Freshwater Levels Research Articles

Metabolic costs associated with seawater acclimation in a euryhaline teleost, the fourspine stickleback (Apeltes quadracus).

The cost of osmoregulation in teleosts has been debated for decades, with estimates ranging from one to 30 % of routine metabolic rate. The variation in the energy budget appears to be greater for euryhaline fish due to their ability to withstand dynamic salinity levels. In this study, a time course of metabolic and physiological responses of the euryhaline fourspine stickleback (Apeltes quadracus) acclimated to freshwater (FW) and then exposed to seawater (SW) was examined. There was 18% mortality in the first 3 days following exposure to SW, with no mortalities in the FW control group. Gill Na+/K+-ATPase (NKA) activity, an index of osmoregulatory capacity, increased 2.6-fold in SW fish peaking on days 7 and 14. Gill citrate synthase activity, an index of aerobic capacity, was 50-62% greater in SW than FW fish and peaked on day 7. Tissue water content was significantly lower in the SW fish on day 1 only, returning to FW levels by day 3. Routine metabolic rate was decreased within 24h of SW exposure and was maintained slightly (8-22%) but significantly lower in SW compared to FW water controls throughout the 2-week experiment. These results indicate that elevated salinity resulted in increased SW osmoregulatory and aerobic capacity in the gill, but with a reduced whole animal metabolic rate to this euryhaline species.

Bioaccumulation and human health risk assessment of heavy metals in food crops irrigated with freshwater and treated wastewater: a case study in Southern Cairo, Egypt.

Food safety has often attracted attention worldwide. Few studies have investigated the heavy metal (HM) pollution and health risk assessment of crops and vegetables. The current work was conducted to evaluate the human risk assessment of HM (Cu, Cd, Cr, Pb, and Zn) in radish, lettuce, tomato, onion, turnip, squash, okra, sunflower, Jews mallow, and garden rocket cultivated in treated wastewater (TWW)-irrigated sites as compared with those cultivated in freshwater (FW)-irrigated sites. Irrigation water, soil, and different plants were collected from 6 farmlands irrigated with TWW and two agricultural sites irrigated with FW (Nile river). Heavy metal transfer factor (HMTF), chronic daily intake of metals (CDIM), health hazard risk (HR), and health hazard index (HI) were estimated. The results showed that the tested HM levels in FW and TWW were below the Food and Agriculture Organization (FAO) and Egyptian standards recommended for irrigation. In soil samples, HM levels were below the permissible limits for both tested sites. The HM in soil and plants grew in TWW-irrigated sites possessed multiple levels higher than those grown in FW-irrigated sites. Among different plants, HM levels in the edible parts of plants grown in TWW-irrigated sites followed in decreasing order: tomato > sunflower >Jew's mallow = turnip = squash > lettuce > okra = radish > onion > garden rocket. The mean CDIM and HR values of plants irrigated using TWW were higher than those irrigated using FW. Furthermore, HR values for all plants grown in polluted and unpolluted sites were < 1 except Cd in plants grown in the TWW-irrigated farmlands. The mean HI for radish, lettuce, tomato, onion, turnip, squash, okra, sunflower, Jews mallow, and garden rocket grown in TWW-irrigated sites were 2.08, 2.39, 1.76, 1.53, 2.08, 1.80, 2.03, 1.91, 1.82, and 1.44 (for adult), and 2.39, 2.75, 2.71, 1.75, 2.38, 2.06, 2.33, 2.69, 2.10, and 1.65 (for children). Plants irrigated with TWW showed a higher HMTF than plants irrigated with FW. Jew's mallow and okra irrigated with TWW had a maximum HMTF. Consequently, different practical measures can be taken to minimize the HM levels in agricultural foodstuffs. These measures include preventing the excessive application of pesticides and fertilizers for crop production and continuous monitoring of different foodstuffs in the market.

Preparatory Mechanisms for Salinity Tolerance in Two Congeneric Anuran Species Inhabiting Distinct Osmotic Habitats.

Anurans occupy a wide variety of habitats of diverse salinities, and their osmoregulatory ability is strongly regulated by hormones. In this study, we compared the adaptability and hormonal responses to osmotic stress between two kajika frogs, Buergeria japonica (B.j.) and B. buergeri, (B.b.), which inhabit coastal brackish waters (BW) in the Ryukyu Islands and freshwater (FW) in the Honshu, respectively. Both hematocrit and plasma Na+ concentration were significantly higher in B.j. than in B.b. when both were kept in FW. After transfer to one-third seawater (simulating the natural BW environment), which is slightly hypertonic to their body fluids, their body mass decreased and plasma Na concentration increased significantly in both species. After transfer, plasma Na+ concentration increased significantly in both species. We examined the gene expression of two major osmoregulatory hormones, arginine vasotocin (AVT) and atrial natriuretic peptide (ANP), after partial cloning of their cDNAs. ANP mRNA levels were more than 10-fold higher in B.j. than in B.b. in FW, but no significant difference was observed for AVT mRNA levels due to high variability, although the mean value of B.j. was twice that of B.b. Both AVT and ANP mRNA levels increased significantly after transfer to BW in B.b. but not in B.j., probably because of the high levels in FW. These results suggest that B.j. maintains high plasma Na+ concentration and anp gene expression to prepare for the future encounter of the high salinity. The unique preparatory mechanism may allow B.j. wide distribution in oceanic islands.

