Biochemical identification of a brackish water type ofPungitius pungitius,and its morphological and ecological features in Hokkaido,Japan

A brackish water type ofPungitius pungitius (Gasterosteidae) is newly reported from Japan. Genetic differences, based on 13 loci of 6 enzymes and muscle protein, were examined between fresh and brackish water types in the Biwase River in the eastern part of Hokkaido. The alleles onGpd, Pgi andPgm were displaced between them, even though their distributions partially overlapped in the river. The brackish water type was also statistically different from the fresh water type on the basis of three morphological characters, such as number of dorsal spines, gill-rakers and vertebrae, although ranges overlapped. Segregation of spawning sites and differing nuptial colors of the male were found between fresh and brackish water types. These facts suggest that the brackish water type is reproductively isolated from the fresh water type and that both types should be regarded as independent species.     

Biological Differences between Brackish and Fresh Water-Derived Aedes aegypti from Two Locations in the Jaffna Peninsula of Sri Lanka and the Implications for Arboviral Disease Transmission

The mainly fresh water arboviral vector Aedes aegypti L. (Diptera: Culicidae) can also undergo pre-imaginal development in brackish water of up to 15 ppt (parts per thousand) salt in coastal areas. We investigated differences in salinity tolerance, egg laying preference, egg hatching and larval development times and resistance to common insecticides in Ae. aegypti collected from brackish and fresh water habitats in Jaffna, Sri Lanka. Brackish water-derived Ae. aegypti were more tolerant of salinity than fresh water-derived Ae. aegypti and this difference was only partly reduced after their transfer to fresh water for up to five generations. Brackish water-derived Ae. aegypti did not significantly discriminate between 10 ppt salt brackish water and fresh water for oviposition, while fresh water-derived Ae. aegypti preferred fresh water. The hatching of eggs from both brackish and fresh water-derived Ae. aegypti was less efficient and the time taken for larvae to develop into pupae was prolonged in 10 ppt salt brackish water. Ae. aegypti isolated from coastal brackish water were less resistant to the organophosphate insecticide malathion than inland fresh water Ae. aegypti. Brackish and fresh water-derived Ae. aegypti however were able to mate and produce viable offspring in the laboratory. The results suggest that development in brackish water is characterised by pertinent biological changes, and that there is restricted genetic exchange between coastal brackish and inland fresh water Ae. aegypti isolates from sites 5 km apart. The findings highlight the need for monitoring Ae. aegypti developing in coastal brackish waters and extending vector control measures to their habitats.     

Long lasting stickleback sperm; is ovarian fluid a key to success in fresh water?

Although the sperm of externally fertilizing fishes usually has a brief life span of up to a few minutes, this study showed that the sperm of the three-spined stickleback Gasterosteus aculeatus moved for several hours in brackish water and up to at least 10 h in the presence of ovarian fluid. Three-spined sticklebacks were able to spawn in waters ranging from full-strength sea water to fresh water, an ability unusual among fishes. The influence of salinity on sperm motility was examined, using three-spined sticklebacks from sea (salinity 30), brackish (5·5) and freshwater (0) populations. All three populations were found to have sperm with long motility periods in brackish water, lasting 165–270 min. Seawater three-spined sticklebacks had sperm motile for up to 65 min in sea water, whereas sperm from fresh- and brackish-water fish were quiescent in this medium. In fresh water, sperm from all three populations showed a very brief motility period, lasting <60 s. The presence of ovarian fluid, however, prolonged the motility period of sperm from both fresh- and brackish-water three-spined sticklebacks, for up to 7 and 10 h in fresh and brackish water, respectively, with some sperm found to be motile for up to 24 h. The results indicated that ovarian fluid created a favourable environment for the sperm and might have facilitated the three-spined sticklebacks' successful penetration of fresh water.     

Evolutionary bottlenecks in brackish water habitats drive the colonization of fresh water by stingrays

Species richness in freshwater bony fishes depends on two main processes: the transition into and the diversification within freshwater habitats. In contrast to bony fishes, only few cartilaginous fishes, mostly stingrays (Myliobatoidei), were able to colonize fresh water. Respective transition processes have been mainly assessed from a physiological and morphological perspective, indicating that the freshwater lifestyle is strongly limited by the ability to perform osmoregulatory adaptations. However, the transition history and the effect of physiological constraints on the diversification in stingrays remain poorly understood. Herein, we estimated the geographic pathways of freshwater colonization and inferred the mode of habitat transitions. Further, we assessed habitat‐related speciation rates in a time‐calibrated phylogenetic framework to understand factors driving the transition of stingrays into and the diversification within fresh water. Using South American and Southeast Asian freshwater taxa as model organisms, we found one independent freshwater colonization event by stingrays in South America and at least three in Southeast Asia. We revealed that vicariant processes most likely caused freshwater transition during the time of major marine incursions. The habitat transition rates indicate that brackish water species switch preferably back into marine than forth into freshwater habitats. Moreover, our results showed significantly lower diversification rates in brackish water lineages, whereas freshwater and marine lineages exhibit similar rates. Thus, brackish water habitats may have functioned as evolutionary bottlenecks for the colonization of fresh water by stingrays, probably because of the higher variability of environmental conditions in brackish water.     

