Trout Fry Research Articles

Cytotoxic activity of Flavobacterium psychrophilum in skeletal muscle cells of rainbow trout (Oncorhynchus mykiss)

Flavobacterium psychrophilum is the etiologic agent of bacterial coldwater disease (BCWD) and rainbow trout fry syndrome (RTFS), which cause significant worldwide losses in aquaculture. Juvenile rainbow trout are particularly susceptible to F. psychrophilum infection, the main external clinical signs of which are extensive necrotic myositis and ulcerative lesions. Despite the economic relevance of this pathogen in aquaculture, little is known about the molecular mechanisms underlying F. psychrophilum infection and pathogenesis. In this study, cultured skeletal muscle cells from rainbow trout (Oncorhynchus mykiss) were co-incubated with the virulent strain of F. psychrophilum JIP02/86 (ATCC 49511). Trypan blue exclusion analysis at 48h post-incubation revealed decreased cellular viability. Direct bacteria-myoblast contact was found a key factor in inducing F. psychrophilum cytotoxicity. Apoptosis was characterized by nuclear DNA fragmentation, decreased plasma membrane integrity, increased caspase activity, and the proteolytic cleavage of poly(ADP-ribose)polymerase-1 (PARP-1). Moreover, bacterial infection induced an early inhibition of NF-κB signaling, as well as a differential expression of the pro- and anti-apoptotic genes, bax and bcl-2. These findings suggest that F. psychrophilum induces rainbow trout muscle apoptosis through the modulation of the NF-κB signaling as a mechanism for nutrient acquisition and survival.

Effects of euthanasia methods on stable carbon (δ13 C value) and nitrogen (δ15 N value) isotopic compositions of fry and juvenile rainbow trout Oncorhynchus mykiss.

Carbon and nitrogen stable isotope analyses of fish tissues are now commonly used in ecological studies but mostly require the sacrifice of the animal. Ethical considerations recommend the use of anesthetics for tissue sampling. This study examines how anesthetics affect stable isotope ratios of fish compared with other euthanasia methods. Rainbow trout fry and juveniles were sacrificed using ice-freezing (as this common method used to kill fish does not affect natural isotopic ratios), electronarcosis or an overdose of chemical anesthetics (2-phenoxyethanol, benzocaine and clove oil). For fry, we sampled the whole animal whereas, for juveniles, white dorsal muscle, liver, red blood cells, plasma, external tegument and pectoral fin were sampled. Isotopic ratios and the elemental compositions of carbon and nitrogen were then measured. The δ15 N values, and the C and N contents of all considered tissues as well as δ13 C values of muscle, liver, red blood cells and plasma, were not affected by the use of chemical anesthetics. Clove oil and to a lesser extent 2-phenoxyethanol and benzocaine decreased δ13 C values of whole fry and juvenile external tegument and pectoral fin. The use of electronarcosis drastically affects the δ13 C and δ15 N values of all fish tissues. Anesthetics should be avoided for δ13 C analysis when tissues are in contact with the water containing the anesthetic. Ice-immersion has to be preferred when approved by guidelines. If not, benzocaine and 2-phenoxyethanol should be preferred over clove oil. Electronarcosis should not be used to kill fish until further investigations are performed.

The Effect on Growth Performance of Rainbow Trout Fry (Onchorhynchus Mykiss, Walbaum 1792) of Rosemary and Clove Oils Added Diet

