Immunostimulatory effects of non-CpG oligodeoxynucleotides on cobia (<Emphasis Type="Italic">Rachycentron canadum</Emphasis>)

Oligodeoxynucleotides (ODNs) with unmethylated CpG dinucleotides mimic the immunostimulatory activity of bacterial DNA in vertebrates through the recognition of cellular Toll-like receptor 9 (TLR9). We previously demonstrated that CpG motif, 5′-GACGTT-3′ (1668), and 5′-GTCGTT-3′ (2395) were highly immunostimulatory in cobia. In this study, motifs containing CpA, CpC, and CpT were used to evaluate their effects on cobia TLR9 (RCTLR9) gene expression. RCTLR9 expression increased significantly in response to CpT ODN in comparison with other ODN motifs. The optimal dosage of CpT ODN was 10 µg/fish. Furthermore, when the endosomal acidification was blocked using chloroquine immersion, the CpT ODN-induced expression of RCTLR9 in spleen and liver was inhibited in a dose-dependent manner. Our results demonstrated that ODNs containing motifs other than CpG, such as CpT ODN, can stimulate RCTLR9 expression, which is mediated, at least partially, by the interaction between CpT ODN and RCTLR9.     

Effects of early weaning strategies on growth,survival and digestive enzyme activities in cobia (<Emphasis Type="Italic">Rachycentron canadum</Emphasis> L.) larvae

The effects of weaning strategies of cobia (Rachycentron canadum L.) larvae to commercial microdiets, either from rotifers or from Artemia, on growth, survival and enzymatic digestive capacity, were investigated. In the first experiment, cobia larvae were weaned from rotifers by co-feeding with a microdiet (Otohime) from 8, 13 or 20 days post-hatching (dph). The larvae in the control treatment were fed rotifers (2–12 dph), Artemia nauplii from 7 dph, and co-fed with the microdiet from 20 dph. In the second experiment, the larvae were weaned from Artemia, which was fed to the larvae from 7 dph, by co-feeding with a microdiet (NRD) from 8, 13 or 18 dph. The larvae in control treatment were fed rotifers, then Artemia to the end of the experiment (28 dph). Weaning of cobia larvae onto a microdiet directly from rotifers significantly reduced growth, survival and digestive capacity of the larvae and did not lead to larval acceptance of the microdiet, compared to those weaned from Artemia in the first experiment. Early weaning of cobia larvae onto NRD microdiet (on 8 or 13 dph) from Artemia in the second experiment also reduced growth, survival rate and gut maturation index, compared to those fed live feed. With available microdiets, weaning of cobia larvae could start from Artemia at around 18 dph in order to obtain comparable growth, survival and gut maturation to larvae fed live feed.     

Study on regional production and economy of cobia <Emphasis Type="Italic">Rachycentron canadum</Emphasis> commercial cage culture

In recent years, cobia has become an emerging farmed species in Asia due to its quick growth and high economic value. This study collects biological and economic data affecting the economic performance of cobia farming in three countries, namely Taiwan, China, and Vietnam. The data are collected by questionnaire sampling and analyzed by multivariate statistical analysis in order to compare the key factors affecting the production and economy of cobia farming in these three countries. The results show that Taiwan, China, and Vietnam have significant differences in input intensities and profitability. China has the highest input intensity (3372.42 TWD/m³), as its high stocking density increases feed input. Taiwan has the highest unit input cost (103.44 TWD/kg), as the high quality of the product increases the price of cobia in Taiwan, which offsets the high product costs. In terms of profitability, the benefit–cost ratio is over one in all three countries, indicating that the profitability of cobia farming is good in all three countries. Profitability analysis shows that fingerlings in China achieve 36.50, which is the highest among the three countries; whereas Taiwan has the highest feed profitability of 0.78, which reveals that the fingerlings produced in China are competitive in both price and quality, while Taiwan has the best feed management efficiency. The production costs and profitability of Vietnam fall between those of Taiwan and China. Feed cost is the main expenditure in cobia culture; thus, good feed management could effectively reduce production costs and increase business performance. The feed quality and input management model of Taiwan, in conjunction with the fingerling quality and stock model of China, could provide future reference for farming management in such areas as feed input and selection of fingerling.     

