Effects of probiotics (Pseudomonas synxantha and Pseudomonas aeruginosa) on the growth, survival and immune parameters of juvenile western king prawns (Penaeus latisulcatus Kishinouye, 1896)

The effects of two selected probiotics ( Pseudomonas synxantha and Pseudomonas aeruginosa ) on the specific growth rate (SGR), survival and immune parameters of juvenile western king prawns ( Penaeus latisulcatus ) were examined for 84 days. Pseudomonas synxantha and P. aeruginosa were either applied into the rearing medium or supplemented in the formulated feed at a concentration of 10⁵ colony forming unit (CFU) mL⁻¹ for each probiotic. The results showed that applications of the probiotics resulted in no significant difference ( P >0.05) in the SGR and survival of the prawns, but significantly decreased the food conversion ratios ( P <0.05) compared with the control where no probiotics were applied. The supplement of the probiotics with the formulated feed enhanced the probiotic numbers in the intestines of the prawns and low bacteria numbers in the haemolymph. The prawns were significantly ( P <0.05) healthier, as indicated by the immune response obtained when both probiotics were given in combination compared with when individual probiotics were used. When applied singly, P. aeruginosa resulted in a higher health status of the prawns compared with P. synxantha . Overall, equal combinations (10⁵ CFU mL⁻¹) and volumes of P. synxantha and P. aeruginosa used as a supplement in the formulated feed resulted in an improvement in the health of the juvenile western king prawns.     

Effect of Customized Probiotics on the Physiological and Immunological Responses of Juvenile Western King Prawns (Penaeus latisulcatus Kishinouye, 1896) Challenged with Vibrio harveyi

Juvenile western king prawn P. latisulcatus were fed 10⁵ colony-forming units (CFU)/mL of two probiotics Pseudomonas synxantha and P. aeruginosa for 28 days. P. latisulcatus were then challenged with V. harveyi at 0 (control), 10³, 10⁵, and 10⁷ CFU/mL. During the seven days of challenge, disease resistance of the probiotic-fed prawns was compared with that of prawns not fed probiotics. The immunological responses of the prawns did not improve during the challenge period in terms of total haemocyte count, hyalinocyte, semi-ganulocyte, granulocyte, clotting time, bacteraemia, and intestinal bacterial load. Overall, when prawns were challenged with V. harveyi, the LT₅₀ values got shorter as V. harveyi concentration increased. LT₅₀ values of prawns fed probiotics were significantly longer (P < 0.05) than those not fed probiotics. At a V. harveyi concentration of 10³ CFU/mL, the 100% survival of the prawns fed probiotics was three times more likely than those of the prawns not fed probiotics.     

Effects of feeding rates on the growth,water quality and nutrient budget of western king prawn (Penaeus latisulcatus Kishinouye) reared in recirculating aquaculture systems

Feed rate is one of the important variables considered for the improvement in production of prawns. The effects of four feeding rates on growth, water quality and the nutrient budget of Penaeus latisulcatus were investigated for 98 days. The specific growth rate (SGR) at 3.0% feed rate was significantly higher than at all higher feed rates, and there was no significant difference in SGR between 4.5%, 6% and 7.5%. Total ammonia nitrogen concentrations in all treatments remained low in the safe range for prawn culture. Dissolved inorganic nitrogen and nitrate nitrogen were significantly higher (P < 0.05) in prawn tanks received feed rates of 6.0% and 7.5%. Total phosphorus and the soluble reactive phosphorus linearly increased with the progression of the trial and feeding rates. Nutrient conversion rates were significantly higher (P < 0.05) in prawns receiving the feeding rate of 3.0% than feeding rates of 6.0% and 7.5%. The lowest food conversion ratio was recorded at feeding rates of 3% and 4.5%. Nutrient budget showed that 6.01–15.99% nitrogen and 4.53–9.29% phosphorus of the nutrient inputs were retained in the prawn biomass. The results indicated that the growth and nutrient conversion rate may not increase with the increase above 3.0% of biomass per day.     