In this study was aimed to determine the growth performance of rainbow trout ( Onchorhynchus mykiss , Walbaum 1792) fry fed with feed containing essential oils rosemary, cloves and rosemary + clove at different rates. In research, rainbow trout fries were used which initial weight 30 ± 5 g. Experimental groups are that rosemary, clove and rosemary + clove essential oils added the diets. These groups were as follow: Control group (no added any additive), % 0.5 Rosemary oil added R-0.5 , % 1 Rosemary oil added R-1 , % 1.5 Rosemary oil added R-1.5 , % 0.5 Clove oil added C-0.5 , % 1 Clove oil added C-1 , % 1,5 Clove oil added C-1.5 , % 0,5 Rosemary + Cove oil added RC-0.5 , % 1 Rosemary + Cove oil added RC-1 , % 1,5 Rosemary + Cove oil added RC-1.5 group. Experiment lasted 60 day. Taken measured values were recorded in fish during trial. At the e nd of the study it was calculated growth parametres ; average live weight , daily absolute live weight gain, relative growth rates, specific growth rate , condition factor with viserosomatic and hepatosomatic factor. The findings obtained in the study were subjected to analysis of variance and the importance degree of difference between the groups was determined by Duncan 's multiple comparison test. In the light of statistical data, daily absolute live weight gains with the viserosomatic and hepatosomatic index values were found to be significant differences between all groups ( p < 0.05).

Weekly changes in prey availability for and the selective feeding of sea trout (Salmo trutta L.) larvae stocked in small forest streams

Abstract This study examined the availability of food and its selection by sea trout Salmo trutta L. fry in the first four weeks of life after yolk sac resorption. The food base and stomach contents of sea trout fry after release in the wild were determined. The study was performed in two small forest streams from April 25 to May 23, 2014. Macro-zoobenthos that comprised the food base for the fry were collected from the streams weekly using a bottom scraper. On the same day, the fry were captured with electrofishing gear. Analysis of the width of the benthic organisms in the food base and in the fish stomachs indicated the prey size range that the fish were feeding on. Further, the study showed that all food items found in both streams during the study weeks were also represented in the intensity of the fish: Cyclopoida, the larvae of Baetidae, Simuliidae and Nemouridae.

Efficacy of a polyvalent immersion vaccine against Flavobacterium psychrophilum and evaluation of immune response to vaccination in rainbow trout fry (Onchorynchus mykiss L.)

Rainbow trout fry syndrome (RTFS) is a disease caused by the Gram-negative bacterium Flavobacterium psychrophilum, responsible for significant economic losses in salmonid aquaculture worldwide. The diversity of F. psychrophilum isolates and the inherent difficulties in vaccinating juvenile fish has hampered the development of a vaccine for RTFS. Disease episodes tend to occur between 10–14 °C with necrotic lesions often seen on the skin surrounding the dorsal fin and tail. At present no commercial vaccines are available for RTFS in the UK, leaving antibiotics as the only course of action to control disease outbreaks. The current work was performed as a pilot study to assess the efficacy of a polyvalent, whole cell vaccine containing formalin-inactivated F. psychrophilum, to induce protective immunity in rainbow trout fry. Duplicate groups of 30 trout (5 g) were immersed in 1 L of the vaccine for 30 s. Samples were taken 4 h, day 2 and 7 post-vaccination (pv) of skin mucus, tissues for histology and gene expression analysis; serum and histology samples were taken 6 weeks pv. A booster vaccination was given at 315 degree days (dd) also by immersion. Challenge was by immersion with a heterologous isolate of F. psychrophilum 630 dd post primary vaccination. The vaccine provided significant protection to the trout fry with a RPS of 84% (p < 0.0001). Detection of increased numbers of IgT positive cells in systemic organs, up-regulation of IgT expression in hind-gut and an increase in total IgT in serum was observed in vaccinated fish; however a functional role of IgT in the observed protection remains to be demonstrated.

Antioxidant-related parameters, digestive enzyme activity and intestinal morphology in rainbow trout (Oncorhynchus mykiss) fry fed graded levels of red seaweed,Gracilaria pygmaea

