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 % CT_MAX) 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 % CT_MIN) 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 % CS_MAX) 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 % CT_MAX, 50 % CT_MIN, UILS, 50 % CS_MAX, 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.     

Effects of Heavy Metals on the Hatching Rates of Brine Shrimp Artemia salina Cysts

The effects of heavy metals (Hg²⁺, Cd²⁺, Co²⁺, Cr³+, Pb²⁺, Cu²⁺, Fe²⁺. Ni²⁺, Zn²⁺, Mn⁷⁺, Mn⁴⁺ and Mn²⁺) on the relative hatching percentage of brine shrimp, Artemia salina were assessed. Artemia salina cysts (San Francisco Bay Brand) hatched in seawater containing various heavy metal concentrations. After 48 hours, the number of hatched nauplii were counted. A negative linear relationship between hatching rate of Ariemia salina cysts and heavy metal concentration occurred, except copper which conformed to a cubic model. The relative toxicities of heavy metals on Artemia salina cysts were as follows (Cu²⁺, most toxic; Mn²⁺, least toxic): Cu²⁺ > Zn²⁺ > Ni²⁺ > Co²⁺ > Mn⁷⁺ > Cr³⁺ > Cd²⁺ > Fe²⁺ > Hg²⁺ > Mn⁴⁺ > Pb²⁺ > Mn²⁺.     

Dietary administration of sodium alginate and kappa-carrageenan enhances the innate immune response of brown-marbled grouper Epinephelus fuscoguttatus and its resistance against Vibrio alginolyticus

Brown-marbled grouper Epinephelus fuscoguttatus which had been fed diets containing sodium alginate and kappa (kappa)-carrageenan at 5, 10 and 20gkg(-1), respectively after 0, 2, 4, 6, 8 and 14 weeks were examined for survival, growth, innate cellular and humoral responses as compared to the fish that fed control non-supplemented diet. Survival was 100% for the fish that fed all diets after 14 weeks and no significant difference in growth was observed among seven diets. The fish that fed a diet containing sodium alginate at 10gkg(-1), and fed a diet containing kappa-carrageenan at 5gkg(-1) over 2-8 weeks showed significantly increased leucocyte count, respiratory burst, phagocytic activity and phagocytic index, The fish that fed a diet containing sodium alginate at 5gkg(-1), and fed a diet containing kappa-carrageenan at 5gkg(-1) over 2-8 weeks showed significantly increased alternative complement activity (ACH50) and lysozyme activity. In another experiment, E. fuscoguttatus, which had been fed control diet, and all diets containing sodium alginate, and kappa-carrageenan after 14 weeks were challenged with Vibrio alginolyticus at 5.0x10(9) colony-forming units (cfu) fish(-1) and then placed in seawater of 34 per thousand. The survival of fish that fed a diet containing sodium alginate at 10gkg(-1) and kappa-carrageenan at 5gkg(-1) was significantly higher than that of fish which fed the control diet over 96-168h. It was concluded that E. fuscoguttatus which fed a diet containing sodium alginate at 10gkg(-1) or less, or fed a diet containing kappa-carrageenan at 5gkg(-1) enhance the innate immunity, and increase the resistance from V. alginolyticus infection.     

Super Intensive Culture of Red-Tailed Shrimp Penaeus penicillatus

Two continuous grow-out experiments of red-tailed shrimp Penueus penicillatus were conducted for one year in a super intensive system with a stocking density of 171 and 286 postlarvae/m², respectively. The postlarval shrimp (Ph_8–12) were cultured in three 0.14 ha earthen ponds with concrete dikes. The shrimp were fed with a commercially prepared feed four times a day. Water quality parameters from the second trial varied as follows: water temperature from 23.0 to 30.4 C, salinity from 15.33 to 21.00 ppt, dissolved oxygen (DO) from 3.47 to 7.34 mg/L, NH₃-N from 0.002 to 0.869 mg/L, nitrite-N from 0.013 to 0.844 mg/L, and nitrate-N from 0.021 to 1.795 mg/L. Ammonia-N increased from 0.022 to 46.110 mg/L, while the pH declined from 8.12 to 7.32. Yield was 4,650 kg/0.42 ha from the first crop in 131 days and 5,160 kg/0.42 ha from the second crop in 141 days. The productivity of this system producing P. penicillatus was 11–12 tons/ha/crop.     

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 .     

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.     

