Behavioral transcriptomic effects of triploidy and probiotic therapy (Bifidobacterium, Lactobacillus, and Lactococcus mixture) on juvenile Chinook salmon (Oncorhynchus tshawytscha).

Aquaculturists use polyploid fish to maximize production albeit with some unintended consequences including compromised behaviors and physiological function. Given benefits of probiotic therapies (e.g., improved immune response, growth, and metabolism), we explored probiotic supplementation (mixture of Bifidobacterium, Lactobacillus, and Lactococcus), to overcome drawbacks. We first examined fish gut bacterial community composition using 16S metabarcoding (via principal coordinate analyses and PERMANOVA) and determined probiotics significantly impacted gut bacteria composition (p = 0.001). Secondly, we examined how a genomic disruptor (triploidy) and diet supplements (probiotics) impact gene transcription and behavioral profiles of hatchery-reared Chinook salmon (Oncorhynchus tshawytscha). Juveniles from four treatment groups (diploid-regular feed, diploid-probiotic feed, triploid-regular feed, and triploid-probiotic feed; n = 360) underwent behavioral assays to test activity, exploration, neophobia, predator evasion, aggression/sociality, behavioral sensitivity, and flexibility. In these fish, transcriptional profiles for genes associated with neural functions (neurogenesis/synaptic plasticity) and biomarkers for stress response and development (growth/appetite) were (i) examined across treatments and (ii) used to describe behavioral phenotypes via principal component analyses and general linear mixed models. Triploids exhibited a more active behavioral profile (p = 0.002), and those on a regular diet had greater Neuropeptide Y transcription (p = 0.02). A growth gene (early growth response protein 1, p = 0.02) and long-term neural development genes (neurogenic differentiation factor, p = 0.003 and synaptysomal-associated protein 25-a, p = 0.005) impacted activity and reactionary profiles, respectively. Overall, our probiotic treatment did not compensate for triploidy. Our research highlights novel applications of behavioral transcriptomics for identifying candidate genes and dynamic, mechanistic associations with complex behavioral repertoires.

Defective UiO-66/cellulose nanocomposite aerogel for the adsorption of heterocyclic aromatic amines.

Heterocyclic aromatic amines (HAAs), arising as chemical derivatives during the high-temperature culinary treatment of proteinaceous comestibles, exhibit notable carcinogenic potential. In this paper, a composite aerogel (AGD-UiO-66) with high-capacity and fast adsorption of HAAs was made with anchoring defective UiO-66 (D-UiO-66) mediated by lauric acid on the backbone of cellulose nanofibers (CNF). AGD-UiO-66 with hierarchical porosity reduced the mass transfer efficiency for the adsorption of HAAs and achieved high adsorption amount (0.84-1.05 µmol/g) and fast adsorption (15 min). The isothermal adsorption model demonstrated that AGD-UiO-66 belonged to a multilayer adsorption mechanism for HAAs. Furthermore, AGD-UiO-66 was successfully used to adsorb 12 HAAs in different food (roasted beef, roasted pork, roasted salmon and marinade) with high recoveries of 94.65%-104.43%. The intrinsic potential of AGD-UiO-66 demonstrated that it could be widely applicable to the adsorption of HAAs in foods.

Postprandial plasma amino acid and appetite responses with ingestion of a novel salmon-derived protein peptide in healthy young adults.

This study assessed postprandial plasma aminoacidemia, glycemia, insulinemia and appetite responses to ingestion of a novel salmon-derived protein peptide (Salmon PP) compared with milk protein isolate (Milk PI). In a randomised, participant-blind crossover design, eleven healthy adults (M = 5, F = 6; mean ± sd age: 22 ± 3 years; BMI: 24 ± 3 kg/m2) ingested 0·3 g/kg/body mass of Salmon PP or Milk PI. Arterialised blood samples were collected whilst fasted and over a 240-min postprandial period. Appetite sensations were measured via visual analogue scales. An ad libitum buffet-style test meal was administered after each trial. The incremental AUC (iAUC) plasma essential amino acid (EAA) response was similar between Salmon PP and Milk PI. The iAUC plasma leucine response was significantly greater following Milk PI ingestion (P < 0·001), whereas temporal and iAUC plasma total amino acid (P = 0·001), non-essential amino acid (P = 0·002), glycine (P = 0·0025) and hydroxyproline (P < 0·001) responses were greater following Salmon PP ingestion. Plasma insulin increased similarly above post-absorptive values following Salmon PP and Milk PI ingestion, whilst plasma glucose was largely unaltered. Indices of appetite were similarly altered following Salmon PP and Milk PI ingestion, and total energy and macronutrient intake during the ad libitum meal was similar between Salmon PP and Milk PI. The postprandial plasma EAA, glycine, proline and hydroxyproline response to Salmon PP ingestion suggest this novel protein source could support muscle and possibly connective tissue adaptive remodelling, which warrants further investigation, particularly as the plasma leucine response to Salmon PP ingestion was inferior to Milk PI.

