Hemagglutinating activity and electrophoretic pattern of hemolymph serum proteins of Penaeus monodon and Macrobrachium rosenbergii to white spot syndrome virus injections

The giant fresh water prawn Macrobrachium rosenbergii is known to be highly tolerant to white spot syndrome virus (WSSV) infections when compared to the widely cultured marine tiger shrimp Penaeus monodon. At present, the exact mechanism of tolerance by M. rosenbergii to WSSV is not known. In this study, we attempt to study the effect of WSSV injections on the hemagglutination activity of the hemolymph serum of both P. monodon and M. rosenbergii and look for changes if any, on their hemolymph serum protein electrophoretic patterns. Our results show that M. rosenbergii had significantly (p < 0.05) higher hemagglutinating activity against mouse erythrocytes when compared to P. monodon. As the infection progressed to 48 h there was a further increase (p < 0.05) in the hemagglutination activity in M. rosenbergii, while it decreased in P. monodon. 12% SDS-PAGE analysis of the hemolymph serum of M. rosenbergii infected with WSSV did not show any new protein bands, whereas few bands with decreased intensity was observed in moribund P. monodon where the hemagglutinating activity was also observed to be decreased. The results indicate that hemolymph hemagglutinin levels are modulated in crustaceans as a response to viral infections.     

Effects of illumination on early life development and digestive enzyme activities in common pandora Pagellus erythrinus L. larvae

The activities of main digestive enzyme (proteases, amylase and lipase) and animal husbandry (mainly growth and survival) were studied in common pandora Pagellus erythrinus larvae until 30 days after hatching (DAH). Three different illumination levels (10, 30 and 100 lx) were compared in triplicate and green water technique was carried out. At the end of the experiment, illumination did not affect neither survival nor growth except 10 lx treatment. Similarly, specific growth rate (SGR) was different in 10 lx treatment (p < 0.05), although no differences were found in group 30 and 100 lx treatments (p > 0.05). In all groups, trypsin and chymotrypsin specific activities were firstly detected on day 3 related with mouth opening and slightly increased until 20 and 25 DAH respectively, and after this date specific activities of those decreased. Although, there was a significant difference between 10 lx treatment and other experimental groups (p < 0.05), no differences were found in other treatments (p > 0.05). Pepsin was firstly detected on day 25 related with stomach formation and sharp increase was observed until 30 DAH and then slight decrease was measured from this date and no differences were found between all groups. Amylase was firstly determined on day 2 and increased to day 5. After this date, slight decreases were measured in all groups and continued until end of experiments. The highest specific activity of amylase was determined in 30 lx treatment and no significant differences were found between groups (p > 0.05). Lipase was firstly detected on day 4 and increased to day 10. Then, activity of lipase decreased until day 15 and increased again until 25 DAH. Slight decreases were found in all groups until day 30 and continued to end of experiments. No significant differences were detected among groups (p > 0.05).

Finally, the significant improvement in survival, larval development and specific enzyme activities of larvae were determined in 30 lx treated group. It is thought that this phenomenon is related to optimal keeping conditions provided by the medium illumination level for Pagellus erythrinus larvae.     

Catches of swimming crabs using fish mince in “teabags” compared to conventional fish baits in collapsible pots

Fishing trials were conducted in a pond using collapsible pots containing fish mince inside a “teabag” or cut fish baits. Scomber japonicus was minced (100 g) and packed in a teabag or cut in half; non-baited pots served as controls. Baits were placed inside box-shaped pots (2.3 cm mesh size) with two slit entrances. Trials consisted of 100-pot hauls/bait type using a 1-day soaking time. The crab catches of Charybdis japonica and Portunus pelagicus from “fish” and “mince teabag” baits were not significantly different, but differences were found for total catch and damselfish Chromis notata. Trials using 80 g of mince and 20 g of sugar as the “teabag” bait lowered the catch of C. japonica, Thalamita sima and Thalamita prymna but not P. pelagicus when compared with fish bait. Results indicate that the mince in teabags is as suitable as fish bait, and that adding substances to the mince might be used to make selective crab baits.     

