Low optimal temperatures for food conversion and growth in the big-headed turtle, Platysternon megacephalum

We held juvenile big-headed turtles, Platysternon megacephalum, from eastern China, at temperatures from 20 to 29.4 °C to determine effects on feeding, growth and food conversion. Food intake increased significantly from 20 to 22.4 °C, remained high until 27.1 °C, and then decreased dramatically at 29.4 °C. Digestive efficiency for energy decreased as temperature increased, whereas the digestive efficiency of protein increased from 20 to 25 °C, and decreased at higher temperatures. The relationships between specific growth rate (SGR), food conversion coefficient (C_c) and temperature (T) were curvilinear, and could be described by quadratic equations: SGR = −0.01 T² + 0.47 T − 5.24 and C_c = −0.37 T² + 17.20 T − 181.85. Maximum growth was estimated to occur at 23.9 °C, with 90% of the maximum being achieved within the range of 21.9–25.8 °C; maximal food conversion occurred at 23.2 °C, with a 90% range from 21.0 to 25.4 °C. The temperature range (22–25 °C) found to promote best growth and food conversion in juvenile P. megacephalum is lower than for many other freshwater turtles. Temperatures of 22–25 °C are recommended for use in culture of this species to maximize growth and food conversion.     

Effects of photoperiod on egg production and hatching success, naupliar and copepodite development, adult sex ratio and life expectancy of the tropical calanoid copepod Acartia sinjiensis

The tropical calanoid copepod Acartia sinjiensis has good potential for mass culture as live feed for reef fish larvae. The present study was conducted to evaluate the effects of photoperiod on various parameters related to A. sinjiensis productivity in culture.Five photoperiods of Light:Dark = 0:24; 6:18; 12:12; 18:6 and 24:0h were setup. Daily egg production of individual females under each photoperiod was monitored for 8 consecutive days. The females were randomly selected daily from stock cultures kept under respective photoperiods and discarded after experiment. The results showed a clear trend of increasing egg production with longer illumination period. Under constant darkness, acclimatization was evident as egg output increased steadily over the 8 day period. Statistics showed that photoperiod significantly (p < 0.005) affected mean daily egg production, with the highest egg output recorded at 18L:6D and 24L:0D (17.6 ± 1.7 and 17.6 ± 1.8 eggs/female/day respectively), which were significantly higher than all other treatments. Photoperiod also significantly affected 48 h egg hatching success (p < 0.005), a trend of increased hatching success with longer light phase was demonstrated. The highest hatching rate (87.2 ± 1.4%) was recorded at 24L:0D, which was significantly higher than the 0L:24D and 6L:18D treatments but not significantly different from the second highest (85.3 ± 2.6%) hatching rate of 18L:6D treatment.Photoperiod was further confirmed to significantly (p < 0.005) affected naupliar and copepodite development with accelerated development observed with increased illumination period of photoperiods. Mean development time from egg to adult was the shortest at 6.00 ± 0.33 days under constant light (24L:0D), followed by 6.24 ± 0.24 days at 18L:6D, both were significantly shorter than that of 0L:24D and 6L:18D treatments although no significantly difference was detected between themselves. Adult life expectancy was also found significantly (p < 0.005) affected by photoperiod with the shortest adult life span recorded under constant light (24L:0D) (9.4 ± 0.4 days), which was significantly shorter than all other photoperiods tested. Adult sex ratio was the only parameter tested that was not significantly affected by photoperiod, a skewed sex ratio in favor of female was found across all photoperiod treatments.Based on results of current study, it is recommended that a photoperiod of 18L:6D being adopted for A. sinjiensis culture to maximize its productivity for aquaculture hatcheries.     

Broad snouted caiman (Caiman latirostris) growth under different rearing densities

The effect of density on growth in body mass (BM) and total length (TL) of Caiman latirostris hatchlings raised in small pens (area 0.36 m²), for 3 months, was investigated. Twelve animals from each of three clutches were randomly distributed (1, 2 or 3 individuals) to two replicates at low rearing density (LRD: 0.12 m²/individual), medium rearing density (MRD: 0.06 m²/individual) and high rearing density (HRD: 0.04 m²/individual. Growth at LRD [BM: 150.75 ± 43.83 g (SE); TL: 11.58 ± 0.93 cm] and MRD (BM: 141.96 ± 22.55 g; TL: 8.38 ± 1.26 cm) was significantly more than at HRD (BM: 57.19 ± 14.38 g; TL: 4.61 ± 0.89 cm; p < 0.001). Density is important to achieving optimal growth and maintaining the well-being of C. latirostris in commercial ranching programs.     

