Amino acids dissolved in stream water as possible home stream odorants for masu salmon

It is well established that salmon return to their home stream by sensing the odors of the stream water. In this study we have attempted to identify the home stream odorants used by masu salmon in Lake Toya. The salmon in Lake Toya return to the home stream which flows into the lake after lake life for 2-3 years. Besides water from the home stream, waters from two other streams which flow into Lake Toya were also used in the experiments. We analyzed the compositions of amino acids, inorganic cations and bile acids in waters from the three streams. Application of mixtures of inorganic cations or bile acids, reconstituted based on the compositions of the stream waters, to the olfactory epithelium induced only very small responses. On the other hand, application of mixtures of amino acids induced large responses. The response to artificial stream water reconstituted based on the compositions of amino acids and salts closely resembles that to the corresponding stream water. Cross-adaptation experiments with three combinations of the mixtures were carried out. The response pattern for each combination closely resembled that to the corresponding combination of stream waters. Based on the results obtained, we concluded that amino acids dissolved in the home stream water are possible home stream odorants.     

Physiological mechanisms of imprinting and homing migration in Pacific salmon Oncorhynchus spp

After several years of feeding at sea, salmonids have an amazing ability to migrate long distances from the open ocean to their natal stream to spawn. Three different research approaches from behavioural to molecular biological studies have been used to elucidate the physiological mechanisms underpinning salmonid imprinting and homing migration. The study was based on four anadromous Pacific salmon Oncorhynchus spp., pink salmon Oncorhynchus gorbuscha, chum salmon Oncorhynchus keta, sockeye salmon Oncorhynchus nerka and masu salmon Oncorhynchus masou, migrating from the North Pacific Ocean to the coast of Hokkaido, Japan, as well as lacustrine O. nerka and O. masou in Lake Toya, Hokkaido, where the lake serves as the model oceanic system. Behavioural studies using biotelemetry techniques showed swimming profiles from the Bering Sea to the coast of Hokkaido in O. keta as well as homing behaviours of lacustrine O. nerka and O. masou in Lake Toya. Endocrinological studies on hormone profiles in the brain-pituitary-gonad axis of O. keta, and lacustrine O. nerka identified the hormonal changes during homing migration. Neurophysiological studies revealed crucial roles of olfactory functions on imprinting and homing during downstream and upstream migration, respectively. These findings are discussed in relation to the physiological mechanisms of imprinting and homing migration in anadromous and lacustrine salmonids.     

Lacustrine sockeye salmon return straight to their natal area from open water using both visual and olfactory cues

Mechanisms of the amazing ability of salmon to migrate a long distance from open water to natal streams for spawning are still unknown. Lacustrine sockeye salmon (Oncorhynchus nerka) in Lake Toya offers an excellent model system for studying the orientation mechanism in open water, because mature fish return to the natal area with a high degree of accuracy. First we examined the percentage of fish returning to the natal area after they were released 7 km south of the natal area. Forty percent of control male mature fish and 25% of the fish blinded by injection of a mixture of carbon toner and corn oil into the eyeball were captured in the natal area within 5 days. Forty-four percent of fish with brass rings (control) and 31% of fish with NdFe magnetic rings which interfere with the magnetic cue were captured in the natal area within 3 days. These experiments suggested that, although the number of blinded fish captured in the natal area was less than that of the controls, the difference was not statistically significant. In the fish captured in the natal area within 3 or 5 days, fish which found the natal area using their olfactory cue after random swimming for a long time and returned to that area may be included. Hence we tracked fish telemetrically using an ultrasonic tracking system, and found that mature males released at a long distance (3.6 or 6.8 km) from the natal area swam straight to the vicinity of the natal area. Interference of the magnetic cue by the attachment of a magnetic ring did not affect their direct return. Blockage of the visual cue caused them to move randomly. These data suggest that lacustrine sockeye salmon return straight to the vicinity of the natal area using their visual cue and finally reach the exact homing point using their olfactory cue.      

