Communications: Blood Chemistry of Laboratory-Reared Golden Trout

Abstract

Golden trout Oncorhynchus aguabonita obtained from a wild stock as fertilized eggs were reared in the laboratory for 21 months. The laboratory-reared golden trout in our study reached sexual maturity earlier and grew more rapidly than wild golden trout do (according to the scientific literature). Male fish averaged 35.6 cm in total length and 426 g in weight, and females averaged 36.2 cm and 487 g. All golden trout were sexually mature when used for hematological analysis. The hematological profile (hematocrit, red blood cells, white blood cells, and thrombocytes) of golden trout was similar to that reported elsewhere for other trout species. Male and female golden trout did not have significantly different thrombocyte counts; however, the immobilization treatment used on the fish (anesthesia versus a blow to the head) resulted in significant treatment differences in thrombocyte numbers and interaction effect of sex in treatment for hematocrits. Gravid female golden trout had significantly higher plasma protein and calcium levels than did males. The ionic compositions of plasma (sodium, potassium, calcium, magnesium, copper, zinc, iron, and chloride) and gallbladder bile (calcium and chloride) were similar to those reported for other salmonids.     

Comparison of the susceptibility of brown trout (Salmo trutta) and four rainbow trout (Oncorhynchus mykiss) strains to the myxozoan Tetracapsuloides bryosalmonae, the causative agent of proliferative kidney disease (PKD)

Differences in susceptibility to the myxozoan parasite Tetracapsuloides bryosalmonae, the causative agent of proliferative kidney disease (PKD), between four strains of rainbow trout (Oncorhynchus mykiss) and brown trout (Salmo trutta) were evaluated. Fish were exposed to water enzootic for the parasite in the field for 5 days and were subsequently transferred to the laboratory. Relative parasite load was determined after 2, 3 and 4 weeks post-exposure (wpe) by quantitative real-time PCR (qPCR) of kidney samples and number of parasite stages was determined in immunohistochemical stained sections of kidney, liver and spleen tissues. According to qPCR results, the highest amount of parasite DNA per equal amount of host tissue at all time points was measured in brown trout. Two of the rainbow trout strains showed lower relative parasite load than all other groups at the beginning of the experiment, but the parasite multiplied faster in these strains resulting in an equal level of relative parasite load for all rainbow trout strains at 4 wpe. A weak negative correlation of fish size and parasite load was detected. Only in samples of a few fish, single stages of T. bryosalmonae were found in sections stained by immunohistochemistry impeding quantitative evaluation of parasite numbers by this method. The results indicate a differential resistance to T. bryosalmonae between the rainbow trout strains investigated and between rainbow trout and brown trout.     

Movement patterns of Lichia amia (Teleostei: Carangidae): results from a long-term cooperative tagging project in South Africa

Understanding fish movement patterns and migrations is important for fisheries management as well as the identification and conservation of critical habitats. Data collected by the Oceanographic Research Institute's Cooperative Fish Tagging Project (ORI-CFTP) were used to describe the large-scale movement patterns of Lichia amia along the South African coastline. A total of 10 762 L. amia were tagged between 1984 and 2012, of which 744 (7%) were recaptured. Most tagging effort, mainly on juvenile and subadult individuals, took place along the south-eastern (i.e. Eastern Cape) and south-western (i.e. Western Cape) regions of the country during summer. Conversely, more L. amia were tagged and recaptured (mostly adults) during winter and spring (July to November) along the north-east coast (i.e. KwaZulu-Natal). Movements (dispersal distance in kilometres) by juveniles were restricted and largely confined to estuaries within the south-eastern and south-western regions of the country. There was a significant effect of fish size on distance moved (ANOVA: p<0.001), with evidence of a distinct seasonal (winter) longshore migration by adults to KwaZulu-Natal. The consequences of juvenile residency in estuarine habitats and high recapture rate of adults during a directed, seasonal spawning migration are discussed in light of improving current fisheries management.     

