Freshwater Trout Research Articles

Freshwater amphipods (Gammarus pulex/fossarum) and brown trout as bioindicators for PFC contamination with regard to the aquatic ecological status of a small stream

BackgroundPer- and polyfluorinated chemicals (PFC) have been in use for more than 60 years. As a result of their high thermal and chemical stability, they have found numerous applications in industrial processes. However, PFC also commonly show disadvantageous properties such as persistence and ubiquitous presence in the environment. The issue of PFC contamination of surface water is addressed in this publication. One aim of this study was to find a potential bioindicator for PFC contamination of small streams, and a second aim was to determine the aquatic ecological quality of such a stream. Standardized methods were used including structural quality mapping of a watercourse, the PERLODES method and electrofishing in four study sections of the stream. PFC contamination was determined in freshwater amphipods (Gammarus pulex/fossarum) and brown trout.ResultsThis study shows that PFC originating from water contamination can be detected both in amphipods and in internal organs of brown trout. The fingerprints in these two species differ considerably from one another. The highest concentrations of PFC were found in the liver and kidneys of brown trout. The methods used in this study also show that the four study sections of the small stream tested fail to achieve the “good” ecological status required by the Water Framework Directive. In particular, this is due to inadequate benthic invertebrates.ConclusionsEven though it is not possible to determine a causal relationship between the ecological status of the small stream and the detection of PFC in aquatic organisms, appropriate measures must be developed and applied to reduce the spreading of PFC in the environment. In addition to the brown trout, freshwater amphipods proved to be useful as a bioindicator for PFC contamination of streams. In the future it will be necessary to observe whether the number of species in the benthic invertebrates continue to decline.

Threats to at‐risk salmonids of the Canadian Rocky Mountain region

AbstractTrout and charr, members of the salmonid family, have high conservation value but are also susceptible to anthropogenic threats in part due to the specificity of their habitat requirements. Understanding historical and future threats facing these species is necessary to promote their recovery. Of freshwater trout and charr in the Canadian Rocky Mountain region, westslope cutthroat trout (Oncorhynchus clarkii lewisi), bull trout (Salvelinus confluentus; a charr species) and Athabasca rainbow trout (Oncorhynchus mykiss) are of conservation concern. And indeed, range contractions and declining populations are evident throughout much of their ranges. Range contraction was most evident in the southern Alberta designatable unit (DU) of westslope cutthroat trout. Diminished populations were also evident in the downstream watersheds of the Alberta bull trout range, and throughout the Athabasca rainbow trout range. We assessed historical and future threats to evaluate the relative importance of individual threats to each DU and compare their impact among species. Individual threats fall into the broad categories of angling, non‐native species and genes, habitat loss and alteration, and climate change. Severity of each threat varies by DU and reflects the interaction between species’ biology and the location of the DU. Severity of threats facing each DU has changed over time, reflecting extirpation of native populations, changes in management and industry best practices, expansion of non‐native species and progressing climate change. The overall threat impact for each DU indicates a high probability of substantial and continuing declines and calls for immediate action.

Life history and genetic characterisation of sea troutSalmo truttain the Adriatic Sea

