Effects of polyethylene and polyvinyl alcohol on growth and protease activity of Acanthopagrus schlegelii juveniles.

Many plastic products are used in aquaculture. Studying the toxicological effects of plastics differing in water solubility on marine organisms can provide valuable information. In this study, different amounts of polyvinyl alcohol (PVA) and polyethylene (PE) films were embedded in the feed and fed to Acanthopagrus schlegelii. After 1, 2, 4, and 8 weeks, the changes in percentage weight gain (PWG), feed efficiency (FE), pepsin activity (PA), and trypsin activity (TA) were observed. Either fed to PE or PVA, PWG, and FE of the experimental group were lower than those of the control group after eight weeks. Plastics level recovered from digestive tracts, feces, and water were higher in the PE groups than in the PVA groups. PA rose with increased feeding of plastics, but TA showed the opposite trend. TA activity decreased with increased feeding of plastics. Further analysis showed that TA was positively correlated with PWG.

Effects of Lead and Zinc Exposure on Uptake and Exudation Levels, Chlorophyll-a, and Phycobiliproteins in Sarcodia suiae.

The present study aimed to determine the changes in the biosorption, bioaccumulation, chlorophyll-a (chl-a), phycobiliproteins, and exudation in the red seaweed Sarcodia suiae exposed to lead and zinc. The seaweed was exposed to ambient lead and zinc environments for 5 days before being transferred to fresh seawater, and the changes in biodesorption, biodecumulation, chl-a, and phycobiliprotein levels in S. suiae were investigated. Lead and zinc biosorption and bioaccumulation in the seaweed increased with the increase in the lead and zinc concentrations and exposure times. Meanwhile, the biosorption and bioaccumulation of zinc in the seaweed following exposure to zinc were significantly higher (p < 0.05) than the biosorption and bioaccumulation of lead in the seaweed following exposure to lead with the same concentration at each exposure time. The chl-a, phycoerythrin (PE), phycocyanin (PC), and allophycocyanin (APC) contents in the seaweed significantly decreased with the increase in the lead and zinc concentrations and exposure times. The chl-a, PE, PC, and APC contents in S. suiae, which was exposed to 5 Pb2+ mg/L for 5 days, were significantly higher (p < 0.05) than those in the seaweed exposed to zinc at the same concentration and for the same exposure times. In the lead and zinc exudation tests, the highest biodesorption and biodecumulation were observed on the 1st day of exudation after the seaweed was transferred to fresh seawater. The residual percentages of the lead and zinc in the seaweed cells were 15.86% and 73.08% after 5 days of exudation, respectively. The biodesorption rate and biodecumulation rate of the seaweed exposed to lead were higher than those of the seaweed exposed to zinc. However, the effect of lead on chl-a and phycobiliproteins was greater than that of zinc. This might be the result of lead not being a necessary metal for these algae, whereas zinc is.

Microplastics in seawater and two sides of the Taiwan Strait: Reflection of the social-economic development.

Microplastic abundance, distribution and source characteristics were investigated for the surface seawaters from the Taiwan Strait as well as those of sediments along its west and east coasts. The microplastic abundances were in the range of 28-208 (mean 90) and 10-246 (mean 69) items/kg (d.w.) along the west and east coasts respectively. The higher microplastic abundance on the west coast might be related to the different local economic development, population, land-use and other human activities. Combined with microplastic pollution and socio-economic development, regression analysis results showed that urbanization level is negatively correlated with foams while positively correlated with fibers. This study, as the first report of microplastics in the Taiwan Strait, suggested further research on microplastics cross-strait transportation and the relationship with economic developments.

Effects of Cadmium on Bioaccumulation, Bioabsorption, and Photosynthesis in Sarcodia suiae.