Dopamine modulates the physiological response of the tiger shrimp Penaeus monodon

Levels of glucose, lactate, pO₂, pCO₂, HCO₃⁻, TCO₂, Na⁺, K⁺, Cl⁻, protein, and oxyhemocyanin in the hemolymph and its osmolality and pH were measured when tiger shrimp, Penaeus monodon (13.5 ± 1.5 g body weight), were individually injected with saline or dopamine at 10^− 8, 10^− 7, or 10^− 6 mol shrimp^− 1. Results showed that hemolymph glucose, lactate, pCO₂, HCO₃⁻, and TCO₂ values increased from 2 to 4 h; hemolymph osmolality, Na⁺, and total protein had increased at 2 h; and hemolymph K⁺ decreased from 2 to 8 h after the dopamine injection. All physiological parameters returned to the control values 4–16 h after receiving dopamine. The dopamine injection also significantly decreased the oxyhemocyanin/protein ratio of P. monodon which occurred at 2 h, resulting from an elevation of hemolymph protein and a slight decrease of oxyhemocyanin. These results suggest that stress-inducing dopamine caused a transient period of modulation of energy metabolism, osmoregulation, respiration, and the acid–base balance in P. monodon in adapting to this environmental stress.     

Age, growth, and reproduction of silky sharks, Carcharhinus falciformis, in northeastern Taiwan waters

To examine the age, growth and reproduction of silky sharks, Carcharhinus falciformis, in the waters off northeastern Taiwan, 469 specimens (213 females and 256 males) were collected from August 2000 to January 2002 at the Nanfanao fish market, northeastern Taiwan. The relationship between body weight (W) and total length (TL) for both sexes combined was expressed as: W = 2.92 × 10⁻⁶ TL^3.15 (n = 469, p < 0.01). The relationship between TL and vertebral centrum radius (R) for both sexes combined data was estimated as: TL = 25.979 + 18.197R (n = 250, p < 0.01). Growth bands (including translucent and opaque zones) in precaudal vertebrae formed once a year between December and January and were counted up to 11 and 14 for females and males, respectively. The von Bertalanffy growth function (VBGF) was used to model the observed length at age data. The sexes combined VBGF predicted an asymptotic length (L_∞) = 332.0 cm TL, growth coefficient (k) = 0.0838 year⁻¹, age at zero length (t₀) = −2.761 year (n = 250, p < 0.01). Size at 50% maturity for males was estimated to be 212.5 cm based on the logistic curve, which corresponded to 9.3 years. Females matured at 210–220 cm, which correspond to 9.2–10.2 years. The length at birth was estimated to be 63.5–75.5 cm TL. The number of embryos per litter was 8–10 and sex ratio of embryos was 1:1.     

Growth performance of gilthead sea bream Sparus aurata in different osmotic conditions: Implications for osmoregulation and energy metabolism

The influence of three different environmental salinities (seawater, SW: 38 ppt salinity; brackish water, BW: 12 ppt; and low salinity water, LSW: 6 ppt salinity) on the growth, osmoregulation and metabolism of young gilthead sea bream (Sparus aurata L.) was studied over a period of 100 days. 480 inmature fish (20 g mean body weight) were randomly divided into six tanks of 2500 l (80 fish per tank) and maintained under three different salinities (38 ppt, 12 ppt and 6 ppt) in an open system. Every three weeks, 10 fish from each tank were anesthetized, weighed and lenghed. At the end of experiment, 10 fish from each tank were anesthetized, weighed and sampled for plasma, brain, gill and liver. Gill Na⁺, K⁺-ATPase activity, plasma osmolality, ions (sodium and chloride), glucose, lactate, protein and triglyceride, and hepatosomatic index were examined. In addition, levels of glycogen, lactate, ATP and activities of potential regulatory enzymes (hexokinase, pyruvate kinase, glycogen phosphorylase, and glucose 6-phosphate dehydrogenase) were assesed in liver, brain, and gill. BW-acclimated fish showed a better growth with respect to SW- or LSW-acclimated fish (12 > 38 > 6 ppt). The same relationship was observed for weight gain and specific growth rate. Osmoregulatory parameters in plasma (osmolality, Na⁺ and Cl⁻ levels) were similar in SW- and BW-acclimated fish but significantly higher than those of LSW-acclimated fish. Gill Na⁺, K⁺-ATPase activity showed lower values in intermediate salinity (6 > 38 > 12 ppt). No changes were observed in metabolic parameters analyzed in plasma, whereas only minor changes were observed in metabolic parameters of liver, gills and brain that could be correlated with the higher growth rates observed in fish acclimated to BW, which do not allow us to attribute the best growth rate observed at 12 ppt to lower metabolic rates in that salinity.     

