Male Trout Research Articles

Current summer heat waves impair rainbow trout (Oncorhynchus mykiss) spermatogenesis: Implications for future fish farming management practices in South Europe

Freshwater environments provide different ecosystem services for human well-being, and are one of the most important natural sources of healthy animal protein. Climate change and extreme temperature events, among other factors, represent a growing concern for their sustainable use. In contrast to the research effort to elucidate how global warming affects these aquatic environments, few studies pay attention to the potential effects of heat waves, a current phenomenon with direct effects on contemporaneous organisms. Here, the potential effects of a single heat wave event on rainbow trout (Oncorhynchus mykiss) males, mimicking a recorded environmental event, have been evaluated by means of biometric, redox and stress response, plasma testosterone, histopathology and transcriptomic analyses. Environmental monitoring showed how, during the last 4 years, mean water temperature increased (2–4 °C; from April to October), heat wave duration progressively increased (from 15 to 100 days), and mean intensities ranged 1.7–3.8 °C with a maximum of 8.0 °C. An experimentally recreated heat wave lasting only 16 days, with mean and maximum intensities of 2.7 and 6.7 °C, induced a decrease in body weight and total antioxidant status, but increased blood plasma cortisol levels and redox enzyme activity. The induced heat wave decreased testosterone plasma levels and sperm quality, arrested spermatocyte type I and II differentiation and increased spermatozoa apoptosis. These effects were correlated with a differential expression of 1156 genes (116 up- and 1040 down-regulated genes) in exposed fish when compared to Control fish, reflecting an alteration in cell apoptosis and spermatozoa maturation processes. These results not only increase concern about current heat wave events, recreating a real scenario, but also unveil for the first time the particular molecular mechanisms of subsequent spermatogenesis disruption, opening new avenues for designing urgent and necessary strategies to mitigate the effects of extreme climate change conditions on freshwater aquaculture.

Эффективность использования солевого раствора при искусственном осеменении икры радужной форели Oncorhynchus mykiss (Waibaum, 1972) и ладожской палии Salvelinus alpinus lepechini (Gmelin, 1788)

In order to increase the level of fertilization of eggs, the effect of buffer solution on sperm activity and the effectiveness of its use as a medium for insemination eggs of rainbow trout which was grown in conditions of flowing water and in recirculation aquaculture system (RAS), as well as Ladoga palia, were tested. In male trout and palia, the number of motile spermatozoa and their activity were higher in solution than in water. In trout which was grown in running water, the percentage of eggs fertilization increased significantly when use solution only in cases when the eggs were contaminated with the contents of burst eggs. In fish grown in the RAS, the replacement of water with a buffer solution increased the level of fertilization in all test variants, regardless of the presence or absence of contamination of the eggs. In Ladoga palia, the result of insemination was the same in water and in solution.

In sea trout, the physiological response to salmon louse is stronger in female than in males.

The aims of this study were to compare male and female sea trout (Salmo trutta) with respect to their hypo-osmoregulatory ability over a simulated migration to seawater and their clinical and physiological response to salmon louse (Lepeophtheirus salmonis) infection in seawater and over a simulated pre-mature return to freshwater. For this purpose, 2-year-old hatchery-reared male and female brown trout (F1 offspring of wild caught anadromous fish) were infected with salmon lice and measured for changes in plasma ions, glucose, lactate and osmolality and relative heart, liver and gonad sizes during a simulated seawater migration and thereafter a premature return to freshwater after 4weeks in seawater (pre-adult louse). Un-infected trout served as control. Male trout used longer time to develop full hypo-osmoregulatory ability in seawater and showed a stronger response in plasma glucose and lactate following simulated premature return to freshwater, compared to female trout. Response to salmon louse was stronger in female trout, shown by stronger osmotic stress by chalimus (plasma Cl-) and pre-adult louse (plasma osmolality) and elevated relative liver size (hepatosomatic index) by pre-adult louse in female compared to male trout. Moreover, high plasma cortisol in infected female and low plasma cortisol in infected male trout produced a significant treatment-sex interaction on plasma cortisol. Lice infection intensity was initially higher in male (0.18 lice g-1) compared to female trout (0.11 lice g-1) at the chalimus stage, but equal between sexes at the pre-adult stage (male 0.15 and female 0.17 lice g-1). This study showed that female trout were better adapted for changes in water salinity, while male trout were more robust against salmon louse infection. These results suggests that the elevated salmon louse infection pressure generated by salmon farming have strong and unexplored negative effects on wild sea trout populations. Further research on this topic is vital for the conservation of wild sea trout populations.

