Are brown trout Salmo trutta fario and rainbow trout Oncorhynchus mykiss two of a kind? A comparative study of salmonids to temperature‐influenced Tetracapsuloides bryosalmonae infection

Proliferative kidney disease (PKD) of salmonids caused by Tetracapsuloides bryosalmonae causes high mortalities of wild brown trout (Salmo trutta fario) and farmed rainbow trout (Oncorhynchus mykiss) at elevated water temperatures. Here the aim was to compare the temperature‐dependent modulation of T. bryosalmonae in the two salmonid host species, which display different temperature optima. We used a novel experimental set‐up in which we exposed brown trout and rainbow trout to an identical quantified low concentration of T. bryosalmonae for a short time period (1 hr). We followed the development of the parasite in the fish hosts for 70 days. PKD prevalence and parasite kinetics were assessed using qPCR. Exposures were performed at temperatures (12°C and 15°C) that reflect an environmental scenario that may occur in the natural habitat of salmonids. T. bryosalmonae infection was confirmed earliest in brown trout kept at 15°C (day 7 post‐exposure) while, in all other groups, T. bryosalmonae was not confirmed until day 15 post‐exposure. Moreover, significantly greater infection prevalence and a faster increase of parasite intensity were observed in brown trout kept at 15°C than in all other groups. These results indicate that PKD is differentially modulated by water temperature in related host species.     

Distribution and prevalence of T. bryosalmonae in Austria: A first survey of trout from rivers with a shrinking population

The first evidence of proliferative kidney disease (PKD) in an Austrian river (the River Kamp) was documented in 2016, and no information on the PKD infection status of trout in other rivers was available. Since then, brown trout (Salmo trutta fario) and rainbow trout (Oncorhynchus mykiss) have been collected from rivers in Upper and Lower Austria for different diagnostic purposes. In this study, we summarize the recent findings of a first survey concerning the distribution of Tetracapsuloides bryosalmonae, the causative agent of proliferative kidney disease (PKD), from these samples. Between September 2015 and October 2017, a total of 280 brown trout and 39 rainbow trout were collected from 21 rivers in the provinces of Upper and Lower Austria. T. bryosalmonae was detected by PCR of kidney tissue in 17 of 21 sampled rivers and in 138 of 280 brown trout as well as in 11 of 39 rainbow trout. Pathological signs of PKD (e.g., hypertrophy of the kidney) were observed in 33 analysed brown trout and six rainbow trout samples. No correlations between fish infected by T. bryosalmonae and the parameters size and age class, condition factor, geological origin of the streams and distribution within the river course were found, while positively tested fish are significantly increased at sampling sites exceeding water temperatures of 15°C for median periods of 115 days. The prevalence within the affected streams or stream sections is highly variable, and in single rivers, infection rates of up to 90% are confirmed.     

A survey on Tetracapsuloides bryosalmonae infections in Slovene fresh waters

Slovenia has no history of health problems related to proliferative kidney disease (PKD) either in farmed or in wild fish. However, due to the past molecular evidence for the presence of Tetracapsuloides bryosalmonae DNA in tissues of some fish from open waters, a survey was conducted on wild salmonids that were primarily sampled for other purposes. In winter 2010–2011, specimens from a total of 244 rainbow trout, Oncorhynchus mykiss (Walbaum), and brown trout, Salmo trutta L., from 30 bodies of fresh water were examined for T. bryosalmonae using a PCR method. The sampled fish showed no clinical signs or gross pathological lesions characteristic of PKD. Nineteen (7.8%) fish from seven (23.3%) fresh waters were positive for T. bryosalmonae. The identity of PCR amplicons was confirmed by sequencing. With one exception, all the positive fish were found in waters from the regions where the average yearly temperatures and the environmental pollution are higher. This preliminary countrywide survey provided the first insight into the situation regarding T. bryosalmonae infection of wild salmonids in Slovenia.     