Population dynamics of three lizard species from the genus Sceloporus: short‐term changes in demographic parameters

Most demographic studies focus on numerical changes that occur within populations across years. However, typically studies at an interannual scale do not provide information on the particular times of the year (particular months or seasons) when rates of survival, recruitment, or migration increase or decrease due to physiological, behavioral or ecological processes. These monthly or seasonal changes in demographic parameters may lead to substantial variations in population abundance. In this study, we collected capture–mark–recapture data on 3 species of lizards of the genus Sceloporus (Sceloporus torquatus, Sceloporus grammicus and Sceloporus megalepidurus) found in ecologically similar habitats to examine potential changes in demographic rates among 3 different climatic seasons: rainy, cold‐dry and warm‐dry seasons. We tested different hypotheses about the effect of these seasons on survival, recruitment of new adults, and temporary emigration. We found that during the season with severe thermal constraints, the cold‐dry season, survival of S. torquatus decreased markedly. We also detected a considerable increase in the recruitment rate of S. grammicus during the rainy season, when these lizards are establishing territories and finding mates. In contrast, we found no evidence of intra‐annual changes in the rate of temporary emigration. In addition, we calculated abundance and population growth rates for each species and for each season. Our study represents a significant contribution to the understanding of intra‐annual demographic variation in lizards.     

Relation of Water Temperature to Bacterial Cold-Water Disease in Coho Salmon,Chinook Salmon,and Rainbow Trout

Abstract

The effect of water temperature on the progress of experimentally induced Cytophaga psychrophila infection was investigated in juveniles of coho salmon Oncorhynchus kisutch, chinook salmon O. tshawytscha, and rainbow trout O. mykiss (formerly Salmo gairdneri). A virulent strain of C. psychrophila was administered to fish by subcutaneous injection. Infected fish were held in tanks containing pathogen-free well water at temperatures ranging from 3 to 23°C. Mean times from infection to death of the fish were shortest at 12–15°C, which were the temperatures associated with the shortest time for doubling the population of this bacterium in vitro. Juvenile steelhead (anadromous rainbow trout) injected with viable C. psychrophila cells and held in 22°C water did not become diseased.     

Impact of Thiamine Deficiency on T-cell Dependent and T-cell Independent Antibody Production in Lake Trout

Abstract

Lake trout Salvelinus namaycush on thiamine-replete and thiamine-depleted diets were evaluated for the effects of thiamine status on in vivo responses to the T-dependent antigen trinitophenol (TNP)-keyhole limpet hemocyanin (TNP-KLH), the T-independent antigen trinitrophenol-lipolysaccaharide (TNP-LPS), or Dulbecco's phosphate-buffered saline (DPBS; negative control fish). Plasma antibody concentrations were evaluated for possible differences in total anti-TNP activity as well as differences in response kinetics. Associations between anti-TNP activity and muscle and liver thiamine concentrations as well as ratios of muscle-to-liver thiamine to anti-TNP activity were also examined. Thiamine-depleted lake trout that were injected with TNP-LPS exhibited significantly more anti-TNP activity than thiamine-replete fish. The depleted fish injected with TNP-LPS also exhibited significantly different response kinetics relative to thiamine-replete lake trout. No differences in activity or kinetics were observed between the thiamine-replete and -depleted fish injected with TNP-KLH or in the DPBS negative controls. Anti-TNP activity in thiamine-depleted lake trout injected with TNP-KLH was positively associated with muscle thiamine pyrophosphate (thiamine diphosphate; TPP) concentration. A negative association was observed between the ratio of muscle-to-liver TPP and T-independent responses. No significant associations between anti-TNP activity and tissue thiamine concentration were observed in the thiamine-replete fish. We demonstrated that thiamine deficiency leads to alterations in both T-dependent and T-independent immune responses in lake trout.

Received July 25, 2011; accepted July 12, 2012     

Group-level factors associated with chemotherapeutic treatment regimens in land-based trout farms in Ontario, Canada.

The lowest level at which fish farmers ordinarily make management decisions is the individual holding unit. To identify factors associated with chemotherapeutic treatment initiation at the holding-unit level, we created a unit of measurement called the "farm-tank-lot" (FTL), which allowed the movements and mixing of groups of fish to be followed during an entire production cycle. Each FTL was comprised of fish with a common history housed in a specific holding unit. Our 21-month prospective observational study (conducted on 14 land-based trout farms in Ontario, Canada) showed that the FTL was a biologically meaningful unit of concern and a feasible unit of measurement on land-based trout farms.Multivariable logistic and Poisson regressions revealed that fish size and growth rate both were associated negatively with the probability and frequency of treatment. FTLs that existed for longer periods of time were more likely to be treated and treated more often. There was a significant farm effect. Future field studies of disease in cultured fish should incorporate these factors into their design and analysis.     