Olfactory responses to natal stream water in sockeye salmon by BOLD fMRI

Many studies have shown that juvenile salmon imprint olfactory memory of natal stream odors during downstream migration, and adults recall this stream-specific odor information to discriminate their natal stream during upstream migration for spawning. The odor information processing of the natal stream in the salmon brain, however, has not been clarified. We applied blood oxygenation level-dependent (BOLD) functional magnetic resonance imaging to investigate the odor information processing of the natal stream in the olfactory bulb and telencephalon of lacustrine sockeye salmon (Oncorhynchus nerka). The strong responses to the natal stream water were mainly observed in the lateral area of dorsal telencephalon (Dl), which are homologous to the medial pallium (hippocampus) in terrestrial vertebrates. Although the concentration of L-serine (1 mM) in the control water was 20,000-times higher than that of total amino acid in the natal stream water (47.5 nM), the BOLD signals resulting from the natal stream water were stronger than those by L-serine in the Dl. We concluded that sockeye salmon could process the odor information of the natal stream by integrating information in the Dl area of the telencephalon.     

Salmonid olfactory system-specific protein (N24) exhibits glutathione S-transferase class pi-like structure

A salmonid olfactory system-specific protein (N24) that has been identified in lacustrine sockeye salmon (Oncorhynchus nerka) was characterized by biochemical and molecular biological techniques. N24 is a homodimer, and the intact molecular mass is estimated as approximately 43.3 kDa by gel filtration. Furthermore, N24 was located only in the cytosolic fraction of the olfactory tissues as determined by subcellular fractionation. cDNA encoding the lacustrine sockeye salmon N24 was isolated and sequenced. This cDNA contained a coding region encoding 216 amino acid residues and the molecular mass of this protein is calculated to be 242,224.77. The protein and nucleotide sequencing demonstrates the existence of a remarkable homology between N24 and glutathione S-transferase (GST; EC 2.5.1.18) class pi enzymes. Northern analysis showed that N24 mRNA with a length of 950 bases is expressed in lacustrine sockeye salmon olfactory epithelium. Olfactory receptor cells showed strong hybridization signals for N24 mRNA in the olfactory epithelium. N24 demonstrated glutathione binding activity in affinity-purified GST column experiments. The present study describes for the first time cDNA cloning of GST in fish olfactory epithelium.     

Cortisol and Pacific Salmon: A New Look at the Role of Stress Hormones in Olfaction and Home-stream Migration

Pacific salmon (genus Oncorhynchus) exhibit an interesting anduncommon life-history pattern that combines semelparity, anadromy,and navigation (homing). During smoltification, young salmonimprint on the chemical composition of their natal stream water(the home-stream olfactory bouquet or "HSOB"); they then migrateto the ocean where they spend a few years feeding prior to migratingback to their natal freshwater stream to spawn. Upstream migrationis guided by the amazing ability to discriminate between thechemical compositions of different stream waters and thus identifyand travel to their home-stream. Pacific salmon demonstratemarked somatic and neural degeneration changes during home-streammigration and at the spawning grounds. The appearance of thesepathologies is correlated with a marked elevation in plasmacortisol levels. While the mechanisms of salmonid homing arenot completely understood, it is known that adult salmon continuouslyutilize two of their primary sensory systems, olfaction andvision, during homing. Olfaction is the primary sensory systeminvolved in freshwater homing and "HSOB" recognition, and willbe emphasized here. Previously, we proposed that the increasein plasma cortisol during Pacific salmon home-stream migrationis adaptive because it enhances the salmon's ability to recallthe imprinted memory of the "HSOB" (Carruth, 1998; Carruth etal., 2000b). Elevated plasma concentrations of cortisol couldprime the hippocampus or other olfactory regions of the brainto recall this memory and, therefore, aid in directing the fishto their natal stream. Thus, specific responses of salmon tostressors could enhance reproductive success.     