Effect of increasing salinity on physiological response in juvenile scallops Argopecten purpuratus at two rearing temperatures

Argopecten purpuratus can be cultivated using Recirculating Aquaculture Systems (RAS) as a method to increase production. In order to determine physiological response of A. purpuratus under different salinities and temperature conditions, two groups of juvenile scallops (small: h = 6.5 mm and large: h = 25.5 mm) were acclimated and close-cultured at salinities of 34, 38, and 42 g/l, at 16 and 22 °C and fed on Isochrysis galbana and Chaetoceros calcitrans. Survival, shell growth and scope for growth were determined at the end of the trials. Survival showed an inverse relationship with temperature and ammonia levels. In small scallops an increase in salinity at 16 °C increased survival. However, this relationship was not evident at 22 °C. On the other hand, salinity did not affect survival of large juveniles. Small juveniles had a lower survival (approximately 40%) than larger scallops (up to 85%) throughout the trials. Oxygen consumption was not affected by salinity. Small scallops showed similar oxygen consumption at 16 and 22 °C but in large juveniles higher values were registered at 22 °C. In large juveniles routine consumption at 16 °C was higher (up to 35%) than standard consumption. This pattern was not evident at 22 °C, suggesting that oxygen demand is higher regardless of feeding condition. NH₄⁺–N excretion rate is inversely related to salinity. Only small juveniles showed a higher NH₄⁺–N excretion rate at 22 °C. Scope for growth was positive in all treatments, although the upper limit of salinity should not be based only in this index. Higher scope for growth values at 38 and 42 g/l was related with a reduction in ammonia excretion and high absorption efficiency. In addition, an increase in salinity produced a reduction in NH₃–N proportion and under hypersaline conditions scallops tended to decrease excretion as a way of osmoconformation. This explains our findings of higher survival rates at higher salinities. Even though the scope for growth is positive at 42 g/l, the osmotic stress reduces the survival chances. The data obtained can be considered useful information for A. purpuratus culture under controlled conditions.     

Breeding and juvenile culture of the lined seahorse, Hippocampus erectus Perry, 1810

All seahorse species worldwide have been placed under CITES Appendix II since 2004, because they have been over-exploited for traditional Chinese medicine and aquarium trades. Aquaculture has been recognized as a long-term solution for sustaining the seahorse trade while minimizing wild collection. In this study, we evaluated the breeding and juvenile culture of an important aquarium seahorse species, the lined seahorse Hippocampus erectus, Perry 1810. Pairing, mating and copulation behavior were observed. Gestation time and brood size were 17.33 ± 2.94 days and 272.33 ± 66.45 individuals/brood, respectively. Growth rates differed among juveniles from different broods. Effects of temperature on the growth rates and survivorship of the juveniles during the first two weeks were compared. The highest growth rate and survivorship of the juveniles occurred at 28–29 °C among the temperatures tested (24–33 °C). Growth rate and survivorship of the juveniles during the first 9 weeks at 28 °C were investigated. The final standard length and survivorship of the juveniles were 6.32 ± 0.52 cm and 71.11 ± 10.18%, respectively, and the relationship between the wet weight and the standard length of the juvenile seahorses can be expressed as: W = 0.0034 L^2.5535 (r² = 0.9903, n = 12, P < 0.01). These findings suggest that H. erectus is a good candidate for commercial aquaculture.     

Effect of probiotics, Enteroccus faecium, on tilapia (Oreochromis niloticus) growth performance and immune response

The aim of this study was to analyze the effect of a probiotic bacterium, Enterococcus faecium ZJ4 on growth performances and immune responses of tilapia (Oreochromis niloticus). The tilapias were treated with E. faecium ZJ4 at a final concentration of 1 × 10⁷ cfu ml^− 1 in aquaria water every 4 days. Six aquaria with three replicates for treated and controls were used. After 40 days, the tilapias supplemented with the probiotic showed significantly better final weight and daily weight gain (DWG) than those fed the basal diet (Control) (P < 0.05). There was no remarkable difference (P > 0.05) in the total serum protein, albumin content, globulin concentration and A/G ratio between the treated and control tilapias. The result of lysozyme activity assay was similar to these biochemical indexes. However, the complement component 3 content, myeloperoxidase (MPO) activity and the respiratory burst activity of blood phagocytes were higher (P < 0.05) in E. faecium treated tilapias (trial 1) than the controls.     