Дослідження та розробка методів отримання міжродових гібридів лососевих (Salmonidae (Jarocki or Schinz, 1822)) риб з метою досягнення ефекту гетерозису та підвищення їх рибопродуктивності

Purpose. To study the possibility of obtaining highly productive intergeneric salmonid hybrids between rainbow trout and brook trout; brown trout and brook trout; rainbow trout and brown trout, as well as to develop methodological approaches and determine of optimal variants of hybrid crosses. Findings. In order to obtain intergeneric hybrids, we used six variants of hybrid crosses with brood fish of three salmonids belonging to three families (Salmo, Oncorhynhus, Salvelinus). The study used age-4 female rainbow trout with average body weight of 3296.8 g, Fork length was 62.6 cm, and the average working fecundity was 7420 eggs. Age-3 rainbow trout males had an average body weight of 1613 g and an average body length of 49.8 cm; age-3 brown trout females had an average body weight of 453.8 g and average working fecundity of 1540 eggs, and males had an average weight of 458.7 g; age-3 brook trout females had an average weight of 809.7 g and a length of 38.9 cm with working fecundity of 1732 eggs, and age-4 males had an average weight of 1212.8 g and an average body length of 46.0 cm. Twelve variants of fertilization were used: six variants at normal water temperature and six variants after a temperature shock. Under natural conditions, the creation of intergeneric hybrids is almost impossible, except for variants between brown trout and brook trout, which is due to the similarity of their biology. However, the efficiency of this cross is low and economically impractical for fish farmers. When applying the temperature shock during fertilization, hybrids proved to be the most effective, where females were rainbow trout, and males were brook trout and brown trout. The average weight of young-of-the-year intergeneric hybrids was, depending on the species of fish, from 8 to 54 g. The highest results were obtained for the creation of hybrids where following broodstock was used: ♂brook trout Х ♀brown trout; ♂brown trout Х ♀rainbow trout. In these variants of crossbreeding, the survival rate of young-of-the-year during the period of cultivation was 94.8 and 92.8%, respectively. In particular, the above hybrids did not suffer from infectious diseases during the growing period. Originality. New data on the development of methods for obtaining viable offspring of newly created hybrids were obtained, and the optimal variants of crossing between females and males of these salmonids were determined. Practical value. The results can be used for artificial breeding of salmonids in specialized farms that will allow obtaining high quality products and reducing their costs. Key words:rainbow trout, brown trout, brook trout, incubation, free embryos, larvae, fry, young-of-the-year.

Mitogen-activated protein kinases contribute to temperature-induced cardiac remodelling in rainbow trout (Oncorhynchus mykiss)

Rainbow trout (Oncorhynchus mykiss) live in environments where water temperatures range between 4°C and 20°C. Laboratory studies demonstrate that cold and warm acclimations of male trout can have oppositional effects on cardiac hypertrophy and the collagen content of the heart. The cellular mechanisms behind temperature-induced cardiac remodelling are unclear, as is why this response differs between male and female fish. Studies with cultured trout cardiac fibroblasts suggests that collagen deposition is regulated, at least in part, by mitogen-activated protein kinase (MAPK) cell signalling pathways. We, therefore, hypothesized that temperature-dependent cardiac remodelling is regulated by these signalling pathways. To test this, male and female trout were acclimated to 18°C (warm) in the summer and to 4°C (cold) in the winter and the activation of MAPK pathways in the hearts were characterized and compared to that of control fish maintained at 12°C. In addition, cardiac collagen content, cardiac morphology and the expression of gene transcripts for matrix metalloproteinases (MMP) -9, MMP-2, tissue inhibitor of matrix metalloproteinases and collagen 1α were characterized. p38 MAPK phosphorylation increased in the hearts of female fish with cold acclimation and the phosphorylation of extracellular signal-regulated kinase increased in the hearts of male fish with warm acclimation. However, there was no effect of thermal acclimation on cardiac morphology or collagen content in either male or female fish. These results indicate that thermal acclimation has transient and sex-specific effects on the phosphorylation of MAPKs but also how variable the response of the trout heart is to thermal acclimation.