A survey of the distribution of the PKD‐parasite Tetracapsuloides bryosalmonae (Cnidaria: Myxozoa: Malacosporea) in salmonids in Norwegian rivers – additional information gleaned from formerly collected fish

The malacosporean Tetracapsuloides bryosalmonae was detected in kidneys from Atlantic salmon parr in 64 of 91 sampled Norwegian rivers. Using real‐time PCR, this parasite was found to be present in Atlantic salmon parr in rivers along the whole coast, from the northernmost and southernmost areas of the country. In addition, T. bryosalmonae was found in kidneys from brown trout parr in 17 of 19 sampled rivers in south‐east Norway, and in Arctic charr sampled in the River Risfjordelva, located at the northernmost edge of the European mainland. In conclusion, T. bryosalmonae has a widespread distribution in salmonids in Norwegian watercourses. Proliferative kidney disease (PKD) caused by T. bryosalmonae and PKD‐induced mortality has been observed in salmonids in several Norwegian rivers and it can be speculated that more PKD outbreaks will occur as a result of climate change.     

Patterns of Tetracapsuloides bryosalmonae infection of three salmonid species in large,deep Norwegian lakes

Proliferative kidney disease (PKD), caused by the myxozoan endoparasite Tetracapsuloides bryosalmonae, is of serious ecological and economical concern to wild and farmed salmonids. Wild salmonid populations have declined due to PKD, primarily in rivers, in Europe and North America. Deep lakes are also important habitats for salmonids, and this work aimed to investigate parasite presence in five deep Norwegian lakes. Kidney samples from three salmonid species from deep lakes were collected and tested using real-time PCR to detect PKD parasite presence. We present the first detection of T. bryosalmonae in European whitefish in Norway for the first time, as well as the first published documentation of the parasite in kidneys of Arctic charr, brown trout and whitefish in four lakes. The observed prevalence of the parasite was higher in populations of brown trout than of Arctic charr and whitefish. The parasite was detected in farmed, but not in wild, charr in one lake. This suggests a possible link with a depth of fish habitat and fewer T. bryosalmonae-infected and PKD-affected fish. Towards a warmer climate, cold hypolimnion in deep lakes may act as a refuge for wild salmonids, while cold deep water may be used to control PKD in farmed salmonids.     

Factors influencing the success of salmonid egg development in river substratum

Interstitial water conditions in the hyporheic zone of the stream bed are determinants of salmonid egg hatching success. We used standardised egg exposures to develop and validate discriminant analysis and generalised linear model models linking the hatching success of brown trout (Salmo trutta) with physicochemical factors of the interstitial zone (e.g., oxygen, specific conductance, nitrate, nitrite, ammonium, pH and redox potential). Interstitial water quality was identified as a limiting factor for egg development (median of relative hatching rates = 0.23). Hatching success was unimodal in hatchery and field references incubated in free‐flowing water, but bimodal (very high or very low hatching success) in natural sediment exposures. The effects of physicochemical factors on the hatching success of Salmo trutta strongly depended on both the time and spatial scale analysed. The variables retained in the models differed between the macroscale (over all rivers), the river‐specific scale (within a river) and the microscale (at different sediment depths). Egg hatching success decreased with increased substratum depth (decrease of 26% in 150 mm compared with 50 mm). Increasingly more variable interstitial water conditions (e.g., oxygen) throughout the egg incubation period suggest progressive degradation rates in the stream substratum during the incubation period at the microscale level. Consequently, consideration of different spatial and temporal scales is necessary for the evaluation of habitat quality in salmonid conservation and catchment management plans.     

Field study indicating susceptibility differences between salmonid species and their lineages to proliferative kidney disease

Tetracapsuloides bryosalmonae (Myxozoa: Malacosporea) is the causative agent of proliferative kidney disease (PKD), which affects both wild and farmed salmonid fish. The objective of this study was to outline differences in susceptibility to PKD in different salmonid species, hybrids and breeding lineages. Susceptibility to T. bryosalmonae infection was established based on cumulative mortality, pathological findings and detection of T. bryosalmonae in the kidney using immunohistochemistry and molecular methods. Determination of pure and hybrid individuals of different species in the genus Salvelinus, and dissimilarity of rainbow trout lineages, was performed using traditional polymerase chain reaction (PCR) and microsatellite analyses. Rainbow trout displayed higher disease severity compared with brook trout and Alsatian charr. Moreover, the results indicated differences in infection susceptibility, not only among different salmonid species but also among different lineages of charr and rainbow trout. Our study indicated that some salmonid species and even different lineages of the same species are more suitable for farming under PKD pressure.     

