Effect of Nutrient Media on the Growth,Physicochemical and Agar Properties of Gracilaria cliftonii Cultured in Ocean and Inland Saline Water

The effect of different nutrient media on growth, physicochemical, and agar properties of the marine algae Gracilaria cliftonii was investigated in ocean water (OW) and inland saline water (ISW). G. cliftonii fronds were cultured in 500-mL and 25-L containers with different nutrient media: Aquasol, Total, Guillard f2, Walnes, and Modified-Provasoli's Enriched Seawater in OW and ISW at 35 ppt. Specific growth rate (SGR) was higher in OW than ISW irrespective of nutrient media in both volumes. Growth of G. cliftonii in Guillard f2-supplemented OW was significantly higher than other nutrient media in both water types. Nutrient media had no influence on growth and agar properties. However, f2^?supplemented OW resulted in significantly higher agar yield (58.3% db) as compared to other media. Either f2 or Walnes media are preferred nutrient sources for culturing G. cliftonii in ocean and inland saline water.     

Survival, growth and osmolality of greenlip abalone Haliotis laevigata (Donovan 1808) when exposed to different ionic profiles of inland saline water

Two experiments over 21 days were conducted to assess the effect of different ionic profiles (mainly K⁺) of inland saline water (ISW) on the survival, growth, osmoregulation and moisture content of juvenile greenlip abalone Haliotis laevigata. In experiment I, the juveniles were cultured in five different water types: ocean water (OW) as a control, inland saline water (ISW or ISW K⁺₀), ISW fortified with 33%, 66% and 100% K⁺ as in OW (ISW K⁺₃₃, ISW K⁺₆₆ and ISW K⁺₁₀₀ respectively). In experiment II, five different culture media, 100% OW(control), 75% OW mixed with 25% ISW (ISW₂₅), 50% OW mixed with 50% ISW (ISW₅₀), 25% OW mixed with 75% ISW (ISW₇₅) and 100% ISW (ISW or ISW₁₀₀), were tested. Within 21 days, all of the abalone in ISW (ISW K⁺₀ and ISW₁₀₀; Na⁺/K⁺ ratio of 111 L^?1) died compared with 100% survival in OW. One‐hundred per cent K⁺ fortification of ISW increased the specific growth rate (SGR) and serum osmolality of the surviving juveniles. The 96 h acute LC₅₀ value was between ISW₅₀ and ISW₇₅. The increased proportion of ISW in OW resulted in significantly (P<0.05) lower survival, SGR and serum osmolality of juveniles. The moisture content of the juveniles was not influenced by any water type. The results suggested that abalone cannot survive in raw ISW, but survival can increase when it is fortified with K⁺ and ionic ratios of ISW are brought closer to OW by mixing it with OW. Furthermore, abalone survival is influenced by other ions in ISW besides K⁺.     

Productivity of Sargassum linearifolium in potassium fortified inland saline water under laboratory conditions

Growing aquatic species in inland saline water (ISW) is one way to reduce the adverse impact of ISW to agriculture farms. This 84‐day laboratory‐trial was conducted to study the growth of Sargassum linearifolium cultured in ocean water (OW), ISW, ISW fortified with potassium equivalent to 100% (ISW100), 66% (ISW66) and 33% (ISW33) of potassium in OW at 35 g/L. The biomass and cumulative specific growth rate (SGR) of S. linearifolium increased significantly (p < .05) with increased potassium in ISW until 56 days and then declined. The ISW100 and OW resulted in similar growth patterns and yielded peak biomass at day 42, proving static biomass for the next 28 days before declining. The biomass of S. linearifolium cultured in ISW and ISW33 significantly (p < .05) decreased and was lower than in ISW100 and died after day 56. The SGR of S. linearifolium in OW, ISW100 and ISW66 levelled off and showed no difference during the first 56 days. The S. linearifolium biomass and SGR negatively and significantly (p < .05) correlated with the concentrations of nitrate, phosphate in all waters. The increased potassium concentration in ISW similar to its concentration in SW brought the growth of S. linearifolium cultured to a level that was similar in OW.     