Salinity effects on reproduction of giant freshwater prawn Macrobrachium rosenbergii (de Man)

Understanding the role of salinity in breeding and growth has the potential to enhance production of giant freshwater prawn, Macrobrachium rosenbergii. This study investigated the reproduction of females reared in salinities of 0, 6, 12, and 18 g L^− 1. Mean weight of females decreased with increased salinity (31.40 ± 1.54, 25.14 ± 1.16, 20.80 ± 0.81, and 16.62 ± 1.04 g at 0, 6, 12, and 18 g L^− 1, respectively). Larval production was delayed by 2 months in females reared in 12 g L^− 1 compared to 6 and 0 g L^− 1 and the cumulative number of berried females decreased with increased salinity. The number of larvae produced was positively correlated to weight of female. A larval production not only differed between salinity treatments, but also that larval production per gram of female differed between treatments. Females reared in lower salinity of 0 and 6 g L^− 1 produced larger numbers of larvae (12,155 ± 480 and 6519 ± 323, respectively) compared to 12 and 18 g L^− 1 (3751 ± 256 and 0, respectively). The number of larvae produced per gram of female was inversely related to the salinity levels (Y = − 37.54X + 685.65, n = 339, r² = 0.995, p < 0.05). Survival of larvae from females reared at 0 and 6 g L^− 1 was higher than those from females reared in 12 g L^− 1. This study clearly shows that female broodstock reared in lower salinity was larger, reproduced early, and produced more offspring than at higher salinity and this could significantly impact coastal prawn culture where seasonal fluctuation of salinity in the hatchery is common.     

Prediction of apparent protein digestibility of ingredients and diets by in vitro pH-stat degree of protein hydrolysis with species-specific enzymes for juvenile Pacific white shrimp Litopenaeus vannamei

Aquafeed production faces global issues related to availability of feed ingredients. Feed manufacturers require greater flexibility in order to develop nutritional and cost-effective formulations that take into account nutrient content and availability of ingredients. The search for appropriate ingredients requires detailed screening of their potential nutritional value and variability at the industrial level. In vitro digestion of feedstuffs by enzymes extracted from the target species has been correlated with apparent protein digestibility (APD) in fish and shrimp species. The present study verified the relationship between APD and in vitro degree of protein hydrolysis (DH) with Litopenaeus vannamei hepatopancreas enzymes in several different ingredients (n = 26): blood meals, casein, corn gluten meal, crab meal, distiller's dried grains with solubles, feather meal, fish meals, gelatin, krill meals, poultry by-product meal, soybean meals, squid meals and wheat gluten. The relationship between APD and DH was further verified in diets formulated with these ingredients at 30% inclusion into a reference diet. APD was determined in vivo (30.1 ± 0.5 °C, 32.2 ± 0.4‰) with juvenile L. vannamei (9 to 12 g) after placement of test ingredients into a reference diet (35 g kg^− 1 CP; 8.03 g kg^− 1 lipid; 2.01 kcal g^− 1) with chromic oxide as the inert marker. In vitro DH was assessed in ingredients and diets with standardized hepatopancreas enzymes extracted from pond-reared shrimp. The DH of ingredients was determined under different assay conditions to check for the most suitable in vitro protocol for APD prediction: different batches of enzyme extracts (HPf5 or HPf6), temperatures (25 or 30 °C) and enzyme activity (azocasein): crude protein ratios (4 U: 80 mg CP or 4 U: 40 mg CP). DH was not affected by ingredient proximate composition. APD was significantly correlated to DH in regressions considering either ingredients or diets. The relationships between APD and DH of the ingredients could be suitably adjusted to a Rational Function (y = (a + bx)/(1 + cx + dx²), n = 26. Best in vitro APD predictions were obtained at 25 °C, 4 U: 80 mg CP both for ingredients (R² = 0.86; P = 0.001) and test diets (R² = 0.96; P = 0.007). The regression model including all 26 ingredients generated higher prediction residuals (i.e., predicted APD − determined APD) for corn gluten meal, feather meal, poultry by-product meal and krill flour. The remaining test ingredients presented mean prediction residuals of 3.5 points. A model including only ingredients with APD > 80% showed higher prediction precision (R² = 0.98; P = 0.000004; n = 20) with average residual of 1.8 points. Predictive models including only ingredients from the same origin (e.g., marine-based, R² = 0.98; P = 0.033) also displayed low residuals. Since in vitro techniques have been usually validated through regressions against in vivo APD, the DH predictive capacity may depend on the consistency of the in vivo methodology. Regressions between APD and DH suggested a close relationship between peptide bond breakage by hepatopancreas digestive proteases and the apparent nitrogen assimilation in shrimp, and this may be a useful tool to provide rapid nutritional information.     