Polychlorinated biphenyl and polybrominated diphenyl ether profiles vary with feeding ecology and marine rearing distribution among 10 Chinook salmon (Oncorhynchus tshawytscha) stocks in the North Pacific Ocean.

Chinook salmon (Oncorhynchus tshawytscha) along the west coast of North America have experienced significant declines in abundance and body size over recent decades due to several anthropogenic stressors. Understanding the reasons underlying the relatively high levels of persistent organic pollutants (POPs) in Chinook stocks is an important need, as it informs recovery planning for this foundation species, as well for the Chinook-dependent Resident killer whales (Orcinus orca, RKW) of British Columbia (Canada) and Washington State (USA). We evaluated the influence of stock-related differences in feeding ecology, using stable isotopes, and marine rearing ground on the concentrations and patterns of polychlorinated biphenyls (PCBs) and polybrominated diphenyl ethers (PBDEs) in Chinook salmon. A principal components analysis (PCA) revealed a clear divergence of PCB and PBDE congener patterns between Chinook with a nearshore rearing distribution ('shelf resident') versus a more offshore distribution. Shelf resident Chinook had 12-fold higher PCB concentrations and 46-fold higher PBDE concentrations relative to offshore stocks. Shelf resident Chinook had PCB and PBDE profiles that were heavier and dominated by more bioaccumulative congeners, respectively. The higher δ13C and δ15N in shelf resident Chinook compared to the offshore rearing stocks, and their different marine distributions explain the large divergence in contaminant levels and profiles, with shelf resident stocks being heavily influenced by land-based sources of industrial contamination. Results provide compelling new insight into the drivers of contaminant accumulation in Chinook salmon, raise important questions about the consequences for their health, and explain a major pathway to the heavily POP-contaminated Resident killer whales that consume them.

Preparation of PCL/lecithin/bacteriocin CAMT6 antimicrobial and antioxidant nanofiber films using emulsion electrospinning: Characteristics and application in chilled salmon preservation.

Multi-functional packaging materials are an important development for food preservation. Emulsion electrospinning is a novel and simple method that can be used to prepare multi-functional packaging materials, which can effectively protect the loaded active substances during the preparation process. In this study, PCL/lecithin/bacteriocin CAMT6 nanofiber films with antimicrobial and antioxidant activity were prepared by emulsion electrostatic spinning. The morphology and homogeneity of the prepared nanofibrous membranes could be improved by optimising the formulation of the emulsion for electrospinning. Analytical testing of the prepared nanofiber films revealed that the nanofibers had a core-shell structure, with bacteriocin CAMT6 effectively encapsulated in the core layer and the PCL and phospholipids homogeneously mixed to form the shell layer. Additionally, the nanofiber films had acceptable tensile properties and water absorption capacity. In chilled salmon meat, the nanofiber film effectively inhibited the growth of bacteria, slowed the oxidation of oil and slowed water loss, which was a good protective effect. This study provides a reference for the encapsulation application of food-active packaging materials and bacteriocins.

Evaluation of healing progression at surgical incision sites and the use of antiseptics for enhancing post-operative survival in subyearling Chinook salmon (Oncorhynchus tshawytscha).