Hatchery seed production of giant freshwater prawn,Macrobrachium rosenbergii using inland ground saline water in India

The suitability of inland saline water (ISW) from the Lahli‐Baniyani Fish Farm, Rohtak was investigated for the larval rearing of giant freshwater prawn (GFP), Macrobrachium rosenbergii. Six experiments were conducted. In Experiment‐I, 54% of the larvae metamorphosed to postlarvae (PL) in constituted seawater (CSW) whereas total mortality occurred at larval stages (LS)‐II and LS‐III in ISW with salinity of 12 g L^?1. Larvae survived to LS‐IV in Experiment‐II, when ISW was supplemented with K⁺~ SW. In Experiment‐III, total hardness in ISW was reduced serially, but K⁺ ~ SW was supplemented. The larvae did not survive beyond LS‐V. In Experiment‐IV, ISW was amended with different ratios of Mg²⁺/Ca²⁺ and K⁺ ~ SW. The larvae successfully metamorphosed to postlarvae with highest survival of 51.6% in Mg²⁺/Ca²⁺ ratio of 2.5. In Experiment‐V, eight larval cycles were run with water quality used in Experiment V, where all the cycles produced PL's with a survival rate of 20–67%. In Experiment VI, the larvae were reared in Mg²⁺/Ca²⁺ ratio of 2.5 and different levels of K⁺ to optimize its requirement. The ISW amended with K⁺ 80% ~ SW and Mg²⁺/Ca²⁺ ratio of 2.5 was found to be commercially suitable for the seed production of GFP.     

Effects of salinity on growth characteristics and osmoregulation of juvenile cobia,Rachycentron canadum (Linnaeus 1766), reared in potassium‐amended inland saline groundwater

The suitability of inland saline groundwater as a medium to culture juvenile cobia, Rachycentron canadum, was assessed. In the first experiment, juvenile cobia stocked in raw (unamended) saline groundwater at salinities of 5, 10, and 15 g/L exhibited complete mortality after 108, 176, and 195 hr, respectively. The second experiment evaluated the rearing of juvenile cobia (mean weight ~9.23 ± 0.12 g) in potassium (K⁺)‐amended saline groundwater (100% K⁺ fortified) and reconstituted seawater at salinities of 5, 10, and 15 g/L to assess growth and osmoregulation in distinct culture media. Following 60 days of culture, all fish survived the experimental period. Final mean bodyweight of cobia reared in K⁺‐amended saline groundwater (103.2–115.8 g) and seawater (111.2–113.8 g) of different salinities did not vary significantly (p > .05). No differences (p > .05) were observed in specific growth rate, weight gain (%), and feed conversion ratio between treatment groups. Serum osmolality increased with salinity and was significantly higher (p < .05) for fish in K⁺‐amended saline groundwater (353–361 mOsmol/Kg) than in reconstituted seawater (319–332 mOsmol/Kg), although differences were not observed between salinities by water type. Cobia stocked in saline groundwater of different salinities were osmoregulating normally, and the higher values observed may be because of variations in ionic composition and other interfering ions in saline groundwater. Trial results suggest that juvenile cobia can achieve optimal growth in K⁺‐amended saline groundwater of low and intermediate salinities.     

Effects of different levels of dietary protein on the growth performance,nutrient utilization,digestive enzymes and physiological status of white shrimp,Litopenaeus vannamei juveniles reared in inland saline water

A 60‐day feeding trial was conducted to illustrate the effect of dietary protein levels on the growth and physio‐metabolic responses of juvenile Litopenaeus vannamei reared in inland saline water (ISW). Six isoenergetic (15 MJ/kg) and isolipidic (60 g/kg) diets with graded level of crude protein viz., 240 (T240), 260 (T260), 280 (T280), 300 (T300), 320 (T320) and 340 (T340) g/kg diet were formulated. Significantly higher (p < .05) weight gain (%), specific growth rate, with lower food conversion ratio were found in T320 and T340 groups. The protein utilizing efficiency and whole‐body protein content were significantly higher (p < .05) in the T320 group. Trypsin activity increased with the increasing dietary CP level but amylase activity decreased with the increasing dietary CP level. Transaminase enzymes, haemolymph protein and haemocyanin were elevated in T320 and T340 groups. The lactate dehydrogenase and malate dehydrogenase activities were significantly higher (p < .05) in the T240 group. Shrimp of T240 group had the lowest (p < .05) glycogen and total haemocyte count with highest (p < .05) haemolymph glucose and antioxidant enzymes activities than the other groups. Based on the results, feeding 320 g CP/kg is found to be optimum for supporting maximum growth and health status of L. vannamei reared in ISW at 8 g/L salinity. The finding of the present study will help in developing a low‐cost feed for L. vannamei reared in ISW.     