Effect of water temperature on the immune response of white shrimp Litopenaeus vannamei to Vibrio alginolyticus

White shrimp Litopenaeus vannamei held in 25‰ seawater at 27 °C or 28 °C were injected with TSB-grown Vibrio alginolyticus at 1 × 10⁴ colony-forming units (cfu) shrimp^− 1 or 1 × 10⁵ cfu shrimp^− 1, and then cultivated onward at water temperatures varying from 20 to 34 °C. Over 24–144 h, mortality of V. alginolyticus-injected shrimp held at 34 °C or 32 °C was significantly higher than that of shrimp held at lower temperatures. In a separate experiment, shrimp held in 25‰ seawater at 28 °C and then cultured onward at 20 to 32 °C were examined for immune parameters at 24–96 h. THC, phenoloxidase activity, respiratory burst, and SOD activity decreased significantly at 24 h after transfer to 32 °C. Shrimp held in 25‰ seawater at 27 °C and then cultured onward at 20 to 34 °C showed a significant reduction in phagocytic activity and clearance efficiency for V. alginolyticus at 24 h after transfer to 34 °C. It was concluded that transfer of shrimp from 27 or 28 °C to higher temperatures (32 and 34 °C) reduced their immune capability and decreased resistance to V. alginolyticus infection.     

Hemolymph Oxyhemocyanin and Protein Levels and Acid-Base Balance in the Tiger Shrimp Penaeus monodon Exposed to Copper Sulfate

Abstract.— Subadult tiger shrimp Penaeus monodon (30.15 ± 2.19 g) were exposed individually in sea water of 25%o to 0 (control), 1.11, 4.76, 10.06 and 19.09 mg/L copper for 24 h. Hemolymph pH, PO₂ (partial oxygen pressure), PCO₂ (partial carbon dioxide pressure), HCO₃, oxyhemocyanin and protein levels were determined. Hemolymph PO₂ increased, whereas hemolymph pH, PCO₂, HC0₃-, oxyhemocyanin. protein and the ratio of oxyhemocyanidprotein levels decreased with increasing concentrations of ambient copper in the range of 1.11 to 19.09 ma/L. It is concluded that P. monodon following 24-h exposure to ambient copper as low as 4.76 mg/L shows a reduction of oxyhemocyanin, protein, and the ratio of oxyhemocyanin to protein, and develops metabolic acidosis in the hemolymph.     

Lethal Effect of Nitrite on Metapenaeus ensis Larvae

Metupenaeus ensis larvae at different stages were exposed to a series of nitrite-N (nitrite as nitrogen) concentrations in static renewal toxicity tests. Larvae at zoea stage were the most susceptible, and postlarvae were the most tolerant to nitrite among the larvae tested. The 24 h LC₅₀s were 31.29, 16.05, 47.60 and 70.06 mg/L nitrite-N for third nauplius substage (N3). second zoea substage (Z2), second mysis stage (M2), and first postlarva substage (PL1), respectively, in 33 ppt seawater at a pH of 8.20 and a water temperature of 30 C. The 48 h LC₅₀ for M2 and PL1 were 20.67 and 27.10 mg/L nitrite-N, respectively. The "threshold time" was 108 h, and the "incipient LC₅₀" for M. ensis PL1 was 7.06 mg/L nitrite-N. A "safe level" for rearing M. ensis larvae was estimated to be 0.71 mg/L nitrite-N in the hatchery.     

Accumulation of Heavy Metals in the Nauplii of Artemia salina

Artemia salina cysts (San Francisco Bay Brand) were exposed in seawater containing multiple concentrations of either a single heavy metal (Hg, Cu, Zn, Pb, Cd, Co, Ni, Cr, Mn and Fe) or combination of heavy metals. The heavy metal accumulation in Artemia nauplii increased linearly with an increase in the heavy metal concentration in water. The metals could be detected in nauplii exposed to a single heavy metal at dosages of 5 μg/L for Hg, Zn, Mn, Fe and Cu; 25 μg/L for Cd and Cr; 50 μg/L for Co; 100 mUg/L for Ni; and 250 μg/L for Pb. Cysts exposed to several heavy metal combinations had minimum effective dosages equal to or lower than the single heavy metals- 5 μg/L for Cu, Cr, Zn, Mn and Fe; 25 μg/L for Cd and Ni; 50 μg/L for Co; and 250 μg/L for Pb. The accumulation of Cu, Ni and Co by Artemiu nauplii did not differ for animals exposed to single or combined heavy metal solutions. Cr, Zn and Pb had a synergistic effect of heavy metal uptake in combined heavy metal element solutions, however, Fe, Cd and Mn had an antagonistic effect.