Growth rates and survival of western rock lobster (Panulirus cygnus) at two temperatures (ambient and 23 °C) and two feeding frequencies

Wild caught post-pueruli, year one and year two post settlement juvenile western rock lobster, Panulirus cygnus, were held at ambient temperatures (15.6 °C to 23.1 °C; mean 19.0 ± 0.07 °C) or at 23 °C, and fed the same ration of a formulated pellet diet either once per night, or 3 times per night, over 12 months, to determine whether elevated temperatures and multiple feeds per night would stimulate growth through increased metabolism and feed utilisation without significant negative impacts on survival. Survival of post-pueruli (mean 63%) did not differ between ambient and 23 °C. Survival of year 1 and 2 juveniles was higher at ambient temperatures (p < 0.01 ambient: year 1 juveniles, 68%; year 2 juveniles, 88%; 23 °C: 57% and 74%, respectively). Feeding frequency did not affect survival of post-pueruli and year 2 juveniles (mean 63%, 81% respectively), but survival was 9% higher for year 1 juveniles fed three times per night (58% versus 67%; p < 0.01). All lobsters grew faster at 23 °C than at ambient temperatures (p < 0.05), with the growth of post-pueruli almost doubled at 23 °C (weight gain at 23 °C versus ambient: post-pueruli, 18 438 % versus 9 915 %; year 1 juveniles 259% versus 165%; year 2 juveniles 23% versus 21%). Feed frequency did not influence the growth of year 1 and 2 juveniles. However, there was an interaction effect of temperature and feed frequency on post-pueruli where weight and carapace length were significantly higher at ambient temperatures when post-pueruli were fed three times a day, whereas at 23 °C weight and carapace length were significantly greater when fed once per day (p < 0.05). Feed intake (g pellet dry matter lobster^− 1 day^− 1) of pellet was higher at 23 °C for all lobsters (p < 0.05), but was the same between lobsters fed 3 times per night versus once per night. This study has shown that increasing temperatures to 23 °C significantly improved the growth of P. cygnus post-pueruli without any adverse effects on survival. The faster growth rates exhibited by year 1 and 2 juveniles at 23 °C may potentially offset their lower survival by significantly reducing culture period. There is no benefit of feeding P. cygnus multiple times at night in terms of growth and survival. The implications for P. cygnus culture are that temperatures should be maintained close to 23 °C during the entire growout period, with due care taken to minimise mortalities through adequate provision of food and shelter. Feeding P. cygnus once daily to excess just prior to dusk to co-incide with nocturnal feeding behaviour is recommended.     

Olive barb, Puntius sarana (Hamilton) is a potential candidate species for introduction into the grow-out carp polyculture system

The compatibility of olive barb, Puntius sarana (Hamilton) with major carps was studied in grow-out carp polyculture system for one year in a set of nine earthen ponds of 0.08 ha each. Three different species combinations evaluated were Control: catla (Catla catla Ham.), silver carp (Hypophthalmichthys molitrix Valenciennes), rohu (Labeo rohita Hamilton) and mrigal (Cirrhinus mrigala Hamilton) at 0.5:0.5:1:1; T1: catla, silver carp, rohu and olive barb at 0.5:0.5:1:1 and T2: catla, silver carp, mrigal and olive barb at 0.5:0.5:1:1 at combined density of 7500 fingerlings/ha. While survival levels of the carps did not differ significantly in treatments (P > 0.05), silver carp recorded highest survival levels (94–96%) followed by olive barb (87–90%), mrigal (72–74%), rohu (72–73%) and catla (67–69%). The specific growth rate (SGR) and average harvested body weight (ABW) of catla and silver carp did not differ significantly among the treatments revealing their competition with mrigal or olive barb to be minimum. In absence of rohu in T2, both mrigal and olive barb showed higher SGR and ABW revealing minimal competition between these two species, while their lower performance in presence of rohu in Control and T1 indicated inter-specific competition with the latter. Such olive barb–rohu inter-specific competition, however, failed to yield significant effect on growth of rohu as revealed from its non-significant SGR difference in presence and absence of olive barb. The lower FCR (2.54 ± 0.06) and higher treatment biomass production (3418.4 ± 95.0 kg ha^− 1 year^− 1) in T1 with rohu–olive barb combination compared to T2 with mrigal–olive barb (2.84 ± 0.11; 3155.1 ± 104.7 kg ha^− 1 year^− 1) indicated feasibility and advantage of culturing rohu with olive barb rather than mrigal in carp polyculture. Further, similar biomass production in Control and T1 also indicated feasibility of replacing mrigal with olive barb in the grow-out carp polyculture system.     