Effects of dietary protein on growth performance,nutrient utilization,digestive enzymes and physiological status of grey mullet,Mugil cephalus L. fingerlings reared in inland saline water

A 60 days feeding trial was conducted to illustrate the effect of graded levels of protein on the growth and metabolic enzymes of grey mullet (Mugil cephalus L.) fingerlings reared in inland saline water (ISW). Six isoenergetic (16 MJ/kg) and isolipidic (60 g/kg) diets containing 240, 260, 280, 300, 320 and 340 g crude protein (CP)/kg diet were formulated and fed to triplicate. Weight gain %, specific growth rate, protein utilizing efficiency, feed efficiency and RNA:DNA ratio were significantly higher (p < .05) in 320 and 300 g CP/kg diets. Fish fed with 240 g CP/kg diet showed significantly higher (p < .05) feed intake, whole‐body lipid content, hepatosomatic index value and liver glycogen content. Transaminase enzymes and malate dehydrogenase activities were elevated in fish fed 340 g CP/kg diet. Protease activity increased with increasing dietary CP level, but amylase activities showed an inverse relationship. No significant (p > .05) variations were observed for lactate dehydrogenase, oxidative stress enzymes, blood parameters and serum osmolality among all the treatment groups, but red blood cell count increases with increasing dietary CP levels. Based on the results, feeding dietary protein level of 300 g CP/kg is economically viable for rearing of grey mullet in ISW.     

Survival,growth and physiology of the juvenile razor clam (Sinonovacula constricta) under Na+/K+ ratio stress

In this study, we investigated the response of the Chinese razor clam (Sinonovacula constricta) under the Na⁺/K⁺ ratio in inland saline water. Acute and long‐term (30 days) stresses were studied to better delineate the process of the effect. At the end of the test, only the groups of Na⁺/K⁺ ratio from 21.28 to 159.95 contained surviving Juvenile S. constricta, and the Na⁺/K⁺ ratio 31.91 and 47.27 groups showed significantly higher survival rates (p < .001). In test groups of Na⁺/K⁺ ratio, the growth rate (shell length growth rate and weight gain rate) was significantly lower under than that the control group (p < .05 and p < .001 for length and weight). The Na⁺/K⁺‐ATPase activities in the test Na⁺/K⁺ ratio groups (R3, R5, R6, R7 and R8) were significantly higher than that in the control group during 30 days (p < .05). The superoxide dismutase activity in the higher Na⁺/K⁺ ratio groups (R7 and R8) was significantly higher than that in the control group during 30 days. However, in the test groups, the aspartate aminotransferase (R3, R6, R7 and R8), acetylcholinesterase (R3, R7 and R8) and lysozyme (R7 and R8) activities were lower than that control group during 30 days. Under ultrahigh and ultralow Na⁺/K⁺ ratio acute stress, the oxygen consumption rate, phagocytosis rate and metabolic activity fluctuated significantly. Unbalances of Na⁺/K⁺ ratio may affect ion regulation, metabolism, immunity and neuromodulation. Juvenile S. constricta adapted to a range of Na⁺/K⁺ ratios and keep to growth. This suggested the possibility of breeding S. constricta to resist inland saline water.     

Physiological characteristics and stress resistance of great sturgeon (<Emphasis Type="Italic">Huso huso</Emphasis>) juveniles fed with vitamins C,E, and HUFA-enriched <Emphasis Type="Italic">Artemia urmiana</Emphasis> nauplii