Effects of temperature and lipid droplet adherence on mortality of hatchery-reared southern hake Merluccius australis larvae

Effects of exogenous (water temperature) and endogenous (lipid droplet adherence) factors were experimentally tested on early survival of southern hake Merluccius australis reared under controlled conditions. Experiments to determine the effect of temperature (10, 12 and 14 °C) on larval growth rates and yolk-sac absorption rates of unfed southern hake were carried out under laboratory conditions. There was no significant differences in growth rates at the temperature range tested (ANCOVA, F = 0.164, p > 0.25), but yolk-sac absorption rates and mortality increased with temperature (ANCOVA, F = 53.84, p < 0.001). A high percentage (between 31 and 81%) of hake eggs showed a lipid droplet not adhered (i.e., freely moving in the yolk, and not located in the posteriormost portion of the yolk-sac). In a second experiment, fed southern hake larvae with the lipid droplet not adhered during embryonic development did not survive after yolk-sac absorption. This study provides the first data on the influence of the lipid droplet absorption on larval survival of cultured hake, and can be used as an early indication of the quality of the batch.     

Efficacy, tissue distribution, and residue depletion of oxytetracycline in WS-RLP infected California red abalone Haliotis rufescens

Oxytetracycline (OTC) was recently demonstrated to be an effective therapeutant for California red abalone (Haliotis rufescens) infected with the agent of withering syndrome (WS), a Rickettsia-like prokaryote (WS-RLP). This study examined the influence of temperature on the efficacy and pharmacokinetics of oral OTC treatments on WS-RLP infected red abalone, H. rufescens. Medication was administered in an artificial diet containing 1.85% active OTC at a rate of 103.4 mg/kg abalone for 10, 20 and 30 days at both 13.4 °C and 17.3 °C. Drug residue concentrations, WS-RLP burden and WS-associated pathological changes within foot muscle and digestive gland were measured 3, 17, 23, 42, 63, 81, 102, 122, and 160 days after OTC treatment. In all samples the highest concentrations of OTC were detected on the first sample day. Furthermore, all sampling periods showed significantly less OTC in foot muscle samples relative to corresponding digestive gland samples (p < 0.001). While drug depuration occurred rapidly from foot muscle, unusually high concentrations and long depuration times were measured within the digestive gland. The pharmacokinetic trends in both the foot muscle and digestive gland at 13.4 °C were adequately described by a one-compartment model, while the digestive gland depuration models within the 17.3 °C treatments conformed to a two-compartment model. At both temperatures treatment duration gave rise to significant differences in OTC accumulation and depletion (p < 0.001). Despite this, terminal elimination rate constants were similar. Estimated half-lives were similar among all treatments for digestive gland (23.8–27.5 d) and all treatments for the foot muscle at 13.4 °C (10.7–13.4 d; calculated only for the 13.4 °C treatments). At both temperatures all three durations of drug treatment led to significant reductions in WS-RLP prevalence, foot muscle atrophy, and mortality. WS-RLP re-challenge trials initiated 44, 88, and 122 days following the 10-day treatment at 17.3 °C showed reduced susceptibility to re-infection for up to 88 days after treatment, suggesting that digestive gland OTC residues confer long-term resistance to pathogen. Atomic absorption spectroscopy conducted on both foot muscle and digestive gland found significantly higher concentrations of iron, zinc, and manganese in the digestive gland, offering an explanation behind the underlying mechanism of OTC retention for extended periods within this tissue.     

