Phosphorus Fractions in Soil and Water of Aquaculture Ponds Built on Clayey Ultisols at Auburn, Alabama

Phosphorus fractions in soil and water were determined for a 22-year-old, 400m² fish pond constructed on clayey Ultisols at Auburn, Alabama. This pond had been used in pond fertilization and fish feeding experiments each year. Total phosphorus concenlrations in bottom soil were greater in deep water than in shallow water areas. Highest concentrations of phosphorus occurred in the 5–10 cm soil layer, but phosphorus had accumulated above its original concentration to depths of 20 to 45 cm ( x = 36.8 cm). The soil phosphorus accumulation rate was 2.68 g/m² per year. Less than 1% of the total phosphorus in soils from three ponds was extractable in distilled water or 0.5 M NaHCO₃. Sequential extraction with 1 M NH₄Cl, 0.5 N HCl, and 0.1 N NaOH removed less than 25% of the total phosphorus. The loosely-bound phosphorus fraction (1 M NH₄Cl extractable) was 0.4 to 5.2% of the extractable phosphorus. The ability to adsorb phosphorus decreased and the capacity to release phosphorus increased in pond soils as total phosphorus concentration increased. After 22 years of aquacultural production, phosphorus adsorption sites in a pond soil were only about half-saturated. Although soluble phosphorus accounted for 37% of the phosphorus in pond water, only 7% of the total phosphorus in pond water was soluble reactive phosphorus. The phosphorus pool in pond soil was over 500 times greater than that of pond water, but most of the soil phosphorus was strongly adsorbed and unavailable.     

Purification and characterization of transglutaminase from squid gill

ABSTRACT: The present study used squid gill as a source of transglutaminase (TGase) because it has extremely high TGase activity compared with other tissues. The enzyme was purified using successive chromatographies of Sephacryl S-300 and hydroxyapatite columns. The yield and purification-fold of the enzymatic activity was 12.6% and 14.1-fold, respectively. The molecular mass of the purified enzyme was estimated to be 94 kDa by using sodium dodecylsulfate–polyacrylamide gel electrophoresis analysis. Enzyme activity was enhanced 15-fold with an increase in NaCl concentration. Although the activity was dependent on Ca²⁺ concentration, it was not sufficiently activated even by 50 mM CaCl₂ in the absence of NaCl, but could be fully activated with 10 mM CaCl₂ in 0.7 M NaCl. However, in the absence of substrates, the enzyme was rapidly inactivated. The pH and temperature optima of the enzyme were approximately pH 8.0 and 20°C, respectively. It was stable in the absence of Ca²⁺ at pH 7.5–9.0 and had a rate constant (K _D ) of 1.6 × 10^–5 s^–1 for thermal inactivation at 50°C. These results in which squid gill TGase could be activated at higher concentrations of Ca²⁺ and NaCl than at a physiological concentration, suggest that contact with seawater or body fluid seems to activate the enzyme if the tissue is disrupted.     

Effects of environmental Ca2+ levels on branchial chloride cell morphology in freshwater-adapted killifish Fundulus heteroclitus

ABSTRACT: To examine the involvement of chloride cells in the uptake of Ca²⁺ in freshwater (FW) killifish, chloride cell morphology was compared in fish acclimated to different defined FW environments with Ca²⁺ concentrations of either 0.1 mM, 0.5 mM, or 2.5 mM. Numerous chloride cells were detected in whole-mount preparations of the gill filaments, which were stained with an antiserum specific for Na⁺, K⁺-ATPase. Chloride cells, located mostly on the afferent–vascular edge of the filaments, were larger at lower Ca²⁺ concentrations. Electron microscopic observations showed that in the 0.1 mM and 0.5 mM Ca²⁺ experimental groups, the apical membrane of chloride cells were flat or slightly projecting and equipped with numerous microvilli. In the 2.5 mM Ca²⁺ group, some chloride cells formed an apical pit, whereas other cells were similar to those observed in the 0.1 mM and 0.5 mM Ca²⁺ groups. Plasma osmolality decreased with decreasing ambient Ca²⁺ concentration, suggesting that environmental Ca²⁺ affects the permeability of the body surfaces. Gill Na⁺, K⁺-ATPase activity in the 0.1 mM and 0.5 mM Ca²⁺ groups were significantly higher than that in the 2.5 mM Ca²⁺ group, implying the involvement of the Na⁺–Ca²⁺ exchanger in Ca²⁺ uptake in the gills. These findings suggest that chloride cells function as the site for Ca²⁺ uptake in killifish acclimated to low Ca²⁺ environments.     