Olfactory Imprinting of Amino Acids in Lacustrine Sockeye Salmon

Juvenile salmon have an olfactory ability to imprint their natal stream odors, but neither the odor properties of natal stream water nor the imprinting timing and duration have been clarified as yet. Here we show, using electrophysiological and behavioral experiments, that one-year-old lacustrine sockeye salmon (Oncorhynchus nerka) can be imprinted around the stage of parr-smolt transformation (PST) by a single amino acid, 1 µM L-proline (Pro), or L-glutamic acid (Glu). We also show by real-time PCR that changes occur in mRNA levels of the salmon olfactory imprinting-related gene (SOIG) around PST. The electro-olfactogram (EOG) responses of test fish exposed to Pro in March (before PST) and April–June (during PST) for 2 weeks were significantly (1.7-fold) greater than those of non-exposed control fish, but not those of test fish exposed in July (after PST). When Pro and control water were added to the water inlets of a two-choice test tank during the spawning season 2 years after the test water exposure, 80% of maturing and matured test fish exposed before and during PST showed a preference for Pro, whereas those exposed after PST did not. The EOG response of test fish exposed to Pro or Glu for 1 hour, 6 hours, 1 day, 7 days, or 14 days in May revealed that only the response after 14 days of exposure was significantly (1.8-fold) greater than the control. The expression levels of SOIG mRNA increased before and during PST, and decreased after PST. We conclude that one-year-old lacustrine sockeye salmon can be imprinted by a single amino acid before and during PST, and that imprinting requires exposure for at least 14 days.     

Deep water stratification in deep caldera lakes Ikeda, Towada, Tazawa, Kuttara, Toya and Shikotsu

From six deep caldera lakes in Japan, namely lakes Ikeda, Towada, Tazawa, Toya, Kuttara and Shikotsu (in Japanese referred to as Ikedako, Towadako, Tazawako, Toyako, Kuttarako and Shikotsuko), fine resolution profiles of temperature, electrical conductivity, dissolved oxygen and pH have been measured. Measurements were conducted just after deep circulation from the end of March to the end of May 2005. Lake Ikeda and Lake Towada did not undergo a complete turnover. Both showed meromictic features with a clear interface separating the recirculated mixolimnion from the deeper monimolimnion. Lake Tazawa and Lake Toya showed a complete turnover in winter 2004/05, while in Lake Kuttara and Lake Shikotsu, a deep water body remained throughout winter due to pressure effects on the temperature of maximum density. Although these deep waters were never fully recycled into the mixolimnion, they presented themselves as well supplied with oxygen and well circulated within themselves. Where overturn had not been complete small gradients in the oxygen profile, pH profiles and electrical conductivity profiles could be detected. However only in Lake Towada were concentrations of dissolved substances and gradients high enough to have decisive impact on the circulation pattern.     

A review of some characteristics of freshwater and coastal ecosystems in Ghana

Data collected over a ten year period have been used to review some characteristics of freshwater and coastal ecosystems in Ghana. Studies were conducted on two recently formed man-made lakes, one river, five coastal lagoons and waters along the ocean front.Freshwaters had near neutral pH while coastal waters were alkaline. In both ecosystems the pH of sediments were lower than those of corresponding waters. While the transparency of coastal waters varied within narrow limits, large variations were observed in freshwaters. The lowest concentration of nutrients occurred in the river and the highest in coastal waters.In general, differences in these physical and chemical characteristics do not depend on whether the aquatic systems were fresh, brackish or saline. They seem to be more influenced by such factors as whether a water body: (1) was lotic or lentic, (2) was influenced by industrial or domestic activities, (3) was affected by impoundment, (4) received turbid water inflows, or (5) was eutrophic.     

An electrophysiological study on the effects of pH on olfaction in mature male Atlantic salmon (Salmo salar) parr

The olfactory responses of mature male Atlantic salmon ( Salmo salar L.) parr to known odorants were studied after exposure of the olfactory epithelia to water of varying pH. Electrophysiological recordings from the olfactory epithelia indicated that the responses of fish to both testosterone and urine from ovulated female Atlantic salmon were significantly reduced at pH 5·5 and 4·5 and abolished at pH 3·5. Concentration response studies indicated that at pH 5·5 and 6·5 significantly higher concentrations of testosterone and urine were required to produce the same amplitude responses as controls. Both testosterone and urine have previously been shown to be important chemical cues eliciting behavioural and physiological responses in Atlantic salmon. The results are therefore discussed in relation to the possible sublethal effects of acidification on reproduction and behaviour of Atlantic salmon and the effects on salmonid stocks.     