In an attempt to develop more effective surgical implantation methods for fish, surgical incisions typical of those made for implanting micro-acoustic transmitters into the peritoneal cavity were evaluated on a weekly basis for healing progression using a suite of metrics. Additionally, four chemicals were evaluated at concentrations commonly used in aquaculture for their ability to prevent surgical site infection and thus to promote incision healing and survival. Chemical treatments included hydrogen peroxide (25, 50, and 100 mg l-1), salt (10 and 30 ppt), Argentyne (1:1, Argentyne:water), and PolyAqua (1/2 tsp 36 l-1). For all study fish, the presence of two intact sutures seven days post-surgery (generally associated with good suturing technique) was negatively correlated with survival. A generalized linear mixed effects model indicated that suture presence, increasing tagging temperature, and the presence of foreign material on sutures decreased survival by 0.56, 0.72 and 0.60 respectively. Conversely, evidence of suture tearing and increasing fork length at tagging increased survival by 0.24 and 0.17. The antiseptic treatments tested promoted neither faster healing of surgical incisions nor higher survival for fish held for 28 days compared to a reference group and two of the chemicals may be contraindicated for prophylactic use at published doses. These findings underscore the need for researchers to adopt a decidedly cautious approach to planning and interpretation of study outcomes that rely on telemetry tagging, carefully considering the study subjects, potential effects of the techniques used, and implications of the environmental conditions experienced.

Arginine-rich peptides as crystallization inhibitors for sodium urate.

Inhibiting the formation of urate crystals is the key to prevent hyperuricemia from developing into gout. Although many studies have focused on the influence of biomacromolecules in the crystallization behavior of sodium urate, the role of peptides with specific structures may contribute to unprecedented regulatory effects. Here, for the first time, we studied the effects of cationic peptides on the phase behavior, crystallization kinetics, and size/morphology of urate crystals. The addition of protamine (PRTM, a typical natural arginine-rich peptide) prolongs the nucleation induction time of sodium urate and inhibits crystal nucleation effectively. PRTM binds to the surface of amorphous sodium urate (ASU) through the hydrogen bond and electrostatic attraction between guanidine groups and urate anions, which is conducive to maintaining the state of ASU and inhibiting crystal nucleation. Moreover, PRTM preferentially binds to the MSUM plane and leads to a significant reduction in the aspect ratio of MSUM filamentous crystals. Further studies showed that there are significant differences in the inhibiting effects of arginine-rich peptides with different chain lengths on the crystallization behavior of sodium urate. Both guanidine functional groups and peptide chain length determine the crystallization inhibiting effect of peptides simultaneously. The present work highlights the potential role of arginine peptides in inhibiting the crystallization of urate and provides new insights into the inhibition mechanism in the pathological biomineralization of sodium urate, demonstrating the possibility of using cationic peptides to treat gout.

Electrospun chitosan-based nanofibers loading tea tree oil for fresh salmon fillet shelf-life extension.

Bioactive packaging can improve the shelf-life of food products and enhance consumer health. It can also alleviate environmental stress on the planet by reducing food waste. Here, the electrospinning of tea tree oil-loaded 2-hydroxypropyltrimethyl ammonium chloride chitosan nanofibers was investigated. The fabricated nanofiber films were characterized by scanning electron microscopy, thermal gravimetric analysis, Fourier transform infrared spectroscopy, and contact angle meter analysis. The prepared nanofibers have a well-defined diameter of about 200 nm and a smooth shape. They have good antibacterial properties against Staphylococcus aureus and Escherichia coli in vitro. Tea tree oil-loaded chitosan-based nanofibers were found to be effective in delaying spoilage and extending the shelf life of salmon by sensory evaluation, texture analysis, color, total viable counts, thiobarbituric acid, and total volatile basic nitrogen during storage in the freshness experiments, thus indicating their health benefits in bioactive packaging.

A novel material for the microbiological, oxidative, and color stability of salmon and chicken meat samples: Nanofibers obtained from sesame oil.