This study was conducted to investigate the effects of dietary red seaweed, Gracilaria pygmaea level on growth, antioxidant-related parameters, digestive enzyme activity and intestinal morphology of rainbow trout (Oncorhynchus mykiss) fry. Five isocaloric and isonitrogenous diets were formulated to contain four G. pygmaea levels (30 (GL-30), 60 (GL-60), 90 (GL-90) and 120 g/kg (GL-120)), and a control diet was used, without inclusion of seaweed. Results of the feeding trial indicated that final body (FBW: 2.8–3.5 g) and specific growth rate (SGR: 4.8–5.2) were significantly improved when 60 g/kg G. pygmaea was supplemented in the diets. However, there was a significant reduction (p < .05) of final body weight (FBW) and specific growth rate (SGR) in fish fed 120 g/kg G. pygmaea. Protease activity was significantly lower in fish fed GL-120 diet when compared to GL-30 or control (p < .05). Lipase was significantly affected by dietary seaweed regardless its level of supplementation (p < .05). The inclusion of the Gracilaria in the diet led to evident changes in the fish antioxidant status with significant reduction of superoxide dismutase, glutathione peroxidase activity and lipid peroxidation products. The histological study demonstrated that all groups of fish displayed normal morphology of anterior intestine and pyloric caeca. Villi absorptive area in the anterior intestine of treatment groups GL-90 and GL-120 significantly decreased (p < .05). Overall, the results obtained in this study indicate that dietary G. pygmaea supplementation up to 90 g/kg improves growth performance in rainbow trout fry without compromising antioxidant responses and digestive function.

Using ultrahigh-resolution mass spectrometry and toxicity identification techniques to characterize the toxicity of oil sands process-affected water: The case for classical naphthenic acids.

Previous assessments of oil sands process-affected water (OSPW) toxicity were hampered by lack of high-resolution analytical analysis, use of nonstandard toxicity methods, and variability between OSPW samples. We integrated ultrahigh-resolution mass spectrometry with a toxicity identification evaluation (TIE) approach to quantitatively identify the primary cause of acute toxicity of OSPW to rainbow trout (Oncorhynchus mykiss). The initial characterization of OSPW toxicity indicated that toxicity was associated with nonpolar organic compounds, and toxicant(s) were further isolated within a range of discrete methanol fractions that were then subjected to Orbitrap mass spectrometry to evaluate the contribution of naphthenic acid fraction compounds to toxicity. The results showed that toxicity was attributable to classical naphthenic acids, with the potency of individual compounds increasing as a function of carbon number. Notably, the mass of classical naphthenic acids present in OSPW was dominated by carbon numbers ≤16; however, toxicity was largely a function of classical naphthenic acids with ≥17 carbons. Additional experiments found that acute toxicity of the organic fraction was similar when tested at conductivities of 400 and 1800 μmhos/cm and that rainbow trout fry were more sensitive to the organic fraction than larval fathead minnows (Pimephales promelas). Collectively, the results will aid in developing treatment goals and targets for removal of OSPW toxicity in water return scenarios both during operations and on mine closure. Environ Toxicol Chem 2017;36:3148-3157. © 2017 SETAC.

Enhancement of immune responses of rainbow trout (Oncorhynchus mykiss) fed a diet supplemented with Aloe vera extract

The effects dietary supplementation with Aloe vera extract (AE) on the immunity responses and hematological parameters of rainbow trout (Oncorhynchus mykiss) fry were evaluated in eight weeks trial. 600 rainbow trout (O. mykiss) fry with an average initial body weight of 13±0.05g were randomly allocated into two treatment groups including placebo-treated group (control) and A. vera extract-treated group, each with three replicates. The fish were hand-fed once a day with diet medicated AE or placebo (70 % lactose, 10 % starch and 20 % talc) at a rate of 1% of feed weight in the first feeding for 8 weeks. At the end of the identical every two weeks (2, 4, 6, and 8 weeks) 24 h after feeding, some of immunological and hematological parameters were analyzed. The results showed that serum total protein, albumin and globulin, respiratory burst activity, phagocytic activity and serum lysozyme activity vary among the two treatment groups which were found to be higher in the AE-treated group (p <0.05). However, there were no significant differences in none of hematological parameters between two groups. It was concluded that supplementation of AE at a rate of 1% in feed registered higher immunological responses in compared to placebo group. Therefore, supplementation of AE in fish diet would enhance the immunity responses in fish. It may use in fish diets particularly at time of outbreaks.