In situ and laboratory studies on the behaviour and survival of Pacific salmon (genus Oncorhynchus)

Juvenile Pacific salmon display a marked surface water orientation during downstream migration, estuarine and nearshore coastal rearing phases. Many estuaries in British Columbia are vertically stratified with a shallow, well-defined halocline which can restrict the dispersion of wastes discharged into less saline surface waters and impose constraints upon aquatic organisms. In situ experiments in an estuary receiving a surface discharge of treated pulp mill wastes, revealed conditions which were lethal to underyearling salmon at, and below the halocline (4.0–6.5 m depth). Behavioural bioassays determined that juvenile chinook salmon were biased towards the water surface and avoided waters at depth. Dissolved oxygen was the variable which affected this distribution most significantly. Surface waters receiving effluent from another pulp mill were lethal to juvenile salmon within 350 m, and a significant vertical avoidance response occurred within 350–950 m of the outfalls. The behavioural response was significantly correlated with in situ temperature, pH and colour (effluent).As a complement to field experiments we developed a 4500 l water column simulator (WCS) to examine salmon behaviour in the laboratory. We investigated the surface water orientation behaviour of juvenile salmon in relation to variations in salinity and dissolved oxygen. Under simulated vertically stratified estuarine conditions, the fish moved freely between overlying fresh water and salt water. Induction of hypoxic conditions in fresh water elicited a downward distribution shift towards the halocline and oxygenated, but more saline, waters. Avoidance reactions (50% level) occurred consistently up to 7–8 mg · l^–1 dissolved oxygen. Salmon continued to examine the hypoxic freshwater zone despite sub-optimal conditions.     

Infection status of pond smelts, Hypomesus olidus, and other freshwater fishes with trematode metacercariae in 6 large lakes

In order to determine the current infection status of pond smelts, Hypomesus olidus, and other freshwater fishes with trematode metacercariae, a total of 4,861 pond smelts and 18 other freshwater fishes, collected from the Soyang and Uiam Lakes in Gangwon-do, the Unam Lake in Jeollabuk-do, the Jangseong Lake in Jeollanam-do, the Uirim-ji (lake) in Chungcheongbuk-do, and the Andong Lake in Gyeongsangbuk-do, were individually digested with 1% pepsin-HCl and examined under a dissecting microscope. In all pond smelts caught from the 6 lakes, we were unable to detect any known human infectious trematode metacercariae in Korea. However, in other freshwater fishes, such as, Squalidus japonicus coreanus (Unam Lake), and Zacco platypus (Jangseong Lake) and Opsariichthys uncirostris amurensis (Jangseong Lake), metacercariae of human-infecting trematodes, i.e., Clonorchis sinensis and Metagonimus sp. were detected, respectively.     

Patterns of composition and species richness of crustaceans and aquatic insects along environmental gradients in Mediterranean water bodies

Differences in the dynamics of ecological processes between Mediterranean and colder temperate aquatic systems could imply different patterns in faunal communities in terms of composition and biodiversity (i.e. species richness and rarity). In order to identify some of these patterns the crustacean and aquatic insect composition and biodiversity of four water body types, classified according to their salinity and water permanence, were compared. Moreover, the relationships between species richness and water, pond and landscape variables were analysed. A total number of 91 water bodies located throughout Catalunya (NE Iberian Peninsula) were sampled. Three species assemblages were observed: one for permanent freshwaters, another for temporary freshwaters, and a third one for saline waters (SW), since permanent and temporary saline water bodies had similar composition. Differences in salinity were associated with proportion of crustaceans versus insects and with singularity. Thus, saline ponds had a higher proportion of crustaceans, and lower values of singularity. Conductivity was significantly related to total (crustaceans plus insects) richness, and also related to insect richness. The main difference between the models obtained for crustacean species richness and insect species richness is the significance of landscape variables in the latter, and this fact could be related to the different dispersion types of these two faunal groups: active for insects versus passive for crustaceans. Electronic supplementary material The online version of this article (doi:) contains supplementary material, which is available to authorized users. Guest editors: R. Céréghino, J. Biggs, B. Oertli & S. Declerck The ecology of European ponds: defining the characteristics of a neglected freshwater habitat     