Effects of stocking density on the nutrient budget and growth of the western king prawn (Penaeus latisulcatus Kishinouye) in a recirculating aquaculture system

Intensification in the commercial culture of prawns can have a significant impact on the water quality and hence on the survival, growth and the surrounding environment. The present study aims to evaluate the effects of stocking density on the water quality and performance of the western king prawns (Penaeus latisulcatus) and the nutrient budget of the culture environment. Four stocking densities of 4, 8, 16 and 32 prawns m^?2 were tested in 12 recirculating systems. Prawn weight and specific growth rate increased with decreasing stocking density, while the survival rate showed the reverse trend. The mean total ammonia nitrogen, nitrate nitrogen, total phosphorus and soluble reactive phosphorus were significantly higher (P<0.05) at the higher stocking density. The nutrient budget revealed that the prawns could assimilate only 9.34–20.13% nitrogen and 4.97–11.25% phosphorus of the total nutrient inputs. The drained water at harvest was the major sink of phosphorus and nitrogen at stocking densities of 4, 8 and 16, which accounted for 45.59–64.82% and 44.28–65.62% of the total inputs, respectively, while a significant proportion of nitrogen sunk into the sediment at 32 prawns m^?2. The study suggested that the stocking densities of western king prawns can be up to 16 prawns m^?2 in the recirculating water environment.     

The Effect of Potassium Concentration in Inland Saline Water on the Growth and Survival of the Western King Shrimp,Penaeus latisulcatus Kishinouye, 1896

Abstract

The survival, growth, and condition of the western king shrimp, Penaeus latisulcatuswere measured for 42 days when grown in marine water and inland saline water with four different concentrations of potassium: 100%, 80%, 60% or 40% of the concentration of potassium in marine water (IS100, IS80, IS60, and IS40). Shrimp survival and cannibalism frequency decreased with decreasing potassium concentration, with final shrimp survival in IS100 significantly higher (P < 0.05) than in IS60 and IS40 and significantly higher (P < 0.05) in MW and IS80 than in IS40. Specific growth rate (SGR), moult increment, and diet ingestion rate of shrimp were significantly higher (P < 0.05) in shrimp grown in marine water than the other water types. At the conclusion of the trial, exoskeleton mineral content of shrimp in IS100 and IS80 was significantly higher (P < 0.05) than in IS60 and IS40. Tail muscle moisture content of shrimp in IS60 and IS40 was significantly higher (P < 0.05) than in the other water types, suggesting a depletion of tail muscle energy reserves in IS60 and IS40. This trial demonstrates the necessity of fortifying potassium-deficient inland saline water to allow the culture of western king shrimp. Furthermore, a potassium concentration greater than approximately 76% of that in marine water and a Na/K ratio of less then 39 appear to be necessary to maintain a comparable level of shrimp survival, as in marine water. However, the slower growth and reduced molt increment of shrimp in inland saline water with 100% and 80% potassium compared to shrimp in marine water demonstrates the presence of other limiting factors.     

Effects of enriched Artemia nauplii on production, survival and growth of the mysid shrimp Mysidopsis almyra Bowman 1964 (Crustacea: Mysidacea)

The effects of enriched Artemia nauplii on larvae production and survival and growth of the mysid Mysidopsis almyra Bowman 1964 are compared. There were no significant differences (P > 0.05) in production between mysids fed the Artemia nauplii (133 ± 69 mysids day⁻¹) and mysids fed the enriched nauplii (139 ± 82 mysids day⁻¹). No differences in size of newly hatched mysids or mysid growth to 15 days (P > 0.05) were found between the two diets. Survival was significantly higher (P < 0.05) for mysids fed the enriched nauplii (59.1%) compared with mysids fed Artemia nauplii (41.4%).     

Use of Cyclop-eeze as a substitute for Artemia nauplii in larval rearing of giant freshwater prawn Macrobrachium rosenbergii (De Man 1879)