Sesame oil nanofibers (diameter min: 286 max: 656nm), starting thermal degradation at 60 °C, were successfully obtained using the electrospinning technique in Türkiye. The distance, high voltage, and flow rate in electrospinning parameters were defined as 10 cm, 25 kV, and 0.065 mL/min. Mesophilic, psychrophilic bacteria, and yeast & molds counts of control group samples were higher (up to 1.21 log CFU/g) than those of salmon and chicken meat samples treated with sesame oil nanofibers. Thiobarbituric acid (TBA) value in control salmon meat samples stored for 8 days was defined between 0.56 and 1.48 MDA/kg (increase: 146%). However, the rise in TBA for salmon samples treated with sesame oil nanofibers was 21%. Also, nanofiber application for chicken samples limited the rapid oxidation up to 51.51% compared to control samples on the 8th day (p < 0.05). b* value (decline: 15.23 %) associated with rapid oxidation of the control group in salmon samples was more rapidly decreased than that of fish samples treated with sesame-nanofibers (b*: 12.01%) (p < 0.05). Chicken fillets b* values were more stable compared to control chicken meat samples for 8 days. Sesame oil-nanofiber application did not adversely affect the L* value color stability of all meat samples.

Enhancing the gastrointestinal stability of salmon calcitonin through peptide stapling.

Salmon calcitonin (sCT) is a polypeptide hormone available in the clinic. sCT is degraded in the gastrointestinal tract in minutes. In this work, a stapled analogue of salmon calcitonin, KaY-1(R24Q), was developed using the cooperative stapling between Lys and Tyr, with R24Q substitution. The analogue exhibited an improved stability in simulated gastric and intestinal fluid and retained the ability to activate the calcitonin receptor. This work will serve as a starting point for the development of an oral sCT drug.

Assessing the role of Piscine orthoreovirus in disease and the associated risk for wild Pacific salmon.

This paper is a response to Polinski, M. P. et al. Innate antiviral defense demonstrates high energetic efficiency in a bony fish. BMC Biology 19, 138 (2021). https://doi.org/10.1186/s12915-021-01069-2.

An Analysis of Seafood Recalls in the United States, 2002 Through 2022.

This review analyzes the seafood recalls registered by the United States Food and Drug Administration (USFDA) from October 2002 through March 2022. There were more than 2,400 recalls for seafood products over this 20-year period. Biological contamination was the listed root cause for about 40% of these recalls. Almost half were designated as Class I recalls, due to the high risk of the recalled seafood to cause disease or death. Independent of the recall classification, 74% of the recalls were due to violations of the Current Good Manufacturing Practices (cGMPs) regulations. The most common cause for these seafood recalls was due to undeclared allergens (34%). More than half of the undeclared allergen recalls were for undeclared milk and eggs. Recalls for Listeria monocytogenes accounted for 30% of all recalls and were all Class I. Finfish comprised 70% of the recall incidents, and salmon was the single most recalled species (22%). Improper cold smoking treatment that resulted in Listeria monocytogenes contamination was the most common reason reported for the salmon recalls. The goal of this review was to evaluate the main causes of food safety failures within the seafood manufacturing and distribution sectors. Human errors and failures to control food safety risks during the processing of food are the main driving factors for most reported recalls in the U.S. Properly applying the Hazard Analysis Critical Control Points (HACCP) approach and procedures are needed to identify the potential food safety risks. The key to reducing the risks of human error and loss of process control is the development and implementation of an effective food safety culture program at the manufacturing facility, which must require strong senior management support at corporate and enterprise levels.

Coproduction and modeling spatial contact networks prevent bias about infectious hematopoietic necrosis virus transmission for Snake River Basin salmonids.