Signal molecule changes in the gills and lungs of the African lungfish Protopterus annectens, during the maintenance and arousal phases of aestivation

African lungfishes are obligate air breathers, with reduced gills and pulmonary breathing throughout their life. During the dry season they aestivate on land, with the collapse of secondary lamellae of their gills and the establishment of an exclusive aerial ventilation through the vascularization and expansion of their lungs. To date, the mechanisms underlining the respiratory organ remodeling in aestivating lungfishes are unknown. This study aimed to identify key switch components of the stress-induced signal transduction networks implicated in both rapid and medium–long term remodeling of the gills and lungs of the African lungfish Protopterus annectens during aestivation. Through immunofluorescence microscopy and Western blotting, the localization and the expression of nitric oxide synthase (NOS), Akt, Hsp-90 and HIF-1α were evaluated in both gills and lungs exposed to three experimental conditions: freshwater (FW), 6 months of experimentally induced aestivation (6mAe), and 6 days after arousal from 6 months of aestivation (6mAe6d). After 6mAe, the expression of NOS (p-eNOS antibody), Akt (p-Akt antibody), and Hsp-90 decreased in the gills, while NOS and Hsp-90 expression increased with Akt remained unchanged in the lungs. Upon 6mAe6d, NOS, Akt and Hsp-90 expression in the gills returned to the respective FW values. In the lungs of the aroused fish, NOS and Akt decreased to their respective FW levels, while Hsp-90 expression was enhanced with respect to aestivation. In both respiratory organs, the qualitative and quantitative patterns of HIF-1α expression correlated inversely to those of NOS. Overall, our findings suggest that the molecular components of the NOS/NO system changed in a tissue-specific manner in parallel with organ readjustment in the gills and lungs of P. annectens during aestivation and arousal.

Differential effects of cortisol and 11-deoxycorticosterone on ion transport protein mRNA levels in gills of two euryhaline teleosts, Mozambique tilapia (Oreochromis mossambicus) and striped bass (Morone saxatilis)

The role of cortisol as the only corticosteroid in fish osmoregulation has recently been challenged with the discovery of a mineralocorticoid-like hormone, 11-deoxycorticosterone (DOC), and necessitates new studies of the endocrinology of osmoregulation in fish. Using an in vitro gill explant incubation approach, DOC-mediated regulation of selected osmoregulatory target genes in the gill was investigated and compared with that of cortisol in two euryhaline teleosts, Mozambique tilapia (Oreochromis mossambicus) and striped bass (Morone saxatilis). The effects were tested in gills from both fresh water (FW)- and seawater (SW)-acclimated fish. Both cortisol and DOC caused an up-regulation of the Na(+),K(+)-ATPase α1 subunit in SW-acclimated tilapia but had no effect in FW-acclimated fish. Cortisol conferred an increase in Na(+),K(+),2Cl(-) cotransporter (NKCC) isoform 1a transcript levels in FW- and SW-acclimated tilapia, whereas DOC had a stimulatory effect only in SW-acclimated fish. Cortisol had no effect on NKCC isoform 1b mRNA levels at both salinities, while DOC stimulated this isoform in SW-acclimated fish. In striped bass, cortisol conferred an up-regulation of Na(+),K(+)-ATPase α1 and NKCC transcript levels in FW- and SW-acclimated fish, whereas DOC resulted in down-regulation of these transcripts in FW-acclimated fish. It was also found that both corticosteroids may rapidly (30 min) alter the mitogen-activated protein kinase signalling pathway in gill, inducing phosphorylation of extracellular signal-regulated kinase 1 (ERK1) and ERK2 in a salinity-dependent manner. The study shows a disparate organisation of corticosteroid signalling mechanisms involved in ion regulation in the two species and adds new evidence to a role of DOC as a mineralocorticoid hormone in teleosts.