Exposure to carbamate fungicide iodocarb does not affect reproductive behavior or milt volumes in precocious male brown trout (Salmo trutta L.) parr

Previous studies with olfactory-disturbing pesticides resulted after exposure in disturbed behavior and physiology in fish. In the present experiment, reproductive behavior and milt volumes of precocious brown trout (Salmo trutta L.) male parr were studied in a large stream aquarium after exposure to the olfactory-disturbing fungicide 15 μg l−1 IPBC (iodocarb; 3-iodo-2-propynyl butyl carbamate) for 96 h. The statistical analyses did not reveal any significant differences for time attending females between controls and IPBC-exposed males. Furthermore, there were no significant differences in milt volumes. However, when taking all fish into consideration, there were significant differences in milt volumes between parr that had been attending females and those had not been attending females. Controls that had attended females had significantly higher milt volumes than controls or IPBC-exposed males that had not attended females. Taking all control and IPBC parr into consideration, there was a statistically significant positive correlation between time attended females and volume of milt and gonadosomatic index (GSI), respectively. In summary, 15 μg l−1 IPBC did not have any significant effects on mature male parr reproductive behavior and milt volumes.

Environmental influences on life history strategies in partially anadromous brown trout (Salmo trutta, Salmonidae)

AbstractThis paper reviews the life history of brown trout and factors influencing decisions to migrate. Decisions that maximize fitness appear dependent on size at age. In partly anadromous populations, individuals that attain maturity at the parr stage typically become freshwater resident. For individual fish, the life history is not genetically fixed and can be modified by the previous growth history and energetic state in early life. This phenotypic plasticity may be influenced by epigenetic modifications of the genome. Thus, factors influencing survival and growth determine life‐history decisions. These are intra‐ and interspecific competition, feeding and shelter opportunities in freshwater and salt water, temperature in alternative habitats and flow conditions in running water. Male trout exhibit alternative mating strategies and can spawn as a subordinate sneaker or a dominant competitor. Females do not exhibit alternative mating behaviour. The relationship between growth, size and reproductive success differs between sexes in that females exhibit a higher tendency to migrate than males. Southern populations are sensitive to global warming. In addition, fisheries, aquaculture with increased spreading of salmon lice,introduction of new species, weirs and river regulation, poor water quality and coastal developments all threaten trout populations. The paper summarizes life‐history data from six populations across Europe and ends by presenting new research questions and directions for future research.

A genetic system for biasing the sex ratio in mice.

Biasing the sex ratio of populations of different organisms, including plants, insects, crustacean, and fish, has been demonstrated by genetic and non-genetic approaches. However, biasing the sex ratio of mammalian populations has not been demonstrated genetically. Here, we provide a first proof of concept for such a genetic system in mammals by crossing two genetically engineered mouse lines. The maternal line encodes a functional Cas9 protein on an autosomal chromosome, whereas the paternal line encodes guide RNAs on the Y chromosome targeting vital mouse genes. After fertilization, the presence of both the Y-encoded guide RNAs from the paternal sperm and the Cas9 protein from the maternal egg targets the vital genes in males. We show that these genes are specifically targeted in males and that this breeding consequently self-destructs solely males. Our results pave the way for a genetic system that allows biased sex production of livestock.

Fishery-exteriors characteristics of the brooders of brown trout, rainbow trout and brook trout