Four feeding experiments, replacing 25% (T₁), 50% (T₂), 75% (T₃) and 100% (T₄), by dry weight, of the live feed Artemia nauplii for Cyclop‐eeze, a new larval feed that was claimed to contain the highest known levels of astaxanthin and omega‐3 polyunsaturated fatty acids, were compared against a control that was fed with Artemia and egg custard alone, to the larvae of giant freshwater prawn Macrobrachium rosenbergii (De Man 1879). Analysis of different production characteristics of the larvae revealed that the highest survival up to postlarvae (PL) stage was obtained for T₂ in which 50% of the Artemia nauplii were replaced by Cyclop‐eeze [freeze‐dried (FD) deep frozen (DF)], and the highest astaxanthin content of the larval tissue obtained in T₄ in which the larvae were fed 100% Cyclop‐eeze, although the survival rate was the lowest in this treatment. The costs of different treatments were also compared. The Artemia consumption million⁻¹ larvae was the highest in control (11490 g), followed by T₁ (8240 g), T₂ (4990 g), T₃ (3730 g) and T₄, which completely replaced Artemia from stage 5 onwards (1830 g). The highest consumption of Cyclop‐eeze million⁻¹ larvae was in T₄ (1670 and 10 880 g), followed by T₃ (850 and 5560 g), T₂ (410 and 2690 g) and T₁ (230 and 1490 g) of FD and DF, respectively. The astaxanthin contents of the late‐stage larvae fed under the four treatments were 24.90, 27.40, 28.60 and 35.60 μg g⁻¹ tissue for T₁, T₂, T₃ and T₄, respectively, while that of the control was 23.70 μg g⁻¹. The lowest cost of live feeds million⁻¹ PL was obtained for T₂ (US$ 428.60), followed by T₁ (US$ 490.46), control (US$ 529.07) and T₃ (US$ 583.26), while it was the highest for T₄ (US$ 890.93). The results indicated that Cyclop‐eeze could economically replace Artemia nauplii at 50% level that could significantly improve the survival and carotenoid composition of the larvae of M. rosenbergii.     

Growth,survival and fatty acid composition of Rhamdia quelen (Quoy and Gaimard, 1824) larvae fed on artificial diet alone or in combination with Artemia nauplii

Black catfish (Rhamdia quelen) is a species of interest for aquaculture in Brazil, Argentina and Uruguay. A feeding trial was conducted to evaluate the effect of feeding R. quelen larvae on either only an artificial diet or in combination with Artemia nauplii (AN) on larval performance and fatty acid composition. For 12 days, larvae were fed from first feeding (3 days after hatching, TL = 5.88 ± 0.23 mm) with artificial food only or a combination of artificial food and AN (co‐feeding). At the end of the trial, total length of co‐fed larvae was significantly higher than that of larvae fed solely artificial food (P < 0.001). No significant differences were found in survival rates. Co‐feeding microdiet with a small amount of AN significantly affected larval fatty acid composition. Lipid and fatty acid composition of food and larvae revealed the importance of n‐3 fatty acids for growth of black catfish larvae and that, as most freshwater fish, R. quelen larvae can elongate and desaturate linolenic acid to n‐3 highly unsaturated fatty acids. Results suggest that R. quelen larvae can be fed from first feeding on microdiets as unique food source, although better larval performances are obtained by co‐feeding with a small amount of AN.     

Ovoviviparously produced Artemia nauplii are a suitable live food source for the larvae of the African catfish (Clarias gariepinus: Burchell, 1822)

Brine shrimp Artemia, the most common live food organism used in larviculture, can reproduce either oviparously (production of dormant cysts) or ovoviviparously (direct production of nauplii), depending on environmental conditions. Ovoviviparous Artemia nauplii have seldom been considered as a source of live food in aquaculture, partly due to the convenience and the developed techniques associated with the production and use of the dormant cysts. In many countries in Africa, however, hatchery managers do not have access to a reliable supply of affordable good quality cysts. In this study, we therefore demonstrated the potential of a system designed for the continuous ovoviviparous production of nauplii at low salinity, using Great Salt Lake Artemia franciscana and micronized agricultural material as feed. The suitability of the produced nauplii was tested by feeding them directly to Clarias gariepinus larvae in comparison with oviparous nauplii and decapsulated cysts. Higher survival (100%), better protein efficiency ratio (2.6 ± 0.1) and food conversion ratio (1.0 ± 0.1) was observed in larvae fed with the ovoviviparous nauplii (p < 0.05). Overall, we conclude that the ovoviviparous nauplii could serve as an alternative live food for larval fish. If optimized, the system could be validated for integration in hatcheries.     