Much remains unknown about variation in pathogen transmission across the geographic range of a free-ranging fish or animal species and about the influence of movement (associated with husbandry practices or animal behavior) on pathogen transmission. Salmonid hatcheries are an ideal system in which to study these processes. Salmonid hatcheries are managed for endangered species recovery, supplementation of threatened or at-risk fish stocks, support of fisheries, and ecosystem stability. Infectious hematopoietic necrosis virus (IHNV) is a rhabdovirus of significant concern to salmon aquaculture. Landscape IHNV transmission dynamics previously had been estimated only for salmonid hatcheries in the Lower Columbia River Basin (LCRB). The objectives of this study were to estimate IHNV transmission dynamics in a unique geographic region, the Snake River Basin (SRB), and to quantitatively estimate the effect of model coproduction on inference because previous assessments of coproduction have been qualitative. In contrast to the LCRB, the SRB has hatchery complexes consisting of a main hatchery and ≥1 satellite facility. Knowledge about hatchery complexes was held by a subset of project researchers but would not have been available to project modelers without coproduction. Project modelers generated and tested multiple versions of Bayesian susceptible-exposedinfected models to realistically represent the SRB and estimate the effect of coproduction. Models estimated the frequency of transmission routes, route-specific infection probabilities, and infection probabilities for combinations of salmonid hosts and IHNV lineages. Model results indicated that in the SRB, avoiding exposure to IHNV-positive adult salmonids is the most important action to prevent juvenile infections. Migrating adult salmonids exposed juvenile cohort-sites most frequently, and the infection probability was greatest following exposure to migrating adults. Without coproduction, the frequency of exposure by migrating adults would have been overestimated by 70 cohort-sites, and the infection probability following exposure to migrating adults would have been underestimated by∼0.09. The coproduced model had less uncertainty in the infection probability if no transmission route could be identified (Bayesian credible interval (BCI) width = 0.12) compared to the model without coproduction (BCI width = 0.34). Evidence for virus lineage MD specialization on steelhead and rainbow trout (both Oncorhynchus mykiss) was apparent without model coproduction. In the SRB, we found a greater probability of virus lineage UC infection in Chinook salmon (Oncorhynchus tshawytscha) compared to in O. mykiss, whereas in the LCRB, UC more clearly exhibited a generalist approach. Coproduction influenced estimates that depended on transmission routes, which operated differently at main hatcheries and satellite sites within hatchery complexes. Hatchery complexes are found outside of the SRB and are not specific to salmonid hatcheries alone. There is great potential for coproduction and modeling spatial contact networks to advance understanding about infectious disease transmission in complex production systems and surrounding free-ranging animal populations.

Surface-enhanced Raman spectroscopy for studying the interaction of N-propyl substituted imidazole compound with salmon sperm DNA.

BACKGROUND: Surface Enhanced Raman Spectroscopy (SERS) is a very promising and fast technique for studying drugs and for detecting chemical nature of a molecule and DNA interaction. In the current study, SERS is employed to check the interaction of different concentrations of n-propyl imidazole derivative ligand with salmon sperm DNA using silver nanoparticles as SERS substrates. OBJECTIVES: Multivariate data analysis technique like principal component analysis (PCA) and partial least squares-discriminant analysis (PLS-DA) are employed for the detailed analysis of the SERS spectral features associated with the mode of action of the imidazole derivative ligand with DNA. METHODOLOGY: Silver nanoparticles were used as a SERS substrate in DNA-drug interaction. Five different concentrations of ligands were interacted with DNA and mix with Ag-NPs as substrate. The SERS spectra of were acquired for all seven samples and processed using MATLAB. Additionally, PCA and PLS-DA were used to assessed the ability SERS to differentiate interaction of DNA-drug. RESULTS: Differentiating SERS features having changes in their peak position and intensities are observed including 629, 655, 791, 807, 859, 1337, 1377 and 1456 cm-1. These SERS features reveal that binding of ligand with DNA is electrostatic in nature, and have specificity to major groove where it forms GC-CG interstrand cross-linking with the DNA double helix. CONCLUSIONS: SERS give significant information regarding to Drug-DNA interaction mechanism, SERS spectra inferred the mode of action of anticancer compound that are imidazole in nature.

Shotgun-Based Mass Spectrometry Analysis of Phospholipid and Triacylglycerol Molecular Species and Eicosanoids in Salmon Muscle Tissue on Feeding Microbial Oil.

The continuous growth of aquaculture places a growing demand on alternative sources of fish oil (FO). Certain microorganisms provide a sustainable replacement for FO due to their content of EPA and DHA, which are essential for fish health. Appreciable evidence shows that changes in feeding sources may alter the nutritional components of salmon; however, the influence of diets on lipid species remains unclear. In this study, the identification and semi-quantification of lipid molecular species in salmon muscle during feeding with a microbial oil (MO) were carried out by focusing on triacylglycerol (TAG) and diacyl-phospholipid using shotgun-based mass spectrometry analysis. DHA in the MO diet was efficiently incorporated into phospholipid structures on feeding, followed by accumulation in salmon muscle. The MO diet elevated the level of certain EPA-containing TAGs, such as TAG C52:5 (16:0_16:0_20:5) and TAG C54:6 (16:0_18:1_20:5), indicating that the MO diet may be an excellent source for enhancement of the abundance of ω3 lipids. Further, prostaglandins (PGs) PGE2 and PGF3α were identified and quantified for the first time in salmonid tissue.