Differentiation of Flavobacterium psychrophilum from Flavobacterium psychrophilum-like species by MALDI-TOF mass spectrometry

Rainbow trout fry syndrome (RTFS) is an important infectious disease caused by Flavobacterium psychrophilum affecting farmed salmonids worldwide. Other Flavobacterium psychrophilum-like species (F. plurextorum, F. oncorhynchi, F. tructae, F. collinsii and F. piscis) have been isolated from diseased rainbow trout fry suspected of RTFS although the epidemiological and clinical relevance of these pathogens are unknown. The objective of this study was to evaluate the potential use of MALDI-TOF (Matrix-Assisted Laser Desorption/Ionization-Time of Flight) Mass Spectrometry as method for specific identification of F. psychrophilum and its differentiation from other F. psychrophilum-like species isolated from diseased fish. Fifty-three isolates were analyzed after the creation of the Main Spectrum Profile (MSP) of reference strains of each of abovementioned species. F. psychrophilum exhibited a mass spectra very different from those of F. psychrophilum-like species, with five peaks (m/z 3654, 4585, 5388, 6730 and 7310) present only in F. psychrophilum isolates, and three peaks (m/z 6170, 7098 and 9241) absent in F. psychrophilum but present in all F. psychrophilum-like species. All F. psychrophilum isolates were correctly identified and differentiated from the F. psychrophilum-like species by MALDI-TOF. Although this approach showed a limited ability to differentiate among F. psychrophilum-like species, its complementation with a few simple biochemical tests may represent an alternative approach for the routine identification of the Flavobacterium psychrophilum-like species.

A RNAi-based therapeutic proof of concept targets salmonid whirling disease in vivo.

Myxobolus cerebralis is a cnidarian-myxozoan parasite that causes salmonid whirling disease. M. cerebralis alternates between two hosts: (1) a vertebrate salmonid and (2) an invertebrate oligochaete, Tubifex tubifex. There is no successful treatment for salmonid whirling disease. MyxSP-1 is a M. cerebralis serine protease implicated in whirling disease pathogenesis. We hypothesized that short-interfering RNA (siRNA)-induced RNA interference (RNAi) can silence MyxSP-1 in the invertebrate host and abrogate the M. cerebralis life cycle. This would preclude whirling disease infection in the salmonid host. To test this hypothesis, we first developed a siRNA delivery protocol in T. tubifex. Second, we determined the effective dose for siRNA treatment of M. cerebralis-infected T. tubifex. M. cerebralis-infected T. tubifex were treated with different concentrations of MyxSP-1 or negative control siRNAs (1μM, 2μM, 5μM or 7μM) at 15°C for 24h, 48h, 72h and 96h, respectively. We monitored MyxSP-1 knockdown using real-time quantitative PCR (qPCR). siRNA treatment with MyxSP-1 siRNA at 2μM concentration for 24h at 15°C showed maximum significant MyxSP-1 knockdown in T. tubifex. Third, we determined the time points in the M. cerebralis life cycle in T. tubifex at which siRNA treatment was most effective. M. cerebralis-infected T. tubifex were treated with MyxSP-1 or negative control siRNAs (2μM concentration for 24h at 15°C) at 24 hours post-infection (24hpi), 48hpi, 72hpi, 96hpi, 1 month post-infection (1mpi), 2mpi and 3mpi, respectively. We observed that siRNA treatment of T. tubifex was most effective at 1mpi, 2mpi and 3mpi. Fourth, we immersed specific-pathogen-free rainbow trout fry in water inhabited by MyxSP-1 siRNA-treated T. tubifex (at 1mpi, 2mpi and 3mpi). The salmonids did not develop whirling disease and showed significant MyxSP-1 knockdown. We also observed long-term RNAi in T. tubifex. Together these results demonstrate a novel RNAi-based therapeutic proof of concept in vivo against salmonid whirling disease.

The malleable gut microbiome of juvenile rainbow trout (Oncorhynchus mykiss): Diet-dependent shifts of bacterial community structures.