This study aims to determine the reproductive indices of three types of salmon fish: brown trout (Salmo trutta morfa fario Linnaeus (1758), rainbow trout (Oncorhynchus mykiss Walbaum, 1792) and brook trout (Salvelinus fontinalis Mitchill, 1814) in aquaculture. For this purpose, 10 females and 10 males of each species were selected on an analogous basis and the main fishery-exteriors and reproductive characteristics were identified. The researching of relative fertility permit to establish that this indicator in brook trout females was 18.36% higher compared to rainbow trout, however, by 7.5% lower compared to the brown trout. It was found that brook trout eggs had less weight and size in comparison with other studied species. In particular, the weight and diameter of the eggs were 2.89 and 9.7% respectively lower than of rainbow trout, as well as 8.1 and 3.44% respectively lower compared to the brown trout. When studying the quantity of ejaculate, it was found that on average, brook trout males had 16.95% less of ejaculates compared to rainbow trout males, but 79.6% more than of brown trout males. When comparing the main indices of the brooders body of the studied species, it was found that the condition factor of the brook trout and rainbow trout females was quite high at 1.55–1.57, the brown trout one was 1.35, and the profile index respectively was 3.5, 3.52 and 4.0. The males' condition factor of brook trout, rainbow trout and brown trout was respectively 1.64, 1.52 and 1.35. Fishery-exteriors indicators met the requirements of brooders species and age specificity norms of the studied fish species.

Levels of Thyroid and Sex-Steroid Hormones in Brown Trout Salmo trutta L. in the Rivers of Karelia

The correlation between the levels of thyroxine and sex-steroid hormones in the blood of brown trout and its life cycle in different rivers and age has been analyzed. It has been revealed that the concentrations of the studied hormones in the blood of juvenile specimens and mature male brown trout from the Alats-oya River (Lake Ladoga basin) are significantly lower than in the blood of fish from the Orzega and Shoksha rivers (Lake Onega basin). It is shown that the concentrations of thyroid hormones in the blood of brown trout change insignificantly with age. On the contrary, the level of sex-steroid hormones increases significantly by the beginning of sexual maturity.

Pharmaceuticals, hormones, pesticides, and other bioactive contaminants in water, sediment, and tissue from Rocky Mountain National Park, 2012–2013

Pharmaceuticals, hormones, pesticides, and other bioactive contaminants (BCs) are commonly detected in surface water and bed sediment in urban and suburban areas, but these contaminants are understudied in remote locations. In Rocky Mountain National Park (RMNP), Colorado, USA, BCs may threaten the reproductive success and survival of native aquatic species, benthic communities, and pelagic food webs. In 2012–2013, 67 water, 57 sediment, 63 fish, 10 frog, and 12 quality-control samples (8 water and 4 sediment) were collected from 20 sites in RMNP. Samples were analyzed for 369 parameters including 149 pharmaceuticals, 22 hormones, 137 pesticides, and 61 other chemicals or conditions to provide a representative assessment of BC occurrence within RMNP. Results indicate that BCs were detected in water and/or sediment from both remote and more accessible locations in RMNP. The most commonly detected BCs in water were caffeine, camphor, para-cresol, and DEET; and the most commonly detected BCs in sediment were indole, 3-methyl-1H-indole, para-cresol, and 2,6-dimethyl-naphthalene. Some detected contaminants, including carbaryl, caffeine, and oxycodone, are clearly attributable to direct local human input, whereas others may be transported into the park atmospherically (e.g., atrazine) or have local natural sources (e.g., para-cresol). One or more pharmaceuticals were detected in at least 1 sample from 15 of 20 sites. Most of the 29 detected pharmaceuticals are excreted primarily in human urine, not feces. Elevated net estrogenicity was observed in 18% of water samples, and elevated vitellogenin in blood was observed in 12% of male trout, both evidence of potential endocrine disruption. Hormone concentrations in sediment tended to be greater than concentrations in water. Most BCs were observed at concentrations below those not expected to pose adverse effects to aquatic life. Results indicate that even in remote locations aquatic wildlife can be exposed to pharmaceuticals, hormones, pesticides, and other bioactive contaminants.