Thermal denaturation and autolysis profiles of myofibrillar proteins of mantle muscle of jumbo squid Docidicus gigas

ABSTRACT:    Jumbo squid was very similar to Japanese common squid in terms of myofibrillar Ca²⁺-, Mg²⁺- and K⁺(EDTA)-ATPase activities. Myofibrils of jumbo squid were significantly stabilized upon addition of Ca²⁺ and destabilized by increasing KCl concentration for heating. Incubation of muscle homogenate of jumbo squid induced a selective cleavage of myosin into two major fragments and the cleavage was inhibited by EDTA. Autolysis was prominent at and above 0.3 M NaCl where myosin filaments dissolve. The enzyme involved in the autolysis was proved to be unstable showing maximal autolysis rate at 25°C. Washing the homogenate partially reduced the autolysis activity.     

Effect of pH and Decapsulation on the Toxicity of Ammonia to the Brine Shrimp Artemia franciscana

Artemia franciscana were hatched and tested for tolerance to ammonia at pH 6.5 and 8.5 in artificial seawater with a salinity of 17 ‰. Nauplii seemed to be less tolerant of ammonia at pH 6.5 despite the fact that more of the ammonia is in the ionized form (NH₄⁺). It is suggested that this greater sensitivity is a result of the NH₄⁺ competing with the Na⁺ during gut transfer. Nauplii hatched from decapulated cysts were more tolerant of ammonia than those hatched from whole cysts, probably because of a greater energy reserve.     

Purification and characterization of the amylase from a small abalone <Emphasis Type="Italic">Haliotis sieboldii</Emphasis>

ABSTRACT:   Amylase, with MW of 59 kDa, was purified from small abalone Haliotis sieboldii by ammonium sulfate fractionation, CM Sepharose Fast Flow and Sephacryl S-100 HR chromatographies. The optimal pH and temperature of purified amylase were 6.0 and 37°C, respectively. The purified enzyme was stable at pH 6.0–8.0 and low temperatures. It was activated by Ba²⁺, Mg²⁺, Ca²⁺, Co²⁺, Ni²⁺, Mn²⁺, K⁺, Ag⁺, Na⁺ and Li⁺, but completely or partially inhibited by Al³⁺, Cu²⁺, Cd²⁺, Hg²⁺ and Zn²⁺. EDTA could completely inhibit, while iodoacetamide, N-ethylmaleimide and urea partially inhibit the purified amylase. According to the digestion mode of various polysaccharides, the purified enzyme was considered to be an α-amylase.     

Differences in performance, flesh composition and water output quality in relation to density and feeding levels in rainbow trout, Oncorhynchus mykiss (Walbaum), farming

Abstract. The aim of this research was lo evaluate the effect of density and feeding level (initially 8 or 16kg/m³ and 1·3% or 2% of present biomass respectively) on productive traits, meat composition and water quality output in farmed rainbow trout, Oncorhynchus mykiss (Walbaum). Individual fish growth shows significant differences in relation to density and feeding level; feed conversion rate is positively Influenced by the lower feeding level and low rearing density. At low density, specific growth rate, protein efficiency ratio and coefficient of variation of individual final weights are more favourable than at high density. Dry matter and lipid composition of meat are positively affected by feeding level; whereas no differences are recorded on the amino acid and fatty acid profiles. Water quality shows differences due to density (NO₃ and NH₄⁺) and feeding level (NO₂, NH₄⁺ and P).     