Temperature and Salinity Tolerances of the Tropical Spiny Lobster,Panulirus ornatus

Interest in the development of aquaculture of the tropical spiny lobster, Panulirus ornatus, has increased markedly over the past 10 yr because of strong market demand and high prices. In Australia, economic conditions will necessitate that a semi‐intensive approach be taken, possibly involving managed environmental conditions. Identification of optimal temperature and salinity levels will be necessary, and therefore two experiments were performed to examine these two parameters. Juvenile lobsters were grown in tanks at five temperatures (19, 22, 25, 28 and 31 C). Growth was significantly affected by temperature (P < 0.01), and maximal growth occurred at 25–31 C. Examination of the temperature effect on molt increment and intermolt period indicated that 27 C was the optimal temperature, at which molt increment was greatest and intermolt period the least. Temperature also had a significant (P < 0.01) positive effect on apparent feed intake (AFI). Juvenile lobsters were also exposed to four different salinities (20, 25, 30 and 35 ppt) over a period of 91 d. Significant differences (P < 0.01) were apparent for both survival and growth. Lowest survival occurred at 35 ppt which may be attributable to higher cannibalism at that salinity. Growth was highest at 35 ppt and progressively less at lower salinities. Although full marine salinity (35 ppt) will generate best performance of P. ornatus, its capacity to tolerate reduced salinity will provide greater opportunity to develop commercial aquaculture.     

A soft technology to improve survival and reproductive performance of Litopenaeus stylirostris by counterbalancing physiological disturbances associated with handling stress

The consequences of handling stress (fishing, transfer, eyestalk ablation) on shrimp broodstock are poorly documented. The weakness of farmed shrimp, Litopenaeus stylirostris, during winter is a major problem in New Caledonia, because of seasonal climate (tropical–sub-temperate). The transfer of broodstock in winter from earthen outdoor ponds to indoor maturation tanks in the hatchery (T = 20 °C, Salinity = 35‰, fed shrimp) usually leads, after 48 h, to high mortality (up to 70%). Eyestalk ablation to induce ovarian maturation in females leads to further mortality.

Starting from a background analysis of physiological disturbances (initial osmoregulatory imbalance) associated with handling stress (Wabete, N., Chim, L., Lemaire, P., Massabuau, J.-C., 2004. Caractérisation de problèmes de physiologie respiratoire et d'échanges ioniques associés à la manipulation chez la crevette pénéide Litopenaeus stylirostris à 20 °C. Styli 2003. Trente ans de crevetticulture en Nouvelle-Calédonie. Ed. Ifremer. Actes Colloq. 38, 75-84.), we developed a protocol using a soft technology, based on modifications of water salinity, temperature and feeding regime. The aim was to minimize problems of osmoregulatory imbalance and associated mortalities. The protocol we developed, called the LSD OT protocol (Low Salinity and Diet, Optimal Temperature), was first evaluated on sub-adult shrimp (20–25 g) and then applied to broodstock. Survival after transfer and following eyestalk ablation, as well as reproductive achievement (spawning rate, nauplii number) was considerably improved when shrimps were transferred under “physiological comfort” i.e. warmed isosmotic water (26 °C and 26‰) and unfed for 3 d. This new handling protocol, based on a better control of salinity, temperature and feeding conditions, has been transferred successfully to private hatcheries and already contributes to an increased profitability of New-Caledonian shrimp industry.     

Substitution of Chaetoceros muelleri by Spirulina platensis meal in diets for Litopenaeus schmitti larvae

The nutritional response of Litopenaeus schmitti larvae to substitution of Chaetoceros muelleri by Spirulina platensis meal (SPM) was evaluated. The substitution levels (S) were 0%, 25%, 50%, 75% and 100%, dry weight basis. Final larval length (FL) ranged from 1.98 to 3.16 mm for the different substitution levels. There was a significant relationship between S and FL, described by the following quadratic equation: FL = 2.853 + 0.01598S − 0.000233S². The substitution level (S) yielding maximum FL was 34.2%. Development index (DI) values ranged from 2.84 to 3.93 and were dependent on substitution level. The corresponding equation was DI = 3.799 + 0.00945S − 0.000189S² (P < 0.01). Maximum DI was obtained at 25.0% substitution. Survival was high (82–87%) and no significant differences were found between treatments. Protein digestibility of either microalgae was high, with 92% for SPM and 94% for C. muelleri, with no significant differences between them. The results in this study indicate that an adequate balance of nutrients in relation to the requirements of the species is critical. To simultaneously improve FL and DI, a 30% substitution of C. muelleri by SPM is suggested. This is equivalent to feeding 0.15 mg larvae^− 1 day^− 1 dry weight basis of a 70% C. muelleri/30% SPM diet, representing 0.078 mg protein larvae^− 1 day^− 1, 0.026 mg lipids larvae^− 1 day^− 1 and 2.732 J larvae^− 1 day^− 1.     