This study was carried out to examine the effect of Artemia urmiana nauplii enriched with HUFA, and vitamins C and E on stress tolerance, hematocrit, and biochemical parameters of great sturgeon, Huso huso juveniles. Cod liver oil (EPA 18% and DHA 12%), ascorbyl-6-palmitate and α-tocopherol acetate were used as lipid, and vitamin C and E sources, respectively. Beluga juveniles at the stage of first feeding (69.7 ± 5.9 mg body weight) were randomly divided into five treatments and three tanks were assigned to each diet. All fish groups were fed non-enriched Artemia for the initial 5 days and then fed enriched Artemia for 7 days. Juveniles were fed with Artemia enriched with HUFA + 20% vitamin C (C group); HUFA + 20% vitamin E-enriched Artemia nauplii (E group); HUFA + 20% vitamin C + 20% vitamin E (C and E group); HUFA without vitamins (HUFA) and non-enriched Artemia (control). After the period of enrichment, Juveniles were fed with Daphnia sp. from the 13th to the 40th day. At day 40, the fish were transferred directly from fresh water (0.5 ppt) to brackish water (6 ppt for 4 days and 12 ppt for 2 days) and warm water (from 27 to 33°C) to evaluate juvenile resistance to salinity and thermal shocks. Moreover, all treatments were separately exposed to freshwater in tanks with the same capacity as used for osmotic and thermal tests (as fresh water control). The addition of vitamins C, E, and C + E to HUFA significantly increased fish resistance to 12 ppt salinity and temperature stress tests, whereas survival was not significantly different among challenges at 6 ppt. There was no significant difference in the hematocrit index under stress conditions. Enrichment had significant influence on plasma Na⁺ level in the C group on the 4th day at 6 ppt. Na⁺ and Ca²⁺ concentrations in C, E, and C and E groups on the 1st day at 12 ppt, and Ca²⁺ level in E group on the 2nd day at 12 ppt were lower than the other groups. The glucose level in the C and C and E groups was lower than the other treatments on the 1st day at 12 ppt and the 2nd day at 33°C. Regardless of Artemia enrichment, plasma ions (Na⁺, K⁺, Ca²⁺, and Mg²⁺) and glucose concentrations in fish exposed to salinity stress tests were higher than fish in fresh water. Glucose concentration in plasma also increased after 2 days at 33°C. Although most of our results were not significantly different, the use of vitamins C, E, and HUFA in Artemia enrichment can improve Juveniles tolerance under stress conditions, and regardless of enrichment, these data show that beluga juveniles are partly sensitive to high salinity and temperature.     

Effects of Salinity and Na+/K+ Ratio on Osmoregulation and Growth Performance of Black Tiger Prawn,Penaeus monodon Fabricius, 1798, Juveniles Reared in Inland Saline Water

A 60‐d growth trial was conducted with the black tiger prawn, Penaeus monodon (ca. 0.8 g juveniles) at CIFE Rohtak Centre to evaluate the effects of salinity and Na⁺/K⁺ ratio of inland saline water on shrimp growth, survival, and osmoregulation. Three different salinities (5, 10, and 15 ppt) and five different Na⁺/K⁺ ratios (25:1, 45:1, 65:1, 85:1, and 27.9:1), for a total of 15 treatments were prepared by ionic manipulation. The medium with Na⁺/K⁺ ratio 27.9 was reconstituted seawater and was used as the reference treatment. At the end of the 60‐d trial both salinity and Na⁺/K⁺ ratio significantly influenced the survival and growth of shrimp in inland saline water (P < 0.05). Final mean individual weight, weight gain (%) (WG [%]) increased with decreasing Na⁺/K⁺ ratios. Survival rates were significantly higher (P < 0.05) for Na⁺/K⁺ ratio 45 and 27.9 at salinities 10 and 15 ppt, respectively. Minimum growth and survival (0–24%) were observed in mediums with Na⁺/K⁺ ratio 85 at all salinities. Serum osmolality and osmoregulatory capacity were similar across all treatments at identical salinities except for sodium to potassium ratio (Na⁺/K⁺) 85. The serum sodium and potassium levels did not show any significant difference (P > 0.05) for mediums with Na⁺/K⁺ ratio 25, 45, 65, and 27.9 at all salinities. Significantly different (P < 0.05) serum sodium levels were observed in mediums with Na⁺/K⁺ ratio 85 at all salinities at the end of the trial. The serum potassium levels were significantly low in treatments with Na⁺/K⁺ ratio 85. There was no significant difference in the serum magnesium levels between treatments and the serum calcium levels were significantly lower for shrimp reared in the reference mediums. Results of this study confirm that P. monodon can be successfully cultured in low salinity waters with Na⁺/K⁺ ratio ranging between 25 and 45:1.     