Toxicity of Un-ionized Ammonia and High pH to Post-larval and Juvenile Freshwater Shrimp Macrobrachium rosenbergii

Post-larval and juvenile Macrobrachium rosenbergii were exposed for 72 h at 29 C to four pH levels (8.5, 9.0, 9.5, 10.0) and four concentrations of un-ionized ammonia-nitrogen (0, 1, 2, and 3 mg/L NH_j-N). Results indicated potentiation between NH₃ and high pH. Juveniles were more tolerant of high pH and NH₃ than post-larvae.
For post-larvae, estimates of 72 h LC50 for pH were 9.43, 9.21, and 8.71 at 0, 1, and 2 mg/L NH₃-N, respectively; 72 h LC50 estimates for NH₃-N were 2.18 and 1.45 mg/L at pH levels of 8.5 and 9.0, respectively. For juveniles, estimates of 72 h LC50 for pH were 9.91, 9.56, 9.04, and 8.76 at 0, 1, 2, and 3 mg/L NH₃-N, respectively; 72 h LC50 estimates for NH₃-N were 2.02 and 0.54 mg/L at pH 9.0 and 9.5, respectively.
In pond culture of M. rosenbergii , high pH levels can cause mortality at stocking. The 72 h data can be used as an indication of safe stocking levels of pH and ammonia. These data suggest that post-larvae should not be exposed to pH > 9.0 nor to NH₃-N > 1 mg/L in the pH range 8.5–9.0 and juveniles should not be exposed to pH > 9.5 nor to NH₃-N > 0 mg/L at pH 9.5, > 1 mg/L at pH 9.0, or > 2 mg/L at pH 8.5.     

Effects of Aeration, Alum Treatment, Liming, and Organic Matter Application on Phosphorus Exchange between Soil and Water in Aquaculture Ponds at Auburn, Alabama

Application of readily-oxidizable organic substrate to laboratory soil-water systems and fish ponds caused anaerobic conditions in bottom soil and water, and concentrations of soluble reactive phosphorus (SRP) increased. Aeration of ponds increased total phosphorus (TP) concentrations by suspending soil particles in the water, but SRP concentrations declined because of increased oxy- genation of bottom water and soil, Alum [Al₂(SO₄)₃·14H₂O] treatment of ponds reduced SRP and TP concentrations in ponds, but the low concentration of alum used, 20 mg/L, had little residual effect on phosphorus concentration. Application of agricultural limestone at 0.2 kg/m² to ponds with soil pH of 5.5 and Ca²⁺ concentration of 5 mg/L did not affect SRP and TP concentration. Unless pond soils were anaerobic at their surfaces, a condition not acceptable in thermally-unstratified fish ponds, soils released little phosphorus to the water. Strong adsorption of phosphorus by soils in intensive ponds with feeding is beneficial, because removal of phosphorus by aerobic soils is a control on excessive phytoplankton growth. In fertilized ponds, phosphorus must be applied at frequent intervals to replace phosphorus removed from the water by soils.     

Olfactory nerve response of masu salmon (Oncorhynchus masou Brevoort) and rainbow trout (O. mykiss Walbaum) to clove oil and MS-222

Two anaesthetics, clove oil and methane sulphonate (MS-222), were examined for their effects on the olfactory nerve response of masu salmon ( Oncorhynchus masou Brevoort) and rainbow trout ( O. mykiss Walbaum). Exposing both species to clove oil for 3 min at concentrations of 50 and 100 mg L⁻¹, or for 10 min at 50 mg L⁻¹, did not significantly reduce their olfactory response. Directly applying clove oil anaesthesia to the olfactory epithelium significantly reduced olfactory response though after 20 min, olfactory response recovered to 70% and 52% of pre-treatment levels in masu salmon and rainbow trout respectively. Compared with the post-anaesthetic recovery of responses after clove oil (50 mg L⁻¹), buffered MS-222 (100 mg L⁻¹) with NaHCO₃ (100 mg L⁻¹), and unbuffered MS-222 (100 mg L⁻¹) treatment for 3 min, the response after MS-222 treatment declined gradually and significantly, but not after clove oil and MS-222+NaHCO₃ treatments. Clove oil appears to be an effective and relatively safe anaesthetic for salmonids with little long-term impact on their olfactory response, which plays a crucial role in their life history.     