AbstractThe brown troutSalmo truttais characterised by both anadromous (sea trout) and resident populations, naturally occurring in Atlantic and Ponto‐Caspian rivers. Sea trout are currently considered absent from rivers of the Mediterranean area, probably because of the non‐optimal chemical–physical characteristics of the Mediterranean Sea. However, the occasional bycatch of smoltifiedS. truttain the Adriatic Sea is well known among fishermen and the biological explanation of this phenomenon is still controversial. The aim of this study was to compare the genetic diversity of freshwater and marine brown trout to try to understand the factors underlying the presence of putative anadromous brown trout in the Adriatic Sea.In this study, we analysed the genetic diversity of: (1) wild brown trout collected from the Esino River (central Italy); (2) a domestic strain of brown trout used for stocking the study area; and (3) a sample of Adriatic sea trout collected near the outlet of the Esino River. Together with genetic analysis, we carried out scale analysis in order to track the freshwater/marine stages of the life cycle in the sea trout samples. The genetic characterisation was carried out by polymerase chain reaction–restriction fragment length polymorphism analysis of the mtDNA fragmentND‐5/6and the nuclear locusLDH‐C1*and by genotyping 15 microsatellite loci. The genetic polymorphism obtained was used to investigate intra‐ and inter‐population genetic diversity, rates of genetic introgression between wild and domestic samples and the origin of sea trout specimens by using assignment tests.Our genetic analyses demonstrated that the sea trout analysed in this study are from the domestic strain of Atlantic origin used in central Italy for stocking activities. The level of genetic introgression between native and domestic samples is high in the Esino River. The populations more resilient to introgressive hybridisation appeared to be those living in the portion of the river network dominated by carbonate rocks. Assignment tests (GeneClass) suggest the existence of a link between stocking efforts and the freshwater origin of the sea trout. In addition, data obtained from the analysis of scales, size measurement, and sex determination showed a pattern of smolt age, size, and sex ratio very similar to those observed in other anadromous populations.In conclusion, the present study highlighted that sea trout from the central Adriatic Sea originated from brown trout of Atlantic origin inhabiting the Esino River. Their seaward migratory behaviour could represent a consequence of an active migration instead of a passive displacement by water flow. Our results also showed that traditional stocking practices represent a negative activity for the conservation of the last Mediterranean nativeS. truttapopulations.

Immunoassays are not immune to errors: Examples from two studies of steroid output from freshwater trout farms

A “reproducibility crisis” is widespread across scientific disciplines, where results and conclusions of studies are not supported by subsequent investigation. Here we provide a steroid immunoassay example where human errors generated unreproducible results and conclusions. Our study was triggered by a scientific report citing abnormally high concentrations (means of 4–79 ng L−1) of three natural sex steroids [11-ketotestosterone (11-KT), testosterone (T) and oestradiol (E2)] in water samples collected from two UK rivers over 4 years (2002–6). Furthermore, the data suggested that trout farms were a major source because reported steroid concentrations were 1.3–6 times higher downstream than upstream. We hypothesised that the reported levels were erroneous due to substances co-extracted from the water causing matrix effects (i.e. “false positives”) during measurement by enzyme-linked immunoassay (EIA). Thus, in collaboration with three other groups (including the one that had conducted the 2002–6 study), we carried out field sampling and assaying to examine this hypothesis. Water samples were collected in 2010 from the same sites and prepared for assay using an analogous method [C18 solid phase extraction (SPE) followed by extract clean-up with aminopropyl SPE]. Additional quality control (“spiked” and “blank”) samples were processed. Water extracts were assayed for steroids using radioimmunoassay (RIA) as well as EIA. Although there were statistically significant differences between EIA and RIA (and laboratories), there was no indication of matrix effects in the EIAs. Both the EIAs and RIAs (uncorrected for recovery) measured all three natural steroids at <0.6 ng L−1 in all river water samples, indicating that the trout farms were not a significant source of natural steroids. The differences between the two studies were considerable: E2 and T concentrations were ca. 100-fold lower and 11-KT ca. 1000-fold lower than those reported in the 2002–6 study. In the absence of evidence for any marked changes in husbandry practice (e.g. stock, diet) or environmental conditions (e.g. water flow rate) between the study periods, we concluded that calculation errors were probably made in the first (2002–6) study associated with confusion between extract and water sample concentrations. The second (2010) study also had several identified examples of calculation error (use of an incorrect standard curve; extrapolation below the minimum standard; confusion of assay dilutions during result work-up; failure to correct for loss during extraction) and an example of sample contamination. Similar and further errors have been noted in other studies. It must be recognised that assays do not provide absolute measurements and are prone to a variety of errors, so published steroid levels should be viewed with caution until independently confirmed.

Antibiotic Resistance Genes in Freshwater Trout Farms in a Watershed in Chile.