Plant-derived protein sources are the most relevant substitutes for fishmeal in aquafeeds. Nevertheless, the effects of plant based diets on the intestinal microbiome especially of juvenile Rainbow trout (Oncorhynchus mykiss) are yet to be fully investigated. The present study demonstrates, based on 16S rDNA bacterial community profiling, that the intestinal microbiome of juvenile Rainbow trout is strongly affected by dietary plant protein inclusion levels. After first feeding of juveniles with either 0%, 50% or 97% of total dietary protein content derived from plants, statistically significant differences of the bacterial gut community for the three diet-types were detected, both at phylum and order level. The microbiome of juvenile fish consisted mainly of the phyla Proteobacteria, Firmicutes, Bacteroidetes, Fusobacteria and Actinobacteria, and thus fits the salmonid core microbiome suggested in previous studies. Dietary plant proteins significantly enhanced the relative abundance of the orders Lactobacillales, Bacillales and Pseudomonadales. Animal proteins in contrast significantly promoted Bacteroidales, Clostridiales, Vibrionales, Fusobacteriales and Alteromonadales. The overall alpha diversity significantly decreased with increasing plant protein inclusion levels and with age of experimental animals. In order to investigate permanent effects of the first feeding diet-type on the early development of the microbiome, a diet change was included in the study after 54 days, but no such effects could be detected. Instead, the microbiome of juvenile trout fry was highly dependent on the actual diet fed at the time of sampling.

Oral DNA vaccines based on CS-TPP nanoparticles and alginate microparticles confer high protection against infectious pancreatic necrosis virus (IPNV) infection in trout

Infectious pancreatic necrosis virus (IPNV) is the etiological agent of a contagious viral disease causing remarkable mortalities in different fish species. Despite the availability of commercial vaccines against IPN, the disease still constitutes one of the main threats to the aquaculture industry worldwide. In this study, we developed a DNA vaccine encoding the VP2 gene of IPNV and evaluated its ability to induce protective immunity in rainbow trout fry (3 g) at doses of 10 and 25 μg/fish and boosting with the same doses two weeks later through the oral route using chitosan/tripolyphosphate (CS-TPP) nanoparticles and alginate microparticles incorporated into fish feed. The distribution of the administered vaccines in different organs and transcription of VP2 gene were confirmed by RT-PCR assay at day 30 post boost-vaccination. Transcript levels of IFN-1, Mx-1, IgM, IgT and CD4 genes was dependent on vaccine dose and was significantly up-regulated in head kidney of all orally vaccinated fish groups compared to controls (pcDNA3.1). Cumulative mortalities post-challenge with virulent isolate of the virus were lower in the vaccinated fish and a relative percentage survival (RPS) of 59% and 82% were obtained for the 10 and 25 μg/fish pcDNA3.1-VP2 groups, respectively. Vaccination with the same amount of pcDNA3.1-VP2 encapsulated with CS-TPP nanoparticles resulted in RPS of 47 %and 70%, respectively. Detectable anti-IPNV antibodies were shown until 90 days postvaccination. The orally administrated vaccines significantly decreased VP4 transcripts thus contributing to reducing viral load in surviving fish on day 45 post-challenge. In conclusion, these results show good to high protection post-vaccination alongside with significant up-regulation of key immune genes and detectable levels of circulating antibodies after oral administration of the DNA vaccine formulated in CS-TPP nanoparticles and alginate microparticles in fish feed.