Effects of manganese on juvenile mulloway (Argyrosomus japonicus) cultured in water with varying salinity—Implications for inland mariculture

The effects of 5 mg/L of dissolved manganese on juvenile mulloway at salinities of 5, 15 and 45 ppt were determined by comparing their survival, growth and blood plasma and organ chemistry with those of fish grown at the same salinities without manganese addition. Survival of mulloway at 45 ppt in the presence of 5 mg/L of manganese (73 ± 13%) was significantly lower than all other treatments, which achieved 100% survival. Those fish grown in water without manganese exhibited rapid growth, which was not affected by salinity (SGR = 4.05 ± 0.29%/day). Those fish grown at 5 ppt and 45 ppt in the presence of manganese lost weight over the 2-week trial (SGR − 0.17 ± 0.42 and − 0.44 ± 0.83%/day, respectively), whilst those at 15 ppt gained some weight (SGR 1.70 ± 0.20%/day). Manganese accumulated in the gills, liver and muscle of the fish and significant differences in blood plasma chemistry were observed. Blood plasma sodium and chloride of fish exposed to manganese were significantly elevated in hyperosmotic salinity (45 ppt) and depressed at hyposmotic salinity (5 ppt) compared with unexposed fish at the same salinity; consistent with manganese causing apoptosis or necrosis to chloride cells. We did not, however, observe any gill epithelial damage under light microscopy. Blood plasma potassium was significantly elevated at all salinities in the presence of manganese and liver potassium and glycogen reduced. These findings are consistent with manganese interfering with carbohydrate metabolism.     

Comparative assessment of purine nucleotides adenosine,guanosine and inosine monophosphates as functional supplements on growth and health performances of red sea bream,Pagrus major juvenile

Purine nucleotides regulate the cellular functions in the animal body. The current study evaluates the comparative efficacy of dietary purine nucleotides, that is, adenosine monophosphate (AMP), guanosine monophosphate (GMP) and inosine monophosphate (IMP) in red sea bream Pagrus major. Semi‐purified basal diet was formulated (Control, D1) containing 550 g/kg protein, supplemented with purine nucleotides AMP, GMP and IMP at their optimum supplementation level (2, 4 and 4 g/kg) to formulate the experimental diet groups D2, D3 and D4, respectively. Initial weight of 3.5 ± 0.01 g fish was randomly fed test diets in triplicate. After 56 days, % weight gain (p = .003), specific growth rate (p = .003) and apparent lipid digestibility (p = .04) were significantly higher in fish fed diet group D4 followed by D3 and D2. Supplemented groups showed significantly higher feed intake in comparison to control (p = .001). Supplemented groups showed significantly increased and decreased NBT (p = .003) and CAT (p = .003) activity, respectively. Fish fed IMP supplemented diets had significantly lower blood urea nitrogen (p = .04), glutamyl oxaloacetic transaminase (p = .04) and glutamic‐pyruvate transaminase (p = .001) followed by other supplemented groups. Supplemented diet groups showed enhanced stress resistance. Interestingly, D2 and D4 groups showed best oxidative stress status of fish. Therefore, these results indicated that among purine nucleotides, supplementation of IMP could be a more effective nucleotide as functional supplement in red sea bream diet.     