The effect of vitamins C and E in (n-3) highly unsaturated fatty acids-enriched Artemia nauplii on growth, survival, and stress resistance of fresh water walleye Stizostedion vitreum larvae

This study aimed to evaluate the effect of enriching Artemia nauplii with vitamin C (ascorbyl-6 palmitate) or vitamin E (α-tocopherol acetate), 20% w/w, together with a mixture of concentrated eicosapentaenoic acid (EPA, 20:5 n-3) and docosahexaenoic acid (DHA, 22:6 n-3) on the growth, survival, and stress resistance of fresh water walleye Stizostedion vitreum larvae. Either cod liver oil (CLO) or EPA/DHA ethyl esters concentrate was used as lipid sources in the Artemia enrichment. Walleye larvae were fed ad libitum for 40 days. At day 40, submersion in salt water (25 g L⁻¹) was performed to evaluate larvae resistance to stress. EPA and DHA levels in walleye juveniles fed EPA/DHA-enriched Artemia increased significantly, by an average of 650% compared with fish fed non-enriched Artemia . A significant increase was found for vitamins C (71.8 ± 1.0 and 42.7 ± 1.2 μg g⁻¹ wet weight (WW)) and E (17.0 ± 3.7 and 6.5 ± 0.9 μg g⁻¹WW) concentrations in fish fed enriched and unenriched Artemia , respectively. Growth was comparable throughout treatments, whereas survival was significantly higher in fish fed CLO-enriched Artemia nauplii compared with fish fed Artemia nauplii enriched with EPA/DHA concentrate. The addition of vitamin C increased fish survival by 1.4-fold compared with fish fed Artemia enriched with only EPA/DHA concentrate. The survival of the latter was similar to control fish ( Artemia without enrichment). The supplementation of vitamin E did not affect fish survival significantly. Stress tests revealed that the resistance of walleye larvae to salinity changes increased when Artemia enrichment was supplemented with vitamin C. However, walleye larvae fed CLO-enriched Artemia had the best performances in the stress test.     

The use of probiotics during the nursery rearing of the pink shrimp Farfantepenaeus brasiliensis (Latreille, 1817) in a zero exchange system

The present work evaluated the use of probiotics during the nursery rearing of the pink shrimp Farfantepenaeus brasiliensis, in a zero exchange aerobic heterotrophic culture system during 30 days. Three replicate tanks were randomly assigned to the following treatments: (1) Bacillus spp. mixture (Sanolife Pro‐‐W^®), (2) Bacillus sp., Enterococcus sp., Lactobacillus spp. mixture (Biomin Start‐grow^®), (3) Bacillus cereus var. toyoi and (4) control treatment (without probiotic addition). Bacteriological analysis monitored the abundance of presumptive Vibrio spp. in the water of experimental tanks. For the immunological analysis, shrimp haemolymph was collected to determine the granular haemocyte count and total protein concentration. Results showed that mean final weight and specific growth rate of shrimp in the probiotic treatments were significantly higher. Furthermore, shrimp reared in the probiotic treatments showed higher levels of total protein and granular haemocyte. The bacteriological analysis showed that the concentration of Vibrio spp. measured in probiotic treatment tanks was lower than that recorded in the control tanks.     

Treatment of vibriosis in European sea bass larvae, Dicentrarchus labrax L., with oxolinic acid administered by bath or through medicated nauplii of Artemia franciscana (Kellogg): efficacy and residual kinetics

European sea bass larvae were challenged by bath with Listonella anguillarum strain 332A, 2.5×10(7) CFUmL(-1) for 1h. Fish either received no treatment or oral treatment with Artemia franciscana (Kellog) nauplii enriched with oxolinic acid, or bath treatments with oxolinic acid. Medication commenced 1day following challenge and was performed on days 1, 3 and 5 post-challenge at a dosage of 20mgL(-1) for 2h for bath treatments, while two doses each of 750 nauplii per fish were administered daily for five consecutive days in oral treatments. Cumulative mortality reached 96% for the unmedicated challenged group, 32% in the group receiving bath treatments and 17% in the group receiving medicated nauplii. Pharmacokinetic parameters of oxolinic acid were calculated in sea bass larvae, for both treatments. Steady-state concentrations of oxolinic acid of 48.0 and 75.2μgg(-1) were achieved for bath treatment and oral treatment, respectively, while the elimination half-life was calculated to be 25.1h for bath treatment and 21.7h for oral treatment.     