Plastic responses by wild brown trout (Salmo trutta) to plant-based diets

Decreasing fishmeal availability and increasing prices promote the usage of plant-derived feedstuff as a substitution for fishmeal in commercial salmonid diets. However, little is known about the impact of plant-derived feedstuff on juvenile brown trout (Salmo trutta), a species that exhibits strong phenotypic plasticity with various genetic sub-structures and high overall genetic diversity. Thus, the production of brown trout for restocking purposes preferentially uses wild fish as broodstock to avoid loss of genetic variability. Because of nutritional programming, the strictly carnivorous feeding habit of wild brown trout broodfish could nevertheless have a negative impact on the digestive physiology of fry and fingerlings that are fed with commercial plant-protein containing trout diets. The present study, therefore, investigated whether the feeding of plant-based diets from first feeding onwards induced a permanent improvement in the utilisation of plant-derived protein sources in wild brown trout juveniles. Any plastic responses to the experimental diets resulting in a long-term physiological effect were hypothesised to be not only observed in growth performance, but also in altered pepsin and amylase activities. We demonstrated that (i) the feeding of wild brown trout fry with inclusion levels of up to 50% of dietary plant proteins is beneficial during the first weeks of life and (ii) continuous feeding of at least 50% plant-derived dietary protein resulted in the same rate of growth when compared to the growth resulting from fishmeal as the exclusive dietary protein source. Pepsin and amylase activities were only partly affected by diet-type and it can be concluded that intestinal pepsin and amylase activities in juvenile brown trout are primarily regulated by intrinsic mechanisms. In the present experiment, we were not able to induce a permanent nutritional programming effect of the first feeding diet; instead, a cross-over diet change applied 89days post first feeding demonstrated that wild brown trout fry exhibit highly plastic responses to different feeding strategies during the first months of life.

A effective DNA vaccine against diverse genotype J infectious hematopoietic necrosis virus strains prevalent in China

Infectious hematopoietic necrosis virus (IHNV) is the most important pathogen threatening the aquaculture of salmonid fish in China. In this study, a DNA vaccine, designated pIHNch-G, was constructed with the glycoprotein (G) gene of a Chinese IHNV isolate SD-12 (also called Sn1203) of genotype J. The minimal dose of vaccine required, the expression of the Mx-1 gene in the muscle (vaccine delivery site) and anterior kidney, and the titers of the neutralizing antibodies produced were used to evaluate the vaccine efficacy. To assess the potential utility of the vaccine in controlling IHNV throughout China, the cross protective efficacy of the vaccine was determined by challenging fish with a broad range of IHNV strains from different geographic locations in China. A single 100ng dose of the vaccine conferred almost full protection to rainbow trout fry (3g) against waterborne or intraperitoneal injection challenge with IHNV strain SD-12 as early as 4days post-vaccination (d.p.v.), and significant protection was still observed at 180d.p.v. Intragenogroup challenges showed that the DNA vaccine provided similar protection to the fish against all the Chinese IHNV isolates tested, suggesting that the vaccine can be widely used in China. Mx-1 gene expression was significantly upregulated in the muscle tissue (vaccine delivery site) and anterior kidney in the vaccinated rainbow trout at both 4 and 7d.p.v. Similar levels of neutralizing antibodies were determined with each of the Chinese IHNV strains at 60 and 180d.p.v. This DNA vaccine should play an important role in the control of IHN in China.

Different Phenotypes of Mature Biofilm in Flavobacterium psychrophilum Share a Potential for Virulence That Differs from Planktonic State.

Flavobacterium psychrophilum is the etiological agent of bacterial coldwater disease and the rainbow trout fry syndrome in salmonid aquaculture worldwide. However, there have been few studies into the capacity of F. psychrophilum to form biofilms and how these cellular accretions differ from planktonic cells or how they affect potential virulence. We evaluated the biofilm formation by three Chilean isolates of F. psychrophilum (LM-02-Fp, LM-06-Fp, and LM-13-Fp) and two non-Chilean strains (JIP02/86 and NCMB1947T), and compared biofilm and planktonic states to obtain insights into expression differences of virulence- and biofilm-related genes (VBRGs). Our findings are based on scanning confocal laser microscopy (SCLM) and LIVE/DEAD staining, enzymatic reactions, reverse transcription-quantitative PCR (RT-qPCR) of genes encoding putative virulence factors, and transcriptomes (RNA-Seq). The LM-02-Fp and NCMB1947T strains were the strongest and weakest biofilm producers, respectively. The strong-biofilm producer showed different physiological cell states distributed in different layers of mature biofilms, whereas the NCMB1947T biofilms consisted of cells arranged in a monolayer. WGA-binding exopolysaccharides would be the main components of their corresponding extracellular matrices. Transcriptomes of F. psychrophilum NCMB1947T and LM-02-Fp were clustered by state (biofilm vs. planktonic) rather than by strain, indicating important state-dependent differences in gene expression. Analysis of differentially expressed genes between states identified putative VBRGs involved in polysaccharide biosynthesis, lateral gene transfer, membrane transport (e.g., for drugs and Fe3+), sensory mechanisms, and adhesion, and indicated that about 60–100% of VBRGs involved in these processes was significantly upregulated in the biofilm state. Conversely, upregulated motility-related genes in the biofilm state were not identified, whereas a lower fraction of proteolysis-related genes (33%) was upregulated in biofilms. In summary, F. psychrophilum strains that produce different biofilm phenotypes show global transcriptional activity in the mature biofilm state that differs significantly from their planktonic counterparts. Also, different biofilm phenotypes share a genetic potential for virulence that is transcriptionally enhanced with respect to free-living cells. Our results suggest that the F. psychrophilum biofilm lifestyle acts as a reservoir for a given set of putative virulence factors, and recommend a deeper understanding of which could help prevent recurring infections in salmonid farms.