146 Effect of pH on Rainbow trout (Oncorhynchus mykiss) Sperm Motility Using Five Extender Solutions

Rainbow trout (Oncorhynchus mykiss) sperm extender protocols can differ considerably with regards to composition of the extenders and handling. The objective of this study was to determine the effect of pH for 5 extenders on rainbow trout sperm motility and activation during storage at 5°C. Two-year-old sexually mature rainbow trout males (n = 46) weighing 1.5 to 2.5 kg were caught at Aquaculture Farm Tatakay in Jilotzingo, Estado de México, México. One semen sample per male was collected by abdominal massage. For the experiments, 100 mL of each extender (306, 512, Mounibs, Erdahl and Graham, and Hanks’ Balanced Salt Solution) was made, with pH adjusted to 6 levels (7.0, 7.2, 7.4, 7.8, 8.0, and 8.2). To evaluate pH dynamics in each extender, pH was tested daily over an 8-day storage interval. Trout sperm was added to each of the extenders in a ratio of 1:1 and motility recorded. Afterwards, each activator solution [DIA 532, saline solution (0.85%) and pond water] was added separately to a sample of the extended sperm, to initiate sperm activation. Motility was evaluated subjectively at 400× and monitored until ~99% of the sperm stopped moving (on average, in 40 s). A one-way ANOVA was used and statistical differences were set at α < 0.05. Extender 512 activated sperm motility when pH was >7.4, whereas extender 306 activated motility at every pH. Hanks’ Balanced Salt Solution activated motility at very low percentages for various pH levels, whereas Mounibs and Erdahal and Graham solutions did not activate motility for any pH tested. The 512 extender with a pH of 7 performed best (P < 0.05) as a storage solution, producing a sperm motility of 54% after activation using DIA 532. These results demonstrated the importance of evaluating effects of sperm extender pH over both short- and medium-term storage.

Seasonal variation in the profile of sex steroids and histological testicular development of brook trout (Salvelinus fontinalis Mitchill) during the annual reproductive cycle in Tasmania

Early maturation in brook trout males has been a constraint for commercial production of this species in Tasmania. However, control of maturation in males can be achieved by manipulating photoperiod (duration of light phase). To design an appropriate light treatment regime, an assessment of the annual reproductive cycle was initially required under ambient photoperiod as the annual reproductive cycle of male brook trout under ambient Tasmanian conditions of light and temperature was unknown. Here we describe the seasonal variations in testicular development and plasma profiles of testosterone (T), 11-ketotestosterone (11-KT) and oestradiol-17β (E2) during the second year in a male brook trout cohort held under ambient light conditions. Brook trout males were recruited for maturation in December when a reduction in daylength presumably stimulated testicular development. Increases in levels of T, 11-KT and gonadosomatic index (GSI) values were observed soon after the summer solstice, corresponding with maturation. The highest GSI mean value (3.44 ± 0.11%) was observed in April when mature males could be manually stripped of milt and such males were present until July. Profiles of sex steroids and histological development observed in this study were finely synchronised with variations in seasonal photoperiod changes. Photoperiod before recruitment during December should be controlled to manipulate maturation in brook trout males during their second year.

Seasonal and Morphological Variations of Brown Trout (Salmo trutta f. fario) Kidney Peroxisomes: A Stereological Study.

Literature about fish kidney peroxisomes is scarce. To tackle this caveat, a stereological approach on renal peroxisome morphological parameters was performed for the first time in a fish, establishing correlations with maturation stages as it was previously done in brown trout liver. Three-year-old brown trout males and females were collected at the major seasons of their reproductive cycle. Trunk kidney was fixed and processed for catalase cytochemistry. Classical stereological methods were applied to electromicrographs to quantitate morphological parameters. Different seasonal variation patterns were observed between genders, and between renal proximal tubule segments I and II. In males, peroxisomes from proximal tubule segment II had a relatively higher volume and number in May, being individually bigger in February. Females presented similar trends, though with less marked variations. Overall, males and females did not show exactly the same seasonal patterns for most peroxisomal parameters, and no correlations were found between the latter and the gonado-somatic index (GSI). Hence, and despite the variations, the morphology of renal peroxisomes is not strictly correlated with gonad maturation kinetics, therefore suggesting that kidney peroxisome morphology is not seasonally modulated by sex steroids, like estradiol, as it seems to happen in liver peroxisomes.

Occurrence of intersex in wild freshwater fish in Slovenian rivers: a histological evaluation.