This study investigated the changes in bioaccumulation, bioabsorption, photosynthesis rate, respiration rate, and photosynthetic pigments (phycoerythrin, phycocyanin, and allophycocyanin) of Sarcodia suiae following cadmium exposure within 24 h. The bioabsorption was significantly higher than the bioaccumulation at all cadmium levels (p < 0.05). The ratios of bioabsorption/bioaccumulation in light and dark bottles were 2.17 and 1.74, respectively, when S. suiae was exposed to 5 Cd2+ mg/L. The chlorophyll a (Chl-a) concentration, oxygen evolution rate (photosynthetic efficiency), and oxygen consumption rate (respiratory efficiency) decreased with increasing bioaccumulation and ambient cadmium levels. The levels of bioaccumulation and bioabsorption in light environments were significantly higher than those in dark environments (p < 0.05). In addition, the ratios of phycoerythrin (PE)/Chl-a, phycocyanin (PC)/Chl-a, and allophycocyanin (APC)/Chl-a were also higher in light bottles compared to dark bottles at all ambient cadmium levels. These results indicated that the photosynthesis of seaweed will increase bioaccumulation and bioabsorption in a cadmium environment.

Metals in Fishes from Yongshu Island, Southern South China Sea: Human Health Risk Assessment.

In order to assess the bioaccumulation of metals associated with gender, tissues, and their potential ecological risk, four species of fish were collected from the Yongshu Island in the Southern South China Sea. Metals and stable Pb isotopes in their tissues (muscle, gill, liver, intestine, and ovary) were determined. The concentrations of metals (mg/kg, dry weight) in these species were ND-21.60 (Cd), 1.21-4.87 (Cr), 0.42-22.4 (Cu), 1.01-51.8 (Mn), 0.30-3.28 (Ni), 6.04-1.29 × 103 (Zn), 14.89-1.40 × 103 (Fe), and 0.22-3.36 (Pb). In general, the liver and intestine absorbed more metals than the other tissues. Metals accumulation can be influenced by gender and feeding behavior and in fact, female fish and dietary exposure are more prone to accumulate metals. In addition, Pb isotopic ratios indicated that all species had significant biological fractionation, which may not make them good tracers for source identification. The metal concentrations of most samples were lower than the national standard values of the FAO (USA), which suggested that human consumption of these species may not cause health risks. However, since the surrounding areas are developing rapidly, the potential environmental risk of metals will intensify and should receive more attention.

Improved source apportionment of PAHs and Pb by integrating Pb stable isotopes and positive matrix factorization application (PAHs): A historical record case study from the northern South China Sea.

To obtain the historical changes of pyrogenic sources, integrated source apportionment methods, which include PAH compositions, diagnostic ratios (DRs), Pb isotopic ratios, and positive matrix factorization (PMF) model, were developed and applied in sediments of the northern South China Sea. These methods provided a gradually clear picture of energy structural change. Spatially, Σ15PAH (11.3 to 95.5ng/g) and Pb (10.2 to 74.6µg/g) generally exhibited decreasing concentration gradient offshore; while the highest levels of PAHs and Pb were observed near the southern Taiwan Strait, which may be induced by accumulation of different fluvial input. Historical records of pollutants followed closely with the economic development of China, with fast growth of Σ15PAH and Pb occurring since the 1980s and 1990s, respectively. The phasing-out of leaded gasoline in China was captured with a sharp decrease of Pb after the mid-1990s. PAHs and Pb correlated well with TOC and clay content for core sediments, which was not observed for surface sediments. There was an up-core increase of high molecular PAH proportions. Coal and biomass burning were then qualitatively identified as the major sources of PAHs with DRs. Furthermore, shift toward less radiogenic signatures of Pb isotopic ratios after 1900 revealed the start and growing importance of industrial sources. Finally, a greater separation and quantification of various input was achieved by a three-factor PMF model, which made it clear that biomass burning, coal combustion, and vehicle emissions accounted for 40±20%, 41±13%, and 19±12% of PAHs through the core. Biomass and coal combustion acted as major sources before 2000, while contributions from vehicle emission soared thereafter. The integrated multi-methodologies here improved the source apportionment by reducing biases with a step-down and cross-validation perspective, which could be similarly applied to other aquatic systems.