Failure of predator conditioning: an experimental study of predator avoidance in brown trout (Salmo trutta)

Many studies have documented that hatchery‐reared salmonids generally have inferior survival after being stocked compared with wild conspecifics, hatchery and wild salmonids have been observed to differ in their antipredator responses. The response of brown trout (Salmo trutta) juveniles (0+) of differing backgrounds to a live predator was compared in two experiments. First, the antipredator behaviour of predator‐naïve hatchery‐reared brown trout and wild‐exposed brown trout were assessed in behavioural trials which lasted for eight days. Second, predator‐naïve and predator‐conditioned hatchery‐reared brown trout were assessed in identical behavioural trials. Brown trout were ‘predator‐conditioned’ by being held in a stream‐water aquarium with adult Atlantic salmon (Salmo salar) and adult brown trout for two days prior to behavioural trials. Predator‐conditioned hatchery‐reared brown trout spent more time in shelters in the trial aquaria than predator‐naïve hatchery‐reared fish, but did not differ in time spent in the predator‐free area. Predator conditioning may account for the increased time spent in the shelter, but does not appear to have affected time spent in the predator‐free area. However, even if significant alteration in behaviour can be noted in the laboratory, the response might not be appropriate in the wild.     

Toxic effects of ammonia on the embryonic development of the cuttlefish Sepia pharaonis

The objective of this study was to investigate the effects of ammonia nitrogen on the embryonic development of the cuttlefish Sepia pharaonis . Embryos were exposed to different concentrations (0, 1, 3, and 5 mg/L) of total ammonia–nitrogen (TAN) during incubation. The developmental rate, malformations, mortality, hatching rate, incubation period, yolk utilization efficiency ratio, and weight of the newly hatched cuttlefish were determined. The results showed that ammonia nitrogen significantly inhibited the development of S. pharaonis embryos and induced malformations and even death. Hatching was delayed, the hatching rate was reduced, mortality and the incubation period increased, and the yolk utilization efficiency ratio and weight of the newly hatched cuttlefish significantly decreased in a dose‐dependent manner after the embryos were exposed to more than 1 mg/L TAN for prolonged period. These variables could be used as an integrative biomarker or indicator of aquatic environmental ammonia contamination. In summary, our results indicated that ammonia caused toxicity in the embryos. When the concentration of TAN is greater than 1 mg/L, ammonia levels should be reduced to prevent toxic effects on embryonic development.     

Purified deoxynivalenol or feed restriction reduces mortality in rainbow trout,Oncorhynchus mykiss (Walbaum), with experimental bacterial coldwater disease but biologically relevant concentrations of deoxynivalenol do not impair the growth of Flavobacterium psychrophilum

Diets containing deoxynivalenol (DON) were fed to rainbow trout Oncorhynchus mykiss (Walbaum) for 4 weeks followed by experimental infection (intraperitoneal) with Flavobacterium psychrophilum (4.1 × 10⁶ colony‐forming units [CFU] mL⁻¹). Mortality of rainbow trout fed either 6.4 mg kg⁻¹ DON or trout pair‐fed the control diet was significantly reduced (P < 0.05) in comparison with trout fed the control diet to apparent satiation (<0.1 mg kg⁻¹ DON). In a second experiment, trout were fed one of three experimental diets; a control diet, a diet produced with corn naturally contaminated with DON (3.3 mg kg⁻¹ DON) or a diet containing purified DON (3.8 mg kg⁻¹); however, these fish were not experimentally infected. The presence of DON resulted in significant reduction (P < 0.0001) in feed intake as well as weight gain after 4 weeks. Respiratory burst of head‐kidney leucocytes isolated from rainbow trout fed diets containing purified DON (3.8 mg kg⁻¹) was significantly higher (P < 0.05) at 35 day post‐exposure compared with controls. The antimicrobial activity of DON was examined by subjecting F. psychrophilum in vitro to serial dilutions of the chemical. Complete inhibition occurred at a concentration of 75 mg L⁻¹ DON, but no effect was observed below this concentration (0–30 mg L⁻¹).     