Toxicity of Ammonia and Nitrite to Penueus monodon Juveniles

Pcnaeus monodon juveniles (35.4 ± 2.2 mm TL) were exposed to seawater (20 ppt) having different concentrations of total ammonia (NH₃+ NH,+) and nitrite. Median lethal concentration (LC₅₀) was determined with static-renewal tests. The LC₅₀, of total ammonia-N, NH₃-N and niMte-N on shrimps decreased with increase of exposure time. The 24, 48, 72, % and 120 h LC₅₀ were 94.96, 61.09, 47.47 45.58 and 38.00 mg/L total ammonia-N (2.68, 1.73, 135, 1.29 and 1.08 mg/L NH₃-N) and 215.85, 185.33, 88.54, 54.76 and 37.97 mg/L nitrite-N, respectively. The 144h LC₅₀ of total ammonia-N and NH₃-N was 36.71 mg/L and 1.04 mgL. The "threshold" of ammonia and nitrite was found at 144h and 120h respectively, from the toxicity curve approaching asymptote. A "safe value" was 3.7 mg/L total ammonia-N, 0.1 mg/L NH₃-N (20 ppt, pH 7.70, 27C) and 3.8 mg/L nitrite-N for P. monodon juvenile .     

Role of prostaglandin in the control of ovulation in the Japaneseeel Anguilla japonica

ABSTRACT:   The in vitro effects of 17,20β-dihydroxy-4-pregnen-3-one(DHP) and prostaglandins (PGE₁, PGE₂, PGF_1α,PGF_2α) on ovulation in the Japanese eel Anguillajaponica were examined. Oocytes with follicle layers at themigratory nucleus stage (approximately 850–900 µmdiameter) were removed using a polyethylene cannula from artificiallymatured fish. At concentrations of 10 and 100 ng/mL,DHP was found to induce both germinal vesicle breakdown and ovulation.The prostaglandins, except for PGE₁, effectively inducedovulation of previously matured oocytes by DHP treatment in vitro .Prostaglandin F_2α was the most effective. Asignificant increase in ovulation rate was observed even at a concentrationof 0.01 µg/mL PGF_2α.Indomethacin blocked the in vitro ovulation induced by DHPand addition of PGF_2α reversed indomethacin-blockedovulation. Actinomycin D and cycloheximide blocked DHP-induced ovulationand PGF_2α reversed the effects of both inhibitors. Theseresults indicate that DHP induces ovulation through endogenous prostaglandinsynthesis in the follicle layers of the Japanese eel.     

Influence of photoperiod, frequency of stripping and presence of females on sperm output in turbot, Scophthalmus maximus (L.)

Abstract. The effect of four environmental conditions was investigated upon sperm output in turbot, Scophthalmus maximus (L.), submitted to three different rhythms of stripping. Males kept under a natural light cycle and under a 6-month contracted light programme released a similar sperm output in terms of total volume of semen produced per fish during the experimental period (4·9 ± 0·9ml), mean sperm concentration (29·4 ± 2·8 × 10⁹ spermatozoa/ml) and total sperm number (163·2 ± 40·5 × 10⁹ spermatozoa). Attempts to stimulate spermiation for a second time just after the end of the natural reproduction period resulted in the release of low sperm output (total volume of semen: 1·6 ± 0·4 ml; mean sperm motility: 2 min 36s ± 0 min 47s; mean sperm concentration: 47·6 ± 10·2 × 10⁹ spermatozoa/ml; total sperm number: 84·5 ± 25·3 × 10⁹ spermatozoa). Stripping frequency had no effect on total volume of semen, mean sperm motility and total sperm number. Monthly collection did not modify sperm samples in relation to stripping rank. However, decreasing volume, motility and sperm concentration were observed when males were stripped fortnightly and weekly. During the natural spawning period, the presence of females in the tank enhanced mean sperm motility (from 3 min 27s + 0 min 52s to 6 min 38s ± 1 min 38s).     