Point sources such as wastewater treatment plants, terrestrial agriculture, and aquaculture may release antibiotic residues, antibiotic resistant bacteria, and antibiotic resistance genes (ARGs) into aquatic ecosystems. However, there is a lack of quantitative studies attributing environmental ARG abundance to specific sources. The goal of this study was to evaluate the role of freshwater trout farms in the release and dissemination of ARGs into the environment. Sediment samples upstream and downstream from five rainbow trout farms were collected over time in southern Chile. A microfluidic quantitative polymerase chain reaction approach was used to quantify an ARG array covering different mechanisms of resistance, and data were analyzed using principal component analysis (PCA) and linear mixed regression models. Surveys were also conducted to obtain information about management practices, including antibiotic use, at the farms. Florfenicol and oxytetracycline were used at these farms, although at different rates. A total of 93 samples were analyzed. In the PCA, , , , , (A), (B), (C), (W), and grouped together. A statistically significant increase in abundance of , , , and several genes was found downstream from the farms compared with upstream sites, and retention ponds had the highest ARG abundance at each site. Antibiotic resistance gene levels returned to baseline at an average distance of 132.7 m downstream from the farms. Although results from this study indicate an influence of trout farms on the presence of ARGs in the immediate environment, the extent of their contribution to ARG dissemination is unknown and deserves further investigation.

Comparison of electrochemically determined metallothionein concentrations in wild freshwater salmon fish and gammarids and their relation to total and cytosolic metal levels

Application of metallothionein (MT) as an early warning sign of metal exposure in aquatic organisms is common in biomonitoring, but there is a huge variability in MT concentrations among different studies. Present research aims to assess MT responses in freshwater fish brown trout (Salmo trutta Linnaeus, 1758) and gammarids (Gammarus balcanicus Schäferna, 1922 and Echinogammarus acarinatus Karaman, 1931) as indicators of metal exposure within the freshwater karst environment (Krka River, Croatia). Sampling was performed upstream (reference site) and downstream (anthropogenically impacted site) of the wastewater discharges in autumn and spring seasons. Brown trout intestine was applied as a bioindicator tissue due to its role in dietborne metal uptake while gammarids were chosen as fish food and potential metal uptake source. Moreover, there is a lack of data on intestinal MT levels, so our results on MT and metal/metalloid concentrations, measured as total and metabolically available cytosolic levels, represent the first data of this kind for the selected indicator species. The results indicated that the ecotoxicological impact of technological and municipal wastewaters on the biota of the karst Krka River was moderate, although higher metal levels at the affected site were evident in both, fish and gammarids. The modified Brdička reaction applied in this study was confirmed as reliable electrochemical technique for MT quantification in both vertebrates and invertebrates, and it indicated higher MT levels in gammarids (1.9–4.1 mg g−1 w.w) than in fish intestine (0.5–2.8 mg g−1 w.w). Due to the lack of the data on MT concentrations in S. trutta and gammarid species G. balcanicus and E. acarinatus, presented results can serve as a preliminary data to establish MT background levels in intestine of wild freshwater fish and gammarids. Obtained MT levels showed species-, tissue- and method-specific differences, so comparison between MT levels should always involve the same species, tissue and measurement method.

Hematological and serum biochemical parameters of five freshwater snow trout fish species from river Jhelum of Kashmir Himalaya, India

Fish blood is a pathophysiological indicator of the whole body function and is thus considered as an important tool in diagnosing the structural and functional status of fish. However, the blood parameters vary between species to species and it mainly depends upon the favorable environmental conditions where the species live. The purpose of the current study is to evaluate the hematological and serum biochemical indices of five Schizothorax species: Schizothorax labiatus, S. plagiostomus, S. esocinus, S. curvifrons and S. niger in order to establish the resemblances and variations between these Schizothorax species which are inhabiting in river Jhelum. The hematological profile including hemoglobin (Hb), total red blood cell (RBC) count, hematocrit (Hct), white blood cell (WBC) count, erythrocyte sedimentation rate (ESR), and erythrocyte indices: mean corpuscular hemoglobin concentration (MCHC), mean corpuscular hemoglobin (MCH), and mean corpuscular volume (MCV) was analyzed from each Schizothorax sp. Statistical analysis showed that there were significant differences (p < 0.05) in blood parameters among five Schizothorax spp. The results showed lowest values of hematological parameters in S. niger with respect to other species, while the highest values of hematological parameters were recorded in S. plagiostomus. Significant differences (p < 0.05) in serum biochemical levels of glucose, protein, cholesterol and urea were also noted in Schizothorax spp. The differences found in the hematological profile and serum biochemical composition in these fishes can be attributed to the individual feeding behavior, tolerance and environmentally adjustable capability of the fish. However, further study is required to correlate the present study with some other parameters such as the nutrient status of the river Jhelum where these fishes live.