Osmoregulation in juvenile Chinese sturgeon (<Emphasis Type="Italic">Acipenser sinensis</Emphasis> Gray) during brackish water adaptation

The osmoregulation capabilities of 7-month-old juvenile Chinese sturgeon (Acipenser sinensis Gray) (128.8 ± 15 g) transferred directly from fresh water (0‰, 46 mOsmol kg⁻¹) to brackish water (10‰, 273 mOsmol kg⁻¹) were studied over a 20-day period. Changes in serum osmolarity, chloride (Cl⁻), sodium (Na⁺), potassium (K⁺) and calcium (Ca²⁺) ion concentrations, as well as gill and spiral valve Na⁺,K⁺-ATPase activities were measured at 3, 12, 24, 72, 216 and 480 h after transfer to BW. The serum osmolarity and ion concentrations (Na⁺, Cl⁻ and Ca²⁺) increased immediately after the transference to BW, reaching maximum at 24 h and returned to a new steady state at 216 h, while the FW control group maintained basal levels which showed lower (P < 0.05) than the BW group. Gill Na⁺,K⁺-ATPase activity of BW group exhibited an abrupt decrease in the first 3 h after transfer, but began to increase at 3 h, reaching a peak value at 24 h, and returned to a new steady state at 216 h. The differences between gill Na⁺,K⁺-ATPase activity of BW and FW fish were significant (P < 0.05) after 12 h. In contrast, Na⁺,K⁺-ATPase activity of the spiral valve showed transient increase after transference from FW to BW, and then decreased rapidly at 3 h, reaching the lowest at 24 h after transference. At 216 h after exposure to BW, Na⁺,K⁺-ATPase activities of the spiral valve increased slowly to the levels of FW control. The results of our study indicate the existence of hyposmoregulatory adaptive mechanisms in 7-month-old juvenile Chinese sturgeon which enable this fish to acclimate itself successfully to brackish water.     

Some Aspects of the Growth of Gracilaria birdiae (Gracilariales, Rhodophyta) in an Estuary in Northeast Brazil

The agarophyte Gracilaria birdiae Plastino & Oliveira was cultivated under field conditions in an estuary over a 6-month period. Biomass production varied significantly (p<0.01) and ranged from 900.0 to 3537.0 g m⁻² with a mean of 2124.4±1004.36 g m⁻² over the cultivation period. The epiphyte-biomass varied from 64.7 to 313 g m⁻², with maximum values observed in July. Relative growth rate (RGR) of G. birdiae varying from −0.59 to 4.67% day⁻¹, with a maximum mean observed in July. Correlation analysis showed that RGR was positively correlated with epiphyte-biomass (R-Pearson=0.69; p< 0.01) and negatively correlated with salinity (R-Pearson=−0.41; p<0.05). These correlations were used in a mathematical regression model to estimate the growth of Gracilaria in an estuary. The regression model explained 61% of RGR variability demonstrating that growth rates of Gracilaria were directly related to the variables salinity and epiphytes. The model was developed to help to predict production at a cultivation site in order to evaluate its suitability for Gracilaria cultivation.     

Responses of antioxidant enzymes,lipid peroxidation,and Na<Superscript>+</Superscript>/K<Superscript>+</Superscript>-ATPase in liver of the fish <Emphasis Type="Italic">Goodea atripinnis</Emphasis> exposed to Lake Yuriria water

Lake Yuriria, located in the heavily populated and polluted Mexican Central Plateau, receives domestic sewage, industrial effluents, and municipal wastewaters that are still directly discharged without treatment into the tributaries and the lake. Pollutants in water and sediments include heavy metals, aromatic hydrocarbons, and organochlorine pesticides. Activities of the antioxidant enzymes superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx), as well as Na⁺/K⁺-adenosine triphosphatase (Na⁺/K⁺-ATPase) activity, and lipid peroxidation (LPO) were evaluated in the livers of the fish Goodea atripinnis after 96 h of exposure to water collected in March and June 2005 from three sites: Y (limnetic zone), L (Lerma tributary), and C (la Cinta tributary). Physical and chemical parameters of the lake water were also analyzed. Increases in CAT activity and LPO levels at all three sites were detected compared with control fish (P < 0.05), while GPx and SOD activities decreased significantly (P < 0.05). Na⁺/K⁺-ATPase activities were similar to the control in fish exposed to limnetic water from both March and June but were higher than control at the two tributary sites in March (P < 0.05); fish exposed to water from the Lerma tributary in June exhibited lower Na⁺/K⁺-ATPase than the control (P < 0.05). During March, CAT and Na⁺/K⁺-ATPase activities were increasing more than in June in Y and L, respectively, while in June, SOD and GPx were depleted more than March in L and Y and L, respectively. Despite the antioxidant defenses of the fish liver, exposure to all water samples from Lake Yuriria exerted alterations in hepatic LPO levels, antioxidant enzymes, and Na⁺/K⁺-ATPase activities that could substantially impair the mechanisms of fish defenses against oxidative stress.     