Long-term separate and combined effects of environmental hypercapnia and hyperoxia in Atlantic salmon (Salmo salar L.) smolts

Atlantic salmon (Salmo salar L.) parr (mean start weight 50 g) were reared in freshwater (FW) and exposed to three levels of oxygen saturation measured in effluent water; control group (93% O₂, LO₂), medium (111% O₂, MO₂) and high (123% O₂, HO₂). Further three groups were exposed to similar water oxygen levels in combination with elevated carbon dioxide levels (17–18 mg L^– 1 CO₂), named LO₂–CO₂, MO₂–CO₂ and HO₂–CO₂, respectively. The experiment was run in duplicate tanks for 42 days, and the fish were subsequently transferred to the same seawater (SW) regime for 45 days for an assessment of post-smolt growth. As a consequence of the CO₂ addition, tank pH levels in the FW period were reduced from 6.7 to 5.9 for the hypercapnia groups compared to for the normcapnia groups. Water temperature in FW ranged between 6.4 and 9.0 °C. Citrate was added to the water to complex labile aluminium.In the CO₂ groups observed ventilation frequencies were significantly increased compared to the control (p < 0.05). This difference declined towards the end of the FW period, suggesting acclimation to elevated CO₂. The degree of oxygenation appeared to contribute to the acclimation as the lowest mean ventilation frequency on day 36 was found in the HO₂–CO₂ group and the highest in the LO₂–CO₂ group. Lower plasma chloride and sodium levels were observed in the CO₂ groups relative to the respective oxygenation groups during the FW period, while plasma chloride and sodium levels were normalised to equal levels for all groups after 44 days in SW. No significant differences were found among treatments for blood concentrations of red blood cells, haemoglobin, potassium and glucose during the experiment.By termination of the FW period, the HO₂ group had significantly higher body weight than all other groups (p < 0.05), with specific growth rate significantly higher than the CO₂ groups (p < 0.05). Further, the condition factor was significantly lower in all the CO₂ groups at the end of the FW period compared to the control and normcapnia groups (p < 0.05). Although variable among replicates, occurrence of nephrocalcinosis was 10 times higher in the hypercapnia groups than in the control and normcapnia groups. Mortality was negligible (< 2.0%) during the trial, and most of the mortality occurred following SW transfer.     

Effects of fish origin probiotics on growth performance,immune response and intestinal health of shrimp (Litopenaeus vannamei) fed diets with fish meal partially replaced by soybean meal

This study was conducted to evaluate the application of four fish origin probiotics to relieve the side effects induced by SBM in shrimp. Shrimps were fed with high fish meal diet (C0, positive control containing 500 g/kg FM and 0 g/kg SBM), low fish meal diet (C, control containing 250 g/kg FM and 340 g/kg SBM) or the control diet (C) supplemented, respectively, with 1.0 × 10⁸ CFU/g Bacillus pumilus SE5 (T1), Psychrobacter sp. SE6 (T2), Enterococcus faecium MM4 (T3) or Bacillus claussi DE5 (T4). After 6 weeks of feeding, the growth performance, hepatopancreatic digestive enzymes, intestinal histological structure and immune parameters were determined. The results showed that compared with high FM group (C0), significantly lower WGR and SGR as well as higher FCR were observed in the low FM group (C) (p < .05). Compared with the control, significantly lower FCR was observed in treatment T1 (p < .05), but not in treatments T2–T4. Protease, amylase and lipase activities in all the treatments (T1‐T4) were significantly higher than the control (p < .05). GOT and GPT activities in the control were significantly lower than the C0 (p < .05), while higher MDA level was recorded in the control (p < .05). Meanwhile, higher GPT activities were observed in treatments T1‐T4 compared with the control (p < .05). Compared with the C0, lower SOD, ACP and AKP activities were observed in the control. Significantly improved SOD and AKP activities were observed in all probiotic feeding groups compared with the control (p < .05). The control diet led to significant reduction of intestinal wall thickness and villus height compared with the C0 (p < .05), while significantly higher intestinal wall thickness and villus height were exhibited in all the probiotic groups compared with the control (p < .05), except intestinal wall thickness in treatment T3. Thus, these results demonstrated that the four fish origin probiotic strains could relieve the side effects induced by high level of SBM in shrimp, while probiotic B. pumilus SE5 showed the best performance.     

