry on salmonids production, the Chilean government promoted the Aquaculture diversification program by 2009, which includes new species such as Merluccius australis, Cilus gilberti and Genypterus chilensis, however, specific research regarding the immune system and vaccine development are issues that still need to be addressed and must be considered as important as the farm production technologies for new fish species. Based on the experience acquired from the salmonid fish farming, should be mandatory an effort to study the immune system of the new species to develop knowledge for vaccination approaches, aiming to protect these aquaculture species before diseases outbreaks may occur.This review focuses on the current status of the Chilean aquaculture industry, the challenges related to emerging and re-emerging microbial pathogens on salmonid fish farming, and the resulting needs in the development of immune protection by rational designed vaccines. We also discussed about what we have learn from 25 years of salmonid researches and what can be applied to the new Chilean farmed species on immunology and vaccinology.

Genome analysis provides insights into the epidemiology of infection with Flavobacterium psychrophilum among farmed salmonid fish in Sweden.

The pathogen Flavobacterium psychrophilum is a major problem for the expanding salmonid fish farming industry in Sweden as well as worldwide. A better understanding of the phylogeography and infection routes of F. psychrophilum outbreaks could help to improve aquaculture profitability and the welfare of farmed fish while reducing the need for antibiotics. In the present study, high-throughput genome sequencing was applied to a collection of F. psychrophilum isolates (n=38) from outbreaks on fish farms in different regions of Sweden between 1988 and 2016. Antibiotic susceptibility tests were applied to a subset of the isolates and the results correlated to the presence of genetic resistance markers. We show that F. psychrophilum clones are not regionally biased and that new clones with a higher degree of antibiotic resistance have emerged nationwide during the study period. This supports previous theories of the importance of live fish and egg trade as a route of infection. Continuous monitoring of recovered isolates by high-throughput sequencing techniques in the future could facilitate tracing of clones within and between countries, as well as the detection of emergent virulent or antibiotic-resistant clones. This article contains data hosted by Microreact.

Whole-Genome Sequence of Infectious Pancreatic Necrosis Virus Isolated from Farmed Brook Trout (Salvelinus fontinalis) in Pennsylvania.

ABSTRACTInfectious pancreatic necrosis (IPN) is an acute contagious systemic disease affecting several fish species and a critical disease in the salmonid fish farming industry. Here, we report the complete genome sequence of IPN virus (IPNV) RNA segments A and B, isolated from a farmed brook trout in Pennsylvania.

Immunohistochemical study of inducible nitric oxide synthase and tumour necrosis factor alpha response in turbot (Scophthalmus maximus) experimentally infected with Aeromonas salmonicida subsp. salmonicida.

Aeromonas salmonicida subsp. salmonicida represents one of the major threats in aquaculture, especially in salmonid fish and turbot farming. In order to fight bacterial infections, fish have an immune system composed by innate and specific cellular and humoral elements analogous to those present in mammals. However, innate immunity plays a primordial role against bacterial infections in teleost fish. Among these non-specific mechanisms, the production of nitric oxide (NO) by inducible nitric oxide synthase (iNOS) pathway and the tumour necrosis factor-alpha (TNFα) produced by mononuclear phagocytes, are two of the main immune effectors to eliminate bacterial pathogens. In this study, the distribution and kinetic of iNOS and TNFα-producing cells of kidney and spleen of turbot experimentally inoculated with A.salmonicida was assessed by immunohistochemistry. In control and challenged fish, individual iNOS(+) and TNFα(+) cells, showing a similar pattern of distribution, were detected. In challenged fish, the number of immunoreactive cells was significantly increased in the evaluated organs, as well as the melanomacrophage centres showed variable positivity for both antigens. These results indicate that A.salmonicida induced an immune response in challenged turbot, which involved the increase of the activity of iNOS and TNFα in the leukocytic population from kidney and spleen.

Mortality and weight loss of Atlantic salmon, Salmon salar L., experimentally infected with salmonid alphavirus subtype 2 and subtype 3 isolates from Norway.

Pancreas disease (PD) caused by salmonid alphavirus (SAV) has a significant negative economic impact in the salmonid fish farming industry in northern Europe. Until recently, only SAV subtype 3 was present in Norwegian fish farms. However, in 2011, a marine SAV 2 subtype was detected in a fish farm outside the PD-endemic zone. This subtype has spread rapidly among fish farms in mid-Norway. The PD mortality in several farms has been lower than expected, although high mortality has also been reported. In this situation, the industry and the authorities needed scientific-based information about the virulence of the marine SAV 2 strain in Norway to decide how to handle this new situation. Atlantic salmon post-smolts were experimentally infected with SAV 2 and SAV 3 strains from six different PD cases in Norway. SAV 3-infected fish showed higher mortality than SAV 2-infected fish. Among the SAV 3 isolates, two isolates gave higher mortality than the third one. At the end of the experiment, fish in all SAV-infected groups had significantly lower weight than the uninfected control fish. This is the first published paper on PD to document that waterborne infection produced significantly higher mortality than intraperitoneal injection.