Physiological responses during stress and subsequent recovery at different salinities in adult pejerrey Odontesthes bonariensis

Mortality and physiological responses (plasma cortisol, glucose, osmolality, ions Na⁺ and Cl⁻ and hematocrit values) were examined in adult pejerrey Odontesthes bonariensis subjected to transport or crowding and subsequent recovery for 1 week at 0, 5 and 20 ppt NaCl. Two experiments were conducted using fish from different hatcheries; the following responses were consistent for both stocks. Mortality was observed only during post-stress recovery at 0 and 20 ppt NaCl but not at 5 ppt. During recovery, blood cortisol and glucose were generally higher, whereas osmolality and blood electrolytes (Na⁺ and Cl⁻) were lower at 0 ppt than at the other salinities. A salinity of 5 ppt NaCl could not prevent the rise in cortisol and glucose levels caused by the stress of transport, but during recovery, it prevented further increases in these features and/or accelerated their return to basal levels. This salinity also helped maintain stable blood electrolyte levels. During recovery at 20 ppt NaCl, osmolality and blood ions increased, whereas plasma cortisol and glucose generally decreased. Hematocrit values were lower at 20 ppt than at the other salinities. These results suggest that freshwater is not an adequate medium for post-stress recovery and that the presence of NaCl in the water either decreases the secretion of cortisol or promotes its clearance in O. bonariensis.     

Replacement of fish meal by meat and bone meal in diets for large yellow croaker, Pseudosciaena crocea

A growth experiment was conducted to investigate the effects of replacement of fish meal (FM) by meat and bone meal (MBM) in diets on the growth and body composition of large yellow croaker (Pseudosciaena crocea). Six isonitrogenous (43% crude protein) and isoenergetic (20 kJ g^− 1) diets replacing 0, 15, 30, 45, 60 and 75% FM protein by MBM protein were formulated. Each diet was randomly allocated to triplicate groups of fish in sea floating cages (1.0 × 1.0 × 1.5 m), and each cage was stocked with 180 fish (initial average weight of 1.88 ± 0.02 g). Fish were fed twice daily (05:00 and 17:30) to apparent satiation for 8 weeks. The water temperature ranged from 26.5 to 32.5 °C, salinity from 32 to 36‰, and dissolved oxygen content was approximately 7 mg l^− 1 during the experimental period. Survival decreased with increasing dietary MBM and the survival in the fish fed the diet with 75% protein from MBM was significantly lower than other groups (P < 0.05). There were no significant differences in specific growth rate (SGR) among the fish fed the diets with 0 (the control group), 15, 30 and 45% protein from MBM. However, SGR in the fish fed the diets with 60 and 75% protein from MBM were significantly lower than other groups (P < 0.05). No significant differences in feeding rate were observed among dietary treatments. The digestibility experiment showed that the apparent digestibility coefficients (ADC) of dry matter, protein, lipid and energy of MBM were significantly lower compared with those of FM (P < 0.05). Essential amino acid index was found to be correlated positively with SGR in the present study, suggesting that essential amino acid balance was important. Body composition analysis showed that the carcass protein and essential amino acids were not significantly affected by dietary MBM. The lipid and n-3 highly unsaturated fatty acid (n-3 HUFA) in fish muscle, however, significantly decreased with increasing dietary MBM. These results showed that 45% of FM protein could be replaced by MBM protein in diets of large yellow croaker without significantly reducing growth. It was suggested that the reduced growth with higher MBM was due to lower digestibility and imbalance of essential amino acids.     