Growth performance and physiological responses in striped knifejaw, Oplegnathus fasciatus, held under different photoperiods

The growth performance and stress response in striped knifejaw, Oplegnathus fasciatus (body weight 100–300 g) reared under four photoperiods (6L:6D, 12L:12D, 16L:8D and 24L:0D) were investigated. Fish were fed a commercial diet to apparent satiation, two times a day for 8 weeks. A trial of acute handling and confinement stress was also carried out to investigate the stress-induced levels of different stress indicators in O. fasciatus. Blood was also collected from undisturbed fish which was considered as control. Although there were no significant differences in weight gain, specific growth rate (SGR) and feed conversion efficiency (FCE) in fish exposed to 6L:6D, 16L:8D and 24L:0D photoperiods, all parameters in these photoperiods were significantly higher than those of 12L:12D photoperiod (P < 0.05). There was no significant difference in protein retention efficiency (PRE) between fish exposed to 16L:8D and 24L:0D photoperiods, but PRE in both photoperiods was significantly higher than that of 12L:12D photoperiod.Acute stress significantly increased the plasma levels of cortisol (110.3 ng ml^− 1) and glucose (195.4 mg dl^− 1), and decreased plasma levels of total protein (0.8 g dl^− 1); however, all parameters were returned back to the levels indistinguishable from those of control, undisturbed fish within 24 h. The levels of cortisol, glucose and total protein in fish exposed to different photoperiods during the study were far from the stress-induced levels (P < 0.05). The results demonstrated that the growth performance of O. fasciatus reared from 100 to 300 g can be stimulated significantly by using the manipulated photoperiods where feeding time may be playing an important role to increase food intake and feed conversion efficiency. It also revealed that the artificial photoperiods did not cause significant stress response in fish.     

Environmental factors affecting the susceptibility of barramundi to Streptococcus iniae

A long-term environmental study was conducted to evaluate the significance of water temperature, pH and salinity on the prevalence of Streptococcus iniae mortalities in barramundi sea-cage facilities. It was determined that there is a strong association between temperature and increased mortality, specifically between 25 and 28 °C. Temperatures outside of that range result in decreased mortalities attributed to S. iniae. There was no statistical significance between pH or salinity and S. iniae induced mortality (p > 0.05), although acidic conditions (< pH 6) occurring due to anthropogenic disturbances did result in acute mortalities and S. iniae was recovered from approximately 70% these fish. Laboratory challenge studies confirmed the temperature dependence of S. iniae infections as well as the increased susceptibility of barramundi to S. iniae during acid water conditions.     

Effect of feeding apidaecin on common carp (Cyprinus carpio) growth performances and immune function

Effect of apidaecin on common carp (Cyprinus carpio) based on growth performance, feed utilization (Feed Conversion Ratio, FCR) and immune parameters were investigated. Twelve aquaria with three replicates for G1-3 and control were used. The synthesized apidaecin peptide was added to common carp basal diets (Control) as additives in three concentrations: 10.0 mg kg^− 1 (G1), 15.0 mg kg^− 1 (G2) and 30.0 mg kg^− 1 (G3) by wet weight of basal diet. After an 80-day feeding experiment, G2 and G3 showed significantly better (P < 0.05) results of growth performances and FCR than the control. However, there was no remarkable difference (P > 0.05) between G2 and G3. As for G1, the final weight, SGR (Specific Growth Rate) and DG (Daily Gain) were increased and the FCR was decreased, but not significantly (P > 0.05) compared with the control. The serum lysozyme activity of G2 and G3 was significantly increased (P < 0.05) than the control. As for the alternative complement pathway activity, G2 and G3 were also higher (P < 0.05) than the control. However, no significant difference (P > 0.05) was found whichever between G2 and G3 or between G1 and G2 in both lysozyme and the alternative complement pathway activities. Furthermore, there was no remarkable difference between G1 and the control in immune parameters.     

Development of cryopreservation for maintaining yellow catfish Pelteobagrus fulvidraco sperm

Yellow catfish (Pelteobagrus fulvidraco) is a candidate freshwater fish for aquaculture in China with its high consumer demand. The aim of this study was to examine the possibility of storage of the sperm of yellow catfish by cryopreservation in liquid nitrogen. Experiments were designed to investigate the effects of the different combinations of three extenders (Ringer extender, Kurokura-1 extender and D-15 extender) and three cryoprotectants (DMSO, Glycerol and Methanol) on the cryopreservation of yellow catfish sperm. Post-thaw sperm motility, fertilization and hatching rate were detected to evaluate the reliability of sperm cryopreservation. The results demonstrated that Ringer extender and 10% methanol was the best combination for protecting the sperm during freezing in liquid nitrogen by a three-step method and thawing in a water bath at 37 °C for 60 s. In this combination for cryopreservation, sperm maintained the highest post-thaw motility (65 ± 5%), fertilization (90.47 ± 3.67%) and hatching rate (88 ± 4%). And more interestingly, the fertilization and hatching rate were similar to those of fresh sperm (97.55 ± 2.74% and 92 ± 5%). Successful sperm cryopreservation techniques for yellow catfish have been developed for hatchery purpose.     