Pathogenicity of Mexican isolates of Aeromonas sp. in immersion experimentally-infected rainbow trout (Oncorhynchus mykiss, Walbaum 1792)

Ten species of Aeromonas have been previously identified in farmed rainbow trout from Mexico. The aim of the current study was to investigate the pathogenicity of 10 Aeromonas isolates belonging to 10 different Aeromonas species in immersion experimentally-infected rainbow trout fry. Isolates of A. bestiarum, A. hydrophila, A. salmonicida, and A. veronii produced significant mortality (8.8%, 12.2%, 18.8%, and 8.8%, respectively). Isolates of A. caviae and A. sobria produced no significantly mortality (3.3% and 1.1%, respectively). No mortality was recorded in fish infected with A. allosaccharophila, A. lusitana, A. media, or A. popoffii. Microscopic lesions and bacterial reisolation were registered in liver of fish infected with the ten different Aeromonas isolates. Our results suggest that all Aeromonas species included in the study have the ability to colonize the liver. The results have confirmed that species A. bestiarum, A. hydrophila, A. salmonicida, and A. veronii affected fish as elsewhere reported. In conclusion, the variation in pathogenicity of Aeromonas isolates included in the study, emphasizes the importance of active, on-going monitoring of Aeromonas in the Mexican rainbow trout farming.

In vivo adherence of Flavobacterium psychrophilum to mucosal external surfaces of rainbow trout (Oncorhynchus mykiss) fry.

The adherence of Flavobacterium psychrophilum to surfaces of epithelial tissues has been inconclusively suggested as a mechanism, which enables the bacterium to invade the host. Hence, the present study aimed to examine the adherence of the cells of two colony phenotypes, smooth and rough, of F.psychrophilum to mucosal tissues of rainbow trout fry and to test the skin mucus as a nutrient for the growth of F.psychrophilum. Fish were immersed in water containing 106 CFU mL-1 F.psychrophilum for each colony phenotype. Mucosal tissue samples from fins, gills, skin and eyes, and swab samples from spleen and kidney were taken and inoculated onto TYES agar plates. Colony phenotypes of F.psychrophilum were identified and number of colonies counted. The results showed that cells of both phenotypes initially (0h) adhered to all mucosal surfaces, but only the rough cells were still present on tissues 1h post-immersion. Both phenotypes showed a tissue tropism with the fin tissue being the most adhered. Furthermore, skin mucus promoted the growth of both colony phenotypes. We suggest that the growth of F.psychrophilum cells in skin mucus apparently facilitates the bacterial adherence to mucosal surfaces, and the subsequent invasion into the host.

Inhibition of an Aquatic Rhabdovirus Demonstrates Promise of a Broad-Spectrum Antiviral for Use in Aquaculture.