The aim of this preliminary research was to establish if there are intersex occurrences in wild freshwater fish in Slovenian rivers and streams. In the first study we evaluated all fish species of both sexes obtained from the river Ljubljanica from its source to mouth. In the second study we focused on the rainbow trout (Oncorhynchus mykiss) and brown trout (Salmo trutta m. fario) males from 30 rivers and streams in different parts of Slovenia. The male gonads were histologically assessed for the presence of oocytes to determine the frequency and degree of intersex. Oocytes were found in the testicular tissue of a single grayling (Thymallus thymallus) and in the adipose tissue adjacent to the testis of a single common barbel (Barbus barbus), both from the Ljubljanica. Several cyst-like structures that resemble degenerated presumptive oocytes were also present in several trout testes. This preliminary report is the first of its kind in Slovenia. To gain a better insight into the intersex issue in Slovenia, we plan to regularly biomonitor freshwater pollution by histologically examining fish gonads and, if possible, by determining vitellogenin plasma levels in fish.

A small number of anadromous females drive reproduction in a brown trout (Salmo trutta) population in an English chalk stream

SummaryBrown trout,Salmo trutta, exhibit one of the most highly variable and polytypic life‐history strategies of all salmonids. Populations may be wholly freshwater‐resident or almost exclusively migratory (anadromous), or fish of a single population may exhibit varying proportions of the two life‐history strategies. Both anadromous and freshwater‐resident trout freely interbreed to produce fertile offspring.We quantify maternal reproductive provisioning by anadromous and freshwater‐resident brown trout to their offspring and assess relative parental fitness (in terms of number, size and time of emergence of offspring). Newly emerged juvenile trout (fry) were sampled (n = 119) over the emergence period in March–April 2007 in a lowland English chalk stream; samples of adult trout [anadromous (6F : 12M) and freshwater‐resident (22F : 56M)], river‐resident trout parr and macroinvertebrate prey were also collected.Using a novel combination of stable isotope analysis and microsatellite genotyping we demonstrate the overwhelming contribution of anadromous parents (both female and male) to fry production, despite the obvious presence and numerical dominance of resident adults. We unambiguously identify the maternal origins of 78% of juveniles sampled and show that maternal reproductive contribution to juvenile production in the river was higher for anadromous females (76%) than freshwater‐resident fish (2.5%). Offspring of anadromous females emerged earlier and at a larger body size than offspring of resident females. Similarly, while the relative contribution of resident males (37%) was higher than that of resident females, anadromous males sired considerably more offspring (63%) than resident males. This is the first study of its kind to accurately assess the reproductive contribution of anadromous male trout.Overall, this study suggests that anadromous maternal traits provide offspring with an adaptive advantage and greater fitness in early ontogeny, and that a small number of anadromous females (six of 96 adults sampled) are the main drivers of reproduction in this system.

Cytological state of gonads and level of thyroid and sex steroid hormones in black sea trout Salmo trutta labrax Pall.

Cytological state of the gonads and hormonal state of hatchery Black Sea trout before differentiation into resident and anadromous forms (parr) at an age of 15 months have been examined. It has been shown that the hormonal changes associated with the choice of life strategy in the Black Sea trout females and males are pronounced to different degrees. As compared with the resident and anadromous individuals; the female parr display a low rate of oogenesis and similar hormonal status, while characteristic of the male parr are an intermediate rate of spermatogenesis, a low level of thyroid hormones and estradiol, and a medium testosterone level. As has been found, the undifferentiated Black Sea trout individuals predominantly develop into the resident form.

Boule gene expression underpins the meiotic arrest in spermatogenesis in male rainbow trout (Oncorhynchus mykiss) exposed to DEHP and butachlor

Boule, the ancestor of the DAZ (Deleted in AZoospermia) gene family, in most organisms is mainly involved in male meiosis. The present study investigates the effects of the plasticizer DEHP (50mg/kg body weight) and herbicide butachlor (0.39mg/L) on male rainbow trout (Oncorhynchus mykiss) for a 10-day period in two independent experiments. The results showed that plasma testosterone (T) concentrations were significantly lower in fish exposed to either DEHP or butachlor compared to the control fish (P<0.05). Fish showed a significantly elevated hepatosomatic index (HSI) in the butachlor treatment (P<0.05). However, no significant difference was observed in HSI values in the DEHP treatment (P>0.05). In addition, no significant differences were found in the gonadosomatic index (GSI) in both DEHP and butachlor treatments (P>0.05). Histologically, testes of male trout in the control groups were well differentiated and filled with large numbers of cystic structures containing spermatozoa. In contrast, the testes of male trout contained mostly spermatocytes with few spermatozoa in both treated group, suggesting that DEHP and butachlor may inhibit the progression of meiosis. Also, boule gene expression was significantly lower in the testes of male trout affected by DEHP and butachlor in comparison with their control groups (P<0.05), which confirmed the meiotic arrest in affected trout. Based on the results, the present study demonstrated that DEHP and butachlor can inhibit the progression of spermatogenesis in male trout, potentially by causing an arrest of meiosis, maybe due to down-regulation of boule gene expression through T and/or IGF1 via ERK1/2 signaling in T-independent pathways. In addition, these results confirmed that boule can be considered as a predictive marker to assess meiotic efficiency.