Fate of polycyclic aromatic hydrocarbons from the North Pacific to the Arctic: Field measurements and fugacity model simulation.

Polycyclic aromatic hydrocarbons (PAHs) have accumulated ubiquitously inArctic environments, where re-volatilization of certain organic pollutants as a result of climate change has been observed. To investigate the fate of semivolatile organic compounds in the Arctic, dissolved PAHs in the surface seawaters from the temperate Pacific Ocean to the Arctic Ocean, as well as a water column in the Arctic Ocean, were collected during the 4th Chinese National Arctic Research Expedition in summer 2010. The total concentrations of seven dissolved PAHs in surface water ranged from 1.0 to 5.1 ng L-1, decreasing with increasing latitude. The vertical profile of PAHs in the Arctic Ocean was generally characteristic of surface enrichment and depth depletion, which emphasized the role of vertical water stratification and particle settling processes. A level III fugacity model was developed in the Bering Sea under steady state assumption. Model results quantitatively simulated the transfer processes and fate of PAHs in the air and water compartments, and highlighted a summer air-to-sea flux of PAHs in the Bering Sea, which meant that the ocean served as a sink for PAHs, at least in summer. Acenaphthylene and acenaphthene reached equilibrium in air-water diffusive exchange, and any perturbation, such as a rise in temperature, might lead to disequilibrium and remobilize these compounds from their Arctic reservoirs.

Environmental and Ecological Risk Assessment of Trace Metal Contamination in Mangrove Ecosystems: A Case from Zhangjiangkou Mangrove National Nature Reserve, China.

Zhangjiangkou Mangrove National Nature Reserve is a subtropical wetland ecosystem in southeast coast of China, which is of dense population and rapid development. The concentrations, sources, and pollution assessment of trace metals (Cu, Cd, Pb, Cr, Zn, As, and Hg) in surface sediment from 29 sites and the biota specimen were investigated for better ecological risk assessment and environmental management. The ranges of trace metals in mg/kg sediment were as follows: Cu (10.79-26.66), Cd (0.03-0.19), Pb (36.71-59.86), Cr (9.67-134.51), Zn (119.69-157.84), As (15.65-31.60), and Hg (0.00-0.08). The sequences of the bioaccumulation of studied metals are Zn > Cu > As > Cr > Pb > Cd > Hg with few exceptions. Cluster analysis and principal component analysis revealed that the trace metals in the studied area mainly derived from anthropogenic activities, such as industrial effluents, agricultural waste, and domestic sewage. Pollution load index and geoaccumulation index were calculated for trace metals in surface sediments, which indicated unpolluted status in general except Pb, Cr, and As.

Changes in hemolymph oxyhemocyanin, acid-base balance, and electrolytes in Marsupenaeus japonicus under combined ammonia and nitrite stress.

Ammonia and nitrite are the most common toxic nitrogenous compounds in aquaculture ponds. We evaluated the effects of a combined treatment with these two compounds on the hemolymph acid-base balance, electrolytes and oxyhemocyanin content in kuruma shrimp, (Marsupenaeus japonicus). The shrimp (6.37±1.29 g) were individually exposed to 9 different ammonia and nitrite regimes {ammonia at 0 (control), 0.39, and 1.49 mM combined with nitrite at 0 (control), 0.38, and 1.49 mM} in a 30‰ saline solution at 22°C. Hemolymph oxyhemocyanin (OxyHc), protein content, acid-base balance, osmolality, and electrolyte levels were measured in treated shrimp after 48 h of treatment. Hemolymph OxyHc, protein content, the OxyHc/protein ratio, pH, pCO2, HCO3(-), TCO2, OH(-)/H(+), osmolality, and Cl(-), Na(+), K(+), Ca(2+), and Mg(2+) levels were inversely related to the dose of ammonia and nitrite. However, hemolymph pO2 levels directly increased with the ammonia and nitrite concentrations. Following exposure to 1.49 mM ammonia+1.49 mM nitrite, the hemolymph pO2 increased by 89.5%, whereas the hemolymph OxyHc, protein content, OxyHc/protein ratio, pH, pCO2, HCO3(-), TCO2, OH(-)/H(+), osmolality, Cl(-), and Na(+) decreased by 51.2, 28.2, 34.9, 2.9, 51.1, 71.5, 70.8, 42.8, 4.9, 32.1, and 38.6%, respectively, compared with control shrimp. Combined ammonia and nitrite stress may therefore exert a synergistic effect on shrimp relative to the stress induced by ammonia or nitrite alone.