Peritrophin-like Genes Are Associated with Delousing Drug Response and Sensitivity in the Sea Louse Caligus rogercresseyi.

Caligus rogercresseyi is the main ectoparasite that affects the salmon industry in Chile. The mechanisms used by the parasite to support its life strategy are of great interest for developing control strategies. Due to the critical role of insect peritrophins in host-parasite interactions and response to pest control drugs, this study aimed to identify and characterize the peritrophin-like genes present in C. rogercresseyi. Moreover, the expression of peritrophin-like genes was evaluated on parasites exposed to delousing drugs such as pyrethroids and azamethiphos. Peritrophin genes were identified by homology analysis among the sea louse transcriptome database and arthropods peritrophin-protein database obtained from GenBank and UniProt. Moreover, the gene loci in the parasite genome were located. Furthermore, peritrophin gene expression levels were evaluated by RNA-Seq analysis in sea louse developmental stages and sea lice exposed to delousing drugs deltamethrin, cypermethrin, and azamethiphos. Seven putative peritrophin-like genes were identified in C. rogercresseyi with high homology with other crustacean peritrophins. Differences in the presence of signal peptides, the number of chitin-binding domains, and the position of conserved cysteines were found. In addition, seven peritrophin-like gene sequences were identified in the C. rogercresseyi genome. Gene expression analysis revealed a stage-dependent expression profile. Notably, differential regulation of peritrophin genes in resistant and susceptible populations to delousing drugs was found. These data are the first report and characterization of peritrophin genes in the sea louse C. rogercresseyi, representing valuable knowledge to understand sea louse biology. Moreover, this study provides evidence for a deeper understanding of the molecular basis of C. rogercresseyi response to delousing drugs.

Influence of laurel (Laurus nobilis) essential oil on gut function of Black Sea salmon (Salmo labrax) juveniles.

The present work investigated the effects of dietary incorporation of laurel (Laurus nobilis) essential oil on the zootechnical performance and digestive physiology of juvenile Black Sea salmon (Salmo labrax). In this trial, 15 fiberglass tanks (39 × 39 cm square and 33 cm high) were used. Forty-five fish (3.52 ± 0.01 g) were placed randomly per tank. Fish were fed for 90 days with the diet containing 50, 100, 200, or 400 mg kg-1 laurel (Laurus nobilis) essential oils, respectively. The work was performed in the recirculating aquaculture system (RAS) operating with freshwater. Fish were manually fed 3% level of live weight during the experiment period. Final weight (FW), weight gain (WG), feed conversion rate (FCR), and specific growth rate (SGR) were not affected by laurel essential oil supplementation. Dietary laurel essential oil (50 mg kg-1) affected positively the surface area of fish intestinal villus that required for digestion. Both villus height and villus width were affected positively in fish fed with 50 mg laurel essential oil kg-1. While incorporation with 100 mg laurel essential oil kg-1 increased the total α-amylase enzyme, 50 mg laurel kg-1 increased lipase enzyme. Moreover, 50 mg laurel essential oil kg-1 increased lactic acid bacteria (LAB) count in fish. Besides, 50 mg laurel essential oil kg-1 reduced the number of total coliform and E. coli.

A volatile basic nitrogens-responsive tag based on aggregation-induced emission luminogen for real-time monitoring and in situ visualization of salmon freshness.