Phylogeography and taxonomic status of trout and salmon from the Ponto-Caspian drainages, with inferences on European Brown Trout evolution and taxonomy.

Current taxonomy of western Eurasian trout leaves a number of questions open; it is not clear to what extent some species are distinct genetically and morphologically. The purpose of this paper was to explore phylogeography and species boundaries in freshwater and anadromous trout from the drainages of the Black and the Caspian Seas (Ponto‐Caspian). We studied morphology and mitochondrial phylogeny, combining samples from the western Caucasus within the potential range of five nominal species of trout that are thought to inhabit this region, and using the sequences available from GenBank. Our results suggest that the genetic diversity of trout in the Ponto‐Caspian region is best explained with the fragmentation of catchments. (1) All trout species from Ponto‐Caspian belong to the same mitochondrial clade, separated from the other trout since the Pleistocene; (2) the southeastern Black Sea area is the most likely place of diversification of this clade, which is closely related to the clades from Anatolia; (3) The species from the Black Sea and the Caspian Sea drainages are monophyletic; (4) except for the basal lineage of the Ponto‐Caspian clade, Salmo rizeensis, all the lineages produce anadromous forms; (5) genetic diversification within the Ponto‐Caspian clade is related to Pleistocene glacial waves; (6) the described morphological differences between the species are not fully diagnostic, and some earlier described differences depend on body size; the differences between freshwater and marine forms exceed those between the different lineages. We suggest a conservative taxonomic approach, using the names S. rizeensis and Salmo labrax for trout from the Black Sea basin and Salmo caspius and Salmo ciscaucasicus for the fish from the Caspian basin.

Can biosecurity and local network properties predict pathogen species richness in the salmonid industry?

Salmonid farming in Ireland is mostly organic, which implies limited disease treatment options. This highlights the importance of biosecurity for preventing the introduction and spread of infectious agents. Similarly, the effect of local network properties on infection spread processes has rarely been evaluated. In this paper, we characterized the biosecurity of salmonid farms in Ireland using a survey, and then developed a score for benchmarking the disease risk of salmonid farms. The usefulness and validity of this score, together with farm indegree (dichotomized as ≤ 1 or > 1), were assessed through generalized Poisson regression models, in which the modeled outcome was pathogen richness, defined here as the number of different diseases affecting a farm during a year. Seawater salmon (SW salmon) farms had the highest biosecurity scores with a median (interquartile range) of 82.3 (5.4), followed by freshwater salmon (FW salmon) with 75.2 (8.2), and freshwater trout (FW trout) farms with 74.8 (4.5). For FW salmon and trout farms, the top ranked model (in terms of leave-one-out information criteria, looic) was the null model (looic = 46.1). For SW salmon farms, the best ranking model was the full model with both predictors and their interaction (looic = 33.3). Farms with a higher biosecurity score were associated with lower pathogen richness, and farms with indegree > 1 (i.e. more than one fish supplier) were associated with increased pathogen richness. The effect of the interaction between these variables was also important, showing an antagonistic effect. This would indicate that biosecurity effectiveness is achieved through a broader perspective on the subject, which includes a minimization in the number of suppliers and hence in the possibilities for infection to enter a farm. The work presented here could be used to elaborate indicators of a farm’s disease risk based on its biosecurity score and indegree, to inform risk-based disease surveillance and control strategies for private and public stakeholders.