Effect of hyperoxygenation and low water flow on the primary stress response and susceptibility of Atlantic salmon Salmo salar L. to experimental challenge with IPN virus

Intensive salmon smolt production normally includes reduced water flow and hyperoxygenation (added oxygen) of remaining water. There is little information on how different water quality parameters influence the fish health and the susceptibility to infectious diseases. The current experiment was carried out to evaluate if the combination of hyperoxygenation and reduced water flow (hyperoxic) can act as a chronic stressor to salmon in freshwater (FW) in such a way that it increases the susceptibility to IPN virus (IPNV) following seawater transfer. In FW, after 22 days of hyperoxic exposure plasma ion, TBARS and cortisol were measured. The cortisol levels were significantly (p = 0.011) higher in the hyperoxic group compared to controls maintained under normal oxygen saturation and water flow (normoxic), indicating chronic stress. Hyperoxygenation in FW caused decreased plasma [Cl⁻] compared to the normoxic group (p = 0.037), while [K⁺] tended to be higher in the hyperoxic group (p = 0.088). No significant differences were observed in plasma [Na⁺], total osmolality, TBARS or hematocrit, but there was a tendency towards a lower hct in the hyperoxic compared to the normoxic group. In SW the mortality was higher in the hyperoxic group challenged with IPNV (34%) compared to the normoxic group challenged with IPNV (20%) (p = 0.02), and no mortality was observed in the PBS injected fish. The challenged fish showed an overall increase in plasma cortisol day 8, 10, 12 and 14 post-challenge (p = 0.015, p = 0.000, p = 0.046 and p = 0.022 respectively). After SW transfer and challenge, plasma [K⁺] was elevated in both challenged groups, but no consistent trends were found for plasma [Cl⁻], [Na⁺] or total osmolality during the SW phase. There were no significant differences in the gene expression level of IFN 1, Mx and IL 1β prior to challenge, suggesting that the basic expression level of these genes were not affected by hyperoxygenation. IPNV was detected in kidney and pylorus, by immunohistochemistry, cell culture, and RT-PCR in head kidney. This experiment indicates that chronic stress induced by a combination of low water flow and hyperoxygenation increases the susceptibility to IPNV challenge.     

Effects of Cu-exchanged montmorillonite on growth performance, microbial ecology and intestinal morphology of Nile tilapia (Oreochromis niloticus)

A total of 360 Nile tilapia at an average initial body weight of 3.9 g were randomly allocated to 4 treatments, each of which had three replicates of 30 fish/tank and used to investigate the effects of Cu²⁺-exchanged montmorillonite (Cu-MMT) on growth performance, microbial ecology of the skin, gill and intestine, and intestinal morphology. The dietary treatments were: 1) basal diet, 2) basal diet + 1.5 g/kg MMT, 3) basal diet + 30 mg/kg copper as CuSO₄ (equivalent to the copper in the Cu-MMT treatment group), or 4) basal diet + 1.5 g/kg Cu-MMT. The results showed that supplementation with Cu-MMT significantly improved growth performance as compared to control and fish fed with Cu-MMT had higher growth performance than those fed with MMT or CuSO₄. Supplementation with Cu-MMT reduced (P < 0.05) the total intestinal aerobic bacterial counts and affected the composition of intestinal microflora with a tendency of Aeromonas, Vibrio, Pseudomonas, Flavobacterium, Acinetobacter, Alcaligence, Enterobacteriaceae decreasing as compared with control. Measurements of villus and microvillus heights at different sites of the intestinal mucosa indicated that dietary addition of MMT or Cu-MMT improved intestinal mucosal morphology. However, the microbial ecology of the skin and gill was not affected by the addition of CuSO₄, MMT or Cu-MMT. Supplementation with CuSO₄ had no (P > 0.05) effect on the growth performance, microbial ecology and intestinal morphology. The results showed that Cu-MMT exhibited antibacterial activity in vivo and protected intestinal mucosa from invasion of pathogenic bacterium and toxins, resulting in a positive effect on the growth performance.     