Effects of short-term acid and aluminum exposure on the parr-smolt transformation in Atlantic salmon ( Salmo salar): Disruption of seawater tolerance and endocrine status

Episodic acidification resulting in increased acidity and inorganic aluminum (Al i) is known to interfere with the parr-smolt transformation of Atlantic salmon ( Salmo salar), and has been implicated as a possible cause of population decline. To determine the extent and mechanism(s) by which short-term acid/Al exposure compromises smolt development, Atlantic salmon smolts were exposed to either control (pH 6.7–6.9) or acid/Al (pH 5.4–6.3, 28–64 μg l −1 Al i) conditions for 2 and 5 days, and impacts on freshwater (FW) ion regulation, seawater (SW) tolerance, plasma hormone levels and stress response were examined. Gill Al concentrations were elevated in all smolts exposed to acid/Al relative to controls confirming exposure to increased Al i. There was no effect of acid/Al on plasma ion concentrations in FW however, smolts exposed to acid/Al followed by a 24 h SW challenge exhibited greater plasma Cl − levels than controls, indicating reduced SW tolerance. Loss of SW tolerance was accompanied by reductions in gill Na +,K +-ATPase (NKA) activity and Na +,K +,2Cl − (NKCC) cotransporter protein abundance. Acid/Al exposure resulted in decreased plasma insulin-like growth factor (IGF-I) and 3,3′,5′-triiodo- l-thyronine (T 3) levels, whereas no effect of treatment was seen on plasma cortisol, growth hormone (GH), or thyroxine (T 4) levels. Acid/Al exposure resulted in increased hematocrit and plasma glucose levels in FW, but both returned to control levels after 24 h in SW. The results indicate that smolt development and SW tolerance are compromised by short-term exposure to acid/Al in the absence of detectable impacts on FW ion regulation. Loss of SW tolerance during short-term acid/Al exposure likely results from reductions in gill NKA and NKCC, possibly mediated by decreases in plasma IGF-I and T 3.

Identification of eel ghrelin in plasma and stomach by radioimmunoassay and histochemistry

The structure of ghrelin has been determined in the Japanese eel, Anguilla japonica. In this study, we identified immunoreactive ghrelin in extracts from plasma and stomach of the eel by radioimmunoassay (RIA) using an antiserum against octanoylated rat ghrelin [1–11]. Using the antiserum, we examined localization of ghrelin-immunopositive cells in the eel stomach. Detection of ghrelin mRNA-expressing cells was also attempted in the eel stomach using a cRNA probe specific for the eel ghrelin gene. Furthermore, we examined ghrelin expression patterns in plasma and stomach after transfer of freshwater (FW) eels to seawater (SW). Multiple types of immunoreactive ghrelin were detected using RIA. These were octanoylated eel ghrelin and other ghrelins that may have different fatty acid modifications, suggesting that this RIA can detect acylated ghrelin of eels as seen previously in the case of rat. Ghrelin-immunopositive cells were observed in the mucosal layer of the stomach, especially in the neck of the fundic gland. Ghrelin mRNA-expressing cells showed similar distribution and characteristics to the immunopositive cells. Plasma ghrelin levels in FW eels starved for one week before experimentation were approximately 40 fmol/ml. Plasma ghrelin levels in control-transferred FW eels did not change for 7 days, but significantly increased on day 14. Plasma ghrelin levels transiently increased fivefold 6 h after SW transfer and then declined to the FW level by 24 h after transfer. Ghrelin content and ghrelin mRNA levels in the stomach did not change after SW transfer, except for a transient decrease in ghrelin content seen 24 h after transfer. The present results suggest that ghrelin may participate in osmoregulation in eels.

Plasma and erythrocyte solute properties of juvenile bull sharks, Carcharhinus leucas, acutely exposed to increasing environmental salinity

Plasma and erythrocyte solute properties were examined in freshwater (FW) acclimated juvenile Carcharhinus leucas following acute transfer to 75% seawater (SW), and 100% SW. Blood samples were taken at 0, 12 and 96 h following transfer to 75% SW and 24 and 72 h after transfer to 100% SW. A control group in FW was subjected to the same sampling regime. Upon transfer of C. leucas to 75% and 100% SW, plasma Na +, Cl −, K +, Mg 2+, Ca 2+, urea and TMAO concentrations all increased significantly but disproportionately. Plasma Na + and Cl − increased immediately, followed by an increase in plasma urea. Erythrocyte urea and TMAO concentrations increased significantly following transfer to 75% and 100% SW; however, changes in erythrocyte inorganic ion concentrations were insignificant. Haematocrit, haemoglobin and mean cell haematocrit did not differ significantly after transfer to seawater; however, plasma water was slightly reduced after 24 and 72 h in 100% SW. Red blood cell (RBC) water content was elevated 24 h after transfer to 100% SW but returned to FW levels after 72 h. These results demonstrate that the transfer from 75% to 100% SW presents C. leucas with a greater osmoregulatory challenge than transfer from FW to 75% SW, despite the larger concentration gradient in the latter. In summary, C. leucas tolerate rapid and significant increases in salinity by rapidly increasing plasma osmolality to be hyperosmotic to the environment whilst maintaining a tight regulation of their intracellular fluid environment.