Replacement of fishmeal in cobia (<Emphasis Type="Italic">Rachycentron canadum</Emphasis>) diets using poultry by-product meal

An 8-week feeding trial was conducted to evaluate the use of local poultry by-product meal (PBM) in replacement of imported fishmeal in the diets of cobia, Rachycentron canadum. Six isolipidic (12%) and isoproteic (45%) experimental diets were formulated using PBM to replace fishmeal at 20, 40, 60, 80 and 100% dietary protein. Eleven juvenile cobia (initial mean weight of 30.7 ± 0.78 g) were randomly stocked in 300-L circular fibreglass tanks and hand-fed based on the total biomass of each tank, twice a day at 0900 h and 1500 h. The fish were group weighed at 2-week intervals to monitor their growth performance in order to adjust the feeding ratio. At the end of the feeding trial, weight gains (WGs) ranging from 221 to 322% were obtained. The specific growth rate (SGR), WG and protein efficiency ratio (PER) for fish fed with PBM-based diets were not significantly different (P > 0.05) when compared to fish fed the control diet. The best SGR was recorded for fish fed with 60PBM diet, which was significantly higher (P < 0.05) than those fed the control diet with values at 2.40 ± 0.01 and 1.97 ± 0.26%/day, respectively. The FCR of 1.83 ± 0.05 for fish fed the 60PBM diet was not significantly different (P > 0.05) when compared to those fed the control diet. The PBM source and dietary level did not significantly (P > 0.05) affect the hepatosomatic index of the fish. The results from this study suggested that PBM could replace 100% dietary fishmeal without adversely affecting the growth performance, but an optimal replacement level at approximately 60% was recommended for better growth performance and efficient feed utilization.     

<Emphasis Type="Italic">Neoheterobothrium hirame</Emphasis> (Monogenea) alters the feeding behavior of juvenile olive flounder <Emphasis Type="Italic">Paralichthys olivaceus</Emphasis>

We monitored feeding behavior and survival of starved juvenile olive flounder experimentally infected with the gill monogenean Neoheterobothrium hirame. Infected flounder increased amount of the time spent in the water column by 117% when trying to capture live mysids, Neomysis sp. They also showed different feeding patterns from those of uninfected fish and made fewer attacks towards prey during one feeding attempt. Although the average numbers of mysids captured by individuals were similar between infected and uninfected fish, heavily infected fish tended to catch less prey. These results indicate that N. hirame reduces the feeding efficiency of the host for capturing live prey and possibly makes them more vulnerable to predation during feeding. We could not detect any obvious difference in survival rates between uninfected, lightly and heavily infected fish during 3 months of starvation. There was no evidence that starvation makes fish more susceptible to N. hirame. The present study provides first experimental evidence that N. hirame affects feeding behavior of juvenile olive flounder and supports the idea that this parasite indirectly reducing the host’s survival and may be responsible for the recent reduction of the flounder population in Japan.     

Repeatable immersion infection with <Emphasis Type="Italic">Photobacterium damselae</Emphasis> subsp. <Emphasis Type="Italic">piscicida</Emphasis> reproducing clinical signs and moderate mortality

Pseudotuberculosis is a bacterial septicaemia caused by Photobacterium damselae subsp. piscicida in several marine fish species. Yellowtail Seriola quinqueradiata is the most sensitive fish species to this disease. The internal organs of naturally infected yellowtail exhibit whitish spots, tubercle-like tissue structures, consisting of bacterial accumulations. There have been many trials for experimental infection, however adequate method of infection that reproduces moderate mortality and primary clinical signs has not yet established. Present investigation evaluated an immersion infection method by using logarithmic culture-phase bacteria resulting in higher mortality than that using stationary culture-phase bacteria. Typical white spots on the spleen and kidney were also observed constantly in dead fish. Transmission electron microscopy and fluorescent antibody microscopy showed bacterial clusters not only in the spleen and kidney but also in the blood channels in the secondary gill filaments. These results were confirmed repeatedly by plural experiments. The use of logarithmic-phase bacteria in immersion infection is an appropriate technique to reproduce moderate mortality and primary clinical signs, which will be a reliable infection method also for the challenge test of pseudotuberculosis vaccine.     