Supplementation of potassium, magnesium and sodium chloride in practical diets for the Pacific white shrimp, Litopenaeus vannamei, reared in low salinity waters

The culture of Litopenaeus vannamei in inland low salinity waters is currently being practiced in various countries around the world. These environments are often deficient in key ions essential for normal physiological function, including potassium (K⁺) and magnesium (Mg²⁺). Farmers have sometimes been able to counteract ionic deficiencies in the water profile by adding mineral salts containing sources of K⁺ and Mg²⁺. The purpose of this study was to explore the possibility of correcting deficiencies of K⁺ and Mg²⁺ in the water profile with dietary supplementation of these minerals. Two separate 7‐week experiments were conducted in 4.0 g⁻¹ artificial low salinity water to evaluate the effects of mineral supplements (K⁺, Mg²⁺ and NaCl) to diets of L. vannamei reared in low salinity waters. In trial 1 seven diets were formulated (10 g NaCl kg⁻¹, 20 g NaCl kg⁻¹, 150 mg kg⁻¹ Mg²⁺, 300 mg kg⁻¹ Mg²⁺, 5 g K⁺ kg⁻¹, 10 g K⁺ kg⁻¹, and a basal diet to serve as a control). Minerals were added in the form of purified potassium chloride (KCl), magnesium chloride (MgCl₂·6H₂O) and NaCl. Trial 2 evaluated the use of a coating agent for the Mg²⁺ and NaCl treatments, while a K⁺ amino acid complex was utilized in the K⁺ treatments to reduce mineral leaching. Trial 2 was performed using similar treatment levels as trial 1. Shrimp survival and growth were assessed in both experiments. Results from trial 1 indicated no significant differences in survival, growth or percent weight gain. Results from trial 2 revealed no significant differences in survival and growth in the NaCl and Mg²⁺ treatments. However, significant differences in growth (P < 0.05) were observed when using the 10 g K⁺ kg⁻¹ treatment, suggesting that dietary supplementation of a K⁺ amino acid complex may help improve growth of the species in low salinity waters.     

Sperm of feral carp <Emphasis Type="Italic">Cyprinus carpio</Emphasis>: optimization of activation solution

The spermatozoa of oviparous fish, such as feral carp (Cyprinus carpio), are immotile in the presence of semen plasma or isotonic solutions, and to obtain good motility, they must be diluted with suitable medium. The objective of this study was to identify the best activating solution for feral carp sperm. Sperm motilities were compared in the new activating solution (a): (50 mM NaCl, 30 mM KCl, 30 mM Tris, pH = 8.5) and activating solution (b): (50 mM NaCl, 40 mM KCl, 30 mM Tris, pH = 8.5) based on effect of pH with everyone of Na⁺ and K⁺ ions versus four other activating solutions Billard’s saline solution, Poupard’s saline solution, distilled water and hatchery water that is routinely used for extending carp semen. Our results showed that maximum total motility period and percentage of motile sperm were seen in selected saline solution (a). The present study describes an activating solution that prolongs feral carp sperm motility.     

Mitochondrion-rich cells distribution,Na<Superscript>+</Superscript>/K<Superscript>+</Superscript>-ATPase activity and gill morphometry of the Amazonian freshwater stingrays (Chondrichthyes: Potamotrygonidae)