The Effects of Air Exposure on Hemolymph Osmoregulatory Capacity of Brown Tiger Shrimp,Penaeus esculentus,and Western King Shrimp,Penaeus latisulcatus,Reared at Different Salinities

Abstract

Hemolymph osmolality and osmoregulatory capacity (OC) of brown tiger shrimp, Penaeus esculentus (0.94±0.04 g mean initial weight) and western king shrimp, P. latisulcatus (5.37±0.10 g mean initial weight) from four salinities (10, 22, 34 and 46 ppt) were determined following 7, 14 and 21 minutes of air-exposure. Hemolymph osmolality of both species increased with increasing salinity. Isosmotic points of brown tiger shrimp calculated from regression lines between hemolymph and medium osmolality were 30.9, 31.9, 32.1 and 31.1 ppt at 0, 7, 14, and 21 minutes of air-exposure, respectively. Isosmotic points of western king shrimp were 33.8, 33.3, 32.8 and 33.1 ppt at 0, 7, 14, and 21 minutes air-exposure, respectively. OC of brown tiger shrimp at salinity of 34 ppt did not change when exposed to air for any length whereas OC of shrimp at other salinities (10, 22 and 46 ppt) were significantly different (P < 0.05) when exposed to air for 21 minutes. OC of western king shrimp at salinity 10 ppt was reduced (P < 0.05) when exposed to 14 and 21 minutes of air when compared to control and 7 minutes of air exposure. The results indicate that both species spent less energy (P < 0.5) for osmoregulation from 30 to 34 ppt. Furthermore, salinities 10 and 46 ppt were unsuitable for rearing brown tiger shrimp and salinity 10 ppt was unsuitable for growing western king shrimp.     

Metamorphosis in summer flounder: manipulation of rearing salinity to synchronize settling behavior, growth and development

In the aquaculture of summer flounder (Paralichthys dentatus), the inherent variation in growth and settling behavior during metamorphosis may lead to cannibalism and necessitate increased labor due to size grading. Our goal was to use an environmental salinity change as a cue to synchronize settling behavior and produce a larger, more uniformly sized cohort of juvenile summer flounder. Early metamorphic flounder were exposed to either a 5-day fluctuating (30–20–30–20–30 ppt; “Flux”) or a single (30–20 ppt; “20 ppt”) drop in rearing salinity. A control (continuous 30 ppt) was used for comparison. Average values for a peak settlement interval (PSI; defined as the interval beginning on the day by which the first 20% had settled until the day 80% had settled) were not affected by salinity manipulation, though the 20-ppt treatment did significantly increase percent settled per day by 54 dah. Average fish size was increased by the 20-ppt treatment (19.3±0.5 mm), but not the flux treatment (17.2±0.4 mm), compared to the control (17.6±0.5 mm). Developmental stage was significantly increased in the 20-ppt treatment (3.2±0.1) in comparison to the Flux (2.9±0.1), but not the control (3.1±0.1). However, the 20-ppt treatment reduced variance in development. To confirm the positive effects of the 20-ppt treatment, a second experiment was performed. A single salinity drop (“20 ppt”), a previously successful ([Gavlik, S., Albino, M., Specker, J.L., 2002. Metamorphosis in summer flounder: Manipulation of thyoid status to synchronize settling behavior, growth, and development. Aquaculture 203 (3-4), 359-373]thyroid hormone manipulation treatment (“TH”) and a combination of the two (“TH+20 ppt”) were compared to a control (continuous 30 ppt, no thyroid manipulation). The mean PSI was significantly reduced by both TH (7±1 days) and TH+20 ppt (8±0 days) treatments, compared to the control (11±1 days). The PSI for the 20-ppt treatment (9±0 days) was not significantly different than the control. The percent flounder settled per day was significantly increased by 20 ppt salinity and significantly modified (decreased, then increased) by TH manipulation. Flounder exposed to 20-ppt salinity were both larger (20 ppt: 18.6±0.3 mm; TH+20 ppt: 18.3±0.3 mm) and more developmentally advanced (20 ppt: 3.1±0.04; TH+20 ppt: 3.2±0.03) than flounder in 30 ppt (TH: 17.8±0.3 mm/3.1±0.3; Control: 17.9±0.3 mm/3.0±0.05). Finally, 20-ppt treatment reduced variance in development, while TH treatment reduced variance in both growth and development. Percent survival was unaffected by treatment in both experiments. In summary, a decrease in rearing salinity, from 30 to 20 ppt, increased growth, settling behavior and development of metamorphosing summer flounder. A decrease in rearing salinity, in combination with a TH manipulation, should result in larger, more uniformly sized flounder cohorts. We expect this synchronization will reduce the cannibalism and labor costs associated with size grading of cultured, metamorphosing summer flounder.