Salinity and temperature tolerance of brown-marbled grouper Epinephelus fuscoguttatus.

Grouper have to face varied environmental stressors as a result of drastic changes to water conditions during the storm season. We aimed to test the response of brown-marbled grouper to drastic and gradual changes in temperature and salinity to understand the grouper's basic stress response. The results can improve the culture of grouper. Brown-marbled grouper, Epinephelus fuscoguttatus (6.2 ± 0.8 g) were examined for temperature and salinity tolerances at nine different environmental regimes (10, 20, and 33 ‰ combined with 20, 26 and 32 °C), in which the fish were subjected to both gradual and sudden changes in temperature and salinity. The critical thermal maximum (50 % CTMAX) and the upper incipient lethal temperature (UILT) were in the ranges of 35.9-38.3 and 32.7-36.5 °C, respectively. The critical thermal minimum (50 % CTMIN) and the lower incipient lethal temperature (LILT) were in the ranges of 9.8-12.2 and 14.9-22.3 °C, respectively. The critical salinity maximum (50 % CSMAX) and the upper incipient lethal salinity (UILS) were in the ranges of 67.0-75.5 and 54.2-64.8 ‰, respectively. Fish at temperature of 20 °C and a salinity of 33 ‰ tolerated temperatures as low as 10 °C when the temperature was gradually decreased. Fish acclimated at salinities of 10-33 ‰ and a temperature of 32 °C tolerated salinities of as high as 75-79 ‰. All fish survived from accumulating salinity after acute transfer to 20, 10, 5, and 3 ‰. But all fish died while transferred to 0 ‰. Relationships among the UILT, LILT, 50 % CTMAX, 50 % CTMIN, UILS, 50 % CSMAX, salinity, and temperature were examined. The grouper's temperature and salinity tolerance elevated by increasing acclimation temperature and salinity. On the contrary, the grouper's temperature and salinity tolerance degraded by decreasing acclimation temperature and salinity. The tolerance of temperature and salinity on grouper in gradual changes were higher than in drastic changes.

The effects of a sudden salinity change on cortisol, glucose, lactate, and osmolality levels in grouper Epinephelus malabaricus.

Grouper Epinephelus malabaricus (weighing 46.37 ± 5.10 g) previously maintained in 24‰ seawater were transferred to 14, 19, 24 (control), 29, and 34‰ seawater. Serum cortisol, glucose, lactate, and osmolality levels were measured at 7 time points during 240 min. Serum cortisol and glucose levels of fish transferred to 29 and 34‰ seawater significantly increased to the highest after 10 and 20 min, respectively. No significant differences in serum cortisol and glucose levels were observed for the fish after 30 min among all treatments. Serum lactate level of fish transferred to 14, 19, 29, and 34‰ seawater was significantly lower than that of the control fish after 10-30 min. However, no significant differences in serum lactate were observed 60 min among five treatments. The serum osmolality of the fish following 240-min transfer increased directly with salinity, whereas the osmoregulatory capacity value (medium osmolality-plasma osmolality) of the fish following 240-min transfer was inversely related to salinity. It is concluded that grouper showed strong osmoregulation in 14-34‰ seawater. Serum cortisol and glucose levels of fish transferred to 29 and 34‰ seawater increased rapidly in 10-30 min, indicating an early stress response.

