Stress reduces the quality of gametes produced by rainbow trout.

In this study we have used the rainbow trout as a model animal to study the biological consequences of stress in terms of gamete quality and quantity. Groups of 30 mature male and female rainbow trout were subjected to repeated acute stress during the 9 mo prior to spawning. Time of ovulation, fecundity, and egg size were recorded in mature females, and sperm counts were carried out on the milt from the male fish, from both the stressed and control groups. Eggs from ovulated females were fertilized with milt from males subjected to the same treatment regime. Approximately 300 eggs from each female were fertilized with a sperm dilution of 10(-3) in diluent. Subsequent development of the fertilized eggs was then monitored. There were no differences in somatic weight or length between the two groups at the end of the experiment, but exposure of rainbow trout to repeated acute stress during reproductive development resulted in a significant delay in ovulation and reduced egg size in females, significantly lower sperm counts in males, and, perhaps most importantly, significantly lower survival rates for progeny from stressed fish compared to progeny from unstressed control fish. Hence, stress reduces the quality of gametes produced by rainbow trout.     

Thermotolerance of brown trout,Salmo trutta,gametes and embryos to increased water temperatures

The present study investigated the effect of increased water temperature on sperm motility when activated in water, on egg water hardening, on the fertilization process and on embryonic development in brown trout, Salmo trutta. Brown trout gametes had a broad temperature optimum ranging from 3 to 15°C. In this temperature range the processes of egg water hardening and fertilization were unaffected. Motility duration was also not affected. The percentages of initial sperm motility and swimming velocity were lower at 9–15°C than at 3–7°C, while the percentage of locally motile spermatozoa was higher. Exposure of brown trout embryos to ≥11°C resulted in reduced percentages of hatched larvae and in increased percentages of malformed larvae. More advanced ontogenetic stages (stages ≥9 according to Ballard [1973]) showed higher tolerance to increased temperature than did less advanced stages (stages <9) when the percentage of eyed stage embryos was used as the evaluation end point. However, when the percentages of hatched larvae and normal shaped larvae were used as evaluation end points, no differences in thermotolerance could be found between the different otogenetic stages.     

Maternal communication with gametes and embryos

Mechanisms for gametes and embryos to interact with their maternal environment are crucial in achieving reproductive success, both in livestock and the human. Long-range (hormones) and short-range signalling molecules play important roles in mediating cell-cell maternal interactions/communications with gametes and embryos. Slight malfunctions or disturbances of the environment that host this interaction can retard embryonic development. This may lead to creation of a memory for the embryo leading to offspring prone to degenerative diseases in adulthood. Despite an overwhelming amount of research and the literature, not all signalling molecules involved and their relationship with each other are known. Progress in the application of high-throughput genomic and proteomic analytical tools, such as microarrays and quantitative proteomic technologies has had a positive impact on our understanding of various aspects of maternal communication with gametes and embryos. Recent advances point to the presence of a local mechanism in the female reproductive tract capable of recognising the arrival of gametes and embryos and modulating the tract's environment accordingly for the next stage. Further investigations are underway to characterise the details of this system. It is important to consider spatial or temporal components of maternal communication with gametes and embryos that may confer consequences for developmental potential. Finally, it seems that the application of a systems biology approach for creation of an interactome map of maternal communication with gametes and embryos is essential and provides an excellent opportunity for an inter-disciplinary collaboration with engineers and mathematical modellers.     

Morphological and functional development of the thyroid tissue in rainbow trout (Oncorhynchus mykiss) embryos

The development of the thyroid tissue in rainbow trout (Oncorhynchus mykiss) embryos between 20 and 88 days postfertilization (dpf) was studied using traditional histological methods in combination with immunostaining using antibodies raised against L-thyroxine (T4) and triiodo-L-thyronine (T3). In 20 dpf embryos, the thyroid primordium appeared as a simple tube aligned with the dorsal surface of the ventral aorta. The tubular primordium becomes progressively bifurcated at each end and follicles are formed by pinching off balls of cells from the tips of the bifurcations. The secretory activity of the thyroid tissue appears first as a synthesis phase, evident at 30 dpf, characterized by T4 and T3 immunostaining present only in the follicle lumen. A later hormone release or secretory phase was first evident for T4 immunostaining at 43 dpf and for T3 immunostaining at 46 dpf and was characterized by the presence of immunostaining both within the follicle lumen and also in the cytoplasm of some (but not all) thyrocytes. This stage was also coincident with a marked increase in the number of pituitary thyrotrophs. The proportion of embryos exhibiting immunostained thyrocytes increased progressively from 43 to 52 dpf, at which stage all embryos had T4- and T3-immunostained thyrocytes.     