Given the ever-growing food safety issues, the establishment of efficient approaches for monitoring food freshness attracts increasing attention. Volatile basic nitrogens (VBNs), including biogenic amines and ammonia, serve as an important biomarker for monitoring food freshness. In this study, a novel VBNs-responsive tag using glutathione capped copper nanoclusters (GSH-CuNCs) aggregates was developed as a fluorescent probe for in situ and real-time visual monitoring of salmon freshness, and the prepared GSH-CuNCs aggregates were characterized and their sensitivity for detecting VBNs using biogenic amines and ammonium hydroxide as the model targets was evaluated. Based on their remarkable response in liquid status, the GSH-CuNCs aggregates-based tag as a gas indicator was then fabricated, which exhibited visible colour changes under UV light as a function of ammonia vapour concentrations. More importantly, through exploring the sensing mechanism of GSH-CuNCs aggregates in VBNs detection, the existence of ligand exchange between the GSH-CuNCs and VBNs was observed and verified for the first time, confirming the effect of hydrogen bonding reported in the literature. Moreover, the GSH-CuNCs aggregates-based tag was applied for quantitative analysis of salmon freshness during different storage periods, which was validated by the standard method for detection of total VBNs in salmon. In addition, a colour card was developed and its feasibility for application in monitoring salmon freshness was validated, which could be used for consumers to obtain the freshness level directly with the naked eye, demonstrating the feasibility of the fabricated tag for real-time and visual monitoring of salmon freshness, thus showing great potentials for its practical applications in the food industry.

PAMPs of Piscirickettsia salmonis Trigger the Transcription of Genes Involved in Nutritional Immunity in a Salmon Macrophage-Like Cell Line.

The innate immune system can limit the growth of invading pathogens by depleting micronutrients at a cellular and tissue level. However, it is not known whether nutrient depletion mechanisms discriminate between living pathogens (which require nutrients) and pathogen-associated molecular patterns (PAMPs) (which do not). We stimulated SHK-1 cells with different PAMPs (outer membrane vesicles of Piscirickettsia salmonis "OMVs", protein extract of P. salmonis "TP" and lipopolysaccharides of P. salmonis "LPS") isolated from P. salmonis and evaluated transcriptional changes in nutritional immunity associated genes. Our experimental treatments were: Control (SHK-1 stimulated with bacterial culture medium), OMVs (SHK-1 stimulated with 1µg of outer membrane vesicles), TP (SHK-1 stimulated with 1µg of total protein extract) and LPS (SHK-1 stimulated with 1µg of lipopolysaccharides). Cells were sampled at 15-, 30-, 60- and 120-minutes post-stimulation. We detected increased transcription of zip8, zip14, irp1, irp2 and tfr1 in all three experimental conditions and increased transcription of dmt1 in cells stimulated with OMVs and TP, but not LPS. Additionally, we observed generally increased transcription of ireg-1, il-6, hamp, irp1, ft-h and ft-m in all three experimental conditions, but we also detected decreased transcription of these markers in cells stimulated with TP and LPS at specific time points. Our results demonstrate that SHK-1 cells stimulated with P. salmonis PAMPs increase transcription of markers involved in the transport, uptake, storage and regulation of micronutrients such as iron, manganese and zinc.

Management of primary choroidal lymphoma presenting as extensive serous retinal detachment and salmon patch of bulbar conjunctiva: a case report.

BACKGROUND: We describe the outcome of ultra-low-dose radiotherapy plus intravitreal methotrexate and rituximab injections for a patient with primary choroidal lymphoma who presented with nodular conjunctival salmon patches and extensive serous retinal detachment. CASE PRESENTATION: A 34-year-old Iranian man presented with a nodular patch of bulbar conjunctiva in the right eye, and 1+ vitritis. A nearly complete shallow serous retinal detachment, retinal folds, and multifocal yellow choroidal infiltrates were seen during a fundus examination of the right eye. Enhanced depth imaging optical coherence tomography revealed macular retinal folds and an uneven, undulating, "seasick" appearance of the choroidal surface with choriocapillaris compression, intraretinal and subretinal fluid, and clusters of optically dense material at the outer retinal level. An incisional biopsy of the conjunctival lesion confirmed the diagnosis of primary choroidal lymphoma with epibulbar involvement. The patient was treated with ultra-low-dose "boom-boom" radiation (4 Gy delivered in two fractions over two consecutive days) as well as intravitreal methotrexate and rituximab injections. After a year, the lesions had completely disappeared, with no adverse effects or recurrence. CONCLUSION: Ultra-low-dose (boom-boom) radiotherapy combined with intravitreal chemotherapy and/or immunotherapy may be an effective treatment for primary choroidal lymphoma with anterior epibulbar extension and diffuse subretinal fluid with favorable response and minimal side effects.