Modulation of innate immunity and gene expressions in white shrimp Litopenaeus vannamei following long-term starvation and re-feeding.

The survival rate, weight loss, immune parameters, resistance against Vibrio alginolyticus and white-spot syndrome virus (WSSV), and expressions of lipopolysaccharide- and ß-glucan-binding protein (LGBP), peroxinectin (PX), prophenoloxidase-activating enzyme (ppA), prophenoloxidase (proPO) I, proPO II, α2-macroglobulin (α2-M), integrin ß, heat shock protein 70 (HSP70), cytosolic manganese superoxide dismutase (cytMnSOD), mitochondrial manganese superoxide dismutase (mtMnSOD), and extracellular copper and zinc superoxide dismutase (ecCuZnSOD) were examined in the white shrimp Litopenaeus vannamei (8.18 ± 0.86 g body weight) which had been denied food (starved) for up to 14-28 days. Among shrimp which had been starved for 7, 14, 21, and 28 days, 100%, 90%, 71%, and 59% survived, and they lost 3.2%, 7.3%, 9.2%, and 10.4% of their body weight, respectively. Hyaline cells (HCs), granular cells (GCs, including semi-granular cells), the total haemocyte count (THC), phenoloxidase (PO) activity, respiratory bursts (RBs), and SOD activity significantly decreased in shrimp which had been starved for 1, 1, 1, 5, 14, and 3 days, respectively. The expression of integrin ß significantly decreased after 0.5-5 days of starvation, whereas the expressions of LGBP, PX, proPO I, proPO II, ppA, and α2-M increased after 0.5-1 days. Transcripts of all genes except ecCuZnSOD decreased to the lowest level after 5 days, and tended to background values after 7 and 14 days. Cumulative mortality rates of 7-day-starved shrimp challenged with V. alginolyticus and WSSV were significantly higher than those of challenged control-shrimp for 1-7 and 1-4 days, respectively. In another experiment, immune parameters of shrimp which had been starved for 7 and 14 days and then received normal feeding (at 5% of their body weight daily) were examined after 3, 6, and 12 h, and 1, 3, and 5 days. All immune parameters of 7-day-starved shrimp were able to return to their baseline values after 5 days of re-feeding except for GCs, whereas all parameters of 14-day-starved shrimp failed to return to the baseline values even with 5 days of re-feeding. It was concluded that shrimp starved for 14 days exhibited three stages of modulation of gene expression, together with reductions in immune parameters, and decreased resistance against pathogens.

Orai1-STIM1 formed store-operated Ca2+ channels (SOCs) as the molecular components needed for Pb2+ entry in living cells.

Heavy metal lead (Pb2+) is a pollutant and causes severe toxicity when present in human tissues especially the nervous system. Recent reviews have suggested that Pb2+ can target Ca2+-related proteins within neurons and that Ca2+ channels might be a candidate for Pb2+ entry. This study's main aim was to identify the functional entry pathway of Pb2+ into living cells. We firstly characterized the endogenous expression of Orai1 and STIM1 mRNA together with the level of thapsigargin (TG) stimulated capacitative Ca2+ entry in PC12 and HeLa cells; this was done by RT-PCR and time-lapse Ca2+ imaging microscopy, respectively. Our data supported Orai1 and STIM1 as contributing to store-operated Ca2+ channel (SOC) basal activity. Secondly, using the indo-1 quenching method with the SOC blocker 2-APB, we observed that Pb2+ was able to enter cells directly through unactivated SOCs without TG pretreatment. Thirdly, we further demonstrated that co-expression of Orai1 and STIM1 differentially enhanced SOC functional activity (4-fold with PC12 and 5-fold with HeLa cells) and Pb2+ entry (5- to 7-fold with PC12 and 2-fold with HeLa cells). Furthermore, after a 1 h of Pb2+ exposure, the depolarization- and histamine-induced Ca2+ responses were significantly decreased in both PC12 and HeLa cells in a dose-dependent manner. This result indicated that the decreased Ca2+ responses were, in part, due to Pb2+ entry. In summary, our results suggest that SOCs are responsible for Pb2+ permeation and that the Orai1-STIM1 protein complex formed by functional SOCs is one of the molecular components involved in Pb2+ entry.