The response of North Atlantic diadromous fish to multiple stressors, including land use change: a multidecadal study

Reduction of freshwater habitat quality due to land use change can have major impacts on diadromous fish. Partitioning this impact from other potential drivers, such as changing marine conditions and climate, is hampered by a lack of long-term data sets. Here, four decades of data were used to assess the impact of land use change on Salmo salar and anadromous Salmo trutta in the Burrishoole catchment, Ireland, one of the few index sites for diadromous fish in the North Atlantic. Land use change was found to have no significant impact on the freshwater survival of either salmon or trout. However, climate impacted significantly on the survival of salmon and trout in fresh water, with poor survival in years with wetter, warmer winters, coinciding with positive North Atlantic Oscillation values. Additionally, cold springs were associated with higher survival in trout. The addition of hatchery fish into the salmon spawning cohort coincided with low freshwater survival. Our results highlight the necessity for a broad ecosystem approach in any conservation effort of these species.

Cardiac remodeling and increased central venous pressure underlie elevated stroke volume and cardiac output of seawater-acclimated rainbow trout.

Substantial increases in cardiac output (CO), stroke volume (SV), and gastrointestinal blood flow are essential for euryhaline rainbow trout (Oncorhyncus mykiss) osmoregulation in seawater. However, the underlying hemodynamic mechanisms responsible for these changes are unknown. By examining a range of circulatory and cardiac morphological variables of seawater- and freshwater-acclimated rainbow trout, the present study revealed a significantly higher central venous pressure (CVP) in seawater-acclimated trout (~0.09 vs. -0.02 kPa). This serves to increase cardiac end-diastolic volume in seawater and explains the elevations in SV (~0.41 vs. 0.27 ml/kg) and CO (~21.5 vs. 14.2 ml·min-1·kg-1) when compared with trout in freshwater. Furthermore, these hemodynamic modifications coincided with a significant increase in the proportion of compact myocardium, which may be necessary to compensate for the increased wall tension associated with a larger stroke volume. Following a temperature increase from 10 to 16.5°C, both acclimation groups exhibited similar increases in heart rate (Q10 of ~2), but SV tended to decrease in seawater-acclimated trout despite the fact that CVP was maintained in both groups. This resulted in CO of seawater- and freshwater-acclimated trout stabilizing at a similar level after warming (~26 ml·min-1·kg-1). The consistently higher CVP of seawater-acclimated trout suggests that factors other than compromised cardiac filling constrained the SV and CO of these individuals at high temperatures. The present study highlights, for the first time, the complex interacting effects of temperature and water salinity on cardiovascular responses in a euryhaline fish species.

Predictors of pesticide concentrations in freshwater trout – The role of life history

Concentrations of halogenated pesticides in freshwater fish can be affected by age, size, trophic position, and exposure history. Exposure history may vary for individual fish caught at a single location due to different life histories, e.g. they may have hatched in different tributaries before migrating to a specific lake. We evaluated correlations of pesticide concentrations in freshwater brown trout (Salmo trutta) from the Clutha River, New Zealand, with potential predictors including capture site, age, length, trophic level, and life history. Life history was determined from otolith (fish ear bone) strontium isotope signatures, which vary among tributaries in the region of our study. Variability in pesticide concentrations between individual fish was not well explained by capture site, age, length, or trophic level. However, hexachlorobenzene (HCB) and chlorpyrifos concentrations were distinct in lake-based trout with different life histories. Additionally, one of the riverine life histories was associated with relatively high concentrations of total endosulfans. Linear models that included all potential predictor variables were evaluated and the resulting best models for HCB, chlorpyrifos, and total endosulfans included life history. These findings show that in cases where otolith isotope signatures vary geographically, they can be used to help explain contaminant concentration variations in fish caught from a single location.

Who's your mama? Riverine hybridisation of threatened freshwater Trout Cod and Murray Cod.