Viral diseases in aquaculture are challenging because there are few preventative measures and/or treatments. Broad-spectrum antivirals are highly sought after and studied because they target common components of viruses. In our studies, we used LJ001, a broad-spectrum antiviral compound that specifically inhibits enveloped viruses. We used the fish rhabdovirus infectious hematopoietic necrosis virus (IHNV) as a model to study aquatic enveloped virus diseases and their inhibition. We demonstrated inhibition of IHNV by LJ001 both in cell culture as well as in live fish. Additionally, we showed that LJ001 inhibited the transmission of IHNV from infected fish to healthy fish, which lays the groundwork for using LJ001 as a possible therapeutic for aquatic viruses. Our results also suggest that virus inactivated by LJ001 induces an immune response, showing potential for future preventative (e.g., vaccine) applications.

Cascading effect of exotic fish fry on plankton community in a tropical Andean high mountain lake: a mesocosm experiment

&lt;p&gt;Fishless Andean high mountain lakes may be vulnerable to fish invasion because they tend to be small, oligotrophic and contain low zooplankton diversity. During the first decades of the twentieth century, rainbow trout, &lt;em&gt;Oncorhynchus mikiss&lt;/em&gt;, was introduced in South America, and stocking of juvenile stages (fry) in lakes continues today. However, their effect on plankton in these lakes has been little studied. We performed a mesocosms experiment to assess effects of trout of different ontogenetic stages on zooplankton and phytoplankton in a tropical-Andean high mountain lake. The presence of trout fry resulted in declines in several large zooplankton taxa, increases in chlorophyll &lt;em&gt;a&lt;/em&gt; and dissolved inorganic nitrogen (DIN). Our results showed that small fry consume large copepods (&lt;em&gt;Colombodiaptomus brandorffi&lt;/em&gt;) at a faster rate than larger fry, and also consumed medium sized copepods (&lt;em&gt;Tropocyclops prasinus&lt;/em&gt;) that are not affected by the larger trout fry. Fish of both sizes consumed &lt;em&gt;Ceriodaphnia quadrangula&lt;/em&gt;, a midsize cladoceran. Fish predation had weak effect on the phytoplankton biomass, but we found a correlation between zooplankton biomass and phytoplankton richness, and significantly larger cell of the alga &lt;em&gt;Peridinium&lt;/em&gt; in the presence of fish. Our results indicate that trout introduction produces cascading ecological and phenotypic effects on the plankton communities of tropical mountain lakes, similar to those observed in temperate latitudes.&lt;/p&gt;

An-3 PUFA depletion applied to rainbow trout fry (Oncorhynchus mykiss) does not modulate its subsequent lipid bioconversion capacity

Nutritional strategies are currently developed to produce farmed fish rich in n-3 long-chain PUFA (LC-PUFA) whilst replacing fish oil by plant-derived oils in aquafeeds. The optimisation of such strategies requires a thorough understanding of fish lipid metabolism and its nutritional modulation. The present study evaluated the fatty acid bioconversion capacity of rainbow trout (Oncorhynchus mykiss) fry previously depleted in n-3 PUFA through a 60-d pre-experimental feeding period with a sunflower oil-based diet (SO) followed by a 36-d experimental period during which fish were fed either a linseed oil-based diet (LO) (this treatment being called SO/LO) or a fish oil-based diet (FO) (this treatment being called SO/FO). These treatments were compared with fish continuously fed on SO, LO or FO for 96 d. At the end of the 36-d experimental period, SO/LO and SO/FO fish recovered >80 % of the n-3 LC-PUFA reported for LO and FO fish, respectively. Fish fed on LO showed high apparent in vivo elongation and desaturation activities along the n-3 biosynthesis pathway. However, at the end of the experimental period, no impact of the fish n-3 PUFA depletion was observed on apparent in vivo elongation and desaturation activities of SO/LO fish as compared with LO fish. In contrast, the fish n-3 PUFA depletion negatively modulated the n-6 PUFA bioconversion capacity of fish in terms of reduced apparent in vivo elongation and desaturation activities. The effects were similar after 10 or 36 d of the experimental period, indicating the absence of short-term effects.