Activation of rainbow trout macrophages

Rainbow trout peritoneal macrophages were stimulated in vitro using Concanavalin A (Con A) and in vivo using formalin-killed Aeromonas salmonicida in Freund's incomplete adjuvant (FIA). Whether these cells had been activated was determined by the measurements of oxygen anions (NBT reduction), H₂O₂ production (oxidation of phenol red), RNA synthesis (³H-uridine incorporation), acid phosphatase activity and bactericidal activity.
In vitro -stimulated macrophages showed an increased NBT reduction and ³H-uridine incorporation over a range of Con A concentrations, compared with untreated control macrophages, but no detectable increases in H₂O₂ production or bactericidal activity were observed. On the other hand, in vivo -stimulated peritoneal cells showed increases in all the assays compared with FIA-elicited control cells, and were considered to have been activated.     

The development of catecholamines in embryos and larvae of the rainbow trout (Salmo gairdneri Rich.)

Catecholamines are present in the late embryonal and early larval developmental stages (alevin) of the rainbow trout. Changes in the alevin behaviour can be related to the catecholamine concentrations.     

Cryopreservation of gametes and embryos of non-domestic species.

Many species of mammals are threatened or endangered. Methods of assisted reproduction that are being used with increasing frequency to produce offspring of domestic animals and humans are often viewed as offering innovative ways to reproduce non-domestic species as well. Uncounted millions of live young of domestic or laboratory species have been produced from gametes and embryos stored at -70 degrees C or below, sometimes for as long as 25 to 35 yrs. Such methods of cryopreservation are now being applied with increasing frequency and urgency to preserve gametes and embryos of non-domestic and threatened species to establish "genome resource banks" or "frozen zoos." But levels of success to produce live young from such cryopreserved gametes or embryos vary considerably from species to species, as well as from individual to individual. It is sometimes thought that differences among species in fundamental characteristics of their gametes may determine the efficacy of cryopreservation and the production of live young. However, it may not be that ineffective cryopreservation is responsible for low success rates. Rather, the limiting factor may be insufficient information and knowledge of the most basic reproductive biology of such non-domestic species. Even standard methods of cryopreservation may be completely adequate to act as a "temporary" expedient to preserve germplasm of non-domestic species to permit time to acquire a fuller understanding of the biology and behavior of non-domestic species.     

A pepsinogen from rainbow trout

1. A pepsinogen, Ia on the basis of its electrophoretic mobility, from rainbow trout stomach, has an optimum pH near 2 for activation. 2. The cognate pepsin is denatured at pH values above 7, in contrast to the zymogen, which is slightly more alkali-stable. It has an optimum pH of 3 for proteolysis of denatured hemoglobin. 3. The intrinsic reactivity of the zymogen and pepsin (rates of activation and of proteolysis, respectively) are quite high, but as they operate at the environmental temperature of the fish, are remarkably similar to rates of activation and proteolysis by mammalian pepsinogens and pepsins.     

Overwinter feeding in rainbow trout

The contents of the stomachs of 38 rainbow trout stocked in Llyn Alaw, Anglesey, in August 1969 and caught between October 1969 and February 1970 were analysed. The fish were actively feeding on the bottom fauna throughout the winter and 21 of the stomachs were full or distended. The mean volume of the contents of the stomachs was 2–8 times greater than that of the contents of stomachs of similarly sized brown trout caught at the same time.     

Immunohistochemical localization of rainbow trout ladderlectin and intelectin in healthy and infected rainbow trout (Oncorhynchus mykiss)

In the present study, the pattern of immuno-reactive ladderlectin and intelectin in healthy rainbow trout is compared to rainbow trout infected with a variety of infectious agents. In healthy rainbow trout, both proteins were localized to individual epithelial cells of the gill and intestine and both proteins were clearly demonstrated within cytoplasmic granules of polymorphonuclear leucocytes and macrophages/monocytes found in blood vessels, hepatic sinusoids, renal interstitium, mucosal epithelium and submucosa of normal intestine. In tissue from infected rainbow trout, there was an overall relative increase in both lectins compared to healthy fish and both proteins were detected in extra-cellular spaces surrounding bacteria, fungi and protozoa. Increased distribution and density of both RTLL and RTInt was demonstrated along mucosal surfaces and within inflammatory leucocytes in infected tissues and immune related organs. These findings represent one of the few examples of in vivo association of defence lectins and infectious agents.     