Immune defense of wild‐caught Norway rats is characterized by increased levels of basal activity but reduced capability to respond to further immune stimulation

Studies of wild animals' immunity often use comparison with laboratory‐raised individuals. Using such an approach, various data were obtained concerning wild Norway rat's immunity. Lower or higher potential of immune system cells to respond to activation stimuli were shown, because of analysis of disparate parameters and/ or small number of analyzed individuals. Inconsistent differences between laboratory and wild rats were shown too, owing to great response variability in wild rats. We hypothesized that wild rats will express more intense immune activity compared to their laboratory counterparts which live in a less demanding environment. To test this, we analyzed the circulating levels of inflammatory cytokine interleukin‐6 (IL‐6), a mediator which has a central role in host immune defense. In addition, we examined the activity of the central immune organ, the spleen, including cell proliferation and production of pro‐inflammatory cytokines interferon‐γ (IFN‐γ) and interleukin‐17 (IL‐17), which are major effectors of cellular adaptive immune response. In order to obtain reasonable insight into the immunity of wild Norway rats, analysis was conducted on a much larger number of individuals compared to other studies. Higher levels of plasma IL‐6, higher spleen mass, cellularity and basal IFN‐γ production concomitantly with lower basal production of anti‐inflammatory cytokine interleukin‐10 (IL‐10) revealed more intense immune activity in the wild compared to laboratory rats. However, lower responsiveness of their spleen cells' proinflammatory cytokine production to concanavalin A (ConA) stimulation, along with preserved capacity of IL‐10 response, might be perceived as an indication of wild rats' reduced capability to cope with incoming environmental stimuli, but also as a means to limit tissue damage.     

Effect of ellagic acid on some haematological,immunological and antioxidant parameters of rainbow trout (Oncorhynchus mykiss)

In this study, effect of ellagic acid on some haematological, immunological and antioxidant parameters in the blood and various tissues of rainbow trout (Oncorhynchus mykiss) were examined. Four groups of rainbow trout were fed experimental diets containing either no ellagic acid (control) or supplemented with ellagic acid at 50 mg/kg diet (EA‐50), 100 mg/kg diet (EA‐100) or 150 mg/kg diet (EA‐150) for 21 days. Samples of the blood and tissue (liver, kidney and spleen) were collected at the end of the experiment and analysed for their haematological profile (the red blood cell count, the haemoglobin concentration and the haematocrit level), immune response (the white blood cell count, the oxidative radical production (NBT activity), the total plasma protein and total immunoglobulin level) and oxidant/antioxidant status (the malondialdehyde level, the superoxide dismutase, catalase and glutathione peroxidase activity as well as the reduced glutathione concentration). The findings of this study demonstrated that ellagic acid had a positive effect on the haematological parameters, the immune response and the antioxidant enzyme activities of the fish.     

The dose–response relation for the antinociceptive effect of morphine in a fish,rainbow trout

Jones, S. G., Kamunde, C., Lemke, K., Stevens, E. D. The dose–response relation for the antinociceptive effect of morphine in a fish, rainbow trout. J. vet. Pharmacol. Therap.  35 , 563–570. There have been suggestions that analgesics be used by fish researchers. But in the absence of dose–response data for morphine, this suggestion seems imprudent. The purpose of the present study was to develop a dose–response relationship in fish using six doses of morphine. The response (movement of the fins or tail) to a noxious stimulus (electrical shock to the face region) was monitored before and after a dose of morphine intraperitoneally (i.p.). The i.p. dose of morphine ED₅₀ in rainbow trout was 6.7 ± 0.8 mg/kg (n = 12 at each dose). The plasma morphine concentration EC₅₀ was 4.1 ± 1.5 mg/L. In a second experiment, rainbow trout tested with equal amounts of morphine and naloxone (30 mg/kg) showed that the antinociceptive effect of morphine was blocked by naloxone. It has been suggested that stress‐induced analgesia has been a confounding factor in some fish studies. However, plasma cortisol levels in our study indicated that stress was not a confounding factor in the present experiments. The ED₅₀ for morphine in fish was higher than that reported for humans or other mammals. Our observation showing a dose–response relation for morphine using a noxious stimulus supports arguments for its effectiveness as an antinociceptive drug in fish.     