A Bayesian assessment of the mercury and PCB temporal trends in lake trout (Salvelinus namaycush) and walleye (Sander vitreus) from lake Ontario, Ontario, Canada

Polychlorinated biphenyls (PCBs) and total mercury (THg) are two of the most prevalent contaminants, resulting in restrictive advisories on consuming fish from the Laurentian Great Lakes. The goal of this study is to examine the temporal trends of the two contaminants in walleye (Sander vitreus) and lake trout (Salvelinus namaycush) for Lake Ontario. We employed Bayesian inference techniques to parameterize three different strategies of time series analysis: dynamic linear, exponential decay, and mixed-order modeling. Our analysis sheds light on the role of different covariates (length, lipid content) that can potentially hamper the detection of the actual temporal patterns of fish contaminants. Both PCBs and mercury demonstrate decreasing temporal trends in lake trout males and females. Decreasing PCB trends are evident in walleye, but the mean annual mercury levels are characterized by a “wax and wane” pattern, suggesting that specific fish species may not act as bio-indicators for all contaminants. This finding may be attributed to the shifts in energy trophodynamics along with the food web alterations induced from the introduction of non-native species, the intricate nature of the prey–predator interactions, the periodicities of climate factors, and the year-to-year variability of the potentially significant fluxes from atmosphere or sediments. Finally, a meaningful risk assessment exercise will be to elucidate the role of within-lake fish contaminant variability and evaluate the potential bias introduced when drawing inference from pooled datasets.

Endocrine disruption screening by protein and gene expression of vitellogenin in freshly isolated and cryopreserved rainbow trout hepatocytes.

Xenobiotics may activate the estrogen receptor, resulting in alteration of normal endocrine functions in animals and humans. Consequently, this necessitates development of assay end points capable of identifying estrogenic xenobiotics. In the present study, we screened the potential estrogenicity of chemicals via their ability to induce vitellogenin (VTG) expression in cultured primary hepatocytes from male trout. A routine method for VTG detection measures the secretion of the protein by enzyme-linked immunosorbent assay (ELISA) in freshly isolated trout hepatocytes. However, this lengthy (6 days) culturing procedure requires that hepatocyte isolation is performed each time the assay is run. We optimized this methodology by investigating the utility of cryopreserved hepatocytes, shortening the incubation time, performing a quantitative real-time PCR (qPCR) method for VTG quantification, and verifying the model system with reference chemicals 17β-estradiol, estrone, diethylstilbestrol, hexestrol, genistein, and a negative control, corticosterone. To test the performance of both freshly isolated and cryopreserved hepatocytes, mRNA was collected from hepatocytes following 24 h treatment for VTG gene expression analysis, whereas cell culture media was collected for a VTG ELISA 96 h post-treatment. EC50 values were obtained for each reference chemical except for corticosterone, which exhibited no induction of VTG gene or protein level. Our results show linear concordance between ELISA and qPCR detection methods. Although there was approximately 50% reduction in VTG inducibility following cryopreservation, linear concordance of EC50 values was found between freshly isolated and cryopreserved hepatocytes, indicating that cryopreservation does not alter the functional assessment of estrogen receptor activation and therefore VTG expression. These studies demonstrate that qPCR is a sensitive and specific method for detecting VTG gene expression that can be used together with cryopreserved trout hepatocytes for screening estrogenic chemicals, resulting in a reduction of the time required to perform the assay and enabling greater access to the model system through the approach of cryopreservation.