Production of androgenetic diploid rainbow trout

Haploid androgenesis was induced in rainbow trout (Salmo gairdneri) when eggs were irradiated with 60Co gamma radiation prior to fertilization. Diploidy was restored to the androgenetic haploid zygotes by suppression of first cleavage division using hydrostatic pressure. Peak survival in the androgenetic diploid lots (32.5-38.9 percent of control) occurred when a pressure shock of 9000 pounds per square inch lasting from one to three minutes was applied to the eggs 345 minutes post-fertilization. Chromosomal analysis confirmed diploidy in the androgenetic individuals and suggested that YY rainbow trout are viable to at least the "eyed stage" of development. Inheritance patterns at two loci confirmed all-paternal inheritance. The relatively high yields of completely homozygous androgenetic rainbow trout and the potential for the use of androgenesis in the production of inbred lines and in genetic studies indicate that androgenesis may become a valuable tool in fish research and breeding.     

Hybridization dynamics between Colorado's native cutthroat trout and introduced rainbow trout

Newly formed hybrid populations provide an opportunity to examine the initial consequences of secondary contact between species and identify genetic patterns that may be important early in the evolution of hybrid inviability. Widespread introductions of rainbow trout (Oncorhynchus mykiss) into watersheds with native cutthroat trout (Oncorhynchus clarkii) have resulted in hybridization. These introductions have contributed to the decline of native cutthroat trout populations. Here, we examine the pattern of hybridization between introduced rainbow trout and 2 populations of cutthroat trout native to Colorado. For this study, we utilized 7 diagnostic, codominant nuclear markers and a diagnostic mitochondrial marker to investigate hybridization in a population of greenback cutthroat trout (Oncorhynchus clarkii stomias) and a population of Colorado River cutthroat trout (Oncorhynchus clarkii pleuriticus). We infer that cutthroat-rainbow trout hybrid swarms have formed in both populations. Although a mixture of hybrid genotypes was present, not all genotype combinations were detected at expected frequencies. We found evidence that mitochondrial DNA introgression in hybrids is asymmetric and more likely from rainbow trout than from cutthroat trout. A difference in spawning time of the 2 species or differences in the fitness between the reciprocal crosses may explain the asymmetry. Additionally, the presence of intraspecific cytonuclear associations found in both populations is concordant with current hypotheses regarding coevolution of mitochondrial and nuclear genomes.     

Habitat shifts in rainbow trout: competitive influences of brown trout

Summary We compared habitat use by rainbow trout sympatric (three streams) and allopatric (two streams) with brown trout to determine whether competition occurred between these two species in the southern Appalachian Mountains. We measured water depth, water velocity, substrate, distance to overhead vegetation, sunlight, and surface turbulence both where we collected trout and for the streams in general. This enabled us to separate the effects of habitat availability from possible competitive effects. The results provided strong evidence for asymmetrical interspecific competition. Habitat use varied significantly between allopatric and sympatric rainbow trout in 68% of the comparisons made. Portions of some differences refelected differences in habitats available in the several streams. However, for all habitat variables measured except sunlight, rainbow trout used their preferred habitats less in sympatry with brown trout than in allopatry if brown trout also preferred the same habitats. Multivariate analysis indicated that water velocity and its correlates (substrate particle size and surface turbulence) were the most critical habitat variables in the interaction between the species, cover in the form of shade and close overhead vegetation was second most important, and water depth was least important.     

Genetic analysis of androgenetic rainbow trout.

We analyzed a number of genetic characteristics in androgenetic rainbow trout (Oncorhynchus mykiss) and their progeny. The androgenetic progeny of individual androgenetic males appeared genetically identical to each other based on eight enzyme loci. Their viability was no higher than that of androgenetic progeny of outbred males. Homozygous androgenetic female rainbow trout produced very poor quality eggs. When common eggs and sperm from outbred individuals were used to produce androgenetic and gynogenetic progeny, the yield of gynogenetic progeny was higher but some were heterozygous at protein loci, while no androgenetic progeny were heterozygous. Some androgenetic diploid rainbow trout were successfully produced from cryopreserved sperm. The progeny of some androgenetic males crossed to normal females were virtually all males, while the progeny of other males were virtually all females. This suggests that both XX and YY androgenetic individuals may develop as males. Androgenesis is likely to be useful for generating homozygous clones for research and for recovering strains from cryopreserved sperm.