Optimizing the size of the area surveyed for monitoring a Eurasian lynx (Lynx lynx) population in the Swiss Alps by means of photographic capture–recapture

We studied the influence of surveyed area size on density estimates by means of camera‐trapping in a low‐density felid population (1–2 individuals/100 km²). We applied non‐spatial capture–recapture (CR) and spatial CR (SCR) models for Eurasian lynx during winter 2005/2006 in the northwestern Swiss Alps by sampling an area divided into 5 nested plots ranging from 65 to 760 km². CR model density estimates (95% CI) for models M₀ and M_h decreased from 2.61 (1.55–3.68) and 3.6 (1.62–5.57) independent lynx/100 km², respectively, in the smallest to 1.20 (1.04–1.35) and 1.26 (0.89–1.63) independent lynx/100 km², respectively, in the largest area surveyed. SCR model density estimates also decreased with increasing sampling area but not significantly. High individual range overlaps in relatively small areas (the edge effect) is the most plausible reason for this positive bias in the CR models. Our results confirm that SCR models are much more robust to changes in trap array size than CR models, thus avoiding overestimation of density in smaller areas. However, when a study is concerned with monitoring population changes, large spatial efforts (area surveyed ≥760 km²) are required to obtain reliable and precise density estimates with these population densities and recapture rates.     

Presence–absence surveys of prey and their use in predicting leopard (Panthera pardus) densities: a case study from Armenia

It is important to predict how many individuals of a predator species can survive in a given area on the basis of prey sufficiency and to compare predictive estimates with actual numbers to understand whether or not key threats are related to prey availability. Rugged terrain and low detection probabilities do not allow for the use of traditional prey count techniques in mountain areas. We used presence–absence occupancy modeling and camera‐trapping to estimate the abundance and densities of prey species and regression analysis to predict leopard (Panthera pardus) densities from estimated prey biomass in the mountains of the Nuvadi area, Meghri Ridge, southern Armenia. The prey densities were 12.94 ± 2.18 individuals km^?2 for the bezoar goat (Capra aegagrus), 6.88 ± 1.56 for the wild boar (Sus scrofa) and 0.44 ± 0.20 for the roe deer (Capreolus capreolus). The detection probability of the prey was a strong function of the activity patterns, and was highest in diurnal bezoar goats (0.59 ± 0.09). Based on robust regression, the estimated total ungulate prey biomass (720.37 ± 142.72 kg km^?2) can support a leopard density of 7. 18 ± 3.06 individuals 100 km^?2. The actual leopard density is only 0.34 individuals 100 km^?2 (i.e. one subadult male recorded over the 296.9 km²), estimated from tracking and camera‐trapping. The most plausible explanation for this discrepancy between predicted and actual leopard density is that poaching and disturbance caused by livestock breeding, plant gathering, deforestation and human‐induced wild fires are affecting the leopard population in Armenia.     

The Effects of Whirling Disease on Growth and Survival of Snake River Cutthroat and Colorado River Rainbow Trout Fingerlings

Abstract

Two sizes of fingerling Snake River cutthroat trout Oncorhynchus clarkii behnkei and Colorado River rainbow trout O. mykiss were raised at hatcheries testing negative for Myxobolus cerebralis and stocked into the Dolores and Cache la Poudre rivers from 1999 to 2001. Populations were resampled over a 2-year period to determine which species and size combination had the highest growth and survival rates. Fish were tested for M. cerebralis via polymerase chain reaction and pepsin?trypsin digest analyses. Growth and survival rates between the species and size groups were not significantly different in either river. In the Dolores River, annual survival for both species and sizes of fish combined ranged from 0.063 to 0.12. In the Cache la Poudre River, survival for both sizes of rainbow trout was 0.004; survival for cutthroat trout ranged from 0.182 to 0.53. Larger fish had higher growth rates than smaller fish, and cutthroat trout had higher rates than similar sizes of rainbow trout. In both rivers, a higher percentage of the rainbow trout sample was infected than in the cutthroat trout sample. Rainbow trout also had a higher mean number of spores per head than cutthroat trout, and small rainbow trout had higher spore counts than large rainbow trout. Survival rates for cutthroat trout in the Cache la Poudre River were the highest of any of the groups, suggesting a difference that is biologically significant. Raising fingerlings to sizes greater than 100 mm can improve poststocking survival. If rainbow trout are stocked into contaminated waters, raising fingerlings to a larger size does not appear to improve growth or survival rates. Stocking rainbow trout in the spring could maximize growth rates but will expose fish to greater triactinomyxon densities, resulting in higher intensities of infection.     