The development of probiotics for the control of multiple bacterial diseases of rainbow trout, Oncorhynchus mykiss (Walbaum)

JB-1 and GC2, which were equated with Bacillus sp. and Aeromonas sobria respectively, were recovered from the digestive tract of rainbow trout, Oncorhynchus mykiss and ghost carp, Cyprinus sp. respectively, and demonstrated effectiveness as probiotics for the control of infections caused by Aeromonas salmonicida, Lactococcus garvieae, Streptococcus iniae, Vibrio anguillarum, Vibrio ordalii and Yersinia ruckeri. When administered to rainbow trout (average weight = 12 g) for 14 days in feed dosed at 2 x 10(8) cells g(-1) of feed, JB-1 led to a reduction in mortalities to 0-13% after challenge with a range of bacterial pathogens compared to 80-100% mortalities of the controls. Similarly, use of GC2 reduced mortalities to 0-16% following the challenge compared to 80-100% mortalities of the controls. The mode of action reflected nutrition, production of inhibitory substances and stimulation of the innate immune responses. Specifically, JB-1 and especially GC2 were positive for siderophore and chitinase production, and increased lysozyme, phagocytic and respiratory burst activity.     

Effects of two dietary probiotics (Bacillus subtilis or licheniformis) with two prebiotics (mannan or fructo oligosaccharide) in Japanese eel,Anguilla japonica

A 12‐week feeding trial was conducted to investigate the effects of two dietary probiotics; Bacillus subtilis KCTC 2217 or Bacillus licheniformis KCCM 11775 with two prebiotics; mannan oligosaccharide (MOS) or fructooligosaccharide (FOS) in Japanese eel. Fish averaging 12.8 ± 0.47 g (mean ± SD) were randomly distributed into five treatments with triplicate tanks. A basal control diet (CON) and four synbiotic diets supplementing B. subtilis + MOS (BSM), B. subtilis + FOS (BSF), B. licheniformis + MOS (BLM), and B. licheniformis + FOS (BLF). Weight gain and specific growth rate of fish fed all synbiotic diets were higher than those of fish fed CON. Immune‐related gene expression of heat shock protein 70 and immunoglobulin M of fish fed BSF and BLM were significantly higher than those of fish fed CON. Fish fed BSF and BLM had significantly higher intestinal villi length than those of fish fed BLF and CON (p < .05). Disease resistance against Aeromonas hydrophila of fish fed all synbiotic diets were significantly higher than those of fish fed CON (p < .05). Therefore, these results indicated that dietary B. subtilis with FOS (BSF) and B. licheniformis with MOS (BLM) could have beneficial effects on intestinal morphology, and immune‐related gene expression in Japanese eel.     

Effect of probiotics on the survival, growth and challenge infection in Tilapia nilotica (Oreochromis niloticus)

This study was conducted in order to evaluate the potential benefit of Bacillus pumilus and a commercial product (‘Organic Green'^?) as a probiotic in the culture of the tilapia nilotica. Two doses of B. pumilus (10⁶ and 10¹² g^?1 diet fed) and Organic Green^? (1 and 2 g kg^?1 diet fed) were used as feed additives and administered for periods of 1 (groups 2, 3, 4 and 5 respectively) and 2 (groups 6, 7, 8 and 9 respectively) months; group 1 served as a control. Each group consisted of 4 equal replicates of 320 fish. Body weight, nitroblue tetrazolium (NBT) activity, haematocrit values and total and differential leucocytic counts (TLC and DLC) were recorded in each group after 1 and 2 months. At the end of the experiment (8 months), survival and individual body weights were recorded. Challenge infections were performed after 1, 2 and 8 months using 0.5 mL culture suspension of a pathogenic reference strain of Aeromonas hydrophila (10⁸ bacteria mL^?1). The NBT values were increased significantly in all treated groups, except those administered B. pumillus (groups 3 and 6). There were significant changes in haematocrit values and TLC and DLCs in the treated groups, except group 3, which showed a significant increase in TLC, lymphocytes and monocytes. There was a statistically significant increase in the individual body weight in groups 5, 6, 8 and 9. The survival rates of all treatment groups were higher than the control group, the differences being statistically significant in groups 3 and 9. The challenge infection showed a variable response with the type and dose of treatment and the period of application. The highest relative levels of protection were an average increase of 62.5% after 1 month (groups 3 and 5), 55.6% after 2 months (group 7) and 15.8% after 8 months (groups 8 and 9). Overall, both types of probiotics induced a similar effect. The results show the potential of using probiotics to enhance immune and health status and improve disease resistance in Oreochromis niloticus, thereby improving growth performance. However, further extensive testing, including a full commercial cost–benefit analysis, is necessary before recommending their application in aquaculture.     