Rates of hybridization and introgression are increasing dramatically worldwide because of translocations, restocking of organisms and habitat modifications; thus, determining whether hybridization is occuring after reintroducing extirpated congeneric species is commensurately important for conservation. Restocking programs are sometimes criticized because of the genetic consequences of hatchery-bred fish breeding with wild populations. These concerns are important to conservation restocking programs, including those from the Australian freshwater fish family, Percichthyidae. Two of the better known Australian Percichthyidae are the Murray Cod, Maccullochella peelii and Trout Cod, Maccullochella macquariensis which were formerly widespread over the Murray Darling Basin. In much of the Murrumbidgee River, Trout Cod and Murray Cod were sympatric until the late 1970s when Trout Cod were extirpated. Here we use genetic single nucleotide polymorphism (SNP) data together with mitochondrial sequences to examine hybridization and introgression between Murray Cod and Trout Cod in the upper Murrumbidgee River and consider implications for restocking programs. We have confirmed restocked riverine Trout Cod reproducing, but only as inter-specific matings, in the wild. We detected hybrid Trout Cod–Murray Cod in the Upper Murrumbidgee, recording the first hybrid larvae in the wild. Although hybrid larvae, juveniles and adults have been recorded in hatcheries and impoundments, and hybrid adults have been recorded in rivers previously, this is the first time fertile F1 have been recorded in a wild riverine population. The F1 backcrosses with Murray cod have also been found to be fertile. All backcrosses noted were with pure Murray Cod. Such introgression has not been recorded previously in these two species, and the imbalance in hybridization direction may have important implications for restocking programs.

Establishing a benchmarking for fish farming - Profitability, productivity and energy efficiency of German, Danish and Turkish rainbow trout grow-out systems

The promotion of Blue Growth in aquaculture requires an understanding of the economic drivers influencing the sector at farm level, but the collection of reliable and comparable data at this level is time-consuming and expensive. This study suggests an alternative strategy for qualitative sampling of freshwater trout farms in Germany, Denmark and Turkey, using a combination of existing data, group discussions and interviews with trout farmers, consultants and researchers. Nine ‘typical’ trout farming models are described, focusing on profitability, productivity and energy efficiency and allowing in-depth comparative economic analyses of different production systems at farm level, across regions. Our results show that the majority of the farms investigated have been profitable. Turkish farms benefit from competitive advantages due to low wages, low capital investment and favourable climate conditions. Large German farms profit from local market prices and advanced farm management. Danish farms using recirculating techniques remain competitive thanks to enhanced productivity and economy of scale. However, small traditional farms in Germany and Denmark may struggle to stay competitive in the long term. Organic farms in both countries face challenges of high feed costs and comparatively low productivity with mixed success. Using edible protein energy return on investment (epEROI) as an indicator of ecological sustainability, all surveyed farms compared very favourably with the terrestrial systems of animal meat production were investigated so far.

The liquorice root derivative glycyrrhetinic acid can ameliorate ionoregulatory disturbance in rainbow trout (Oncorhynchus mykiss) abruptly exposed to ion-poor water

To consider the idea that a dietary botanical supplement could act as an adaptogen in a teleost fish, the effect of a liquorice root derivative (18β-glycyrrhetinic acid, 18βGA) on rainbow trout following an acute ionoregulatory stressor was examined. Freshwater (FW) trout were fed a control or 18βGA supplemented diet (0, 5, or 50μg 18βGA/g diet) for 2weeks, then abruptly exposed to ion-poor water (IPW) for 24h. Following IPW exposure, muscle moisture content and serum cortisol levels elevated and serum [Na+] and/or [Cl−] reduced in control and 50μg/g 18βGA-fed fish. However, these endpoints were unaltered in 5μg/g 18βGA-fed fish. Gill tissue was investigated for potential mechanisms of 18βGA action by examining mRNA abundance of genes encoding corticosteroid receptors (CRs), 11β-hydroxysteroid dehydrogenase 2 (11β-hsd2), and tight junction (TJ) proteins, as well as Na+-K+-ATPase and H+-ATPase activity, and mitochondrion-rich cell (MRC) morphometrics. Following IPW exposure, CR and 11β-hsd2 mRNA, MRC fractional surface, Na+-K+-ATPase and H+-ATPase activity were unaltered or decreased in 50μg 18βGA fish, as was mRNA encoding select TJ proteins. In contrast, 5μg 18βGA-fed fish exhibited elevated 11β-hsd2 and CR mRNA abundance versus 50μg 18βGA-fed, and reduced MRC apical area as well as some differences in TJ protein mRNA abundance versus control fish. Data suggest that 18βGA, at low levels, may be adaptogenic in trout and might help to ameliorate ionoregulatory perturbation following IPW exposure. This seems to occur, in part, through 18βGA-induced alterations in the biochemistry and physiology of the gill.