Identification of progesterone and 17α-hydroxyprogesterone in polychaetes (Perinereis sp.) and the effects of hormone extracts on penaeid oocyte development in vitro

A possible factor behind the successful use of polychaetes for prawn broodstock diet was investigated. Some steroid hormones in polychaetes have been suspected as being a potential component for prawn reproductive system development. Progesterone (P4) and 17-alpha hydroxyprogesterone (17α-OHP4) were extracted from polychaetes (Perinereis sp.) and female Penaeus monodon broodstock at various maturation stages. Concentration of both hormones was determined using RIA or HPLC. Immature natural polychaetes (2 months old) displayed the highest level of P4 and 17α-OHP4 compared to more mature ones (4, 6 and 8 months old polychaetes) (47.48 ± 6.70 ng P4/mg Protein and 143.74 ± 28.70 ng 17α-OHP4/mg Protein) whereas opposite results were received with cultured polychaetes in which a reverse relationship between P4 and its derivative with polychaete maturation was obtained. Diverse concentrations of both hormones may be linked with other factors such as types of polychaetes, habitats, seasons, and feed intakes. Twenty-four hours in vitro incubation of prawn previtellogenic oocytes with P4 or 17α-OHP4 extracted from polychaetes or with synthetic hormones, significantly increased percentages of vitellogenic oocytes and oocytes with cortical rod compared to the control with no hormones. P4 was more effective in enhancing the final maturation of oocytes while 17α-OHP4 had more effects on vitellogenic oocytes. Synthetic steroid hormones at equal hormone concentrations produced similar results to steroid hormones extracted from natural polychaetes.     

Lactobacillus spp. bacteria as probiotics in gilthead sea bream (Sparus aurata, L.) larvae: Effects on growth performance and digestive enzyme activities

In this study, the influence of commercial probiotic, Lactobacillus spp., supplementation was investigated on growth parameters and digestive enzyme activities in gilthead sea bream, Sparus aurata, during larval development. All experiments were triplicated and designed in three different administrations of probiotic from 3 days after hatching (DAH) concurrently with starting of exogenous feeding. In the first group, probiotic was added to live food (rotifer and Artemia). In the second group, probiotic was supplemented directly to both live food and water. In the third group, probiotic was added directly to water. Also, no probiotic treatment was maintained in control group. Total bacterial counts among probiotic probiotic-supplemented groups were significantly different from total bacterial counts in controls in water and digestive tract of larvae (p < 0.05). The mean of total bacterial counts in control was approximately 4 × 10⁴-fold increased from the experimental groups in the sea water (p < 0.05). Besides, mean digestive enzyme activities of all probiotics treatment groups were significantly different (p < 0.05) with that of the control. Except probiotic water supplementation group, in all treatments, the specific activities of pancreatic and intestinal enzymes were significantly higher (p < 0.05) in larvae to which probiotic had been supplemented by live food and live food with water. Also, S. aurata larvae that had probiotic administered by live food with water demonstrated significant (p < 0.05) increases in both survival (13–105% higher) and specific growth rate (2–9% higher) as compared to controls. As a result, supplementation of probiotic to directly tank water could not significantly increase growth parameters and digestive enzyme activities and therefore, administration of probiotics by this method would not be effective in terms of husbandry parameters and nutritional condition.     

Effects of bioflocs with different C/N ratios on growth,immunological parameters,antioxidants and culture water quality in Opsariichthys kaopingensis Dybowski