Enrichment of Artemia nauplii in essential fatty acids with different types of liposomes and their use in the rearing of gilthead sea bream (Sparus aurata) larvae

The present study aimed to optimize the enrichment protocols of Artemia nauplii with highly unsaturated fatty acid liposomes. Three different types of liposomes formulated with krill phospholipid extract were tested, using a commercial emulsion as the control treatment. The incubation time of the nauplii in the enrichment medium was chosen as the first variable studied. Total lipid, lipid class and fatty acid profiles were analyzed in enriched nauplii after 18 and 21 h incubation in the enrichment media. A third sampling point at 24 h, where nauplii had been kept in clean seawater at 28 °C for 3 h, was established to emulate potential changes in the nutritional value of the nauplii after some hours in the larval rearing tank. A second objective of the study was the establishment of a suitable liposome-based diet for the rearing of gilthead sea bream larvae, as a model of marine fish.Differences in the enrichment efficiencies through time were found not only between liposome and emulsion based diets, but also among the different types of liposomes. Whereas nauplii enriched with multilamellar vesicles and unilamellar liposomes prepared by extrusion had maximum enrichment performance at 18 h of incubation, nauplii enriched with unilamellar liposomes prepared by the detergent solubilization method followed the enrichment pattern of the control emulsion, with an optimal incubation period closer to 21 h. Analyses of the diets of the larval nutrition trial seemed to confirm the results obtained in the first experiment, although changes in certain variables occurred among the enrichments carried out during the larval feeding trial. Biochemical characterization of body lipids of the larvae indicated differences in total lipid and essential fatty acid content, although no significant differences were found in growth and survival of the larvae. Thiobarbituric acid reactive substances (TBARS), used to determine the peroxide content of the larvae, indicated a significant higher content of these compounds in larvae fed emulsion enriched nauplii compared to larvae fed liposome enriched prey.     

Isolation and screening of probiotic candidates from marron,Cherax cainii (Austin, 2002) gastrointestinal tract (GIT) and commercial probiotic products for the use in marron culture

Six strains of bacteria including Bacillus mycoides (A10) and Shewanella species (A12) isolated from healthy marron intestine, Bacillus species (PM1), Bacillus subtilis (PM3), Bacillus sp. (PM4) and Bacillus sp. (AQ) from commercial probiotic products were investigated for probiotic potential in marron culture. Antibiotic susceptibility tests indicated PM3 and PM4 were susceptible to all nine antibiotics evaluated. A10, A12 and AQ were resistant to class penicillins, whereas PM1 was resistant to class penicillin and macrolides. All strains were non‐pathogenic for marron. Strong inhibition against Vibrio mimicus and Vibrio cholerae non‐01 was exhibited by PM4 and PM3. A10 inhibited V. mimicus at 72 h of growth, but not V. cholerae non‐01, whereas A12 inhibited V. cholerae non‐01 but not V. mimicus, and AQ showed no inhibition activity. A wide range of enzymes were produced by A10 and AQ using the API ZYM test. Protease enzymes were produced by PM3, PM4, AQ and PM1. In order of effectiveness, the following bacteria have probiotic potential: B. subtilis (PM3), Bacillus sp. (PM4) and B. mycoides (A10). Further study is required to determine the bacterium or any combination that gives a multibeneficial effect on marron.     