Distribution of Infectious Pancreatic Necrosis Virus (IPNV) Based on Surveillance Programs in Freshwater Trout Farms of Mexico.

Diagnostic testing was performed between 2000 and 2012 to determine the distribution of infectious pancreatic necrosis virus (IPNV) in the main states of the Mexican Republic with freshwater Rainbow Trout Oncorhynchus mykiss (Walbaum) farms. This virus was positively identified from Rainbow Trout farms in seven of the eight states assessed. Due to nonnormal data distribution, a logistic regression model was applied for statistical analysis, the results of which indicated that virus prevalence was variable between states, with moderate but significant differences. Regarding the time periods evaluated, IPNV prevalence was higher during the first years of the study. The susceptible, infected, removed model was used to examine this phenomenon, which indicated that the decreased prevalence during the latter years of the study could be associated with a real elimination of the infection. The information of the cases analyzed also suggests a relationship with the irregularity in the submission of samples to the laboratory and emphasizes other factors that have contributed to the transmission of IPNV throughout the country. Received November 10, 2014; accepted December 5, 2015.

Restricted process water limits starch gelatinization, and reduces digestibility of starch, lipid, and energy in extruded rainbow trout (Oncorhynchus mykiss) diets

Abstract Five diets were produced with crude protein (CP) from hydrolysed wheat gluten (HWG) replacing graded (0, 12.5, 25, 50, 75%) amounts of CP from LT fish meal. Diets with 0–25% of CP from HWG were extruded with 19–20% water added in the extruder. Water addition was limited to 11% of the extrudate for the 50% HWG diet, and no water was added to the diet with 75% CP from HWG in order to obtain even flow of material in the extruder. Based on the degree of starch gelatinization (DG), estimated by differential scanning calorimetry (DSC), the diets with 19–20% process water were characterized by high gelatinization (HG), medium DG was obtained by limiting water addition to 11% (MG), and low gelatinization (LG) was the result of extruding without water added. Each diet was fed to two groups of rainbow trout in freshwater, and feces were obtained by abdominal stripping. Yttrium oxide (Y 2 O 3 ) was used as inert marker. The average apparent digestibility of starch ranged from 97 to 99% in the HG group, to 76% in the MG group, and 52% in the LG group. The AD of starch increased linearly (R 2 = 0.99) with increasing water addition in the extruder. The AD of lipid was reduced in MG and LG groups, and there was a linear (R 2 = 0.87) relationship between AD of starch and AD of lipid. As a consequence of AD of starch being quantitatively more influenced by water supplementation in the extruder than the AD of lipid, the regression of water supplementation on AD of energy was best explained (R 2 = 0.99) by a 2nd degree polynomial function, in which both the 1st and 2nd degree components were > 0. The findings emphasize the necessity of using sufficient process water during extrusion of salmonid diets both to ensure efficient digestion of starch and lipid. Statement of relevance This paper will increase the awareness in the feed industries of the necessity to secure complete gelatinization in extruded diets, since it is of high importance for digestible energy in the feed.