The impact of a brominated flame retardant on smoltification and olfactory function in Atlantic salmon (Salmo salar L.) smolts

Despite a reduction in exploitation of salmon stocks throughout the NE Atlantic, there continues to be a decline in many populations. The factors regulating these populations remain poorly understood, although there is evidence that environmental conditions experienced in freshwater can effect survival in the marine environment. The aim of this study was to investigate the impact of a brominated flame retardant (hexabromocyclododecane, HBCD) on the parr-smolt transformation in juvenile salmon. It is during this pre-adaptive period to marine life that olfactory imprinting to the natal river is considered to occur. Fish were exposed to low levels of HBCD for 30 days over the peak smoltification period in freshwater, and then transferred to clean seawater for 20 days. Fish were sampled weekly to assess changes in some of the physiological parameters associated with smoltification, and olfactory response to conspecific smolt urine was measured using an electro-olfactogram (EOG). Exposure to HBCD did not affect seawater adaptability, although there was some disruption of plasma thyroid hormone levels, as well as a reduction in olfactory function to conspecific smolt urine. The results are discussed in relation to the marine survival and successful homing of adult salmon.     

The impact of a brominated flame retardant on smoltification and olfactory function in Atlantic salmon (Salmo salar L.) smolts

Despite a reduction in exploitation of salmon stocks throughout the NE Atlantic, there continues to be a decline in many populations. The factors regulating these populations remain poorly understood, although there is evidence that environmental conditions experienced in freshwater can effect survival in the marine environment. The aim of this study was to investigate the impact of a brominated flame retardant (hexabromocyclododecane, HBCD) on the parr-smolt transformation in juvenile salmon. It is during this pre-adaptive period to marine life that olfactory imprinting to the natal river is considered to occur. Fish were exposed to low levels of HBCD for 30 days over the peak smoltification period in freshwater, and then transferred to clean seawater for 20 days. Fish were sampled weekly to assess changes in some of the physiological parameters associated with smoltification, and olfactory response to conspecific smolt urine was measured using an electro-olfactogram (EOG). Exposure to HBCD did not affect seawater adaptability, although there was some disruption of plasma thyroid hormone levels, as well as a reduction in olfactory function to conspecific smolt urine. The results are discussed in relation to the marine survival and successful homing of adult salmon.     

也西湖噬藻体的分离与鉴定

【背景】噬藻体是一类特异性侵染蓝藻的病毒,广泛存在于淡水和海水水体中,参与调控宿主蓝藻的丰度和种群密度,被认为是潜在的蓝藻水华生物防控工具。但目前的研究多集中于海洋噬藻体,对淡水噬藻体的生物学特性和结构生物学等研究较少。【目的】分离更多种类的淡水噬藻体,为研究淡水噬藻体的三维结构、侵染机制、与宿主的共进化关系,及其在蓝藻水华防治中的应用提供理论基础。【方法】采集中国科学技术大学西校区内景观湖也西湖水华暴发水域的水样,利用液体培养基和双层固体平板法对17种宿主蓝藻进行筛选,通过NaCl-PEG沉淀法和氯化铯密度梯度离心分离和纯化噬藻体,并利用负染电镜观察噬藻体的形态,同时采用梯度稀释法测定裂解液的效价。【结果】发现也西湖的水样可特异性侵染本实验室分离自巢湖的一株拟鱼腥藻Pan。侵染后的裂解液中存在4株形态各异的噬藻体,包括1株短尾噬藻体和3株长尾噬藻体,其中包括首次发现的1株含有非典型长轴状头部结构的淡水噬藻体。【结论】也西湖作为巢湖流域的一个小型水体,具有与巢湖类似的水华蓝藻及其噬藻体分布谱,因此可以用于模拟大型湖泊进行相关分子生态学和生物防控的研究。     

Olfactory sensitivity of coho salmon,Oncorhynchus kisutch (Walbaum): can salinity play a role?