Trait‐mediated seed predation,dispersal and survival among frugivore‐dispersed plants in a fragmented subtropical forest,Southwest China

By tracking the fate of individual seeds from 6 frugivore‐dispersed plants with contrasting seed traits in a fragmented subtropical forest in Southwest China, we explored how rodent seed predation and hoarding were influenced by seed traits such as seed size, seed coat hardness and seed profitability. Post‐dispersal seed fates varied significantly among the 6 seed species and 3 patterns were witnessed: large‐seeded species with a hard seed coat (i.e. Choerospoadias axillaries and Diospyros kaki var. silvestris) had more seeds removed, cached and then surviving at caches, and they also had fewer seeds predated but a higher proportion of seeds surviving at the source; medium‐sized species with higher profitability and thinner seed coat (i.e. Phoebe zhennan and Padus braohypoda) were first harvested and had the lowest probability of seeds surviving either at the source or at caches due to higher predation before or after removal; and small‐seeded species with lower profitability (i.e. Elaeocarpus japonicas and Cornus controversa) had the highest probability of seeds surviving at the source but the lowest probability of seeds surviving at caches due to lower predation at the source and lower hoarding at caches. Our study indicates that patterns of seed predation, dispersal and survival among frugivore‐dispersed plants are highly determined by seed traits such as seed size, seed defense and seed profitability due to selective predation and hoarding by seed‐eating rodents. Therefore, trait‐mediated seed predation, dispersal and survival via seed‐eating rodents can largely affect population and community dynamics of frugivore‐dispersed plants in fragmented forests.     

Developmental competence of eggs produced by rainbow trout Doubled Haploids (DHs) and generation of the clonal lines

Poor quality eggs produced by the fully homozygous doubled haploids (DHs) may impair generation of clonal lines in fish species. In the present research, gynogenetic development of rainbow trout (Oncorhynchus mykiss) was induced in eggs originated from the DH females. Eggs were activated with the UV‐irradiated grayling (Thymallus thymallus) spermatozoa and subjected to the high hydrostatic pressure (HHP) shock to provide diploid clonal individuals. Only two of four DH females produced eggs that were successfully activated by the irradiated spermatozoa and subsequently developed into the gynogenetic embryos. Survival rates of rainbow trout from the clonal lines equalled 21.5% and 19.8% during embryogenesis and decreased after hatching to 18.6% and 14.9%, respectively. Some of the dead rainbow trout clones collected between hatching and swim‐up stage were emaciated and exhibited spinal deformities including scoliosis. Provided results confirmed limited developmental competences of eggs produced by rainbow trout DH females. Clonal rainbow trout developing in such eggs exhibited reduced survival and increased frequency of the body abnormalities.     

Home range variability,spatial aggregation,and excursions of Akodon azarae and Oligoryzomys flavescens in Pampean agroecosystems

Rodents are reservoirs of various types of hantavirus, some of which are agents of hantavirus pulmonary syndrome in humans. Each hantavirus is associated with a single rodent host species but successive spill‐over events may eventually lead to host‐switching and new species’ becoming host of a given pathogen. This study aims to gain an understanding of the spatial ecology of two hantavirus‐host species, Akodon azarae, and Oligoryzomys flavescens, by identifying factors modulating their home range sizes and stability, and by evaluating intra‐ and interspecific spatial aggregation for these species and a third one—Oxymycterus rufus—living in sympatry. For this, eleven capture‐mark‐recapture surveys were carried out, spanning 22 months. We found that A. azarae males have larger and more mobile home ranges than females, independently of the season. Consequently, males could likely have a more relevant role in the transmission of hantavirus because of their greater exposure both to a higher number of contacts between individuals and viral contamination of the environment. Contrasting, O. flavescens individuals showed negligible displacements of their home range through time, which could limit the range of hantavirus spread in host populations. Since O. flavescens is host to Lechiguanas hantavirus (pathogenic to humans) this result encompasses epidemiological relevance, for it may imply the existence of local foci of infection. Additionally, individuals of both species performed excursions outside their home ranges. These events could enable hantavirus spread over distances beyond the normal range of movements and lead to new hantavirus outbreaks in formerly non‐infected rodent populations, favoring the persistence of the virus in nature.     