Induction of diploid gynogenesis in turbot <Emphasis Type="Italic">Scophthalmus maximus</Emphasis> with left-eyed flounder <Emphasis Type="Italic">Paralichthys olivaceus</Emphasis> sperm

Turbot Scophthalmus maximus exhibits sexually dimorphic growth, with females growing faster and reaching larger adult sizes than males. Thus, development of techniques for preferentially producing females is necessary to optimize production of these species. In this paper, gynogenetic diploids of turbot were induced by activating egg development with ultraviolet (UV)-irradiated left-eyed flounder Paralichthys olivaceus sperm combined with cold shock to prevent extrusion of the second polar body. The results of UV irradiation experiments showed that survival, motility, and duration of activity of P. olivaceus sperm generally decreased with increase in UV dose. The typical Hertwig’s effect was observed after fertilized turbot eggs with UV-irradiated P. olivaceus sperm and the optimal UV dose for gynogenetic haploid production was 36,000 erg mm⁻². At 15°C, appropriate timing of cold shock for retention of the second polar body in turbot eggs was at 6 min after fertilization. Results of different combinations of two shock temperatures (1 or 3°C) and four shock durations (15, 25, 35 or 45 min) at 6 min after fertilization demonstrated that shock of 25 min at 1°C gave the highest production of diploid gynogens (39.58% relative to its diploid control). The results of this study reveal that the use of UV-irradiated P. olivaceus sperm for activation of turbot eggs and cold shock for polar body retention is an effective method to produce gynogenetic offspring.     

Effect of a probiotic bacterium <Emphasis Type="Italic">Bacillus circulans</Emphasis> PB7 in the formulated diets: on growth,nutritional quality and immunity of <Emphasis Type="Italic">Catla catla</Emphasis> (Ham.)

Bacillus circulans PB7, isolated from the intestine of Catla catla, was evaluated for use as a probiotic supplement in the feeds for the fingerlings of Catla catla. The effect of supplement on the growth performance, feed utilization efficiency, and immune response was evaluated. Catla fingerlings (ave. wt. 6.48 ± 0.43 g) were fed diets supplemented with 2 × 10⁴ (feed C1), 2 × 10⁵ (feed C2), and 2 × 10⁶ (feed C3) B. circulans PB 7 cells per 100 g feed for 60 days at 5% of the body weight per day in two equal instalments in triplicate treatments. The control feed (CC) was not supplemented with the B. circulans. All the feeds were isocaloric and isonitrogenous. Fish fed with feed C2 displayed better growth, significantly (P ≤ 0.05) highest RNA/DNA ratio, a lower feed conversion ratio (FCR), and a higher protein efficiency ratio (PER) than the other experimental diets. Highest carcass protein and lipid was also observed in the fish fed C2 feed compared to the others. Significantly (P ≤ 0.05), highest protease was recorded in fish fed feed C2 (47.9 ± 0.016) and lowest in fish fed feed C3 (32.10 ± 0.009), where α-amylase activity did not differ significantly (P ≤ 0.05) beyond the lowest inclusion level. ALP, ACP, GOT, and GPT in the liver of Catla catla were the highest (P ≤ 0.05) in fish fed C2 feed. The highest TSP, albumin, and globulin was observed in fish treated with C2 feed after 60 days feeding trial, but the lowest glucose level was observed in the same treatment. After the feeding trial, the non-specific immunity levels and disease resistance of fish were also studied. Phagocytic ratio, phagocytic index, and leucocrit value were the highest in fish fed feed C2. After the feeding trial, the fish were challenged for 10 days by bath exposure to Aeromonas hydrophila (AH1) (10⁵c.f.u. ml⁻¹ for 1 h, and, after 7 days, 10⁷c.f.u. ml⁻¹ for 1 h). Highest survival percentage was observed in fish fed with feed C2 compared with only 6.66% in the controls, which indicated the effectiveness of B. circulans PB 7 in reducing disease caused by A. hydrophila.     

Frequent occurrence of the tetrodotoxin-bearing horseshoe crab <Emphasis Type="Italic">Carcinoscorpius rotundicauda</Emphasis> in Vietnam

Tetrodotoxin (TTX) was detected by HPLC and LC-MS/MS in specimens of the horseshoe crab Carcinoscorpius rotundicauda collected from Tan Hai village, Vung Tau province, Vietnam, where cases of poisoning due to the consumption of horseshoe crab have frequently been reported. The soft tissue from 10 out of the 12 specimens analyzed showed considerable levels of TTX, making them unsafe for human consumption (81.2 ± 50.3 MU/g). Paralytic shellfish poisoning (PSP) toxins were also detected in all of the specimens, but the levels were low. These findings show that the frequency of occurrence of C. rotundicauda specimens with a high level of TTX is significantly high, and that the TTX is responsible for the food poisonings caused by the consumption of C. rotundicauda in this area.     