Diet of otters (Lutra lutra) in closely associated freshwater, brackish and marine habitats in south-west Portugal

The diet of otters was studied in closely associated freshwater, brackish and marine habitats, from spraints collected on the Portuguese south-west coast over an 18-month period. In areas where marine prey was the only available resource, diet was dominated by blennies, wrasses and gobies, but other prey was taken in areas near coastal lagoons, marshes and estuaries. Eels and amphibians were considered the typical prey of freshwater habitats, and grey mullet the typical prey of brackish water habitats. If the inland habitats near the coast were large enough, otters preferred to forage there rather than in the sea. In one area where otters alternatively used marine and inland habitats, the former were used most extensively in autumn and winter, and the latter in spring and summer. It is suggested that otters prefer to forage inland rather than in the sea.     

Larval development of Aedes aegypti and Aedes albopictus in peri-urban brackish water and its implications for transmission of arboviral diseases

Aedes aegypti (Linnaeus) and Aedes albopictus Skuse mosquitoes transmit serious human arboviral diseases including yellow fever, dengue and chikungunya in many tropical and sub-tropical countries. Females of the two species have adapted to undergo preimaginal development in natural or artificial collections of freshwater near human habitations and feed on human blood. While there is an effective vaccine against yellow fever, the control of dengue and chikungunya is mainly dependent on reducing freshwater preimaginal development habitats of the two vectors. We show here that Ae. aegypti and Ae. albopictus lay eggs and their larvae survive to emerge as adults in brackish water (water with <0.5 ppt or parts per thousand, 0.5-30 ppt and >30 ppt salt are termed fresh, brackish and saline respectively). Brackish water with salinity of 2 to 15 ppt in discarded plastic and glass containers, abandoned fishing boats and unused wells in coastal peri-urban environment were found to contain Ae. aegypti and Ae. albopictus larvae. Relatively high incidence of dengue in Jaffna city, Sri Lanka was observed in the vicinity of brackish water habitats containing Ae. aegypti larvae. These observations raise the possibility that brackish water-adapted Ae. aegypti and Ae. albopictus may play a hitherto unrecognized role in transmitting dengue, chikungunya and yellow fever in coastal urban areas. National and international health authorities therefore need to take the findings into consideration and extend their vector control efforts, which are presently focused on urban freshwater habitats, to include brackish water larval development habitats.     

Apparent anion imbalance in the fresh water adapted eel

Summary On adapting brackish waterAnguilla anguilla to fresh water it was noted that, while the plasma sodium, magnesium,pCO₂ and pH were held reasonably constant, there was a substantial fall in chloride concentration (–33 mEq). The gradient of the linear correlations between plasma sodium and chloride also fell (brackish water gradient=0.92, fresh water gradient=0.21) indicating that a new pattern of plasma ion interrelationships was being established. Comparison with plasma Na/Cl ion ratios from other fishes suggested that this phenomenon was peculiar toA. anguilla. Corresponding with the very low plasma chloride levels plasma bicarbonate was four to five times that found in other fishes, and this was thought related to the finding that the haematocrit value almost doubled during adaptation to fresh water. In fresh water adapted fish a fall in plasma chloride was associated with a rise in plasma bicarbonate, however the charge compensation effect of this response was only partial as summing the common plasma cations and anions left an anion deficit of about 34 mEq to be accounted for.     

The effect of salty diets and gradual transfer to sea water on osmotic adaptation, gill Na+, K+-ATPase activation, and survival of brook charr, Salvelinus fontinalis, Mitchill

The intensity and duration of the period of osmotic disturbance during introduction of brook charr into sea water were decreased by introducing the fish according to a gradient of salinity over a period of 6 days. Survival in summer increased from 25 to 90% with the use of a salinity gradient. However, kinetics and levels of activation of the gill Na⁺, K⁺-ATPase were not affected by the mode used for introducing brook charr into sea water. Neither was its level of activity modified by the use of a salted diet when the fish were in fresh water. The addition of 8 and 12% of salt to the diet prevented the plasma electrolyte surge of concentrations during the first days in sea water. In very cold water, survival rate was also drastically improved by giving an 8% salted diet during the 6 weeks preceding the introduction into sea water. These results show that both salty diets and exposure to brackish water during 6 days help brook charr face osmotic stress and improve their survival rate when introduced into full-strength sea water. The combined use of these preconditioning strategies might facilitate rearing this species in sea cages or silos.     