Detailed measurements of gill area and constituent variables (total filament number, total filament length and mean filament length), and immunolocalization of the α-subunit of Na⁺/K⁺-ATPase and Na⁺/K⁺-ATPase activity were performed on both hemibranchs of all five arches of freshwater potamotrygonid stingrays (Paratrygon aiereba and Potamotrygon sp.). Both species exhibit similar mass-specific gill area, 89.8 ± 6.6 and 91.5 ± 4.3 mm² g⁻¹ for P. aiereba and Potamotrygon sp., respectively. The density of Na⁺/K⁺-ATPase-rich MRCs and Na⁺/K⁺-ATPase activity was higher in the 4th gill arch in both species. The Na⁺/K⁺-ATPase activity was positively correlated to the Na⁺/K⁺-ATPase-rich (Na⁺/K⁺-ATPase rich) mitochondrion-rich cell (MRC) distribution among the gill arches of P. aiereba but not in Potamotrygon sp. The levels Na⁺/K⁺-ATPase activity were not correlated to the gill surface area among the arches for both rays’ species. Considering that the Na⁺/K⁺-ATPase-rich MRC is the main site for active ion transport in the gill epithelia and Na⁺/K⁺-ATPase activity plays a crucial role in osmoionoregulatory function, we suggesting that 4th gill arch is more relevant for osmoregulation and ion balance in these potamotrygonids.     

Effects of varying levels of aqueous potassium and magnesium on survival,growth, and respiration of the Pacific white shrimp,Litopenaeus vannamei,reared in low salinity waters

Inland shrimp culture is being practiced in several regions of the United States. In Alabama, the culture of shrimp (Litopenaeus vannamei) in inland low salinity well water (approximately 4.0 ppt) faces several challenges. The ionic composition of these waters is deficient in several key minerals, including potassium (K⁺) and magnesium (Mg²⁺). The objective of the present study was to evaluate the effects of several aqueous K⁺ and Mg²⁺ concentrations on survival, growth, and respiration in juvenile L. vannamei. Two experiments, a 14-day trial with postlarvae and a 7-week trial with juvenile (∼ 0.2 g) shrimp were conducted to evaluate effects of K⁺ supplementation to culture water. Four different levels of K⁺ (5, 10, 20, and 40 mg l^− 1) were utilized and a treatment of 4 ppt reconstituted seawater was used as a reference for comparison to ideal ionic ratios. Additionally, a 6-week growth trial (∼ 1 g juvenile shrimp) was performed to evaluate the effects of five concentrations of Mg²⁺ (10, 20, 40, 80, 160 mg l^− 1). Following completion of growth trials, measurements of basal respirometry rates were conducted to assess stress. Results from the 7-week K⁺ growth trial indicated significant differences (P < 0.05) in survival and growth among treatments. Individual weight, specific growth rate, and percent weight gain appeared to increase with increasing K⁺ concentration (decreasing Na:K ratios). Results from the Mg²⁺ experiment reveal a significant difference in survival between the lowest Mg²⁺ treatment (60%) and all other experimental treatments (90–97%). However, no differences in growth were observed. Shrimp respiration in the lowest Mg²⁺ treatment (10 mg l^− 1) was significantly higher than in the 80 mg l^− 1 treatment. These results suggest a potentially higher energetic cost associated with depressed aqueous Mg²⁺ concentrations that are common in low salinity environments.     

Partial characterization of alkaline proteases from viscera of vermiculated sailfin catfish Pterygoplichthys disjunctivus Weber, 1991

Vermiculated sailfin catfish (Pterygoplichthys disjunctivus, Weber, 1991), a member of the Loricariidae family and an invasive species of several inland waters around the world, possess an enormous digestive tract representing about 10% of fish weight. Thus, the aim of this study was to partially characterize proteases from their digestive tracts. Azocasein digestion of the crude extract of intestine at different pH values and temperatures revealed the presence of alkaline proteases with optimum activities at pH 9.0 and 50°C. Incubation assays of the crude extract with inhibitors such as phenyl methyl sulfonyl fluoride, N-α-p-tosyl-l-lysine chloromethyl ketone, N-tosyl-phenyalanine chloromethyl ketone, benzamidine, pepstatin A and ethylenediamine tetra-acetic acid showed that trypsin and chymotrypsin are the main alkaline proteinases present. Zymography showed that the crude extract of Pterygoplichthys disjunctivus viscera contained proteases with molecular masses ranging from 21.5 to 116 kDa. Trypsin and chymotrypsin were inhibited by the following ions in decreasing order: Hg²⁺, Fe²⁺, Cu²⁺, Li⁺, Mg²⁺, K⁺, while Mn²⁺, and Ca²⁺ had no effect. Activities decreased continuously as the NaCl concentration increased from 0 to 30%. These results constitute important background information for future studies and for the potential biotechnological use of the crude digestive extract from this invasive species.     