Alternative routes to piscivory: Contrasting growth trajectories in brown trout (Salmo trutta) ecotypes exhibiting contrasting life history strategies

Large and long‐lived piscivorous brown trout, Salmo trutta, colloquially known as ferox trout, have been described from a number of oligotrophic lakes in Britain and Ireland. The “ferox” life history strategy is associated with accelerated growth following an ontogenetic switch to piscivory and extended longevity (up to 23 years in the UK). Thus, ferox trout often reach much larger sizes and older ages than sympatric lacustrine invertebrate‐feeding trout. Conventional models suggest that S. trutta adopting this life history strategy grow slowly before a size threshold is reached, after which, this gape‐limited predator undergoes a diet switch to a highly nutritional prey source (fish) resulting in a measurable growth acceleration. This conventional model of ferox trout growth was tested by comparing growth trajectories and age structures of ferox trout and sympatric invertebrate‐feeding trout in multiple lake systems in Scotland. In two of the three lakes examined, fish displaying alternative life history strategies, but living in sympatry, exhibited distinctly different growth trajectories. In the third lake, a similar pattern of growth was observed between trophic groups. Piscivorous trout were significantly older than sympatric invertebrate‐feeding trout at all sites, but ultimate body size was greater in only two of three sites. This study demonstrates that there are multiple ontogenetic growth pathways to achieving piscivory in S. trutta and that the adoption of a piscivorous diet may be a factor contributing to the extension of lifespan.     

Developing bioelectrical impedance analysis methods for age‐0 brook trout

Year class strength of many fishes often is determined by survival through the first winter. Increased fat reserves improve survival and overall cohort success. Bioelectrical impedance analysis (BIA) methods are established for estimating proximate composition of adult brook trout, Salvelinus fontinalis (Mitchill), but none have been developed for early life stages. Small‐fish BIA would provide information about age‐0 percent dry weight, a proxy for fat reserves, allowing for better prediction of cohort success. The objective of this study was to develop BIA methods that provide reliable estimates of percent dry weight for age‐0 brook trout. BIA measurements were taken at seven anatomical locations from 48 to 115 mm fish. A model developed using BIA measures taken by subdermal needle electrodes precisely predicted percent dry weight (best model, RMSE = 1.03, R² = 0.86). Although lacking support, as determined by information theoretical analysis, BIA measured with non‐invasive external rod electrodes also precisely predicted percent dry weight (RMSE = 1.09, R² = 0.85). Models developed using two electrode locations performed better than models developed with only one location. For small brook trout, a dorsal to ventral pre‐dorsal fin electrode location should be used in conjunction with a dorsal total length location when measuring BIA to predict percent dry weight.     

Immunohistochemical and PCR studies of wild fish for Tetracapsula bryosalmonae (PKX), the causative organism of proliferative kidney disease

Tetracapsula bryosalmonae, previously referred to as PKX, causes proliferative kidney disease (PKD) in salmonids and is an economically important myxozoan pathogen in salmonid culture. A variety of molecular and immunological tools have been developed to detect the parasite. To determine the specificity of four monoclonal antibodies (MAbs) raised against T. bryosalmonae, archive material of fish infected with various myxosporean species was obtained and immunostained. Wild fish were also collected from enzootic waters and examined for T. bryosalmonae infection using immunohistochemistry and the polymerase chain reaction (PCR). Three of the MAb probes appear to be specific for T. bryosalmonae while only two of the five sets of primers tested appeared to specifically amplify T. bryosalmonae DNA. The results of the immunostaining and the PCR demonstrate that T. bryosalmonae occurs in the tubules of grayling Thymallus thymallus L., brown trout, Salmo trutta L. and Atlantic salmon, Salmo salar L. outside of the PKD season (June‐September) in the UK. This confirms the results of previous studies that these species are the preferred fish hosts for the parasite in the UK.     

A new potential therapeutic remedy against Aeromonas hydrophila infection in rainbow trout (Oncorhynchus mykiss) using tetra,Cotinus coggygria