Food niche variation of European and American mink during the American mink invasion in north-eastern Belarus

Understanding processes allowing the co-existence of ecologically similar species is important but difficult to study in community ecology. Introductions of alien species are unplanned experiments allowing investigation of co-adaptation of both native and invasive species over a short period. We analysed food niche differentiation between native European mink and alien American mink after invasion of the latter species in Belarus. European mink feed mainly on crayfish, frogs and fish whereas American mink prefer small mammals, fish and frogs. The diet of both species varied between seasons and during the period of alien mink invasion. Concurrent with the progress of American mink invasion, the European mink food niche has narrowed to feeding mainly on frogs, with the proportion of aquatic prey (fish and crayfish) in their diet drastically reduced. In contrast, the American mink food niche became wider during invasion. The breadth was stable but included a varied proportion of different prey categories: namely an increased proportion of aquatic prey and a decreased proportion of water vole and waterfowl. The increase in abundance of American mink saw a decrease in the proportion of larger prey in their diet. When American mink preyed more often on frogs, food niche overlap of both predators increased. This result suggests that arrival of an alien competitor reduced food abundance (exploitative competition) and caused a change in native mink diet.     

Brief migration of the grass puffer, Takifugu niphobles, to fresh water from salt water

The grass puffer, Takifugu niphobles, is a peripheral freshwater fish often seen in brackish water. In the Sai River, Miyazu, Japan, we found that the grass puffer migrate to fresh water from salt water in June and July. Most of the grass puffer stayed in fresh water for 3.6 h on average and returned to the sea during the day. A low salinity tolerance experiment indicated that the grass puffer can live in fresh water for 2 days but not for longer than 4 days. Based on these findings, we discuss the physiological effects and ecological implications of the migration.     

Ecology and seasonal distribution of Vibrio parahaemolyticus in aquatic environments of a temperate region

Abstract An environmental survey was done to study the ecology and distribution of Vibrio parahaemolyticus in 5 selected stations in Okayama Prefecture, which included fresh, brackish, and marine aquatic environments. Water and plankton samples were collected monthly for quantitative and qualitative analyses during the period October, 1987 to October, 1989 for V. parahaemolyticus . The pathogen was not detected from fresh water environments. A seasonality of the organism was observed in brackish and marine environments where average salinity ranged between 0.39 and 1.28%.Plankton samples yielded higher densities of V. parahaemolyticus compared with water samples. By applying several enrichment techniques, the pathogen was detected quite frequently during the winter months in the environments with temperatures ranging between 10 and 14°C. The identification following conventional tests, by the API 20E system and by serological methods reveal that the API 20E system is satisfactory to identify V. parahaemolyticus and further confirms that the serological method could be a simpler and more rapid procedure for V. parahaemolyticus identification.     

Changes of the serotonergic system activity in fish tissues during an increase of water salinity

Changes of environment parameters produce essential effects on the serotonergic system activity in the body tissues. This paper describes study of the serotonergic system activity in various tissues of half-migratory fish - the Caspian roach Rutilus rutilus caspicus and the bream Abramis brama orientalis - caught in the fresh and brackish water. Using the method of indirect solid-phase ELISA test, the serotonergic system activity was assessed by determining in the studied fish tissues of the amount of serotonin-modulating anticonsolidation protein (SMAP) that is in the linear correlation with serotonin level. A significant change of the SMAP level in the Caspian roach and bream brain was found under effect on increased water salinity. Analysis of the SMAP amount in the bream liver did not reveal any differences in the animals caught in the brackish and fresh water. At the same time, in the bream gill there was a small rise of the SMAP content in the fish caught in the brackish water. The revealed in this experiment elevation of the SMAP amount in the Caspian roach and bream brain reflecting a corresponding increase of the serotonergic system activity indicates involvement of adaptive readjustments in the animal body.     