Immunolocalization of Na+, K+-ATPase-rich cells in the gill and urinary system of Persian sturgeon, Acipenser persicus, fry

Localization of Na⁺, K⁺-ATPase-rich cells in the gill and urinary system of Acipenser persicus fry was performed through immunofluorescence light microscopy using a mouse monoclonal antibody IgGα₅ raised against the α-subunit of chicken Na⁺, K⁺-ATPase. Different types of epithelia were clearly identified in the gill epithelium: epithelia of branchial arch, interbranchial septum, filament and lamellar epithelium. The Na⁺, K⁺-ATPase-rich cells were found in the epithelia of branchial arch, interbranchial septum, filament, interlamellar region and also in the lamellae. Histologically, the urinary system is divided into head kidney, trunk kidney and short caudal kidney. The head kidney is composed of the pronephric tubules and the haemopoietic tissues, while the trunk kidney is composed of a large number of glomeruli and convoluted nephrons. Each nephron consisted of a large glomerulus and tubules (neck, proximal, distal and collecting tubules) which connected to ureters. Posteriorly, ureters extended and joined together to form a small urinary bladder. In the urinary system, no specific fluorescence staining was observed in the glomerulus, neck segment and proximal tubules. The distal tubule cells and collecting tubule cells showed a strong immunostaining of Na⁺, K⁺-ATPase. Epithelia of ureters and urinary bladder also showed several isolated immunofluorescent cells. Immunofluorescent cells were rich in Na⁺, K⁺-ATPase enzyme which is very important for osmoregulation.     

Urine properties in carp, Cyprinus carpio L., with sekoke disease

Abstract. Changes in the properties of urine and blood of carp with sekoke disease were studied. With the progress of this disease, the parameters of blood and particularly urine changed significantly. After 120 days, urine flow, osmolarity and concentrations of inorganic ions (K⁺, Ca²⁺, Mg²⁺, Cl⁻) and organic compounds (ammonium, creatine and creatinine, protein, glucose) in the experimental group showed higher values than those of the controls. Even after 60 days, when clinical signs of sekoke disease were not evident, abnormally high concentrations of ammonium, protein and glucose in the urine were found in the experimental group.     

Metabolic costs of changing the cation–anion difference in the diet of juvenile African catfish Clarias gariepinus (Burchell)

The influence of dietary cation–anion difference (CAD, Na⁺ + K⁺– Cl^–, mEq kg^–1) on energy metabolism and nitrogen losses in juvenile African catfish Clarias gariepinus (Burchell) was examined in fish exposed to different dietary CAD levels (–146, 116, 497, 713 and 828 mEq kg^–1 diet). The experiment was conducted in open circuit balance respiration chambers over a 3-week period. Five 24-h monitoring periods were carried out at 3-day intervals during the experimental period with O₂ consumption, ammonia and nitrate + nitrite (NO_x) and CO₂ production being measured at 5-min intervals for each chamber. The negative dietary CAD (–146 mEq kg^–1) resulted in the highest energy expenditures (83 kJ kg^–0.8· d^–1). With increasing dietary CAD levels, heat loss gradually decreased to minimum values of 56 kJ kg^–0.8 day^–1 at a dietary CAD level of 713 mEq kg^–1. Consequently, metabolizable energy utilization efficiency (MEU, percentage of retained energy over metabolizable energy) quadratically ( P  < 0.05) increased and reached a maximum at a dietary CAD of 713 mEq kg^–1.     