Flow Cytometric Analysis of Mitogen‐Induced Activation of Rainbow Trout (Oncorhynchus mykiss) Peripheral Blood Leucocytes

Proliferation of rainbow trout peripheral blood leucocytes in vitro is usually assessed by measuring incorporated tritiated thymidine. In this report we monitored the in vitro proliferative response to the mitogen Concanavalin A (Con A) by means of flow cytometry (FCM) and ³H‐thymidine incorporation. When analysed by FCM, blood leucocytes displayed two main cell populations with distinct forward and side scatter (FSC/SSC) characteristics: lymphocytes with low FSC/SSC values and non‐lymphoid leucocytes (NLL) with increased FSC/SSC values. The nature of these cell types were confirmed by microscopy. Interestingly, the FSC/SSC pattern of lymphocytes remained unchanged after in vitro stimulation with Con A, whereas cells from the NLL population showed a marked shift towards increased FSC values. In stimulated cultures, the increase of FSC values of the NLL population significantly correlated with contemporarily measured ³H‐thymidine incorporation (r=0.7, P < 0.001). The mitogenic response of blood leucocytes originating from different individual fish varied over wide ranges. It was found to be related to the numbers of NLL present in the leucocyte sample. The present results show that qualitative and quantitative FCM analysis of morphological parameters (FSC/SSC) of blood leucocytes makes it possible to discriminate between leucocyte populations of the rainbow trout and to monitor cell proliferation experiments.     

Influence of light colours on growth and stress response of rainbow trout (Oncorhynchus mykiss) under laboratory conditions

Summary The influence of light colours on growth and stress response in rainbow trout Oncorhyncus mykiss (15.16 ± 0.29 cm; 32.27 ± 1.18 g) was studied. Fish were reared in 16 glass aquaria (140 × 30 × 80 cm) each with 12 fish under one of four different lighting spectra: yellow (546 nm), red (605 nm), blue (470 nm) and white (full spectrum, control). Experiments lasted 125 days. The stress response was evaluated by measuring cortisol levels. Body weight and total length of the fish reared under yellow light were greater compared with the other colour regimes while feed conversion ratio significantly lowers. Condition factor and specific growth rate, however, were not differentiated among experimental light treatments. Stressed fish showed lower cortisol levels under yellow light compared with other light exposures. The study indicates that under laboratory conditions, rainbow trout grow best under yellow light and that yellow light lowers the stress‐induced cortisol response in this fish species.     

Rearing unit-level factors associated with bacterial gill disease treatment in two Ontario, Canada government salmonid hatcheries

Early-rearing salmonids in Ontario Ministry of Natural Resources (OMNR) fish hatcheries have been consistently affected by bacterial gill disease (BGD) (causative agent: Flavobacterium branchiophilum) for many years. Separate retrospective epidemiological investigations of BGD treatments at two OMNR fish hatcheries (Hatcheries A and B) for the 1999 production year were conducted using on-site hatchery records. Both investigations were carried out at the rearing unit-level, with early-rearing (<9 months of age) “tank-lot” as the unit of analysis to identify unique fish populations over time. Multivariable repeated measures logistic regression models were created for both hatchery datasets, controlling for lot-level and species effects. For Hatchery A, the species brook trout (Salvelinus fontinalis) and brown trout (Salmo trutta) were significantly associated with BGD treatment, as well as lower water exchange rate, and higher feeding and mortality percentages during the 2 weeks previous to BGD treatment. At Hatchery B, the species brook trout (S. fontinalis) and splake (Salvelinus namaycush × S. fontinalis) were significantly associated with BGD treatment, as well as lower individual fish weights and treatment for BGD during the previous week. These results emphasize the importance of water quality, feeding rate, fish size and prior mortality on the development of BGD. Significant hatchery and species effects were evident, and future observational research on BGD must account for these factors in their design and analysis.