Different antibiotic‐based drugs are being used for the treatment of Aeromonas hydrophila infection in rainbow trout, and several studies emphasize the use of medicinal plants as immunostimulants for prophylactic measure against Aeromoniasis disease. However, therapeutic effects of aqueous methanolic extracts of tetra (Cotinus coggygria) against A. hydrophila in rainbow trout were not investigated. Four different concentrations of tetra extract (0 [control], 4, 8 and 12 mg/100 µl) and also two different positive control groups (florfenicol and doxycycline antibiotics) were administered orally using feeding needles to individual rainbow trout, Oncorhynchus mykiss of all experimental groups twice a day after intramuscular inoculation of A. hydrophila. The study period was for 10 days. On 0th, 3rd, 7th and 10th day, blood and tissues were collected from the fish and changes in humoral immune responses, haematology and immune‐related gene expressions were determined. In the study, superoxide radical production was decreased generally in all experimental groups except in 12 mg tetra and florfenicol treatments compared to control (p < .05). Lysozyme activity was generally decreased (p < .05), or no differences were observed in all experimental groups compared to the control. Myeloperoxidase activity was significantly increased in florfenicol‐treated fish group on 7th day (p < .05). Generally, myeloperoxidase activity showed an increase in almost all tetra‐treated groups. Haematological parameters increased but were not significantly high enough in treatments. Almost all immune‐related gene expressions were significantly enhanced on 3rd and 10th day of the study. Survival rate of 53.33% was found in control group. There were no significant differences in survival between control and 4 mg tetra‐treated group (p > .05). All the other groups' survival rate was significantly increased compared to control. The highest survival rate was found in florfenicol group (80%). In 12 mg tetra‐, doxycycline‐ and 8 mg tetra‐treated groups, survival rate was recorded as 74.44%, 70% and 70%, respectively. Our results suggest that tetra methanolic extract is an effective therapeutic remedy against A. hydrophila infection in rainbow trout at the dose of 24 mg/32.34 g body weight/day.     

Distribution patterns of saprolegniosis cutaneous lesions in wild and farmed brown trout (Salmo trutta L.) obtained using a geographic information system (GIS)

A retrospective study was conducted using 250 clinical records of brown trout (Salmo trutta L.) with saprolegniosis by Saprolegnia parasitica, which had been collected from 8 rivers and 1 fish farm in the province of León (Spain). A geographic information system (GIS) was used to obtain skin lesion distribution patterns in males and females. Lesions in wild brown trout affected 15.31 ± 13.33% of the body surface, with a mean of 12.76 ± 6.56 lesions per fish. In addition, 51.23% of wild trout presented lesions with necrosis of the skin or fins. The pattern obtained when not distinguishing between sexes indicated that saprolegniosis lesions are mainly located above the lateral line and most frequently affect the dorsal cephalic region, the adipose fin, the peduncle and the caudal fin. However, differences were observed between males and females. Farmed trout presented a lower percentage of affected body surface (2.06 ± 4.36) and a lower number of lesions with and without necrosis because they received preventive treatment for saprolegniosis.     

Activities of digestive enzymes in relation to ingested natural food in three carp species of the Himalayan River Ladhiya,northern India

The present investigation was carried out to study gut content and accompanying digestive enzyme profiles of the three carp species, golden mahseer Tor putitora, snow trout Schizothorax richardsonii and hill trout Barilius barila. The fishes were captured from the River Ladhiya, Champawat district, Uttarakhand, northern India. The digestive tract length was on average 1.8 and 2.3 times longer than the body, while members of Bacillariophyceae, Chlorophyceae, Cyanophyceae and Zygnematophyceae made up 95.15% and 99.63% of gut content in the golden mahseer and snow trout, respectively. In hill trout, the digestive tract being only half the body length, gut content was insects (72.73%), eggs of aquatic organisms (22.73%) and nematodes (4.55%). Estimated gut enzyme activities demonstrated that amylase activity was significantly (P < 0.05) higher in golden mahseer and snow trout compared to hill trout, while total protease, trypsin and chymotrypsin activities were all significantly (P < 0.05) higher in the hill trout compared to other species. A strong co‐relation was found among relative gut length, food composition and digestive enzyme activities of each species. This may guide the future formulation of feed to these important species of carps kept in farm.     