High diversity of bacterial pathogens and antibiotic resistance in salmonid fish farm pond water as determined by molecular identification employing 16S rDNA PCR, gene sequencing and total antibiotic susceptibility techniques

The aim of this study was to examine the microbiological and related parameters (antibiotic resistance and pathogen identification) of water at two salmonid fish farms in Northern Ireland. Total Bacterial Counts at the Movanagher Fish Farm was 1730 colony forming units (cfu)/ml water (log10 3.24cfu/ml) and 3260cfu/ml (log10 3.51cfu/ml) at the Bushmills Salmon Station. Examination of resulting organisms revealed 10 morphological phenotypes, which were subsequently sequenced to determine their identification. All these organisms were Gram-negative and no Gram-positive organisms were isolated from any water sample. From these phenotypes, eight different genera were identified including Acinetobacter, Aeromonas, Chryseobacterium, Erwinia, Flavobacterium, Pseudomonas and Rheinheimera. One unnamed novel taxon was identified from water at the Movanagher Fish Farm, belonging to the genus Acinetobacter and has been tentatively named Acinetobacter movanagherensis. No other novel taxa were observed. All but one of these environmental organisms (Erwinia) are potential pathogens of fish disease. Total antibiotic resistance was observed to varying degrees in water specimens. The most resistant populations were observed in water taken from the Bushmills Salmon Station inlet, followed by water from the Movanagher Fish Farm. No resistance was observed against tetracycline and there was only one occurrence of resistance against ciprofloxacin. Overall, this study indicates that potential fish pathogens made up the majority of environmental organisms identified, even in the absence of recorded fish disease. There was also relatively high levels of total antibiotic resistance in the bacterial water populations examined, where tetracycline was the only antibiotic with zero resistance. These data indicate that the threat of bacterial disease is relatively close due to the indigenous colonization of farm water and that husbandry standards should be maintained at a high standard to avert bacterial disease outbreaks, rather than relying on the absence of specific pathogens in the immediate farm environment.

Artificial rearing of Sakhalin taimen (Parahucho perryi) on salmonid fish farms in the district of Sakhalin region (Russia)

Artificial rearing of Sakhalin taimen (Parahucho perryi) on salmonid fish farms in the district of Sakhalin region (Russia)

Detecting molecular adaptation at individual codons in the glycoprotein gene of the geographically diversified infectious hematopoietic necrosis virus, a fish rhabdovirus

Salmonid fishes, the principal hosts of the infectious hematopoietic necrosis virus (IHNV), are a candidate species for aquaculture in many countries. IHNV causes an acute disease resulting in severe economic loss in salmonid fish farming. Previous phylogenetic analyses revealed the existence of multiple genogroups of this virus throughout the geographical range of its host. Here, we report the importance of natural selection in shaping the evolution of certain codons at the surface glycoprotein (G-protein) gene of this virus. Maximum likelihood (ML)-based codon substitution analyses revealed that approximately 2.8% of the codons for the entire G-protein are shown to have higher nonsynonymous substitution per nonsynonymous site (dn) than the synonymous substitutions per synonymous site (ds) (dn/ds = ω > 4.335). Thus, the data suggest that positive selection ( ω > 1) is the major driving force in the evolution of certain codons. However, majority of these positively selected sites cannot be mapped to the regions of antigenic determinants of IHNV. Based on the reports of previous studies, epitopes with positively selected sites are immunodominant and viruses can escape from immune responses by producing antigenic variation at positively selected sites, therefore, vaccines directed against these neutralizing epitopes of IHNV that consist of no positively selected sites will be more effective. Some of the positively selected sites showed radical change in amino acids with respect to their charge and polarity; however, it is unclear how these changes affect the fitness of the virus.

Ecology of sea lice parasitic on farmed and wild fish

Sea lice, especially Lepeophtheirus salmonis and Caligus spp., have the greatest economic impact of any parasite in salmonid fish farming and are also a threat to wild salmonids. Here, I review how the biology and ecology of various louse and host species influence their pathogenicity and epidemiology. Recent discoveries of new species and genotypes emphasize the need for more basic research on louse taxonomy and host preferences. Louse development rates are strongly dependent on temperature, and increasing mean sea temperatures are likely to increase infestation pressure on farms and wild fish, as well as affecting the geographical distribution of hosts and parasites. Despite progress in finding L. salmonis larvae in the plankton and in modelling louse production in several countries, more data on larval behaviour and distribution are required to develop dispersal and transmission models for both L. salmonis and Caligus spp. This knowledge could be used to take measures to reduce the risks of lice affecting farmed and wild fish.