Biofloc technology (BFT) is a new ecological aquaculture technology that is intended not only to eradicate pollutants and elevate feed utilization but also to enhance immunity and antioxidant activity in aquatic animals. A 28‐day feeding trial was implemented to evaluate the effects of promoted bioflocs on the water quality, growth performance, immunological parameters and antioxidant status of Opsariichthys kaopingensis Dybowski juveniles in low exchange culture tanks. Three hundred sixty healthy O. kaopingensis juveniles (7.1 ± 0.02 g) were irregularly distributed among 12 tanks. Four C:N ratios in triplicate tanks were tried: C/N = 10.8:1 with a commercial diet (control), C/N = 15:1, C/N = 20:1 and C/N = 25:1. Anhydrous glucose (99.97%) was added to the commercial diet to maintain elevated carbon ratio in the three treatments. Uninterrupted 24‐hr aeration was supplied during the test by using an air‐stone connected to an air pump to develop the bioflocs in the water column. The results indicated that BFT significantly reduced total ammonia nitrogen (TAN), nitrite (NO2^?‐N), total phosphorus (TP) concentrations and water transparency (Secchi depth) in C/N 15, C/N 20 and C/N 25 (p < .05), whereas the chemical oxygen demand (COD) and nitrate nitrogen (NO3^?‐N) concentrations gradually increased over time. In addition, weight gain, specific growth rate (SGR), survival and protein efficiency ratio (PER) were significantly elevated in C/N 20, while food conversion rate (FCR) was considerably decreased (p < .05) in C/N 20, compared with the control. Significant increases in lysozyme (LSZ), glutamic pyruvic transaminase (GPT), glutamic oxaloacetic transaminase (GOT) and alkaline phosphatase (AKP) activities of O. kaopingensis were found in the C/N 20 group after a 28‐day feeding trial (p < .05). Comparing the antioxidant capacity of O. kaopingensis in gills, brains, kidneys, hepatopancreas, intestines and serum of juveniles from the four experimental groups, the activity of superoxide dismutase (SOD), total antioxidant capacity (T‐AOC), catalase (CAT) activity and glutathione peroxidase (GSH‐PX) activity of juveniles in the C/N 20 group were significant higher (p < .05), and the content of malondialdehyde (MDA) was considerably lower, than in the control. Overall, these findings suggest that BFT not only can enhance O. kaopingensis growth performance and strengthen antioxidant status but also can upgrade immune response, ensuring the sustainable development of aquaculture.     

Determination of optimal temperature(s) in juvenile red‐spotted grouper Epinephelus akaara (Temminck & Schlegel) based on growth performance and stress responses

This study sought to determine the optimal temperature(s) for aquaculture of juvenile red‐spotted grouper Epinephelus akaara (Temminck & Schlegel) (mean initial BW: 3.1 g). Growth performance, insulin‐like growth factor 1 (IGF‐1) expression and thermal stress responses (plasma cortisol, glucose, and hepatic heat shock protein 60 expression) were evaluated at three constant temperatures (24°C, 26°C and 28°C) in a 2‐week trial. At the end of the trial, final BW was significantly higher at 26°C and 28°C than at 24°C (p < 0.05); a quadratic regression analysis of final BW showed the optimum temperature for growth was 27.5°C (p < 0.05, R² = 0.806). The highest hepatic IGF‐1 expression was observed at 26°C (p < 0.05). On the other hand, hepatic heat shock protein 60 expression was highest at 28°C (p < 0.05), suggesting thermal stress. In conclusion, temperature optima, which support excellent growth but induce minimal thermal stress, was 26°C. This fine information within a narrow temperature range is expected to give empirical information for red‐spotted grouper farmers to sustain maximal production efficiency with avoiding thermal stress and to determine the future location of production, especially in consideration of arising seawater temperatures.     

Effects of emulsified versus saline administration of GnRHa on induction of ovulation in rainbow trout, Oncorhynchus mykiss

The effects of saline-dissolved or Freund's incomplete adjuvant (FIA)-emulsified GnRHa treatment on the induction of ovulation in rainbow trout, Oncorhynchus mykiss, were examined. The FIA-emulsified GnRHa was first diluted in 0.25 ml physiological saline and then mixed with an equal volume of FIA. Fish were selected in the beginning of the spawning season and were allocated into four groups and were treated intraperitoneally with (a) 0.5 ml of emulsified GnRHa (GnRHa–FIA), (b) 0.5 ml saline-dissolved GnRHa in a single injection (GnRHa-1), (c) 0.5 ml saline-dissolved GnRHa in two injections spaced 1 d apart (GnRHa-2) and (d) 0.5 ml of saline (Control). The GnRHa dose in all hormone treatments was 25 µg kg^− 1. All fish in the FIA–GnRHa and GnRHa-2 groups ovulated within 10 and 11 d after treatment, respectively. In contrast, only 75% in the Control fish and 60% of the fish in the GnRHa-1 group ovulated within 36 d after treatment. None of the treatments caused any pre- or post-spawning mortality in the broodstock. Fertilization, eyeing and hatching percentages of the produced progeny were normal in all the treatment groups and did not differ significantly among them. In conclusion, FIA-emulsified GnRHa can be effective in advancing the onset of and synchronizing the ovulation of rainbow trout within a two-week period, thus shortening the egg collection period, without affecting broodstock survival and egg quality.