A Radioimmunoassay for the Determination of Arginine Vasotocin (AVT): Plasma and Pituitary Concentrations in Fresh- and Seawater Fish

A specific radioimmunoassay was developed and characterized for the measurement of arginine vasotocin (AVT) in teleost fish. Specificity of the antibody for AVT was demonstrated by parallelism of a series of AVT standards with serially diluted pituitary and plasma extracts. Crossreactivity of the antibody with the other teleost neurohypophysial peptide, isotocin, was less than 1% and the sensitivity of the assay was 0.24 fmol/assay tube. AVT was extracted from plasma by reverse-phase liquid chromatography [efficiency of 87.6 ± 9.3% ( n = 5)] and demonstrated as an effective procedure for plasma volumes ranging from 0.4 to 1.2 ml. Plasma AVT concentrations measured in a range of euryhaline and stenohaline teleost fish were between 10 -12 and 2 × 10 -11 M (1-20 pg/ml). There were no consistent differences between plasma AVT levels in euryhaline fish (flounder, trout, and eel) adapted to fresh water (FW) and sea water (SW). In flounder, pituitary AVT levels in FW- and SW-adapted fish were also similar.

Effect of environmental salinity changes on the secretory activity of prolactin cells in ocular pituitary transplants in Poecilia latipinna (teleostei)

Observations were made on plasma sodium levels and on the pituitary prolactin cells in Poecilia latipinna in one-third seawater ( 1 3SW ) and at various times up to 7 days after transfer to freshwater (FW). The pattern of adjustment of plasma sodium, an initial fall during the first 16–18 hr followed by restoration to normal FW levels at 3 and 7 days, was similar in both normal fish and in fish with the pituitary gland autotransplanted to the anterior chamber of the eye. In both groups the prolactin cells displayed histological signs of progressive increase in activity from 6 hr onwards. However, both plasma sodium values and prolactin cell nuclear area indicated that prolactin secretion in FW was rather lower in the autotransplants than in the in situ gland, although probably about normal in fish adapted to 1 3SW . It is suggested that this may indicate some partial inhibition of the prolactin cells by catecholamines in the eye tissues. Our main conclusion is that P. latipinna prolactin cells in vivo can be activated by reduced environmental salinity in the absence of hypothalamic connexions, and that the time-course of this activation follows closely the pattern in intact fish.

Adaptive variations in prolactin secretion in relation to external salinity in the teleost Poecilia latipinna

Measurements of plasma sodium levels, pituitary prolactin content, and detailed cytological studies of the pituitary prolactin cells were made on Poecilia latipinna adapted to different salinities and during adaptation to freshwater and 1 3 seawater. The prolactin band was identified in polyacrylamide gel disc electrophoregrams of pituitary homogenates. The optical density of the stained band was shown to be a measure of the amount of prolactin in the original homogenate. Plasma sodium is lowest in fish adapted to freshwater (FW), higher in 1 3 seawater ( 1 3 SW) and highest in seawater (SW). After transfer from 1 3 SW to FW, sodium falls during the first 18 hr, then rises to the normal FW range by 72 hr. After transfer from FW to 1 3 SW, sodium reaches 1 3 SW levels by 24 hr. Pituitary prolactin content is 6 times greater in FW than in SW, with intermediate levels in 1 3 SW. After transfer from 1 3 SW to FW prolactin content falls during the first 18 hr and then rises to FW levels by 8 days. The content remains high during the first 24 hr after transfer from FW to 1 3 SW. Content of putative growth hormone does not change with salinity. Cytological data show the prolactin cells to be more numerous and more active in FW than in SW; in 1 3 SW they are only slightly more active than in SW. After transfer from 1 3 SW to FW, cytological indices show that prolactin synthesis progressively increases, with loss of storage granules during the first 30 hr and return of granulation by 72 hr. The synthetic rate appears to be reduced 24 hr after transfer from FW to 1 3 SW. Considering all the data, it is concluded that in fish adapted to their surrounding medium, prolactin secretion rate is inversely proportional to salinity and directly proportional to pituitary prolactin content. During adaptation to new salinities, however, prolactin content and secretion rate are temporarily independent. Prolactin content in all circumstances closely parallels the granulation and size of the prolactin cells. The findings are discussed in relation to the known osmoregulatory effects of mammalian prolactin in teleosts, and are taken as confirming the physiological role of endogenous prolactin in adapting sodium metabolism to external salinity in P. latipinna.