Changes in the digestive enzymes and hormones of gilthead seabream larvae (Sparus aurata, L. 1758) fed on Artemia nauplii enriched with free lysine

Variations in digestive enzymes and hormones during the larval development of gilthead seabream (Sparus aurata) fed on live prey (Artemia nauplii) enriched with free lysine were investigated for 16 days (from day 24 to day 40). Prior to initiation of the experiment, newly hatched larvae were transferred from incubators to fiber glass tanks (300 l) with black walls and fed the same diet until day 24. Each experiment was performed in triplicate. The amount of free lysine in the Artemia nauplii was increased by adding a 5.3 mM free amino acid solution to the culture water during a 16-h enrichment period. Larvae were sampled four times at four-day intervals, for 16 days. Larvae fed on the control group had lower growth than those fed on the lysine group at the end of the study but the difference was not significant (P > 0.05). The highest trypsin activity and lap/leu–ala peptidase ratios were observed in the control group (P < 0.05). Significant differences between bombesin activities of the treatment groups were not found in the fifth minute after the start of feeding (P > 0.05) but were significant in the fifteenth minute (P < 0.05). Significant differences between the CCK levels of the treatment groups were found (P < 0.05). In conclusion, three main ideas are presented in this study. First, Artemia nauplii can successfully be enriched with free lysine. Second, bombesin (GRP) and cholecystokinin (CCK) activity can be stimulated by free lysine. Third, the mechanisms controlling adaptation of the activity of the trypsin to the amount of dietary protein were not activated until day 40.     

Establishing larval feeding regimens for the Forktail Blenny Meiacanthus atrodorsalis (Günther, 1877): effects of Artemia strain,time of prey switch and co‐feeding period

This study aimed to establish feeding strategies covering the whole larval period of the forktail blenny, Meiacanthus atrodorsalis, based on the standard hatchery feeds of rotifers and Artemia. Three purposely designed experiments were conducted to determine the appropriate times and techniques to transition larvae from rotifers onto Artemia nauplii of a Great Salt Lake (GSL) strain, and a specialty AF strain, as well as subsequent transition onto enriched metanauplii of GSL Artemia. With a 3‐day co‐feeding period, larvae adapted well to a transition from rotifers to newly hatched GSL Artemia nauplii as early as 5 days posthatching (DPH), and as early as 3 DPH when fed the smaller AF Artemia nauplii. However, prolonging the rotifer‐feeding period up to 11 DPH did not negatively affect survival. Larvae fed Artemia nauplii of the AF strain showed 17–21% higher survival, 24–33% greater standard length and body depth, and 91–200% greater dry weight, after 20 days relative to those fed nauplii of the GSL strain. Meanwhile, enriched Artemia metanauplii of the GSL strain were shown to be an acceptable alternative to AF Artemia nauplii for later larvae, producing similar survival and growth when introduced from 8 DPH. Based on our findings, we recommend feeding M. atrodorsalis larvae rotifers as a first food between 0 and 2 DPH, introducing AF Artemia nauplii from 3 DPH, followed by enriched GSL Artemia metanauplii from 8 DPH onward, with a 3‐day co‐feeding period between each prey change.     

Effects of diets supplemented with either individual or combined probiotics,Bacillus subtilis E20 and Lactobacillus plantarum 7‐40, on the immune response and disease resistance of the mud crab,Scylla paramamosain (Estampador)

Two trials, including firstly, diets incorporating individual or combined species of probiotics, Bacillus subtilis E20 and Lactobacillus plantarum 7‐40, were fed to the mud crab, Scylla paramamosian, for 28 days to evaluate the growth, immunity and disease resistance, and secondly, the optimal level of B. subtilis E20 in the diet by tracking the growth, immunity and disease resistance after 28 days of feeding were analysed. No significant differences in growth, total haemocyte counts, respiratory bursts, superoxide dismutase, or glutathione peroxidase were found in the two trials. Crabs fed the B. subtilis E20‐containing diet (D20) had significantly increased phenoloxidase (PO) activity, phagocytic activity (PA) and disease resistance compared with crabs fed the control and the mixed probiotics diet (MD). The mortality of crabs fed the D20 was significantly lower than that of crabs fed the L. plantarum 7‐40‐containing diet. L. plantarum 7‐40 had a great inhibitory effect on the growth of B. subtilis E20, which may have led to the decreased probiotic effect of the MD. An analysis of the optimal level of B. subtilis E20 in the diet showed that crabs fed the B. subtilis E20‐containing diet at a level of 10^9–10 cfu kg^?1 had better immune response of PO and PA, and disease resistance in the second trial.