Differences in osmotolerance in freshwater and brackish water populations of Theodoxus fluviatilis (Gastropoda: Neritidae) are associated with differential protein expression

The euryhaline gastropod Theodoxus fluviatilis is found in northern Germany in freshwater or in brackish water habitats in the Baltic Sea. Previous studies have revealed that individuals from both habitats are not distinguishable by morphological characters or by sequence comparison of DNA encoding 16S RNA or cytochrome C. As reported in this study, animals collected in the two habitats differ substantially in their physiological ability to adapt to different salinities. Comparison of accumulation rates of ninhydrin-positive substances (NPS) in foot muscle upon transfer of animals to higher medium salinities revealed that brackish water animals were perfectly able to mobilize NPS, while freshwater animals had only limited ability to do so. In an attempt to explore whether this difference in physiology may be caused by genetic differentiation, we compared protein expression patterns of soluble foot muscle proteins using 2D gel electrophoresis and silver staining. Of the 40 consistently detected protein spots, 27 showed similar levels in protein expression in animals collected from freshwater or brackish water habitats, respectively. In 12 spots, however, protein concentration was higher in brackish water than in freshwater animals. In four of these spots, expression levels followed increases or decreases in medium salinities. In a different set of 4 of these 12 spots, protein levels were always higher in brackish water as compared to freshwater animals, regardless of their physiological situation (14 days in artificial pond water or in medium with a salinity of 16‰). The remaining 4 of the 12 spots had complex expression patterns. Protein levels of the remaining single spot were generally higher in freshwater animals than in brackish water animals. These expression patterns may indicate that freshwater and brackish water animals of T. fluviatilis belong to different locally adapted populations with subtle genetic differentiation.     

Head‐shape polymorphism in Japanese eels Anguilla japonica in relation to differences of somatic growth in freshwater and brackish habitats

The age, total length (L_T), head shape and skull shape were investigated for 379 Japanese eels Anguilla japonica sampled in freshwater and brackish areas of the Kojima Bay–Asahi River system, Okayama, Japan, to learn about the differentiation process of head‐shape polymorphism. The relative mouth width (ratio of mouth width to L_T) of A. japonica > 400 mm L_T collected in fresh water was significantly greater than that of fish collected in brackish water. Growth rates of mouth width and the distance from the snout to the midpoint of the eyes (the ratio of width and distance to age, respectively) were not significantly different between freshwater and brackish‐water samples, whereas the somatic growth rate (the ratio of L_T to age) of freshwater samples was significantly lower than that of brackish‐water eel samples. These results suggest that the factors affecting head and somatic growth of A. japonica are not identical. According to these results and feeding patterns in each habitat reported by another study, it is suggested that somatic growth appears to play a significant role in the differentiation process of the head‐shape polymorphism in A. japonica, with the slow‐growing fish in fresh water becoming broad‐headed and the fast‐growing fish in brackish water becoming narrow‐headed.     

The feeding strategies of Liza ramada (Risso, 1826) in fresh and brackish water in the River Tagus, Portugal

The stomach contents of samples of the mugilid Liza ramada (Risso, 1826), captured at Alcochete (brackish water zone of the Tagus Estuary) differed from those from Vala Nova (freshwater zone of the same estuary).
In the freshwater zone of the estuary the mullet ingested preferentially planktonic micro-algae, while in the brackish water they ate benthic micro-algae.
The stomach contents from Vala Nova showed a higher diversity of food items, compared with those from Alcochete. At Alcochete the Naviculales (numerical frequency, 82%) was the most representative order, while at Vala Nova it was the Coscinodiscales (61%) and the Chlorococcales (16%).
In fresh water, the stomach contents contained about 25–4% organic matter, while at Alcochete only 8.7% was found.
In the brackish water zone the ingestion rhythm was related to the tidal cycle, attaining a maximum during the high tide and a minimum at low tide.
At Vala Nova the mullet ate regularly throughout the day. showing a slight increase at dawn and dusk. Apparently, the fish remained inactive by night, at both locations.     

Changes of the serotonergic system activity in fish tissues during an increase of water salinity

Changes of environment parameters produce essential effects on the serotonergic system activity in the body tissues. This paper describes study of the serotonergic system activity in various tissues of half-migratory fish—the Caspian roach Rutilus rutilus caspicus and the bream Abramis brama orientalis—caught in the fresh and brackish water. Using the method of indirect solid-phase ELISA test, the serotonergic system activity was assessed by determining in the studied fish tissues of the amount of serotonin-modulating anticonsolidation protein (SMAP) that is in the linear correlation with serotonin level. A significant change of the SMAP level in the Caspian roach and bream brain was found under effect on increased water salinity. Analysis of the SMAP amount in the bream liver did not reveal any differences in the animals caught in the brackish and fresh water. At the same time, in the bream gill there was a small rise of the SMAP content in the fish caught in the brackish water. The revealed in this experiment elevation of the SMAP amount in the Caspian roach and bream brain reflecting a corresponding increase of the serotonergic system activity indicates involvement of adaptive readjustments in the animal body.     