Otolith size differences during early life of dwarf and cannibal Arctic char (Salvelinus alpinus)

In Arctic and alpine lakes, Arctic char [Salvelinus alpinus (L.)] often form two distinct morphs: invertebrate feeders (‘dwarfs’) and piscivores (‘giant or cannibals’). Here, we test for early life history growth variation in dimorphic Arctic char as a proximate explanation for the observed life history variation between the two forms. Char were sampled in 11 alpine and Arctic Norwegian lakes. Dwarfs (defined as sexually mature char less than 15 cm long; N = 304) had a mean total length of 105 mm, whereas the typical cannibal (body length above 20 cm; N = 153) was 388 mm long. A positive correlation between egg size and otolith hatching ring were ascertained in a separate hatching experiment with brown trout (Salmo trutta) and it is assumed that this relationship also is valid for Arctic char, and otolith size was used as a proxy for length. Otolith hatching ring from Arctic char cannibals was larger (mean ± SD; 187 ± 24 μm) than those from dwarfs (mean ± SD; 164 ± 23 μm). There were only minor size differences between dwarfs and cannibals during the next three years, after which dwarfs usually matured. Two mutually nonexclusive, proximate explanations for the differentiation into separate morphs (dwarfs and cannibals) are therefore maternal effects and/or genetic based differentiation. The high catchability of large piscivorous char and low production in alpine and Arctic lake ecosystems may make these stocks particularly vulnerable to overexploitation.     

Effect of combination of dietary Bacillus subtilis and trans‐cinnamic acid on innate immune responses and resistance of rainbow trout,Oncorhynchus mykiss to Yersinia ruckeri

The present study investigated the effects of combination of dietary Bacillus subtilis and trans‐cinnamic acid on serum biochemical parameters, innate immune responses and resistance of rainbow trout, Oncorhynchus mykiss to Yersinia ruckeri. Six experimental groups of fish with mean weights of 20.58 ± 0.35 g were used in the study. Five experimental groups of fish were fed diets containing Bacillus subtilis (10⁷ per gram) or a mix of the Bacillus subtilis (BS) and trans‐cinnamic acid (25 mg/kg‐25trcBS, 50 mg/kg‐50trcBS, 75 mg/kg‐75 trcBS, 150 mg/kg‐150 trcBS), whereas an additive‐free basal diet served as the control (Cont). In this study, an increase was observed in granulocyte percentage, respiratory burst activity, phagocytic activity, phagocytic index, myeloperoxidase activity and total antiprotease activity especially in fish fed with mix of the BS and trans‐cinnamic acid‐supplemented diets (p < .05). Moreover, at the end of the 20‐day challenge period the survival rates and antibody titre (p < .05), and relative per cent survival were higher in the BS group and all trcBS groups compared with control group. As a conclusion, the results in the present study show that feeding rainbow trout with diets containing a mix of B. subtilis and trans‐cinnamic acid over a 60‐day period might be sufficient for improving fish immune responses and disease resistance against Y. ruckeri.     

Effects of rearing density on growth,fatty acid profile and bioremediation ability of polychaete Nereis diversicolor in an integrated aquaculture system with rainbow trout (Oncorhynchus mykiss)

The objective of this study was to assess the effects of rearing density on the growth and fatty acid profile of Nereis diversicolor and on its capability to bioremediate wastewater in an integrated culture system with rainbow trout (Oncorhynchus mykiss). To this end, a batch of juvenile N. diversicolor (0.03 ± 0.01 mg) was assigned into four different densities (T₁ = 250, T₂ = 500, T₃ = 1,000, T₄ = 2,000, besides T₀ = with no worm) in three replicates. The worm groups were fed with solid waste that was supplied from tanks containing rainbow trout (107.17 ± 13.5 g; 1.39 ± 0.18 kg/m³). During the experiment (60 days), the water temperature was 17.71 ± 0.6°C. The results revealed that SR%, SGR% and WG% of N. diversicolor in T₁ were significantly higher than those of T₂, T₃ and T₄. Both FER rate and biomass gain in T₄ were significantly higher than those in the other groups. The highest removal rate of NO₂‐N (73.72%), NH₃‐N (65.70%), PO₄‐P (59.32%), BOD₅ (69.60%) and TSS (82.33%) were observed in T₄. The major fatty acids presents were palmitic acid, oleic acid, linoleic acid, stearic acid and alpha‐linolenic acid in all worm‐treated groups, with no difference observed in the concentration of these fatty acids among them. Taken together, these findings suggest that organic wastes from rainbow trout farms could be recycled to achieve a sustainable aquaculture goal, and demonstrate that a high percentage of fatty acids in fish feed is not absorbed by rainbow trout but is done by N. diversicolor.