Some Limiting Factors in Superintensive Production of Juvenile Pacific White Shrimp,Litopenaeus vannamei,in No‐water‐exchange,Biofloc‐dominated Systems

Superintensive shrimp culture in zero‐exchange, biofloc‐dominated production systems is more biosecure and sustainable than traditional shrimp farming practices. However, successful application of this technology depends upon optimizing dietary formulations, controlling Vibrio outbreaks, and managing accumulative changes in water quality and composition. A 49‐d study investigated the effect of two commercial feeds of differing protein content and an indoor limited‐exchange, biofloc‐dominated culture environment on Litopenaeus vannamei performance and tissue composition, water quality and ionic composition, and Vibrio dynamics. Juveniles (5.3 g) were stocked at 457/m³ into four 40 m³ shallow raceways containing biofloc‐dominated water and fed one of two commercial feeds with differing protein content, 35 or 40%. Shrimp performance, Vibrio populations, and changes in shrimp and culture water composition were monitored. There were no significant differences (P > 0.05) in shrimp performance (survival, weight, growth, specific growth rate, total biomass, yield, feed conversion ratio, and protein efficiency ratio) or proximate composition between feed types. The 40% protein feed resulted in higher culture water nitrate and phosphate concentrations, alkalinity consumption and bicarbonate use, and higher phytoplankton density. The presence of Vibrio, specifically Vibrio parahaemolyticus, reduced shrimp survival. This survival decrease corresponded with increased culture water Vibrio concentrations. Culture water K⁺ and Mg²⁺ increased significantly (P < 0.05), and Sr²⁺, Br^?, and Cl^? decreased significantly (P < 0.05) over time. While Cu²⁺ and Zn²⁺ did increase in shrimp tissue, no heavy metals accumulated to problematic levels in culture water or shrimp tissue. These results demonstrate the importance of monitoring Vibrio populations and ionic composition in limited‐exchange shrimp culture systems.     

Lack of arterial PO<Subscript>2</Subscript> downregulation in Atlantic salmon (<Emphasis Type="Italic">Salmo salar</Emphasis> L.) during long-term normoxia and hyperoxia

Regulation of arterial partial pressure of O₂ (P_aO₂) in Atlantic salmon (Salmo salar) was investigated during resting conditions in normoxic and hyperoxic water. Dorsal aorta cannulated adult Atlantic salmon (1.2–1.6 kg, n = 8) were exposed to 2 week sequential periods of normoxia [16.7 ± 1.1 kPa (mean ± SD)] and hyperoxia (34.1 ± 4.9 kPa) in individual tanks containing seawater (33.7 ± 0.2 ppt) at stable temperature conditions (8.7 ± 0.7°C) and a light regime of L:D = 12:12. Tank design and sampling procedures were optimized to provide suitable shelter and current for the fish, and to allow repeated, undisturbed sampling of blood from free-swimming fish. Fish were sampled regularly through the experimental period. P_wO₂, P_aO₂, blood ion composition (Na⁺, K⁺, Cl⁻), acid–base status (pH, PCO₂, HCO₃ ⁻), haematocrit and glucose were measured. The most frequently observed P_aO₂ values were in the range of 60–80% of P_wO₂, both during normoxia and hyperoxia, and P_aO₂ values were significantly lower during normoxia than during hyperoxia. Blood pH, PCO₂ and HCO₃ ⁻ were significantly elevated during hyperoxia, while, Na⁺, Cl⁻ and Hct were significantly lower. K⁺ and glucose showed no significant differences. This study demonstrates a lack P_aO₂ regulation in Atlantic salmon to low partial pressures, in contrast to previous reports for many aquatic gill breathing animals. Both during normoxia and hyperoxia, P_aO₂ reflects P_wO₂, and alterations in external PO₂ consequently result in proportional arterial PO₂ changes. Physiological adaptation to hyperoxia, as illustrated by changes in several blood parameters, does not include down-regulation of P_aO₂ in Atlantic salmon. The lack of P_aO₂ regulation may make Atlantic salmon vulnerable to the oxidative stress caused by increased free radical formation in hyperoxic conditions.