A pretreatment method for the determination of nitrate in brackish water and seawater based on the hydrazinium reduction technique

The hydrazinium reduction technique has so far been inapplicable to the determination of nitrate in brackish water and seawater due to interference from magnesium ions. We developed a pretreatment method for brackish water and seawater samples for the determination of nitrate based on the hydrazinium reduction technique. Magnesium ions in water samples were converted to a precipitate of a complex with oxine (8-quinolinol) at pH 9.5, and then the precipitate was centrifuged at 3000 rpm for 20 min. The extra oxine in the resulting sample (the supernatant liquid), which inhibits the reduction of nitrate to nitrite, was removed using a Sep-Pak C18 cartridge. Thus we achieved the preparation of a magnesium-free water sample. Using the hydrazinium reduction technique with the proposed pretreatment method, nitrate in brackish water and seawater as well as fresh water was quantitatively determined with high accuracy. Received: July 21, 1999 / Accepted: September 26, 1999     

Oxygen consumption rates of tilapia in fresh water, sea water, and hypersaline sea water

Whole animal oxygen consumption rates and plasma constituents were determined in the tilapia O. mossambicus , acclimated for 1 month in fresh water, sea water, and 1·6 × sea water. Oxygen consumption rates for the three water salinities were: 177·2 ± 16·86, 78·6 ± 2·32, and 195·4 ± 15·39 mg O₂ kg⁻¹ h⁻¹ (means ± 1 s.e.), respectively. Plasma prolactin (tPRL₁₈₈) concentration was significantly lower in 1·6 × sea water compared to fresh and sea water. There were no significant differences among mean plasma cortisol concentration and lysozyme activity. Ventilation was significantly higher in fish in sea water compared to the fish in fresh and 1·6 × sea water. The lowest oxygen consumption rates were found in fish acclimated to sea water. That salinity is probably closest to the brackish waters from which they were captured in the wild, and this agreement likely reflects the selection for optimal morphological and physiological characteristics to live in that environment.     

Blood pressure, electrolyte composition and renal studies of pike, Esox lucius, following administration of salmon calcitonin

Pike were adapted to both fresh water and brackish water (salinity 2.2‰) conditions prior to measurements of blood pressure, electrolyte composition and renal function both before and after administration of either 3.75 μg calcitonin kg⁻¹ body weight or acetate saline. This protocol was designed to test the ability of calcitonin to affect significantly the fluid and electrolyte status of fish exposed to varying salinity. Whilst in freshwater-adapted animals the plasma content of calcium and magnesium was significantly depressed following calcitonin injection, no corresponding effect was seen following 2.2‰ salinity brackish water-adaptation. Moreover, no significant changes were apparent in renal function of pike adapted to either medium. It is concluded from these experiments that calcitonin in pike may play some part in extra-renal control of plasma calcium and magnesium content in fresh water but that it is unable to affect significantly the measured plasma ion content of brackish water-adapted animals.     

Strain variation and geographic endemism in Streptococcus iniae

Twenty-six Israeli isolates of Streptococcus iniae from both marine and fresh/brackish water sources were compared with each other and with 9 foreign isolates. All the isolates were tentatively identified according to their biochemical profile. Direct sequencing of approximately 600 bp PCR products of the 16S rDNA confirmed their identification as S. iniae at the molecular level and revealed a new (one-nucleotide) variant among Israeli isolates, in addition to 2 variants that had been previously reported. Strain variation was further examined by subjecting the isolates to randomly amplified polymorphic DNA (RAPD) and amplified fragment length polymorphism (AFLP) analyses. The RAPD method allowed separation of the isolates into only 2 groups, one including 5 Israeli fresh/brackish water isolates and one including all the other isolates. The AFLP method grouped the Israeli marine isolates into one homogeneous cluster, although they had been obtained in different years (1995 to 2001) from different species of fish, and from wild (Red Sea) as well as cultured (both Mediterranean and Red Sea) sources. The Israeli fresh/brackish water isolates and foreign isolates separated into distinct entities that clustered at generally high degrees of similarity. The distance between the clusters of the Israeli marine and fresh/brackish water isolates indicates that the S. iniae streptococcosis that has been afflicting the aquaculture industries in the 2 environments in recent years was caused by distinct strains. AFLP showed superior discriminative properties over RAPD in detecting intraspecific variation and proved to be an important tool for the characterization of S. iniae. A correlation between strain variation and geographic endemism was established.