Evaluation of treatment methods using sodium percarbonate and formalin on Australian rainbow trout farms

In aquaculture, effective parasite management often relies on chemical therapy when prevention strategies are ineffective. Fish are often medicated using immersion (bath) treatments. The efficacy of bath treatment relies on maintaining at least the minimum effective dose for the necessary treatment duration. Dose is influenced by the product used, calculation of system volume, application method, flow, mixing, treatment degradation rate and environmental conditions. To maximize efficacy the relationships between system, delivery and effective dose need to be understood or controlled. We tested four application methods using sodium percarbonate (SPC) and two application methods using formalin (FOR, 37% formaldehyde [FA]) in four semi-closed flow-through systems on four Australian freshwater trout farms with different flow and water quality characteristics. Target dose was 64mg/L SPC and 200mg/L FOR. Hydrogen peroxide (HP) released from SPC was measured photometrically and FA levels were measured colorimetrically. Each application method achieved consistent doses across repeated applications but not all methods resulted in the dose reaching the target concentration in all parts of the system for the whole treatment duration. Eliminating the influence of system variables by creating static baths provides the most stable treatment environment. Where this is not possible, minimising system variables by modifying flow assists in retaining treatment in the system and improving accuracy of delivered doses. Treatment methods must be validated in a system prior to being routinely applied and mechanisms to optimise dose-duration identified and implemented.

Impacts of hypersaline acclimation on the acute toxicity of the organophosphate chlorpyrifos to salmonids

Acclimation to hypersaline conditions enhances the acute toxicity of certain thioether organophosphate and carbamate pesticides in some species of euryhaline fish. As the organophosphate chlorpyrifos is commonly detected in salmonid waterways, the impacts of hypersaline conditions on its toxicity were examined. In contrast to other previously examined pesticides, time to death by chlorpyrifos was more rapid in freshwater than in hypersaline water (16ppth). The median lethal time (LT50) after 100μg/L chlorpyrifos exposure was 49h (95% CI: 31–78) and 120h (95% CI: 89–162) for rainbow trout (Oncorhynchus mykiss) in freshwater and those acclimated to hypersaline conditions, respectively. Previous studies with hypersaline acclimated fish indicated induction of xenobiotic metabolizing enzymes that may detoxify chlorpyrifos. In the current study, chlorpyrifos metabolism was unaltered in liver and gill microsomes of freshwater and hypersaline acclimated fish. Acetylcholinesterase inhibition in brain and bioavailability of chlorpyrifos from the aqueous exposure media were also unchanged. In contrast, mRNA expression of neurological targets: calcium calmodulin dependent protein kinase II delta, chloride intracellular channel 4, and G protein alpha i1 were upregulated in saltwater acclimated fish, consistent with diminished neuronal signaling which may protect animals from cholinergic overload associated with acetylcholinesterase inhibition. These results indicate targets other than acetylcholinesterase may contribute to the altered toxicity of chlorpyrifos in salmonids under hypersaline conditions.

Measuring gill paracellular permeability with polyethelene glycol-4000 in freely swimming trout: proof of principle

The influence of swimming activity on gill paracellular permeability has not been measured previously in fishes. We critically assessed the use of tritium-labeled polyethylene glycol ([(3)H]PEG-4000) for this purpose, a substance that is also a classic marker for extracellular fluid volume, glomerular filtration rate and drinking rate. Tests (8 h) on resting freshwater trout showed that when measuring [(3)H]PEG-4000 clearance from the plasma in the efflux direction, correction for a large excretion via glomerular filtration was essential, necessitating urinary catheterization. When measuring [(3)H]PEG-4000 clearance from the water in the influx direction, correction for a significant uptake by drinking was essential, necessitating terminal gut removal, whereas glomerular filtration losses were minimal. After correction for these alternate routes of loss and uptake, [(3)H]PEG-4000 clearance rates by efflux from the plasma and by influx from the water were identical, showing that gill paracellular permeability is not rectified, and can be measured in either direction. The influx technique with terminal gut removal was used to assess gill paracellular permeability in trout without urinary catheters freely swimming at 1.2 body lengths s(-1) for 8 h. Branchial [(3)H]PEG-4000 clearance rate (by influx from the water) increased significantly by ~80% in accord with a similar measured increase in O2 consumption rate. Thus in trout, gill paracellular permeability does increase during exercise, in accord with the traditional concept of the osmorespiratory compromise.