Effect of sulfide on the immune response and susceptibility to Vibrio alginolyticus in the kuruma shrimp Marsupenaeus japonicus.

Kuruma shrimp Marsupenaeus japonicus held in 34 per thousand seawater were injected with tryptic soy broth (TSB)-grown Vibrio alginolyticus (2.7x10(6)cfu shrimp(-1)), and then placed in water containing concentrations of sulfide at 0 (control), 51, 106, 528 and 1050microgl(-1), respectively. After 12-144h, mortality of V. alginolyticus-injected shrimp exposed to 528 and 1102microgl(-1) sulfide was significantly higher than that of shrimp exposed to 51microgl(-1) sulfide and the control solution. In another experiment, M. japonicus which had been exposed to 0, 56, 112, 525 and 1076microgl(-1) sulfide for 6, 12, 24 and 48h were examined for immune parameters, and phagocytic activity and clearance efficiency of V. alginolyticus. Sulfide concentrations at 525microgl(-1) or greater for 12h resulted in decreased total haemocyte count (THC) and phenoloxidase activity, phagocytic activity and bacterial clearance efficiency, whereas a sulfide concentration at 1076microgl(-1) for 24h caused a significant increase in respiratory burst and superoxide dismutase activity of M. japonicus. It is concluded that concentrations of sulfide at 528microgl(-1) or greater increased the susceptibility of M. japonicus against V. alginolyticus infection by a depression in immune ability. The increased production of superoxide anion by M. japonicus exposed to 525microgl(-1) sulfide or greater was considered to be cytotoxic to the host.

An increase of uricogenesis in the Kuruma shrimp Marsupenaeus japonicus under nitrite stress.

Kuruma shrimp Marsupenaeus japonicus Bate, under the stress of 0.36 and 1.39 mM nitrite at 30 per thousand (parts per thousand, g kg(-1)) for 48 h, were examined for nucleotide-related compounds, specific activities of xanthine dehydrogenase (XDH), xanthine oxidase (XOD), and uricase. The levels of total nucleotide-related compounds, including xanthine and hypoxanthine, in the gill increased directly with ambient nitrite, whereas the levels of total nucleotide-related compounds, including xanthine and hypoxanthine, in the hepatopancreas were inversely related to ambient nitrite. Specific activity of XOD in the hepatopancreas increased directly with ambient nitrite, whereas no significant difference in uricase activity in the hepatopancreas was observed among three treatments. In another experiment, M. japonicus, following 48 h exposure to 0.36 and 1.39 mM nitrite, were examined for ammonia, urea, and urate levels in tissues. Hemolymph urea and exoskeleton urate levels increased directly with ambient nitrite, whereas hemolymph urate and exoskeleton urea levels were inversely related to ambient nitrite. It is concluded that M. japonicus exhibited uricogenesis and uricolysis, and an increase of uricogenesis occurred for the shrimp under nitrite stress. Urate produced in the hepatopancreas was transported and accumulated in the epidermis, and removed along with the exoskeleton at the time of molting.

Increase of uricogenesis in the kuruma shrimp Marsupenaeus japonicus reared under hyper-osmotic conditions.