The importance of environmental ions to olfactory sensitivity in fish has not been clarified by electrophysiological experiments. We tested the ability of juvenile coho salmon, Oncorhynchus kisutch (Walbaum), to detect L-serine after acclimation to either fresh water or sea water. The olfactory threshold was identical in the two waters, indicating that the sensitivity of olfaction was not affected by ionic content of the solutions bathing the sensory tissue.     

Cloning and characterization of an odorant receptor in five Pacific salmon

The olfactory system of fish is extremely important as it is able to recognize and distinguish a vast of odorous molecules involved in wide ranges of behaviors including reproduction, homing, kin recognition, feeding and predator avoidance; all of which are paramount for their survival. We cloned and characterized one type olfactory receptors (ORs) from five congeneric salmonids: lacustrine sockeye salmon (Oncorhynchus nerka), pink salmon (O. gorbuscha), chum salmon (O. keta), masu salmon (O. masou) and rainbow trout (O. mykiss). Lacustrine sockeye salmon olfactory receptor 1 (LSSOR1) showed high sequence homology to the OR subfamily, and was expressed only in the olfactory epithelium (as indicated by PCR amplified genomic DNA and cDNA). OR genes from the five salmonids examined all showed strong homology (96-99%) to each other. Hypervariable regions, believed to be ligand-binding pockets, showed homologous completely matched amino acid sequences except for one amino acid in pink salmon olfactory receptor 1 (PSOR1), revealing that these ORs may be well conserved among salmon species. These results suggest that the isolated 5 salmonid ORs might play an important role in salmon life cycles.     

Actinobacterial 16S rRNA genes from freshwater habitats cluster in four distinct lineages

We analysed the phylogenetic relatedness of 16S rRNA genes from freshwater bacteria affiliated with the class Actinobacteria. A polymerase chain reaction assay was developed to identify reliably rare Actinobacteria-related inserts within 16S rRNA gene clone libraries. In 18 libraries constructed from seven freshwater systems, altogether 63 actinobacterial sequence types were collected from a total of > 1800 clones. Sixty of the newly obtained sequences grouped within four distinct phylogenetic lineages. They constitute approximately 75% of the nearly complete sequences within these clusters that are presently available. A comparison with > 300 sequences from various soil habitats revealed that two of these monophyletic actinobacterial clades (acI and acII) almost exclusively harbour 16S rRNA sequence types from freshwaters and estuaries. This may indicate that such bacteria are not inoculated to freshwaters from terrestrial sources, but are autochthonous components of freshwater microbial assemblages. In contrast, sequence types from freshwaters, marine sediments and soils were clearly mixed in another of the actinobacterial lineages (acIV). Sequence divergence within acIV was the highest of all four lineages (88% minimum similarity), which potentially reflects its radiation across several habitat types. Within the freshwater lineages, groups of essentially identical sequence types were retrieved from geographically distant aquatic systems with strikingly different hydrological and limnological characteristics. This points to the necessity to investigate genotypic variability, in situ abundances and activities of these Actinobacteria in freshwater plankton in greater detail by cultivation-independent techniques.     

Annual Abundances, Diversity and Growth Potential of Gymnamoebae in a Shallow Freshwater Pond

ABSTRACT. Abundance and diversity of gymnamoebae were assessed at monthly intervals for one year in the sediment surface layer of a shallow, highly productive, freshwater pond located in northeastern United States using laboratory culture enumeration techniques. The range in total abundance was 81/ml in January to 1,568/ml in June (spring peak) and 1,813/ml in October (autumn peak). The latter is equivalent to over 1.8 million per liter, one of the highest values reported for gymnamoebae from a freshwater pond. Maximum diversity (H = 3.3) occurred in April, July, and August. The growth potential of the gymnamoebae was determined in laboratory cultures (20° C) containing only the pond water (control culture) and pond water emended with malt-yeast extract and glucose (nutrient culture) using pond water collected in December. The total abundance of gymanmoebae increased from 363/ml in the initial pond water sample to approximately 1,800/ml in each of the culture conditions in the first week. Therafter, abundances reached 5,678/ml (control culture) and 8,016/ml (nutrient culture) by the third week indicating the vigorous growth potential from winter standing stock.