Economic analysis of cobia (<Emphasis Type="Italic">Rachycentron canadum</Emphasis>) cage culture in large- and small-scale production systems in Brazil

Economic feasibility studies regarding aquaculture systems are relatively scarce, but they are important to potential investors and for the allocation of resources for research and technological development. This study evaluated the economic viability of cobia cage culture from the actual investment and operational costs of a large-scale operation off Recife, northeastern Brazil (industrial system; IS), and a family-run farm located in a near-shore area of Rio de Janeiro (familiar system; FS). The IS had twenty-four 1607 m³ floating cages deployed at a depth of 24 m, while the FS had six 75 m³ wooden cages installed in a sheltered 6- to 7-m-deep area. Analyses of profitability (gross revenue, operational profit, cost price and break-even production) and investment (net present value—NPV; and payback time) were performed. An analysis of sensitivity was also carried out. The IS required an initial investment of approximately US$ 1.5 million dollars and was more sensitive to FCR, selling price and productivity fluctuations than the FS. The FS required a relatively small initial investment (US$ 16,000 dollars), which makes it more flexible to variations in production parameters and market fluctuations. The NPV was positive for both systems, and the payback times were estimated to be 3.88 years for the IS and 2.07 years for the FS. Therefore, given the assumptions of this study, cage culture of cobia in Brazil may be considered economically feasible in offshore production systems and in near-shore, FSs. Governmental support through decreased import taxes is recommended as a way to accelerate the early development of the cage culture of marine fish in Brazil.     

Effects of soy sauce <Emphasis Type="Italic">koji</Emphasis> and lactic acid bacteria on the fermentation of fish sauce from freshwater silver carp <Emphasis Type="Italic">Hypophthalmichthys molitrix</Emphasis>

The effects of soy sauce koji and the lactic acid bacterium, Tetragenococcus halophilus, were studied on the fermentation of fish sauce prepared from Chinese silver carp. The fish sauce prepared without koji and the lactic acid bacterium contained low levels of organic materials, total nitrogen, and organic acids. The use of koji was effective in increasing these qualitative parameters and further improved the amino acid score of the fish sauce. Addition of T. halophilus had an effect on lowering the pH value during the initial period of fermentation when the soy sauce koji was also supplemented. In contrast, T. halophilus-like bacteria were found to be predominant for all tanks fermented under the different starting conditions. Although it was not examined whether the T. halophilus-like bacteria observed after fermentation were the same as the starter-bacterium or not, it was suggested that T. halophilus plays an important role in the successful fermentation of silver carp fish sauce. Sensory evaluation conducted with Japanese and Chinese panelists also suggested the superiority of the use of koji for fermentation of silver carp fish sauce.     

Compensatory growth of Chinese shrimp, <Emphasis Type="Italic">Fenneropenaeus chinensis</Emphasis> following hypoxic exposure

Compensatory growth following stress is a strategy aquatic animals use to adjust themselves to a variable environment. Studies on the recovery growth of aquatic animals are not only of theoretical value in ecophysiology and evolution, but also important to applications in aquaculture and fisheries resource management. In this experiment, juvenile Fenneropenaeus chinensis with an initial average body weight of about 3.72 g were exposed to hypoxic water (about 2.08 and 3.11 mg/l of dissolved oxygen (DO) content) for 10 days and then switched to normoxic water (about 5.63 and 5.59 mg/l DO). Compared with juveniles in normoxia, juveniles in the hypoxia period allocated a greater proportion of energy to metabolism and exuviations but allocated less energy to daily metabolism per gram shrimp weight (J/g/day). This reduced feed conversion efficiency and feeding rate. Finally, F. chinensis suffered growth depression. The juveniles completely compensated for hypoxia-induced growth depression in 30 days after being switched into normoxic water and the compensation was achieved mainly by hyperphagia and slightly by improvement of feed conversion efficiency. During the recovery period the hypoxic-stressed shrimp showed higher daily metabolic energy (J/g/day) than controls (P < 0.05). Which means the stressed shrimp had more energy for feeding-related activities. So hyperphagia was observed. Energy analysis indicated that F. chinensis improved feed-conversion efficiency mainly by reducing the percentage of energy lost in feces and exuviations. The results showed that short-term non-lethal hypoxia would not affect the growth of juvenile F. chinensis if there were enough time for the stressed shrimp to recover. This suggested F. chinensis was capable of adapting to DO fluctuation to some extent and short-term non-lethal hypoxia would not have an obvious effect on natural, released, and cultured shrimp stock.     