Cryopreservation of Sperm of Farmed European Eel Anguilla anguilla

Sexual maturation and sperm release were induced in farmed European eels Anguilla anguilla kept exclusively in fresh water by using two dosages of human chorion gonadotropin (100 International Unit (IU)-Group one and 250 IU/individual per week-Group two). Sperm release took over 13 wk in both groups. The quality of sperm was investigated on the eighth, ninth, and tenth wk. The average cell densities were 0.94 ± 0.4 × 1010 (Group one) and 0.93 to. ± 0.6 × 10¹⁰ (Group two) spermatozoa/mL. The estimated motility of eel sperm was 33, 55, and 49% on the eighth, ninth, and tenth wk of treatment, respectively. The estimated average motility of samples selected for cryopreservation was 73 ± 10%, while the post-thaw motility of cryopreserved samples was 36 ± 11%. The extender originally developed for common carp sperm crypreservation together with methanol as cryoprotectant was found suitable for the cryopreservation of European eel sperm.     

Survival and Growth of Red Drum Sciaenops ocellatus in Saline Groundwaters of West Texas, USA

Abstract.— West Texas saline groundwaters were assessed as potential media for culturing red drum, Sciaenops ocellatus . In 30-d bioassays with juvenile red drum (0.4–3.0 g), highest survival was 85% in a 5-ppt (parts per thousand = g/L) salinity, high-sulfate (1,723 mg/L SO₄^-2), high-calcium (427 mg/L Ca^±2) groundwater from a windmill catchment pond in Pecos County. The lowest survival was 0% in a 35-ppt saline groundwater from a gravel pit, also in Pecos County. In complementary bioassays of low-salinity (<5 ppt) groundwaters from the same region, calcium chloride (CaCl₂) addition to a 3-ppt. Reeves County ground-water low in Ca^±2 resulted in the greatest increase in survival (0–93). It was unclear whether the beneficial effect of CaCl₂ was a result of increasing Ca^±2 concentration (36–336 mg/L), increasing Cl^- concentration (639–1,296 mg/L). or both. The concentration of total dissolved solids ("salinity") appears to be the single most important consideration in estimating the red drum aquaculture potential of a groundwater, with the optimum salinity being 5–15 ppt. Concentrations of arsenic (As), cadmium (Cd), chromium (Cr), mercury (Hg), lead (Pb), and selenium (Se) tended to be higher in west Texas groundwaters than in typical U. S. surface waters. However, based on current maximum tolerable daily intake recommendations, red drum cultured for 30 d in west Texas groundwaters contained whole-body As, Cd, Cr, Hg, Pb, and Se concentrations that would not pose a substantial health risk to adult human consumers.     

Effect of various medium compositions on survival and hatching rates of embryos of the giant freshwater prawn Macrobrachium rosenbergii cultured in vitro

SUMMARY: The effectiveness of in vitro embryo culture of the giant freshwater prawn Macrobrachium rosenbergii depended on the age of the embryos at the onset of culture and on the concentrations of various compositions in the medium. Embryos that started being cultured on day 0.5 after oviposition were more sensitive to variations in the medium compositions than those that started being cultured on day 10.5 after oviposition. An optimal NaCl level was essential for embryonic development, survival, hatching and survival of the newly hatched larvae. Variations of NaCl or KCl levels dramatically altered embryonic development, and variation of the MgCl₂ + MgSO₄ level significantly lowered survival of the embryos that started being cultured at the early stage of development. In contrast, no significant change in embryonic development was observed upon variation of the CaCl₂ level. Hatching of the embryos required the presence of NaCl and CaCl₂ but not KCl or MgCl₂ + MgSO₄. The ionic requirements of the newly hatched larvae differed from that of the developing embryos. Variations of NaCl, KCl or CaCl₂ but not MgCl₂ + MgSO₄ levels significantly influenced the survival of the newly hatched larvae.