Tissues of kuruma shrimp Marsupenaeus japonicus Bate (5.7+/-1.1 g) reared in salinities of 18, 26, 34 and 42 were examined for levels of nucleotide-related compounds, ammonia, urea and uric acid, and activities of xanthine dehydrogenase (XDH), xanthine oxidase (XOD) and uricase. Levels of total nucleotide-related compounds, including xanthine and hypoxanthine, in gill increased directly with salinity, whereas these same levels in hepatopancreas were inversely related with salinity. Hemolymph ammonia, urea and uric acid levels, and epidermal ammonia, urea and uric acid levels increased directly with salinity, whereas hepatopancreas ammonia and uric acid and gill uric acid levels were inversely related to salinity. Activities of XDH and XOD in hepatopancreas increased directly with salinity level, whereas no significant difference of uricase activity in hepatopancreas was observed among the four salinities. It is concluded M. japonicus exhibited uricogenesis and uricolysis, and an increase of uricogenesis occurred for the shrimp under hyper-osmotic conditions (salinity of 42). Uric acid produced in the hepatopancreas was transported and accumulated in the epidermis, and removed along with the spongy connective tissue at the time of molting.

The immune response of the white shrimp Litopenaeus vannamei and its susceptibility to Vibrio infection in relation with the moult cycle.

The white shrimp Litopenaeus vannamei (8.0-14.4 g) was examined for haemocyte count, phenoloxidase activity, respiratory burst (release of superoxide anion), phagocytic activity, and clearance efficiency to the pathogen Vibrio alginolyticus in relation with moult cycle (postmoult, A, B; intermoult, C; premoult, D(0)/D(1)D(2)/D(3)). Granular cells were the highest at C and D(0)/D(1)stage, and the lowest at A stage. Hyaline cells and THC (total haemocyte count) were higher at C stage, but lower at postmoult stages. Phenoloxidase activity was the highest at C stage, and the lowest at A stage. Respiratory burst was lower at A stage. Phagocytic activity of shrimps against V. alginolyticus decreased significantly at postmoult and premoult stages. Additionally, the clearance efficiency of shrimps to V. alginolyticus was significantly lower for shrimps at A stage than those at C stage. In another experiment, L. vannamei at different moult stages were injected with tryptic soy broth (TSB)-grown V. alginolyticus (1x10(5)cfu shrimp(-1)) and then held in 34% seawater. After 10 h, the mortality of V. alginolyticus-injected shrimps was significantly higher for shrimps at postmoult stage than those at intermoult stage. Over 48-120 h, the mortality of V. alginolyticus-injected shrimps was 50.0%, 33.3% and 40.0% at postmoult, intermoult and premoult stage, respectively. It is concluded that L. vannamei showed a decrease in resistance at A stage through a reduction of its haemocyte count, phenoloxidase activity, respiratory burst, phagocytic activity and clearance efficiency against V. alginolyticus.

Study on the oxyhemocyanin, deoxyhemocyanin, oxygen affinity and acid-base balance of Marsupenaeus japonicus following exposure to combined elevated nitrite and nitrate.

Marsupenaeus japonicus (11.47+/-0.71 g) exposed individually to six different nitrite and nitrate regimes [(0.002 (control), 0.359 and 1.456 mM nitrite combined with 0.005 (control) and 7.458 mM nitrate)] in salinity of 30 ppt (parts per thousand) were examined for hemocyanin oxygen affinity, the fractionation of oxyhemocyanin and deoxyhemocyanin, and the acid-base balance after 24 h. Ambient nitrite at concentration of 0.359 mM caused reduction of oxyhemocyanin and protein by 27 and 11%, respectively, whereas ambient nitrate as high as 7.458 mM caused reduction of oxyhemocyanin and protein by 10 and 7%. Ambient nitrite at concentration of 1.456 mM caused increases of P(50) (indicating reduced oxygen affinity) and pO(2), but caused reduction in hemolymph pCO(2), pH, HCO(3)(-) and TCO(2). Following exposure to combined solutions of 1.456 mM nitrite and 7.458 mM nitrate there were no further changes in oxyhemocyanin, protein, hemolymph P(50), pO(2), pCO(2), HCO(3)(-) and TCO(2), but there was a significant reduction of pH.