Age and growth of two gizzard shads, <Emphasis Type="Italic">Nematalosa come</Emphasis> and <Emphasis Type="Italic">N. japonica</Emphasis>, in coastal waters around Okinawa Island,southwestern Japan

The age and growth of two Nematalosa species around Okinawa Island were examined using sectioned otoliths collected from September 2003 to April 2006. Monthly changes in the frequency of the appearance of a translucent band on the outer margin of the otoliths indicated that ring formation occurred once a year from January to July for Nematalosa come and from January to March for Nematalosa japonica. The von Bertalanffy growth equations for both species were as follows: N. come: L _t  = 365.5{1 − exp[−0.111 × (t + 0.288)]} for females and L _t  = 214.7{1 − exp[−0.700 × (t – 1.110)]} for males; N. japonica: L _t  = 205.1{1 − exp[−1.068 × (t − 1.180)]} for females and L _t  = 195.5 {1 − exp[−1.293 × (t − 1.269)]} for males. The maximum ages observed for N. come and N. japonica were 11 and 6 years old, respectively. The growth of these species was characterized by the slow growth of N. come over many years, resulting in a larger size than N. japonica.     

Pyridoxine deficiency of grouper, <Emphasis Type="Italic">Epinephelus coioides</Emphasis>: physiological and biochemical alteration

Pyridoxine is essential for animals. The purpose of this study was to investigate the pyridoxine deficiency symptoms and the fit dose of pyridoxine to keep normal physiological function in grouper, Epinephelus coioides. Graded levels of pyridoxine (0, 1, 2, 4, 8, 16, 32 mg pyridoxine kg⁻¹ diet) were fed to grouper juveniles (14.83±0.31 g) for 8 weeks, and then some physiological and biochemical parameters were measured. Pyridoxine deficiency symptoms such as anorexia, poor survival, convulsion, helically swimming and hyperirritability were observed in fish fed the pyridoxine-deficient diet, and these fish also showed low whole blood haemoglobin level, liver glutamic-pyruvic transaminase (GPT) and superoxide dismutase (SOD) activities. Slight symptoms of pyridoxine deficiency also appeared among fish fed the diet with very low dose (1 mg pyridoxine kg⁻¹ diet) of pyridoxine in 6 weeks. Broken-line regression analysis showed that the optimal pyridoxine levels for these fish to keep normal survival, whole blood haemoglobin level, liver GPT and SOD activities were approximately 1.75, 1.87, 2.22 and 2.05 mg kg⁻¹, respectively.     

Plasma biochemical responses of the omnivorous crucian carp (<Emphasis Type="Italic">Carassius auratus</Emphasis>) to crude cyanobacterial extracts

Healthy crucian carp (Carassius auratus) were treated by intraperitoneal (i.p.) injection of crude cyanobacterial extracts at two doses, 50 and 200 μg MC-LR equiv kg⁻¹ BW. High mortality (100%) was observed within 60 h post injection in the high-dose group. In the treated fish, activities of four plasma enzymes, alanine aminotransferase (ALT), alkaline phosphatase (ALP), aspartate aminotransferase (AST), and lactate dehydrogenase (LDH), all showed substantial increases, with both dose and time-dependent effects. These increases of enzyme activity indicate severe impairment occurred in the liver of crucian carp over time. Plasma concentrations of energy-related biomolecules including glucose (GLU), cholesterol (CHO), triglyceride (TG), and total protein (TP) showed marked changes in the high-dose group, possibly a nutritional imbalance correlated with the liver injury caused by intraperitoneal exposure to crude cyanobacterial extracts.     

Circadian melatonin release <Emphasis Type="Italic">in vitro</Emphasis> by European sea bass pineal

Melatonin production by the fish pineal is under the control of photoperiod. In addition, in some fish species pineal melatonin production is self-sustained by an intrapineal circadian pacemaker, not in salmonids, though. The aim of this research was to test melatonin release in vitro by the pineal of sea bass held under LD 12:12 and DD conditions to determine the endogenous nature of melatonin rhythm. Circadian rhythmicity ( = 28.3 h) was found since rhythmic melatonin release persisted on DD.